1. Researchers regenerated haploid plants from anther culture of banana (Musa paradisiaca cv. Puttabale).
2. The highest rate of callus formation (90%) was achieved using 3mg/l of 2,4-D plant growth regulator. Embryoids developed from callus after 20 days.
3. Shoot growth was optimized using 4mg/l BAP and 0.4 mg/l IAA, producing an average of 7 shoots per callus. Root formation was achieved using 0.6 mg/l NAA.
4. Flow cytometry analysis confirmed the production of haploid plants, with nuclear DNA content half that of diploid plants. The efficient protocol
The document discusses the production of double haploid (DH) plant lines in cucumber. It describes screening cucumber accessions for resistance to Cucumber mosaic virus (CMV) using DAS-ELISA. Ovule culture techniques were used to establish DH plant lines from selected CMV-resistant accessions. The DH lines were then screened for CMV resistance to develop homozygous cucumber lines with improved virus resistance.
This document discusses in-vitro androgenesis in rice. It provides background on rice production and genetics. Rice anther culture is described as a two-step process of callus induction and plant regeneration. Factors affecting rice androgenesis include genotype, donor plant physiology, anther development stage, pretreatment, culture medium, and growth regulators. Applications of doubled haploids in rice include shortening breeding cycles and fixing traits. Challenges include low response rates and albinism in regenerated plants.
The document discusses the production of double haploid plants through anther and pollen culture techniques. It provides background on the history of double haploid development, the importance of double haploids in plant breeding, and methods used to induce haploids including anther culture, pollen culture, ovary slice culture, and ovule culture. Key factors influencing anther culture success are also reviewed, such as genotype, culture medium, microspore stage, temperature, and donor plant physiology. Advantages and disadvantages of generating double haploid lines are presented.
This document summarizes methods for producing haploid and doubled haploid plants for plant breeding programs. It discusses dihaploid production through halving tetraploid chromosome numbers. It also describes protocols for in vitro haploid production through unfertilized ovule/ovary culture and isolated microspore culture, including donor plant growth, explant collection and sterilization, culture medium, induction methods, and embryo regeneration. The goal is to accelerate the production of homozygous lines for more efficient plant breeding.
Haploids and Double haploids are having considerable importance in plant breeding and genetics. Also, the production of haploids through conventional breeding methods is long term efforts. therefore many scientists investigated the production of haploids through anther culture.
Anther and pollen culture is the production of haploid plants exploiting the totipotency of microscope and the occurrence of single set of chromosome in microscope.
This document discusses anther culture and summarizes 4 experimental studies on anther culture. The first study examines the effect of cultivar and media supplementation on callus induction and root formation in red cabbage. The second study looks at callus induction, embryo induction, and plantlet initiation in red cabbage using different media. The third study analyzes the effect of macronutrient and agar concentration on callus induction and embryo formation in different cucumber varieties. The fourth study evaluates androgenesis induction, callogenesis, regeneration and cytogenetics in tomato haploids.
The document discusses tissue culture techniques and their applications in plant breeding. It provides a historical background of tissue culture dating back to the early 20th century. The major steps involved in tissue culture are explained, including selection of explant tissue, sterilization, establishment in culture medium, multiplication through callus formation, root formation, and hardening of plantlets. Key techniques like protoplast culture, haploid culture, and micropropagation are also summarized along with their achievements in generating somatic hybrids, true breeding lines, and large-scale clonal propagation respectively.
The document discusses the production of double haploid (DH) plant lines in cucumber. It describes screening cucumber accessions for resistance to Cucumber mosaic virus (CMV) using DAS-ELISA. Ovule culture techniques were used to establish DH plant lines from selected CMV-resistant accessions. The DH lines were then screened for CMV resistance to develop homozygous cucumber lines with improved virus resistance.
This document discusses in-vitro androgenesis in rice. It provides background on rice production and genetics. Rice anther culture is described as a two-step process of callus induction and plant regeneration. Factors affecting rice androgenesis include genotype, donor plant physiology, anther development stage, pretreatment, culture medium, and growth regulators. Applications of doubled haploids in rice include shortening breeding cycles and fixing traits. Challenges include low response rates and albinism in regenerated plants.
The document discusses the production of double haploid plants through anther and pollen culture techniques. It provides background on the history of double haploid development, the importance of double haploids in plant breeding, and methods used to induce haploids including anther culture, pollen culture, ovary slice culture, and ovule culture. Key factors influencing anther culture success are also reviewed, such as genotype, culture medium, microspore stage, temperature, and donor plant physiology. Advantages and disadvantages of generating double haploid lines are presented.
This document summarizes methods for producing haploid and doubled haploid plants for plant breeding programs. It discusses dihaploid production through halving tetraploid chromosome numbers. It also describes protocols for in vitro haploid production through unfertilized ovule/ovary culture and isolated microspore culture, including donor plant growth, explant collection and sterilization, culture medium, induction methods, and embryo regeneration. The goal is to accelerate the production of homozygous lines for more efficient plant breeding.
Haploids and Double haploids are having considerable importance in plant breeding and genetics. Also, the production of haploids through conventional breeding methods is long term efforts. therefore many scientists investigated the production of haploids through anther culture.
Anther and pollen culture is the production of haploid plants exploiting the totipotency of microscope and the occurrence of single set of chromosome in microscope.
This document discusses anther culture and summarizes 4 experimental studies on anther culture. The first study examines the effect of cultivar and media supplementation on callus induction and root formation in red cabbage. The second study looks at callus induction, embryo induction, and plantlet initiation in red cabbage using different media. The third study analyzes the effect of macronutrient and agar concentration on callus induction and embryo formation in different cucumber varieties. The fourth study evaluates androgenesis induction, callogenesis, regeneration and cytogenetics in tomato haploids.
The document discusses tissue culture techniques and their applications in plant breeding. It provides a historical background of tissue culture dating back to the early 20th century. The major steps involved in tissue culture are explained, including selection of explant tissue, sterilization, establishment in culture medium, multiplication through callus formation, root formation, and hardening of plantlets. Key techniques like protoplast culture, haploid culture, and micropropagation are also summarized along with their achievements in generating somatic hybrids, true breeding lines, and large-scale clonal propagation respectively.
The document discusses androgenesis, which is the production of haploid plants through microspore culture. Two main techniques are described: anther culture and pollen/microspore culture. Anther culture involves culturing excised anthers while pollen/microspore culture involves isolating and culturing individual pollen grains. Successful androgenesis requires stress treatments, specific culture media formulations, and plant growth regulators. The process can result in direct embryogenesis from microspores or indirect embryogenesis through a callus phase. Factors influencing androgenesis success and the pathways and stages of microspore development are also covered.
Haploid plant formation can occur through several methods, including anther/pollen culture, ovule culture, and distant hybridization. Anther/pollen culture involves culturing anthers or isolated microspores to induce microspores or pollen to undergo embryogenesis rather than gametogenesis. Success requires triggering a shift from the gametophytic to sporophytic development pathway. Temperature shock, culture medium composition, and donor plant conditions influence response. Haploids can be doubled to create homozygous diploid lines using colchicine, valuable for shortening plant breeding cycles.
This document summarizes a study on using colchicine pretreatment of maize anthers to induce doubled haploids. Two maize genotypes were used as donors for anther culture. Anthers were pretreated with different concentrations of colchicine for various durations before culture. Results showed increased embryogenic structure formation with 100 mg/L colchicine in one genotype and 300 mg/L in the other. Both genotypes produced doubled haploid plantlets when anthers were pretreated with 250 mg/L colchicine for 6 days. The study demonstrates the use of colchicine pretreatment to improve chromosome doubling rates in anther culture of maize.
This document discusses methods for producing haploid plants through gynogenesis, or haploid production from female gametes. It describes two main methods - ovary culture and ovule culture - for inducing gynogenesis in vitro. Key steps include excising ovaries from plants and culturing them on nutrient media supplemented with hormones to induce parthenogenesis. Successful gynogenesis has been achieved in several plant families, with the frequency of responsive ovules typically being low, around 1-5%. Producing haploid plants provides benefits for genetic studies.
This document discusses haploid plant production through anther culture. It begins by defining key terms like gametophyte, sporophyte, haploid and diploid plants. It then describes the two main methods of haploid production - anther culture and isolated microspore culture. For anther culture, it outlines the process of culturing immature anthers on nutrient media, including pretreatments, media composition and plant regeneration. Anther culture can result in direct or indirect embryogenesis and four pathways of pollen development are described. The document provides detailed steps of anther culture and potential issues like production of diploid plants.
The document discusses anther culture and haploid production techniques. It begins by explaining that anther culture can be used to produce haploid plants faster than traditional breeding methods. It then provides details on the history and development of anther culture techniques. The document outlines the basic procedure for anther culture, including plant growth conditions, anther selection, culture medium, and isolation of haploid plants. It discusses factors that influence anther culture success and limitations of the technique. Finally, it provides some examples of commercial varieties developed using anther culture.
Marker Assisted production of doubled haploid plants - A boon for feeding the...Ragavendran Abbai
Plant breeding is the science of production of new superior varieties from the existing varieties.But the challenge is that the techniques of conventional plant breeding is not sufficient to feed the entire human population.So, the way out is promoting the Marker Assisted production of DH lines.
PRODUCTION OF HAPLOID PLANTS AND HOMOZYGOUS DIPLOID LINESAmbika Prajapati
This document presents information on producing haploid plants and homozygous diploid lines through plant biotechnology. It discusses two approaches for producing haploid plants, in vivo and in vitro. It also describes techniques for identifying haploids, diploidizing haploids through colchicine treatment or endomitosis to produce homozygous plants, and the significance of haploid plants for plant breeding programs.
Haploid culture are known to be culture the anther/pollen and ovary/ovule of plants.
Make sporophyte with the help of gametophyte.
One set of chromosome
Recessive mutation is easily detectable
This seminar presentation summarizes anther culture and haploid plant regeneration. Anther culture involves culturing microspores or anthers to produce haploid plants. It has been successful in over 250 plant species. There are different pathways that microspores can take to develop into haploid plants, including equal or unequal division. Pretreatment of anthers and culture conditions like medium, temperature and light exposure can influence success rates. Anther culture is useful for producing haploids but requires skill and has risks of chimera or callus formation.
INTRODUCTION
WHAT IS ANDROGENESIS ?
