Micropropagation is a proven means of producing millions of identical plants under a controlled and aseptic condition, independent of seasonal constraints. It not only provides economy of time and space but also gives greater output and allows further augmentation of elite disease free propagules.India is homeland of many important fruit crops such as Indian gooseberry (Emblica officinalis Gaertn), bael (Aegle marmelos Corr.), Guava (, Psidium guajava), jamun or black plum (Syzygium cuminii L. Skeels.), Mango (Mangifera indica) and Papaya (Carica papaya).
Micropropagation and commercial exploitation in horticulture cropsDheeraj Sharma
Micro-propagation – principles and concepts, commercial exploitation in horticultural crops. Techniques - in vitro clonal propagation, direct organogenesis, embryogenesis, micrografting, meristem culture. Hardening, packing and transport of micro-propagules.
Micropropagation and commercial exploitation in horticulture cropsDheeraj Sharma
Micro-propagation – principles and concepts, commercial exploitation in horticultural crops. Techniques - in vitro clonal propagation, direct organogenesis, embryogenesis, micrografting, meristem culture. Hardening, packing and transport of micro-propagules.
This presentation is done by 2010/2011 batch of Export Agriculture students of Uva Wellassa University of Sri Lanka as a requirement for the subject which is “Rice & Field Crop Production”. Note that the information included here is relevant to Sri Lankan condition.
The slides describing about the different techniques of seed production, as the seed is the basic part of any production program. Therefore, please provide review about these techniques.
The Presentation is prepared by N.S Institution of science, Markapur.
It consists of a basic introduction related to hybrid seed production related to rice.
Hybridization between individuals from different species belonging to the same genus or two different genera, is termed as distant hybridization or wide hybridization, and such crosses are known as distant crosses or wide crosses.
Plant tissue culture is used widely in the plant since , forestry and in horticulture .
Plant tissue culture relies on the fact that many plant cells have the ability to regenerate a whole plant .
Tissue culture of Strawberry provides an alternative and novel possibility of enhancing the production of planting materials, including virus-free plants for large-scale planting.
Tissue culture of strawberry could also make a significant contribution in improving the qualitative and quantitative characters of the plant.
This presentation is done by 2010/2011 batch of Export Agriculture students of Uva Wellassa University of Sri Lanka as a requirement for the subject which is “Rice & Field Crop Production”. Note that the information included here is relevant to Sri Lankan condition.
The slides describing about the different techniques of seed production, as the seed is the basic part of any production program. Therefore, please provide review about these techniques.
The Presentation is prepared by N.S Institution of science, Markapur.
It consists of a basic introduction related to hybrid seed production related to rice.
Hybridization between individuals from different species belonging to the same genus or two different genera, is termed as distant hybridization or wide hybridization, and such crosses are known as distant crosses or wide crosses.
Plant tissue culture is used widely in the plant since , forestry and in horticulture .
Plant tissue culture relies on the fact that many plant cells have the ability to regenerate a whole plant .
Tissue culture of Strawberry provides an alternative and novel possibility of enhancing the production of planting materials, including virus-free plants for large-scale planting.
Tissue culture of strawberry could also make a significant contribution in improving the qualitative and quantitative characters of the plant.
Introduction
Advantages of Micropropagation over the conventional methods
History
Stages of Micropropagation
1. Stage 0; Preparative stage
2. Stage 1; Initiation of aseptic cultures
A) Explant
B) Sterilization
C) Browning of medium