HISTORY
TYPES OF ANDROGENESIS TECHNIQUES
ONTOGENY OF ANDROGENIC HAPLOIDS
GYNOGENESIS
FACTORS AFFECTING ANDROGENESIS
APPLICATIONS OF ANDROGENESIS
LIMITATIONS
REFERENCES
Clonal Propagation: Introduction, Techniques, Factors, Applications and Disadvantages
Multiplication of Apical or Axillary bud, Shoot tip or meristem culture
Production of Disease free plants by Micropropagation techniques: their Advantages and Disadvantages
Anther culture is a technique where anthers are excised from flower buds and cultured to produce haploid plants. The first report of haploid tissue from anther culture was in 1964-1966 in Datura pollen grains. Over 250 species have been produced through anther culture, most commonly in families like Solanaceae, Cruciferae, and Poaceae. Haploid plants are useful for identifying recessive traits, eliminating lethal genes, and producing homozygous diploid plants more quickly. There are several pathways that microspores can follow during anther culture, such as symmetric or asymmetric division, to produce haploid plants. Successful anther culture requires optimizing various factors like donor plant genotype, anther
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
This document discusses haploid plant production. Haploid plants have half the normal number of chromosomes. They are important for breeding programs. Methods to produce haploids include anther or pollen culture (androgenic haploids) and ovary or ovule culture (gynogenic haploids). Both in vivo and in vitro techniques have been used, with higher success rates for in vitro methods like anther culture. Gynogenesis involves culturing unpollinated ovaries to trigger female gametophytes to develop into plants. While useful, ovary culture is less common than anther culture due to difficulties in dissecting ovaries.
Androgenesis is the production of haploid plants through the culture of male gametophytes or microspores. There are two main methods - anther culture and isolated pollen/microspore culture. Anther culture involves excising anthers from flower buds and culturing them on nutrient media, while microspore culture isolates microspores from anthers. Several factors influence androgenesis success, including genotype, anther wall components, culture medium, growth regulators, and physical conditions. Androgenic haploids can develop directly from microspores or indirectly through a callus phase, following various developmental pathways. Androgenesis allows for the efficient production of haploid plants for breeding programs.
The document discusses haploid plant culture techniques. It describes anther/pollen culture and ovule culture as important methods to produce haploid plants. Anther/pollen culture involves culturing anthers or isolated pollen grains to develop into haploid embryos or callus. Ovule culture involves culturing isolated ovules. The document outlines factors that affect androgenesis (anther/pollen culture) and discusses using techniques like colchicine treatment or long-term callus culture to induce chromosome doubling and generate homozygous diploid plants from haploids.
Haploid Production - Techniques, Application & Problem ANUGYA JAISWAL
Haploid is applied to any plant originating from a sporophyte (2n) and containing (n) number of chromosomes.
Artificial production of haploids was attempted through distant hybridization, delayed pollination, application of irradiated pollen, hormone treatment and temperature shock.
The artificial production of haploids until 1964 was attempted through:
1. Distant hybridization
2. Delayed pollination
3. Application of irradiated pollen
4. Hormone treatments
5. Temperature shocks
The development of numerous pollen plantlets in anther cultures of Datura innoxia, first reported by two Indian scientists (Guha and Maheshwari, 1964, 1966), was a major breakthrough in haploid breeding of higher plants.
The technique of haploid production through anther culture ('anther - androgenesis') has been extended successfully to numerous plant species, including many economically important plants, such as cereals and vegetable, oil and tree crops.
1) The document describes methods for micropropagating date palm and guava plants through tissue culture techniques. It provides details on direct organogenesis for date palm propagation and somatic embryogenesis for guava propagation.
2) For date palms, methods are described for initiation, multiplication, elongation, rooting, and acclimatization stages. The optimal conditions found were 2 mg/L 2iP plus 1 mg/L BA for initiation, 4 mg/L 2iP with 2 mg/L BA and 1 mg/L NAA for multiplication, 0.5 mg/L GA3 for elongation, and 0.5 mg/L NAA for rooting.
3) For guava, the
A high frequency microcloning protocol for subsequent cryopreservation in Kae...iosrphr_editor
This document describes a high frequency micropropagation protocol for Kaempferia galanga, an endangered medicinal plant. Key findings:
- Shoot cultures were established from rhizome explants on MS medium with BA and multiplied through subculturing. Maximum shoot multiplication (10.6 shoots/explant) was achieved on MS with 4 mg/L BA, 1 mg/L kinetin, and 1 mg/L NAA.
- Repeated subculturing every 4 weeks increased shoot multiplication 3-4x, allowing production of up to 30 shoots/explant within 12 weeks.
- Shoots elongated and rooted simultaneously without needing separate media, reducing production time versus previous protocols.
-
The document discusses androgenesis, which is the production of haploid plants through microspore culture. Two main techniques are described: anther culture and pollen/microspore culture. Anther culture involves culturing excised anthers while pollen/microspore culture involves isolating and culturing individual pollen grains. Successful androgenesis requires stress treatments, specific culture media formulations, and plant growth regulators. The process can result in direct embryogenesis from microspores or indirect embryogenesis through a callus phase. Factors influencing androgenesis success and the pathways and stages of microspore development are also covered.
Haploid plant formation can occur through several methods, including anther/pollen culture, ovule culture, and distant hybridization. Anther/pollen culture involves culturing anthers or isolated microspores to induce microspores or pollen to undergo embryogenesis rather than gametogenesis. Success requires triggering a shift from the gametophytic to sporophytic development pathway. Temperature shock, culture medium composition, and donor plant conditions influence response. Haploids can be doubled to create homozygous diploid lines using colchicine, valuable for shortening plant breeding cycles.
This document summarizes a study on using colchicine pretreatment of maize anthers to induce doubled haploids. Two maize genotypes were used as donors for anther culture. Anthers were pretreated with different concentrations of colchicine for various durations before culture. Results showed increased embryogenic structure formation with 100 mg/L colchicine in one genotype and 300 mg/L in the other. Both genotypes produced doubled haploid plantlets when anthers were pretreated with 250 mg/L colchicine for 6 days. The study demonstrates the use of colchicine pretreatment to improve chromosome doubling rates in anther culture of maize.
This document discusses methods for producing haploid plants through gynogenesis, or haploid production from female gametes. It describes two main methods - ovary culture and ovule culture - for inducing gynogenesis in vitro. Key steps include excising ovaries from plants and culturing them on nutrient media supplemented with hormones to induce parthenogenesis. Successful gynogenesis has been achieved in several plant families, with the frequency of responsive ovules typically being low, around 1-5%. Producing haploid plants provides benefits for genetic studies.
This document discusses haploid plant production through anther culture. It begins by defining key terms like gametophyte, sporophyte, haploid and diploid plants. It then describes the two main methods of haploid production - anther culture and isolated microspore culture. For anther culture, it outlines the process of culturing immature anthers on nutrient media, including pretreatments, media composition and plant regeneration. Anther culture can result in direct or indirect embryogenesis and four pathways of pollen development are described. The document provides detailed steps of anther culture and potential issues like production of diploid plants.
The document discusses anther culture and haploid production techniques. It begins by explaining that anther culture can be used to produce haploid plants faster than traditional breeding methods. It then provides details on the history and development of anther culture techniques. The document outlines the basic procedure for anther culture, including plant growth conditions, anther selection, culture medium, and isolation of haploid plants. It discusses factors that influence anther culture success and limitations of the technique. Finally, it provides some examples of commercial varieties developed using anther culture.
Marker Assisted production of doubled haploid plants - A boon for feeding the...Ragavendran Abbai
Plant breeding is the science of production of new superior varieties from the existing varieties.But the challenge is that the techniques of conventional plant breeding is not sufficient to feed the entire human population.So, the way out is promoting the Marker Assisted production of DH lines.
PRODUCTION OF HAPLOID PLANTS AND HOMOZYGOUS DIPLOID LINESAmbika Prajapati
This document presents information on producing haploid plants and homozygous diploid lines through plant biotechnology. It discusses two approaches for producing haploid plants, in vivo and in vitro. It also describes techniques for identifying haploids, diploidizing haploids through colchicine treatment or endomitosis to produce homozygous plants, and the significance of haploid plants for plant breeding programs.
Haploid culture are known to be culture the anther/pollen and ovary/ovule of plants.
Make sporophyte with the help of gametophyte.
One set of chromosome
Recessive mutation is easily detectable
This seminar presentation summarizes anther culture and haploid plant regeneration. Anther culture involves culturing microspores or anthers to produce haploid plants. It has been successful in over 250 plant species. There are different pathways that microspores can take to develop into haploid plants, including equal or unequal division. Pretreatment of anthers and culture conditions like medium, temperature and light exposure can influence success rates. Anther culture is useful for producing haploids but requires skill and has risks of chimera or callus formation.
INTRODUCTION
WHAT IS ANDROGENESIS ?
HISTORY
TYPES OF ANDROGENESIS TECHNIQUES
ONTOGENY OF ANDROGENIC HAPLOIDS
GYNOGENESIS
FACTORS AFFECTING ANDROGENESIS
APPLICATIONS OF ANDROGENESIS
LIMITATIONS
REFERENCES
Clonal Propagation: Introduction, Techniques, Factors, Applications and Disadvantages
Multiplication of Apical or Axillary bud, Shoot tip or meristem culture
Production of Disease free plants by Micropropagation techniques: their Advantages and Disadvantages
Anther culture is a technique where anthers are excised from flower buds and cultured to produce haploid plants. The first report of haploid tissue from anther culture was in 1964-1966 in Datura pollen grains. Over 250 species have been produced through anther culture, most commonly in families like Solanaceae, Cruciferae, and Poaceae. Haploid plants are useful for identifying recessive traits, eliminating lethal genes, and producing homozygous diploid plants more quickly. There are several pathways that microspores can follow during anther culture, such as symmetric or asymmetric division, to produce haploid plants. Successful anther culture requires optimizing various factors like donor plant genotype, anther
Anther culture:- the in vitro culturing of anthers containing microspores or immature pollen grains on a nutrient medium for the purpose of generating haploid plantlets.
Culturing anthers for the purpose of obtaining Double Haploid is not easy with many field crop species, particularly with the cereals, cotton, and grain legumes.
This document discusses haploid plant production. Haploid plants have half the normal number of chromosomes. They are important for breeding programs. Methods to produce haploids include anther or pollen culture (androgenic haploids) and ovary or ovule culture (gynogenic haploids). Both in vivo and in vitro techniques have been used, with higher success rates for in vitro methods like anther culture. Gynogenesis involves culturing unpollinated ovaries to trigger female gametophytes to develop into plants. While useful, ovary culture is less common than anther culture due to difficulties in dissecting ovaries.
Androgenesis is the production of haploid plants through the culture of male gametophytes or microspores. There are two main methods - anther culture and isolated pollen/microspore culture. Anther culture involves excising anthers from flower buds and culturing them on nutrient media, while microspore culture isolates microspores from anthers. Several factors influence androgenesis success, including genotype, anther wall components, culture medium, growth regulators, and physical conditions. Androgenic haploids can develop directly from microspores or indirectly through a callus phase, following various developmental pathways. Androgenesis allows for the efficient production of haploid plants for breeding programs.
The document discusses haploid plant culture techniques. It describes anther/pollen culture and ovule culture as important methods to produce haploid plants. Anther/pollen culture involves culturing anthers or isolated pollen grains to develop into haploid embryos or callus. Ovule culture involves culturing isolated ovules. The document outlines factors that affect androgenesis (anther/pollen culture) and discusses using techniques like colchicine treatment or long-term callus culture to induce chromosome doubling and generate homozygous diploid plants from haploids.
Haploid Production - Techniques, Application & Problem ANUGYA JAISWAL
Haploid is applied to any plant originating from a sporophyte (2n) and containing (n) number of chromosomes.
Artificial production of haploids was attempted through distant hybridization, delayed pollination, application of irradiated pollen, hormone treatment and temperature shock.
The artificial production of haploids until 1964 was attempted through:
1. Distant hybridization
2. Delayed pollination
3. Application of irradiated pollen
4. Hormone treatments
5. Temperature shocks
The development of numerous pollen plantlets in anther cultures of Datura innoxia, first reported by two Indian scientists (Guha and Maheshwari, 1964, 1966), was a major breakthrough in haploid breeding of higher plants.
The technique of haploid production through anther culture ('anther - androgenesis') has been extended successfully to numerous plant species, including many economically important plants, such as cereals and vegetable, oil and tree crops.
1) The document describes methods for micropropagating date palm and guava plants through tissue culture techniques. It provides details on direct organogenesis for date palm propagation and somatic embryogenesis for guava propagation.
2) For date palms, methods are described for initiation, multiplication, elongation, rooting, and acclimatization stages. The optimal conditions found were 2 mg/L 2iP plus 1 mg/L BA for initiation, 4 mg/L 2iP with 2 mg/L BA and 1 mg/L NAA for multiplication, 0.5 mg/L GA3 for elongation, and 0.5 mg/L NAA for rooting.
3) For guava, the
A high frequency microcloning protocol for subsequent cryopreservation in Kae...iosrphr_editor
This document describes a high frequency micropropagation protocol for Kaempferia galanga, an endangered medicinal plant. Key findings:
- Shoot cultures were established from rhizome explants on MS medium with BA and multiplied through subculturing. Maximum shoot multiplication (10.6 shoots/explant) was achieved on MS with 4 mg/L BA, 1 mg/L kinetin, and 1 mg/L NAA.
- Repeated subculturing every 4 weeks increased shoot multiplication 3-4x, allowing production of up to 30 shoots/explant within 12 weeks.
- Shoots elongated and rooted simultaneously without needing separate media, reducing production time versus previous protocols.
-
International Journal of Pharmaceutical Science Invention (IJPSI)inventionjournals
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
Isolation, identification of antagonistic rhizobacterial strains obtained fro...Shazia Shahzaman
Plant growth promoting rhizobacteria (PGPR), are associated with roots, found in the rhizosphere and can directly or indirectly enhance the plant growth. In this study soil was collected from rhizosphere of chickpea fields of different areas of Rawalpindi division of Pakistan. PGPR were isolated, screened and characterized. Eight isolates of rhizobacteria (RHA, RPG, RFJ, RC, RTR, RT and RK) were isolated from Rawalpindi division and were characterized. The antagonistic activity of these PGPR isolates against root infecting fungi (Fusarium oxysporum and Verticillium spp.,) was done and production of indole acetic acid (IAA), siderophore and P-solubilization was evaluated. The isolates RHA, RPG, RFJ, RC, RRD and RT were found to be positive in producing siderophore, IAA and P-solubilization. Furthermore, most of the isolates showed antifungal activity against Fusarium oxysporum, and Verticillium spp. The rhizobacterial isolates RHA, RPG, RFJ, RC, RRD, RTR, RT and RK were used as bio-inoculants that might be beneficial for chickpea cultivation as the rhizobacterial isolates possessed the plant growth promoting characters i.e. siderophore, IAA production, phosphate solubilization. In in vitro tests, Pseudomonas sp. and Bacillus spp. inhibited the mycelial growth of the fungal root pathogens. The isolates (RHA and RPG) also significantly increased (60-70%) seed germination, shoot length, root length of the chickpea. The incidence of fungi was reduced by the colonization of RHA and RPG which enhanced the seedling vigor index and seed germination. The observations revealed that isolates RHA and RPG is quite effective to reduce the fungal root infection in greenhouse, and also increases seed yields significantly. These rhizobacterial isolates appear to be efficient yield increasing as well as effective biocontrol agent against fungal root pathogen.
In vitro tuberization and colchicine content analysis of Gloriosa superba L.Nagendra Worlds
Gloriosa superba L. is an herbaceous climber distributed in tropical parts of the
world. Pharmaceutically important alkaloid-colchicine, present in its tubers and
seeds and due to overexploitation it becomes vulnerable in the forests. In the
present investigation, in vitro tuber production was carried out for its propagation
and conservation. The plant possesses a very strong apical dominance.
Consequently, any damage to the plant apical meristem is fatal for it which was also
exhibited during in vitro culture. Only apical meristems were able to produce a
single and un-branched shoots and nodal explants were remained dormant even in
the presence of exogenous cytokinin. The in vitro propagation was accomplished
by the microtuber formation technique, in two steps. Maximum number of
microtubers 9.8±0.8 per culture in eight weeks, were produced in vitro on
Murashige and Skoog medium with sucrose (60 g LG1) and in the presence
of 35.5 μM 6-benzyladenine (BA) with citric acid and polyvinyl pyrrolidone-40.
Subsequently the induced microtubers were sub-cultured on to the medium with
lower cytokinin level, 8.88 μM BA. The individual microtubers with shoots were
subjected to a single step rooting and in vitro acclimatization in coco-pit containing
vessels, exhibited 90% survival. In vitro grown tubers contained less percentage of
colchicine than the natural field-grown plant tubers. However, microtubers showed
increased colchicine content, as they grow older.
Key words: Apical dominance release, bud culture, cytokinin, glory lily
In vitro regeneration and transformation studies on Pelargonium graveolens (g...Fathima Benazir
Development of improved protocols for the regeneration of Pelargonium graveolens is important for their commercial utilization. Callus was induced from leaf explants on Murashige and Skoog (MS) medium supplemented with auxins and combinations of auxins and cytokinins. Micropropagation of nodal bud explants was achieved using medium supplemented with cytokinins. The regenerated explants were acclimatized and the transformation potential of geranium was demonstrated using Agrobacterium tumefaciens. Maximum proliferation was obtained in medium supplemented with combinations of 20 μM indole butyric acid (IBA) + 10 μM kinetin (KIN), 20 μM IBA + 10 μM benzyl amino purine (BAP) and 20 μM indole-3-acetic acid (IAA) + 10 μM KIN. Plantlets were regenerated when callus was given a photoperiod of 16 h light and 8 h dark conditions at 25°C. After being transferred to greenhouse conditions, the plantlets were successfully acclimatized and transferred to the field condition. The study has established an efficient protocol for in vitro regeneration and transformation of P. graveolens as an alternative method for vegetative propagation.
This document describes a study that established an efficient protocol for in vitro regeneration and genetic transformation of Pelargonium graveolens (geranium). Maximum callus proliferation was obtained on medium supplemented with combinations of 20 μM IBA + 10 μM kinetin, 20 μM IBA + 10 μM BAP, or 20 μM IAA + 10 μM kinetin. Plantlets were regenerated from callus under 16 hours of light per day. The regenerated plantlets were successfully acclimatized and transferred to soil. Genetic transformation of geranium was demonstrated using Agrobacterium tumefaciens containing a binary vector.
In vitro propagation and conservation of Swertia bimaculata Bir Bahadur Thapa
1) The document describes a study aimed at the in vitro propagation and conservation of Swertia bimaculata, an herbaceous plant used for its medicinal properties.
2) Shoot tips were used as explants and cultured on MS medium supplemented with plant growth regulators to induce shoot multiplication over 10 weeks. Root induction was achieved by culturing shoots on medium with auxins.
3) Chromosomal analysis of regenerated plants showed numerical stability between somatic and reproductive cells, demonstrating the effectiveness of the propagation protocol for the conservation of this important plant species.
In vitro propagation in Bamboo(micropropagation) (plant tissue Culture)Rahul Chandera
This document discusses the in vitro propagation of bamboo through tissue culture techniques. It describes the selection of nodal explants from juvenile bamboo shoots, sterilization procedures, and various initiation, multiplication, rooting, and hardening media compositions that produced the best results. The highest rates of shoot bud induction and multiplication were achieved using MS media supplemented with BAP and KIN plant growth regulators. Root induction was maximized with IBA or NAA in the media. Hardening involved transferring plantlets to soil mixtures in the greenhouse, resulting in 92.5% survival rates. Tissue culture is presented as an efficient method for large-scale bamboo propagation.
Effect of Colchicine Tablets on Morphology of Torenia fournieridrboon
The document summarizes research on inducing polyploidy in the plant species Torenia fournieri through treatment with colchicine solution. Leaves were soaked in various concentrations of colchicine for different durations. Higher concentrations and longer durations decreased survival rates. Putative polyploids were selected based on larger stomata size and darker green leaves. Flow cytometry and chromosome counting confirmed some plants were induced to tetraploid (4n=36 chromosomes), compared to the normal diploid (2n=18 chromosomes). The highest induction rate was 6.67% for treatment with 15 ppm colchicine for 3 days. Morphological differences between induced tetraploids and control diploids
Organogenic Regeneration of an Elite Cultivar of Chinese Jujube (Zizyphus juj...Agriculture Journal IJOEAR
Abstract— An efficient and relatively simple regeneration system was developed for an elite cultivar of Chinese Jujube, a perennial tree, by culturing young twig segments as explants from 8-15 year old trees. The twig segments were disinfected by submerging them in 1% sodium hypochlorite (NaOCl) for 15 min with 3 min vacuum. Calli developed from both ends of the twig segments on half-strength MS medium supplemented with sucrose and BA or BA and NAA in combination. The frequency of shoot formation from calli was higher than 80% when the explants were placed on the half - strength MS medium supplemented with BA (2.581 μM) and NAA (2.685 μM). Roots were produced from adventitious buds for 90% of the regenerated shoots when they were placed on the MS medium supplemented with 4.920 μM IBA and 5.708 μM IAA. After transplanting to soil, 82% of the regenerated seedlings survived when they were covered with glass containers to maintain humidity. The results suggest that Chinese jujube can be reproduced and multiplied using organogenesis with the appropriate explant and culture medium.
Effect of different p h on in vitro seed germination of vanda tessellata (rox...Alexander Decker
The document summarizes a study that investigated the effect of different pH levels on in vitro seed germination of Vanda tessellata, an endangered medicinal orchid. Immature seeds were germinated on basal MS media at pH levels of 3.5, 4.5, 5.5 and 6.5. The highest seed germination rate of 95% was observed at pH 5.5, while pH 3.5 and 6.5 showed poor germination. The study concluded that a pH of 5.5 is best for seed germination of V. tessellata compared to the other pH levels tested.
This document summarizes research on in vitro propagation of the medicinal plant Cocculus orbiculatus through cultivation from leaf explants. The study found that MS medium supplemented with 1.0 mg/L of BAP was optimal for inducing shoots directly from leaf explants. Combinations of BAP and adenine sulfate resulted in increased multiple shoot formation. The highest number of shoots was observed with 1.0 mg/L BAP and 5 mg/L adenine sulfate. The protocol established a reliable method for rapid multiple shoot induction and propagation of C. orbiculatus using leaf explants, cytokinins and adenine sulfate.
Mutation Induction for Improvement of Banana (Musa Spp). "Berangan Cv. Intan-...paperpublications3
Abstract: In vitro mutation induction by using gamma ray at 20, 30, 40 and 60Gy was used to generate variability in triploid banana "Berangan cv. Intan (AAA), so as to provide the opportunity to select plants with desirable characters such as early fruiting and short stature. Mutation frequency increased with increased dosage whereas, survival and capacity to regenerate decreased with increased doses. Time to initiation varied from 4-8 weeks for gamma-irradiated materials compared to 2-3 weeks in the control. It appeared that the higher the dose, the longer it took for shoot initiation. The exposure of shoot-tip meristem pieces to radiation doses produced wide variation in growth and morphogenetic performance. Mutagenic treatments induced 2 to 3- fold increases in variability in both quantitative and qualitative traits at different stages, in vitro; at nursery and field. For the field-grown plants; the proportion varied from 2.9% for the control plants to 16.8% for 60 Gy and 20.1% for 40 Gy-treatments, while for treatments at 20 and 30 Gy variations was at 18% and 19.2%, respectively. The frequency of variants was highest in 40 Gy followed by 30 Gy and 20 Gy, while it was very low in 60 Gy except for plant stature (dwarfism or stunted growth). Earliness to flowering variants were recorded at 20, 30 and 40 Gy at low frequencies (0.6%, 0.7% and 1.7%) respectively, while none was observed for 60 Gy treatments. As in vitro mutation induction could create genetic variability as well as many undesirable variants, it is highly desirable to integrate in vitro mutation with a selection system that can screen for large mutagen treated population. The useful variants recorded for earliness to flowering were selected for 30 and 40 Gy treatments. 40 Gy showed high frequency in earliness as compared with 20 and 30 Gy., which came to flowering as early as 6 – 6.5 months compared to 7-8 months for control. The useful dwarf, which considered as desirable traits, showed a balance of height and girth.
Mutation Induction for Improvement of Banana (Musa Spp.) Berangan Cv. Intan-AAApaperpublications3
Abstract: In vitro mutation induction by using gamma ray at 20, 30, 40 and 60Gy was used to generate variability in triploid banana "Berangan cv. Intan (AAA), so as to provide the opportunity to select plants with desirable characters such as early fruiting and short stature. Mutation frequency increased with increased dosage whereas, survival and capacity to regenerate decreased with increased doses. Time to initiation varied from 4-8 weeks for gamma-irradiated materials compared to 2-3 weeks in the control. It appeared that the higher the dose, the longer it took for shoot initiation. The exposure of shoot-tip meristem pieces to radiation doses produced wide variation in growth and morphogenetic performance. Mutagenic treatments induced 2 to 3- fold increases in variability in both quantitative and qualitative traits at different stages, in vitro; at nursery and field. For the field-grown plants; the proportion varied from 2.9% for the control plants to 16.8% for 60 Gy and 20.1% for 40 Gy-treatments, while for treatments at 20 and 30 Gy variations was at 18% and 19.2%, respectively. The frequency of variants was highest in 40 Gy followed by 30 Gy and 20 Gy, while it was very low in 60 Gy except for plant stature (dwarfism or stunted growth). Earliness to flowering variants were recorded at 20, 30 and 40 Gy at low frequencies (0.6%, 0.7% and 1.7%) respectively, while none was observed for 60 Gy treatments. As in vitro mutation induction could create genetic variability as well as many undesirable variants, it is highly desirable to integrate in vitro mutation with a selection system that can screen for large mutagen treated population. The useful variants recorded for earliness to flowering were selected for 30 and 40 Gy treatments. 40 Gy showed high frequency in earliness as compared with 20 and 30 Gy., which came to flowering as early as 6 – 6.5 months compared to 7-8 months for control. The useful dwarf, which considered as desirable traits, showed a balance of height and girth.
nduced Mutation by Colchicine Treatment of Somatic Embryos in ‘Namwa’ Banana ...drboon
Hexaploids of the ‘Namwa’ banana (Musa sp ABB) were obtained by in vitro colchicine treatment of somatic embryos. Somatic embryos were induced on a medium containing MS medium supplemented with 8 mg/l picloram. Somatic embryos were treated with four different concentrations of colchicine (0, 0.3, 0.5, 1.0, %) in liquid MS medium supplemented with 0.22 mg/l zeatin, and shaken (60 rpm) at 25 0C in darkness for 48, 72 or 96 hours. Higher colchicine concentrations exhibited higher mortality rates ranging from 8–20%, 48–62% and 80–90% mortality on concentrations 0.3, 0.5, and 1.0 % colchicine respectively. Mortality rate generally increased with increased treatment time. Hexaploids were obtained at a frequency of 2 % with treatments 0.05 % colchicine for 96 hours, and 1 % colchicine for 48 hours as determined by flow cytometry.
Banana plantlet production through tissuecultureAbed Hawas
Banana plantlet production through tissueculture
Banana plantlet production through tissuecultureBanana plantlet production through tissuecultureBanana plantlet production through tissuecultureBanana plantlet production through tissuecultureBanana plantlet production through tissuecultureBanana plantlet production through tissuecultureBanana plantlet production through tissueculture
International Journal of Pharmaceutical Science Invention (IJPSI) is an international journal intended for professionals and researchers in all fields of Pahrmaceutical Science. IJPSI publishes research articles and reviews within the whole field Pharmacy and Pharmaceutical Science, new teaching methods, assessment, validation and the impact of new technologies and it will continue to provide information on the latest trends and developments in this ever-expanding subject. The publications of papers are selected through double peer reviewed to ensure originality, relevance, and readability. The articles published in our journal can be accessed online.
In vitro mutagenesis of Cymbidium La bell “Anna Belle” by γ-rays irradiation ...IJEAB
The optimum media for multiplication of protocorm like bodies (PLBs) and shoot buds of Cymbidium La bell “Anna Belle” were studied in order to prepare the in vitro samples for irradiation. The values of LD50 (lethal dose of 50% samples) of PLBs, shoot buds and plantlets of tested Cymbidium after cultivation of 4 months were also determined about 35.0, 41.0 and 83.1 Gy, respectively. The addition of oligochitosan played as an very important trigger for promotion on the generation of shoot bud from PLBs after irradiation. The in vitro variations have been generated by γ-rays irradiation of PLBs with doses in range of 20 - 50 Gy. The highest mutant frequency (3.83‰) of C. La bell was found by the irradiation of PLB samples at 30 Gy. The different properties of obtained in vitro variations compared to wild types were found to be chlorophyll, short leaves, long leaves, and violet pericardium variations. The genetic relationships among generated variant lines in M1V4 and wild type were analyzed using RAPD techniques.
STUDY OF FUNGAL, BACTERIAL AND ACTINOMYCETES POPULATION IN TENDU LEAF LITTER ...Dr Dama
STUDY OF FUNGAL, BACTERIAL AND ACTINOMYCETES POPULATION IN TENDU LEAF LITTER VERMICOMPOST IN COMPARISON WITH PRESS MUDCAKE.
Mushan L.C*. Rao K.R.**, Shagalolu V.V*. and Dama L.B*@.
Similar to Production of Haploids Plants from Anther Culture of Musa Paradisiaca cv. ‘Puttabale’ (20)
Teacher’s Accomplishment Level of The Components of an E-Learning Module: A B...RSIS International
This study determined the extent to which teachers in a private institution in La Trinidad, Benguet, Philippines have accomplished the essential components of an E-learning module, and identified the factors that influenced their level of accomplishment of these components. This study used mixed method explanatory sequential design. Total enumeration was used to determine the population of respondents who were full-time tertiary teachers. Out of the thirty-six full-time teachers, twenty-eight teachers responded and answered the self-assessment survey questionnaire for the quantitative phase, while seven of them who were selected using purposive sampling were interviewed for the qualitative part. Descriptive statistics using weighted mean was used to analyze quantitative data while descriptive approach using thematic analysis for the qualitative data. Quantitative descriptive analysis revealed that the teachers’ overall accomplishment level of the criteria for a quality E-learning module is partial/moderate (2.68 overall weighted mean/WM). Specifically, the teachers partially accomplished the following components of the E-learning module: instructional design (2.84 WM); communication, interaction, and collaboration (2.85 WM); student evaluation and assessment (2.89 WM); instructional materials and technologies (2.75 WM); and accessibility (2.66 WM) while slightly accomplished the components: learner support and resources (2.21 WM), and course evaluation (2.24 WM). For the qualitative descriptive inquiry, absence of capacity-building training in making E-learning module; lack of awareness of the school services, learner support, and resources sites; lack of time; lack of systemic approach to developing an online module; non-consistency of instructions; and amotivation and lack of enthusiasm emerged as factors that influence the teachers’ level of accomplishment of the parts of an E-learning module. The results show that the E-learning modules have fallen short of strict quality standards attributable to certain dire circumstances. This study thus recommends that the institution may adopt or develop its online module criteria to guide the teachers as well as the institution in writing or designing their online modules, and may conduct training in E-module design for teachers.
Development Administration and the Challenges of Neo-liberal Reforms in the E...RSIS International
The quest of every nation state is to attract, sustain and fast-track growth and development in all ramifications. The Nigerian state between 1960 and 2020 has experienced an unabated expansion of universities. Ironically, the war against illiteracy is yet to be won, despite the experimentation of different western developmental ideologies in the Nigerian educational system. The study examined the contributions of development administration in the Nigerian educational sector as well as ascertained if the current neo-liberal reform has aided the expansion or retrogression of the educational sector most especially, the university sub-sector in Nigeria. The theoretical framework of the study was anchored on the post-colonial Nigerian state theory. Methodologically, the study strictly utilized the documentary method and data were sourced through the secondary sources and analyzed in content. The study found that the experimentation of western development ideologies in Nigeria actually aided the expansion of the universities but undermined the realization of free, quality and accessible university education for all and sundry. The study recommended among others: the applications of the action plan of Professor Okonjo as regards creating a universal tertiary education for all Nigerians and sustainably financing it.
The Nexus of Street Trading and Juvenile Delinquency: A Study of Chanchaga Lo...RSIS International
I. INTRODUCTION
Globally, the number of working children has been decreasing around the world in recent years, but child labour has continued to be a widespread problem today, especially in developing countries (Paola, Viviana, Flavia & Furio2007). International Programme on Elimination of Child Labour (IPEC 2016) reported that between 2012 to 2016, about 182 million children in the developing world aged 5-14 years were engaged in work. Against this background, governments, international organizations, and non-governmental organizations (NGOs) have focused their efforts on tackling in particular the worst forms of child labour such as forced and bonded labour, which put children in physically and mentally harmful working conditions (Bunnak 2007).
Determination of Bacteriological and Physiochemical Properties of Som-Breiro ...RSIS International
The study seeks to examine the Bacteriological and
physiochemical properties of Sambrero River in Ahoada East
Local Government Area of Rivers State. Three (3) points were
sampled from different locations designated as location (L1)
location (L2) and location (L3) respectively, samples were
collected in 0.1m of Sterile containers and were transported to
the laboratory for immediate analysis. Ten (10) physiochemical,
three (3) heavy metal sand three microbiological parameters
were observed. Data was analyzed using standard methods
(ALPHA, 1998) 20th edition and Ms-Excel version 2013 software.
The result showed little variation in physiochemical parameters
which are in line with World Health Organization (WHO)
standard of potable water but shows much variation in
microbiological parameters which are not in line with WHO
standard, thereby making the water not wholesome and not
potable for consumption except after proper treatment of the
water. The work therefore recommends that members of Ekpena
Community should ensure basic water treatment such as boiling
and chlorination before consumption.
Power and Delay Analysis of Logic Circuits Using Reversible GatesRSIS International
This paper determines the propagation delay and on
chip power consumed by each basic and universal gates and
basic arithmetic functions designed using existing reversible
gates through VHDL. Hence a designer can choose the best
reversible gates to use for any logic circuit design. The paper
does a look up table analysis of truth tables of the reversible
gates to find the occurrence of the AND OR, NAND, NOR and
basic arithmetic functions, useful to build complex combinational
digital logic circuits.
Innovative ICT Solutions and Entrepreneurship Development in Rural Area Such ...RSIS International
The use of internet and information communication
technology (ICT) infrastructures is an essential aspect of
learning, this is why a lot of information on entrepreneurship
career choices are available online. However, the emerging
growth in the use of information and communication
technologies and services towards entrepreneurship development
is a challenge for efficient information dissemination and
learning especially in rural areas. This paper pointed out an area
in which MCIU can use Information and Communication
Technology (ICT) resources/infrastructure it possess for
entrepreneurship development and poverty alleviation in its
community. Thereby, encouraging social and economic growth,
and overcome the gap between urban and rural areas
entrepreneurship development. An online learning platform,
using video may contribute greatly in rural entrepreneurship
development such as MCIU community. Some examples of some
programmes like learning make over, headgear tying, bead
making, cake designing, etc online for a period of 4 to 6weeks
Indigenous Agricultural Knowledge and the Sustenance of Local Livelihood Stra...RSIS International
Natural disasters in most parts of the world have
resorted to many fatalities, forced migration and involuntary
resettlement of the affected population. Lake Nyos Gas Disaster
of 1986 which killed about 1,746 people and led to forced
migration of over 15,000 affected people and the subsequent
resettlement of survivors in resettlement camps in near by
administrative sub-divisions in the North West Region of
Cameroon is one of such natural catastrophies. The paper
evaluates the use of Indigenous Agricultural Knowledge (IAK) in
agriculture and how it has helped to sustain the livelihood of this
environmentally traumatized resettled population. The study
sampled two of these resettlement villages (Buabua and Kimbi)
to access Indigenous Agricultural Knowledge (IAK) and the
sustenance of local livelihood strategies. Field campaigns
including the administration of semi-structured questionnaires
and focus group discussions (FGDs) facilitated the collection of
data on IAK practices and how this knowledge helps in
sustaining local livelihoods. A total of 24 Indigenous Agricultural
Knowledge (IAK) were identified, with 54.16% of them used in
crop cultivation, and 45.83% in livestock farming including the
raising of small ruminants, poultry and piggery production. IAK
shows successful results after being applied as there is increased
crop and livestock yields. The use of IAK in agriculture has led
to sustainable and efficient land use within the study area.
Despite the rising use of IAK and potential benefits in
agriculture and the sustenance of local livelihoods in Buabua and
Kimbi, survivors still express a strong desire to return to the
former disaster zone. The underlying reasons behind this phobia
is mainly small land sizes ranging from 30-50 square metres that
were allocated to households for both crop cultivation and
grazing, and the fact that the limits between grazing and crop
land are not clearly demarcated. Prospects for agricultural
expansion within the area are therefore slim and need to be
addressed
Wireless radio signal drop due to foliage in illuba bore zone ethiopiaRSIS International
The exponential growth in energy utilization &
consumption in cellular network by the user devices and by
telecom equipment has imposed critical problemsbecause of
adaptation of high range frequency in available spectrum (Ultra
High Frequency-UHF) by government and technology.The other
reason for more power consumption is extensive applications of
mobile data services to video streaming, surveillance, internet
surfing and healthcare monitoring.Other important causes of
energy consumption which has been recognized are powerhungry
processors, poor design of power amplifiers etc. Presence
of different species of foliage in hilly area increases signal
attenuation, consequently in order to maintain the threshold
value of signal, the power is increased. The recent researches
predict that the data traffic is being increased by several-fold
every year. Under such predictions, energy expenditure at its
control is a major challenging task for telecom companies and
for research communities. This paper studies the actual signal
intensity drop because of irregular nature of terrestrial pattern
and foliage in Illuba Bore zone, from theoretical perspective as
well as practical point of view.
The Bridging Process: Filipino Teachers’ View on Mother TongueRSIS International
This document summarizes a study on Filipino teachers' views of using mother tongue as the medium of instruction. The study surveyed 35 elementary teachers at a school in the Philippines. It found that the teachers moderately perceived the mother tongue-based approach as effective and efficient in achieving learning goals. Specifically, teachers agreed that using the mother tongue-based approach, objectives are clear and easy to facilitate. They also agreed that learning tasks can be easily covered within allotted time. The study provides insight into teachers' perspectives on implementing the Philippines' language education policy of using mother tongue in early grades.
Optimization of tungsten inert gas welding on 6063 aluminum alloy on taguchi ...RSIS International
In this paper, the Taguchi method is used for the
Optimization of Tungsten Inert Gas Welding on 6063
Aluminum Alloy. The Taguchi method L27 is used to
optimize the pulsed TIG welding process parameters of 6063
aluminum alloy weldments for maximizing the mechanical
properties. Analysis of Variance is used to find the impact of
individual factors. Then the optimal parameters of the TIG
welding process is determined and the experimental results
illustrate the proposed approach.
Investigation of mechanical properties of carbidic ductile cast ironRSIS International
The objective of the present work is to increase the wear resistance for long life of applications. It is found that increase in the carbides in an alloy which resulted in to enhancement in hardness and wear resistance. The wear resistance was evaluated by testing in accordance with ASTE International Committee G-99 Standard. An improved performance of wear resistanceis obtained by increasing the content of chromium in the carbidic ductile cast iron. The results are discussed based on the influence of chromium content on the casting.
Six Sigma Methods and Formulas for Successful Quality ManagementRSIS International
This paper is about the five phases of Six Sigma which are Define, Measure, Analyze, Improve& Control. The methods used in each phase are discussed in detail and the various tests used in Analyze Phase of Six Sigma are given; Six Sigma can be implemented in an organization by using the methods and formulas used in each phase combined with the help of Statistical Software Minitab 18.
Task Performance Analysis in Virtual Cloud EnvironmentRSIS International
Cloud computing based applications are beneficial for
businesses of all sizes and industries as they don’t have to invest
a huge amount on initial setup. This way, businesses can opt for
Cloud services and can implement innovative ideas. But
evaluating the performance of provisioning (e.g. CPU scheduling
and resource allocation) policies in a real Cloud computing
environment for different application techniques is challenging
because clouds show dynamic demands, workloads, supply
patterns, VM sizes, and resources (hardware, software, and
network). User’s requests and services requirements are
heterogeneous and dynamic. Applications models have
unpredictable performance, workloads, and dynamic scaling
requirements. So a demand for a Simulation toolkit for Cloud is
there. Cloudsim is self-contained simulation framework that
provides simulation and modeling of Cloud-based application in
lesser time with lesser efforts. In this paper we tried to simulate
the task performance of a cloudlet using one data center, one
VM. We also developed a Graphical User Interface to
dynamically change the simulation parameters and show
simulation results.
Design and Fabrication of Manually Operated Wood Sawing Machine: Save Electri...RSIS International
In India power cut is big problem also having many
remote places where electricity not reached and that will affect
many small scale business and ongoing work, like Carpentry,
ongoing work got stop because of power cut. To overcome this
problem manually operated economical; conceptual model of a
machine which would be capable of performing different
operation like sawing/cutting and grinding without use of power
i.e. wood working machine is introduced.
In this paper, design concept and fabrication of manually
operated wood sawing/cutting machine is explained. It is
designed and fabricated so portable that it can be move and used
at various places. It is used for sawing/cutting of wood, plywood,
thin metals (<=2mm), and pvc pipes. The material can be cut
without any external energy like fuel or current. As machine uses
no electric power and fuel, this will help to maintain green
environment. The observations show that power required for
pedaling is well below the capacity of an average healthy human
being.
Effect of Surface Treatment on Settlement of Coir Mat Reinforced SandRSIS International
1) The document discusses a study on the effect of surface treatment on the settlement of sand reinforced with coir mats. Coir mats were treated with sodium hydroxide and epoxy resin to make their surface hydrophobic and increase frictional interaction with soil.
2) Model footing tests were conducted by placing treated and untreated coir mats at different depths in sand beds. Settlement reduction factor was calculated to evaluate the performance of mat reinforcement.
3) The results showed that surface treatment of coir mats significantly increased the settlement reduction factor compared to untreated mats. Surface treatment improved the interaction between the reinforcement and soil, leading to reduced settlement under load.
Augmentation of Customer’s Profile Dataset Using Genetic AlgorithmRSIS International
Data is the lifeblood of all type of business. Clean,
accurate and complete data is the prerequisite for the decisionmaking
in business process. Data is one of the most valuable
assets for any organization. It is immensely important that the
business focus on the quality of their data as it can help in
increasing the business performance by improving efficiencies,
streamlining operations and consolidating data sources. Good
quality data helps to improve and simplify processes, eliminate
time-consuming rework and externally to enhance a user’s
experience, further translating it to significant financial and
operational benefits [1] [2]. All organizations/ businesses strive to
retain their existing customers and gain new ones. Accurate data
enables the business to improve the customer experience. Data
augmentation adds value to base data by enhancing information
derived from the existing source. Data augmentation can help
reduce the manual intervention required to develop meaningful
information and insight of business data, as well as significantly
enhance data quality. Hence the business can provide unique
customer experience and deliver above and beyond their
expectations. The Data Augmentation is immensely important as
it helps in improving the overall productivity of the business. It
is also important in making the most accurate and relevant
information available quickly for decision making.
This work focuses on augmentation of the customer
dataset using Genetic Algorithm(GA). These augmented data are
used for the purpose of customer behavioral analysis. The data
set consists of the different factors inherent in each situation of
the customer to understand the market strategy. This behavioral
data is used in the earlier work of analyzing the data [13]. It is
found that collecting a very large amount of such data manually
is a very cumbersome process. It is inferred from the earlier
work [13] that the more number of data may give accurate
result. Hence it is decided to enrich the dataset by using Genetic
Algorithm.
System Development for Verification of General Purpose Input OutputRSIS International
In SoC no. of IP block inside it depends upon specific
application, increase in the Ip block increases no. of digital
control lines causes increase in the size of the chip. GPIO helps
internal IP blocks to share digital control lines using MUX and
avoids additional circuitry. Since design productivity cannot
follow the pace of nanoelectronics technology innovation, it has
been required to develop various design methodologies to
overcome this gap. In system level design, various design
methodologies such as IP reuse, automation of platform
integration and verification process have been proposed. GPIO
configuration register decides in which mode system has to work
GPIO has four modes i.e input, output, functional, interrupt. As
per operation particular mode is selected and the operation get
performed. Devices with pin scarcity like integrated circuits such
as system-on-a-chip, embedded and custom hardware, and
programmable logic devices cannot compromise with size can
perform well without additional digital control line circuitry.
De-noising of Fetal ECG for Fetal Heart Rate Calculation and Variability Anal...RSIS International
Fetal monitoring is the way of checking the condition
of unborn baby during labor and delivery by continuously
monitoring his or her heart rate. A normal fetal heart rate (FHR)
can reassure safe birth of the baby. Fetal monitoring techniques
are broadly classified into invasive and non-invasive techniques.
Non-invasive techniques are involves monitoring the fetus
through mother’s abdominal region. This can be done in all
gestation weeks and during the delivery also. Abdominal ECG
(AECG) is a composite ECG signal containing both mother’s as
well as fetal ECG. This paper presents an efficient technique to
extract FECG from abdominal ECG. A modified Pan Tompkin’s
method is employed for the QRS detection. It involves series of
filters and methods like band pass filter, derivative filter,
squaring, integration and adaptive thresholding. Further heart
rate of fetus and mother is calculated and heart rate variability
analysis is done using detected R-peaks. The algorithm is tested
on 5 different non-invasively recorded abdominal and direct
FECG signals taken from MIT PhysioNet database and the
results are obtained using MATLAB software. The performance
of the QRS detector is evaluated using parameters like
Sensitivity and Positive Prediction.
This document summarizes a study on active vibration control of composite plates using piezoelectric materials. Finite element models of composite beams with integrated piezoelectric layers were developed in ANSYS to simulate displacement feedback and direct velocity feedback control loops. The models were validated by comparing natural frequencies to experimental results. Vibration suppression was then analyzed for different composite layups under free and forced vibration conditions. The results showed that both feedback control methods successfully reduced vibration levels, with direct velocity feedback achieving steady state more quickly. Higher control gains led to greater vibration attenuation in both free and forced vibration cases.
Batteries -Introduction – Types of Batteries – discharging and charging of battery - characteristics of battery –battery rating- various tests on battery- – Primary battery: silver button cell- Secondary battery :Ni-Cd battery-modern battery: lithium ion battery-maintenance of batteries-choices of batteries for electric vehicle applications.
Fuel Cells: Introduction- importance and classification of fuel cells - description, principle, components, applications of fuel cells: H2-O2 fuel cell, alkaline fuel cell, molten carbonate fuel cell and direct methanol fuel cells.
Introduction- e - waste – definition - sources of e-waste– hazardous substances in e-waste - effects of e-waste on environment and human health- need for e-waste management– e-waste handling rules - waste minimization techniques for managing e-waste – recycling of e-waste - disposal treatment methods of e- waste – mechanism of extraction of precious metal from leaching solution-global Scenario of E-waste – E-waste in India- case studies.
Comparative analysis between traditional aquaponics and reconstructed aquapon...bijceesjournal
The aquaponic system of planting is a method that does not require soil usage. It is a method that only needs water, fish, lava rocks (a substitute for soil), and plants. Aquaponic systems are sustainable and environmentally friendly. Its use not only helps to plant in small spaces but also helps reduce artificial chemical use and minimizes excess water use, as aquaponics consumes 90% less water than soil-based gardening. The study applied a descriptive and experimental design to assess and compare conventional and reconstructed aquaponic methods for reproducing tomatoes. The researchers created an observation checklist to determine the significant factors of the study. The study aims to determine the significant difference between traditional aquaponics and reconstructed aquaponics systems propagating tomatoes in terms of height, weight, girth, and number of fruits. The reconstructed aquaponics system’s higher growth yield results in a much more nourished crop than the traditional aquaponics system. It is superior in its number of fruits, height, weight, and girth measurement. Moreover, the reconstructed aquaponics system is proven to eliminate all the hindrances present in the traditional aquaponics system, which are overcrowding of fish, algae growth, pest problems, contaminated water, and dead fish.
A SYSTEMATIC RISK ASSESSMENT APPROACH FOR SECURING THE SMART IRRIGATION SYSTEMSIJNSA Journal
The smart irrigation system represents an innovative approach to optimize water usage in agricultural and landscaping practices. The integration of cutting-edge technologies, including sensors, actuators, and data analysis, empowers this system to provide accurate monitoring and control of irrigation processes by leveraging real-time environmental conditions. The main objective of a smart irrigation system is to optimize water efficiency, minimize expenses, and foster the adoption of sustainable water management methods. This paper conducts a systematic risk assessment by exploring the key components/assets and their functionalities in the smart irrigation system. The crucial role of sensors in gathering data on soil moisture, weather patterns, and plant well-being is emphasized in this system. These sensors enable intelligent decision-making in irrigation scheduling and water distribution, leading to enhanced water efficiency and sustainable water management practices. Actuators enable automated control of irrigation devices, ensuring precise and targeted water delivery to plants. Additionally, the paper addresses the potential threat and vulnerabilities associated with smart irrigation systems. It discusses limitations of the system, such as power constraints and computational capabilities, and calculates the potential security risks. The paper suggests possible risk treatment methods for effective secure system operation. In conclusion, the paper emphasizes the significant benefits of implementing smart irrigation systems, including improved water conservation, increased crop yield, and reduced environmental impact. Additionally, based on the security analysis conducted, the paper recommends the implementation of countermeasures and security approaches to address vulnerabilities and ensure the integrity and reliability of the system. By incorporating these measures, smart irrigation technology can revolutionize water management practices in agriculture, promoting sustainability, resource efficiency, and safeguarding against potential security threats.
Presentation of IEEE Slovenia CIS (Computational Intelligence Society) Chapte...University of Maribor
Slides from talk presenting:
Aleš Zamuda: Presentation of IEEE Slovenia CIS (Computational Intelligence Society) Chapter and Networking.
Presentation at IcETRAN 2024 session:
"Inter-Society Networking Panel GRSS/MTT-S/CIS
Panel Session: Promoting Connection and Cooperation"
IEEE Slovenia GRSS
IEEE Serbia and Montenegro MTT-S
IEEE Slovenia CIS
11TH INTERNATIONAL CONFERENCE ON ELECTRICAL, ELECTRONIC AND COMPUTING ENGINEERING
3-6 June 2024, Niš, Serbia
KuberTENes Birthday Bash Guadalajara - K8sGPT first impressionsVictor Morales
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Production of Haploids Plants from Anther Culture of Musa Paradisiaca cv. ‘Puttabale’
1. International Journal of Research and Scientific Innovation (IJRSI) | Volume IV, Issue III, March 2017 | ISSN 2321–2705
www.rsisinternational.org Page 1
Production of Haploids Plants from Anther Culture of
Musa Paradisiaca cv. ‘Puttabale’
K. Girish Kumar1
, V. Krishna2
*, Venkatesh3
, R. Shashikumar4
, H. S. Arunodaya5
1, 2, 4, 5
Department of P. G. Studies and Research in Biotechnology, Bioscience Block, Kuvempu University, Jnana Sahyadri,
Shankaraghatta‐577 451, Shivamogga (Dist.), Karnataka, India.
3
Department of Biochemistry Indian Institute of Science Bangalore. Karnataka, India
* Corresponding author: V. Krishna
Abstract: Haploid plants were regenerated from the anther
callus of banana Musa paradisiaca (AB) cv. Puttabale. The
highest frequency of callus induction (90%) was observed
at the concentration of 3mg/l 2, 4-D . After 20 days of
incubation organization of embyroids were organised
from the callus mass. Interaction of 4mg/l BAP and 0.4
mg/l IAA provoked shoot growth of the embryoids and
well organised roots were developed at the concentration
of 0.6 mg/l NAA and the media was agumented with 0.2%
activated charcoal. Flow cytometry study was carried out
to analyse the DNA content of the regenerated haploid
plants. The results of the investigation reported the
efficient production of haploid plants from the anther
culture.
Key words: Musa paradisica, puttabale, anther culture,
haploid plants
I. INTRODUCTION
anana and plantain (Musa spp.) are the world’s major
food crop for millions of people. Many breeders
improved the quality by manipulating the ploidy of this crop
(Pillay et al., 2003). But the conventional breeding method is
laborious and took more generations for the development of
haploid plants. The production of haploids through anther
culture has many advantages since it does not imply the
inbreeding depression caused by selfing and does not involve
pollinator genome contamination. Many factors are involved
and interact in the optimization of the anther culture protocol
to produce haploid plants; therefore it is necessary to consider:
pre‐treatment of anthers, genotype effects, the stage of
pollen development, culture conditions and induction of
embryos and regeneration of plants. Anther or microspore
cultures have been found to be the most efficient techniques
for obtaining a large number of haploid plants (De Buyser and
Henry.,et al 1980). Haploid production is extremely important
for crop improvement and eradication of pathogens (Brown
and Thorpe et al., 1995) and dramatically improves the yield.
(Badoni and Chauhan et al., 2010). Anther or microspore
cultures has proved to be the most efficient techniques for
obtaining a large number of haploid plants (Buyser and Henry
et al., 1980) and valuable in plant breeding and genetics
studies (Custódioa et al., 2005). The establishment of
homozygous lines of new varieties is possible in a short
period of time, hence provide a useful tool to accelerate plant
breeding successions (Devaux and Li et al., 2001; Zhang et
al., 2011). Haploids can obtained through natural
parthenogenesis or androgenesis. Artificial induction of
ovaries (Muren et al., 1989), ovules (Hansen et al., 1994),
anthers (Foroughi et al., 1982), microspores (Kohler and
Wenzel et al., 1985) is also a promising tool for haploid
production.
In the genus Musa, accurate determination of the
ploidy by phenotypic traits, including stomatal size, density,
and pollen size, and by chromosome counting is difficult and
laborious, mainly due to strong genotypic influences (Hamill,
1992; Vandenhout et al., 1995; Van Duren et al., 1996;
Dolezel, 2004). Musa paradisiaca cv. Puttabale (AB group)
belongs to family Musacae and an indigenous banana cultivar
of Karnataka, India. Previously we published the high
frequency plant regeneration protocol from shoot tip, leaf and
immature male floral explants. No reports available on
haploid production from this plant. This paper deals with the
in vitro haploid plant regeneration from anther culture of
banana cultivar Puttabale.
II. MATERIALS AND METHOD
Explants Collection And Sterlization
Banana cv puttubale inflorescence of appropriate
stage was collected from farmyard of Shimoga district,
Karnataka. The inflorescence were disinfected in 2% sodium
hypochloride solution for 15 min, followed by 8-10 rinses in
sterile distilled water.The immature male flowers are detached
from bract, tepals aseptically. Male flowers contained five
stamens with fully developed anthers. Anthers from the
stamens was carefully isolated and transfered into pertriplates.
Anthers with uninucleated microspore stage was determined
using 1% aceto-carmine stain at this stage anthers were
inoculated on to the callusing media.
Plant Regeneration
B
2. International Journal of Research and Scientific Innovation (IJRSI) | Volume IV, Issue III, March 2017 | ISSN 2321–2705
www.rsisinternational.org Page 2
The callusing media consisted of MS media
constituents agumented with 40 g/l sucrose, 8 g/l agar,
160 mg/l adenine sulfate, 100 mg/l tyrosine and the growth
regulator 2, 4-D tested at the range of 1 to 6 mg/l. The
pH was adjusted to 5.8 before autoclaving at 121ºC for
20 min. The cultures were incubated at 25 ± 2ºC with
12 hrs photoperiod and the callogenic frequency
was evaluated after 30 days of incubation. Regenerated
androgenic embryos like structures developed from callus
were transferred to proliferation medium containing different
conentrations of BAP (2 to 6 mg/l) and IAA( 0.2 to 0.6 mg/l)
the media was agumented with 100mg/l of adanine sulphate
and tyrosin. The well grown elongated shoots were
aspectically transferred to the medium containg 0.25 to
o.75mg/l NAA and without hormones fortified with 0.2%
activated charcoal for the proper development of root system.
The plantlets from culture bottles are moved from the
laboratory later they are shifted to green house for primary
hardening where they are first gently washed free of agar
medium. The well rooted plants were washed with 0.2% of
bovastin solution then followed by primary hardening in
cocopeats, maintained in polythene house for a period of two
weeks. During this period, 90-95 % humidity is maintained
for the initial 6-8 days under diffused light, later they were
maintained at 70 %, humidity and 26 °C Then transferred to
green house for secondary hardening in a polythene bags
containing red soil, sand and cattle manure in the ratio of
1:1:2 respectively. The regenarents grew to the height of 10
to 15 cm were transferred to field condition.
Determination of Ploidy Level
The flow cytometric technique was followed to
determine the ploidy level of the anther-derived plants. Leaf
pieces (1×1 cm) from in vitro plants derived from anther
cultures were chopped with a razor blade in 600 μl buffer
solution consisting of 45 mM MgCl2, 30 mM Tri-sodium
citrate (Na3C6H5O7, 2H2O), 20 mM MOPS, 1% triton X-
100, 10 mM sodium-metabisulfite (Na2S2O5) (pH 7) to
obtain a nuclei suspension. The whole sample was sieved
through a 40-μm mesh nylon filter and stained with 6 μg ml-1
DAPI solution. A diploid parental plant was used as an
internal standard. Nuclei analysis was carried out by using
Partec CA II flow cytometer following the method of Assani
et al., 2003.
III. RESULTS AND DISCUSSION
The anthers isolated aseptically were inoculated on to
the callusing media (1 mg/l. 2,4D to 6 mg/l. 2,4D) showed
callogenic response within a week of incubation. Prior to
inoculation anthers were selected at the diod or tetraoid stage
of meiosis (Fig A) Optimum callogenesis noticed at the
concentration of 3 mg/l. 2,4D (Table.1). On callusing media
anthers began to swell and the size increases to three folds.
After ten days of incubation, callogenic mass noticed from the
dorsal side while the anther wall cells turns black (Fig B).
After 20 days of culture organization of embyronic nodules
were appeared from the sprouted callus mass. The formation
of androgenic embryroids from the protruded callus was
similar to the findings of Assani et al., 2002.
The interaction of BAP at higher levels (2.0 – 6.0
mg/l) and IAA at lower levels (0.2 ‐ 0.6 mg/l) provoked the
growth and development of the enbryonic mass in to shoot
buds. The mean number of shoots organized at each
combination of BAP and IAA is shown in the (Table 1). The
shoot multiplication was optimized at the concentration of 4
mg/l BAP and 0.4 mg/l IAA with a mean of 7.10±0.88 shoots
per callus. shoot buds. The caulogenic mass with multiple
shoots were subjected to colchecin treatment with a range of
(0.025% and 0.05%). Then they were transferred to the
development media consisted of 0.6 mg/l NAA and 0.2%
activated charcoal. The well rooted shoots were maintained
on the media for two to three cycles, then they were
transferred to the rhizogenic media.
The induction of roots depends on the composition of
mineral nutrients and growth regulators. Shoot buds were
transferred to MS with 0.6 mg/l NAA, 0.2% activated
charcoal initiated roots from the base of microshoots ( Fig E),
after three weeks of culture. The well rooted plantlets (Fig H)
were hardened primarily in coco peats containing equal
proportion of garden soil, sand, and cattle dung manure in
polythene house for a period of two weeks (Fig I) and then
they were subjected to secondary hardening in perforated
polythene bags in green house (Fig J). The presence of diploid
plants was also observed among the regenerants. While this
could be a consequence of the regeneration of diploid anther
tissues such as anther wall or connective tissue, in monocots
the regeneration of somatic anther tissue has been very rarely
reported. The other possibility is the spontaneous
chromosome doubling in haploid cells. The possible factors
leading to spontaneous doubled-haploid plants could be
nuclear fusion in the early divisions of the microspores,
endomitosis, endoreduplication or multipolar mitosis during
the callus phase (Chen et al.1982; De Buyser and Henry1986).
The polar transport of auxin is essential for the establishment
of bilateral symmetry during embryogenesis in
dicotyledonous and monocotyledonous species (Fisher,c, et
al.1996).
Androgenesis has been traditionally divided in two
main stages, namely induction and regeneration. In the former
one, somatic cells acquire embryogenic characteristics by
means of a complete reorganization of the cellular state,
including physiology, metabolism and gene expression
(Feher, A. et al. 2002). In the present study also androgenic
embryoids developed from the primary callogenic mass and
upon subculture on the cytokinin supplemented media
embryoids grew up in to plantlets.
Many authors have reported that the developmental
stage of anthers can drastically affect plant regeneration
because microspores respond only at a specific development
stage, which ranges from the tetrad, early and mid-uninucleate
to the early binucleate stage. As such anther of the Lilium ×
3. International Journal of Research and Scientific Innovation (IJRSI) | Volume IV, Issue III, March 2017 | ISSN 2321–2705
www.rsisinternational.org Page 3
‘Enchantment’, excised at the uninucleate microspores stage
could form callus and regenerate bulblets (Niimi et al., 2001).
Medium is another important factor affecting anther culture.
Custódio et al. (2005) reported that the best medium for calli
induction of carob tree was MS media supplemented with 2.3
μM 2, 4-D + 8.2 μM TDZ. Plant regeneration from anther of
banana (Assani et al., 2003) was obtained in 4.4 μM BAP +
2.3 μM.
While genotype or days in vitro are factors that favor
the frequency of appearance of DNA polyploid plants or even
DNA aneuploids (Tremblay et al. 1999; Endemann et al.
2001; Currais et al. 2013), Therefore, the changes observed in
nuclear DNA content might be caused by the effect of growth
regulators used in the culture media. We noticed that synthetic
auxin 2, 4-D might be involved in embryoid formation from
the microspore mass, because it was found to be a factor
favouring changes in the genome of in vitro regenerated plants
(May and Sink 1995; Jin et al. 2008; Clarindo et al. 2008).
Additional assays would be useful to confirm the possible role
of 2,4-D as a factor promoting the variations observed, as well
as NAA or BAP, which were also mentioned as possible
mutagenic agents (Lim and Loh 2003; Mishiba et al. 2006;
Barow and Jovtchev 2007).
The histogram developed from the result of
cytometric analysis (Fig. 2 .A and B) showed a prominent
peak of nuclei in PI fluorescent intensity in Gi l and G1 stages
of interphase of the doubled diploid cells. The intensity of the
nuclei in the interphase of doubled diploid plant and it is two
folds as compared to haploid cells. Extensive fluorescent
emissions at higher intensities are the indicative of
populations of nuclei at increased ploidy levels. Flow
cytometric analyses was performed by measuring the leaf
DNA intensity of the in vitro regenerated plants. The report
of Jin et al. (2008) suggested that detection of plants with
lower nuclear DNA intensity value is due to the existence of
aneuploid plants. This assumption is based on the fact that
many of the changes that can be detected by FCM are
associated with variations in the number of chromosomes
(Clarindo et al. 2008; Currais et al. 2013). These variations
might be enough to indicate the complete or partial loss of a
chromosomes during doubling process (Pfosser et al. 1995;
Roux et al. 2003).The flow cytometric studies enlightened
many investigators to detect plants with values of nearly
twice the nuclear DNA content, suggesting a possible in vitro
polyploidization (Barow and Jovtchev 2007).
IV. CONCLUSION
Anther culture is the classical method employed in
the regeneration of haploid plants but in conventional
breeding production of haloid plant require screening of many
generations and it is difficult in pseudo fruit crops like banana.
The results of the investigation reported here showed the
efficient production haploid plants from the anther culture of
Musa paradisica cv. Puttabale
Fig. 1 High frequency regeneration of plantlets from anther explants of Musa
paradisica cv. Puttabale.
A. Uninuleate stage of anther. B. Development of anther callus. C and D.
Androgenic embryos formed from the calli. E and F. Shoot growth of the
embroids. G. Growth of haploid plants. H. Root induction from the haploid
plants. I. Primary hardening plants. J. Secondary hardened haploid plants.
A B
C D
E F
G H
I J
4. International Journal of Research and Scientific Innovation (IJRSI) | Volume IV, Issue III, March 2017 | ISSN 2321–2705
www.rsisinternational.org Page 4
Figure 2
Histograms of relative DNA content obtained after flow cytometric analysis of nuclei isolated from (A) leaf tissue of the diploid banana plant, and (B) leaf tissue
of the haploid banana plant
Table 1 Effects of growth regulators on frequency of anther callogenesis and plant regeneration from Musa paradisiaca cv.
Puttabale
Plant growth regulators mg/L Frequency of callus formation per
explant
%
Number of multiple shoots per propagule
Mean ± S.D
2, 4 -D mg/L BAP mg/L IAA mg/L
1
2
3
4
5
6
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
F Value
-
-
-
-
-
-
2
2
2
3
3
3
4
4
4
5
5
5
6
6
6
-
-
-
-
-
-
0.2
0.4
0.6
0.2
0.4
0.6
0.2
0.4
0.6
0.2
0.4
0.6
0.2
0.4
0.6
16.66
36.66
90.00
56.66
43.33
33.33
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
-
0.90±0.74
1.60±0.70
1.30±0.95
2.00±0.47
3.00±0.67
2.10±0.57
3.20±1.14
7.10±0.88
4.40±1.71
4.00±1.15
5.50±0.97
4.60±1.35
3.30±1.16
5.40±1.58
4.20±1.03
27.3
The value of combination consisted of mean ± S.D. of 10 replicates.
The F-value is significantly different when p< 0.05.
5. International Journal of Research and Scientific Innovation (IJRSI) | Volume IV, Issue III, March 2017 | ISSN 2321–2705
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REFERENCES
[1]. Assani , F. Bakry ,F. Kerbellec, R. Haicour G. Wenzel, B.
Foroughi-Wehr (2003) Afza R, Shen M, Zapata J, Xie J, Fundi H,
Lee K, Bobadilla E, Kodym A(2000)., Effect of spikelet
position on rice anther culture efficiency., Plant Sci. 153:155-159.
[2]. Assani A, Bakry F, Kerbellec F, Haicour R, Wenzel G, Foroughi
B (2003)., Production ofhaploids from anther culture of banana
[Musa balbisiana (BB)]., Plant Cell Rep., 21: 511-516.
[3]. Assani A, Haicour R, Wenzel G, Cote FX, Bakry F, Foroughi-
Wehr B, Ducreux G, AguillarME, Grapin A (2001) Plant
regeneration from protoplasts of dessert banana Grande Naine
(Musa spp., Cavendish sub-group AAA) via somatic
embryogenesis. Plant Cell Rep 20:482–488.
[4]. Assani A, Haïcour R, Wenzel G, Foroughi-Wehr B, Bakry F, Côte
FX, Ducreux G, Ambroise A, Grapin A (2002) Influence of donor
material and genotype on protoplast regeneration in banana and
plantain cultivars (Musa spp.). Plant Sci 162:355–362.
[5]. Barow M, Jovtchev G (2007) Endopolyploidy in plants and its
analysis by flow cytometry.In: Dolezel J, Greilhuber J, Suda J
(eds) Flow cytometry with plant cells. Wiley,
Weinheim,Bronsema F, Van O, Prinsen E, Van L(1998).,
Distribution of (14C) dichlorophenoxyacetic acid in cultured
zygotic embryos of Zea mays L., JPlant Growth Regul.,17:81–88.
[6]. Barow M, Jovtchev G (2007) Endopolyploidy in plants and its
analysis by flow cytometry. In: Dolezel J, Greilhuber J, Suda J
(eds) Flow cytometry with plant cells. Wiley, Weinheim, pp
349–372.
[7]. Chen Z, Qian C, Qin M, Xu X, Xiao Y (1982) Recent advances in
anther culture of Hevea brasiliensis (Muell.-Arg.). Theor Appl
Genet 62:103–108.
[8]. Clarindo WR, de Carvalho CR, Arau´jo FS, de Abreu IS, Otoni
WC (2008) Recovering polyploid papaya in vitro regenerants as
screened by flow cytometry. Plant Cell Tissue Organ Cult 92:207–
214.
[9]. Currais L, Loureiro J, Santos C, Canhoto JM (2013) Ploidy
stability in embryogenic cultures and regenerated plantlets of
tamarillo. Plant Cell Tissue Organ Cult 114:149–159.
[10]. Custódioa L, Carneirob MF, Romanoa A (2005).
Microsporogenesis and anther culture in Ceratonia siliqua L.).
Scientia Horticulturae, 104: 65-77.
[11]. De Buyser J, Henry Y (1980) Induction of haploid and diploid
plants through in vitro anther culture of haploid wheat (n=3x=21).
Theor Appl Genet 57:57–58.
[12]. De Buyser, J. and Henry, Y., Wheat production of haploids,
performance of doubled haploids, and yield trials. Biotechnology
in Agriculture and Forestry, Transgenic Crops I (ed. Bajaj, Y. P.
S.), Springer–Verlag, Berlin, 1986, vol. 2, pp. 73–88
[13]. Devaux H, Li P (2001). Enhancement of microspore culture
efficiency of recalcitrant barley genotypes 20: 475.
doi:10.1007/s002990100368
[14]. Dolezel and Batros,.J. Dolezel, J. Batros (2005) Plant DNA flow
cytometry and estimation of nuclear genome sizeAnnals of
Botany, 95 (2005), pp. 99–110.
[15]. Endemann, M., K. Hristoforoglu, T. Stauber and E. Wilhelm.
2001. Assessment of age-related polyploidy in Quercus robur L.
somatic embryos and regenerated plants using DNA flow
cytometry. Biol. Plant. 44:339–345.
[16]. Feher A, Pasternak T, Otvos K, Miskolczi P, Dudits D(2002).,
Induction of embryogenic competence in somatic plantcells: a
review., Biol Bratislava.,57:5–12.
[17]. Fisher C, Neuhaus G (1996)., Influence of auxin on the
establishment of bilateral symmetry in monocots., Plant J.,9:659–
669.
[18]. Foroughi-Wehr B, Friedt W, Wenzel G (1982) On the genetic
improvement génétique du bananier (Musa spp.). PhD thesis,
Ecole Nationale Agronomique de Rennes, France Jin S, Mushke
R, Zhu H, Tu L, Lin Z, Zhang Y, Zhang X (2008) Detection of
somaclonal variation of cotton (Gossypium hirsutum) using
cytogenetics, flow cytometry and molecular markers. Plant Cell
Rep 27(8):1303–1316.
[19]. Hamill, S.D. Hamill, M.K. Smith, W.A.(1992) Dodd In vitro
induction of banana autotetraploids by colchicine treatment of
micropropagated diploids Australian Journal of Botany, 40 (1992),
pp. 887–896.
[20]. Hansen AL, Plever C, Pedersen HC, Keimer B, Andersen SB
(1994) Efficient in vitro chromosome doubling during Beta
vulgaris ovule culture. Plant Breed 112:89–95.
[21]. Jin S, Mushke R, Zhu H, Tu L, Lin Z, Zhang Y, Zhang X (2008)
Detection of somaclonal variation of cotton (Gossypium
hirsutum) using cytogenetics, flow cytometry and molecular
markers. Plant Cell Rep 27(8):1303–1316.
[22]. Kerbellec F (1996) Etablissement d’une technique d’androgenèse
pour l’amélioration génétique du bananier (Musa spp.).
[23]. Kohler F, Wenzel G (1985) Regeneration of isolated barley
microspores in conditioned media and trials to characterise the
responsible factor. J Plant Physiol 121:181–191.
[24]. Lim WL, Loh CS (2003) Endopolyploidy in Vanda Miss
Joaquim (Orchidaceae). New Phytol 159:279–287.
[25]. May RA, Sink KC (1995) Genotype and auxin influence direct
somatic embryogenesis from protoplasts derived from
embryogenic cell suspensions of Asparagus officinalis L. Plant
Sci108:71–84.
[26]. Mishiba K, Tawada K, Mii M (2006) Ploidy distribution in the
explant tissue and the calluses induced during the initial stage of
internode segment culture of Asparagus officinalis L. In Vitro
Cell Dev Biol Plant 42:83–88.
[27]. Murashige T, Skoog F (1962) A revised medium for rapid growth
and bioassays with tobacco tissue cultures. Physiol Plant 15:473–
497.
[28]. Muren RC (1989) Haploid plant induction from unpollinated
ovaries in onion. Hortic Sci 24:833–834.
[29]. Niimi Y, Han D, Fujisaki M (2001)., Production of virus-free
plantlets by anther culture of LiliumבEnchantment’., Scientia
Horticulturae.,90: 325-334.
[30]. Pfosser M, Amon A, Lelley T, Heberle-Bors E (1995) Evaluation
of sensitivity of flow cytometry in detecting aneuploidy in wheat–
rye addition lines. Cytometry 21:387–393.pp 349–372.
[31]. Roux N, Toloza A, Radecki Z et al (2003) Rapid detection of
aneuploidy using flow cytometry. Plant Cell Rep 21:483–490.
[32]. Tenhola T, Tanhuanpä P, Immonen S (2005)., The effect of cold
and heat treatments on the anther culture response of diverse rye
genotypes., Euphytica.,145: 1-9.
[33]. Tremblay L, Levasseur C, Tremblay F M (1999) Frequency of
somoclonal variations in plant of black spruce (Picea mariana)
and white spruce (P.glauca, pinaceae) derived from somatic
embryogenesis and identification of some factors involved in
genetic instability.Am J Bot86:1373- 1381
[34]. Van Duren et al., M. Van Duren, R. Morpugo, J. Dolezel, R.
Afza.(1996) Induction and verification of autotetraploids in
diploid banana (Musa acuminata) by in vitro techniquesEuphytica,
88 (1996), pp. 25–34.
[35]. Vandenhout et al.,H. Vandenhout, R. Ortiz, D. Vuylsteke, R.
Swennen, K.V. Bai(1995)Effect of ploidy on stomatal and other
quantitative traits in plantain and banana hybrids Euphytica, 83
(1995), pp. 117–122.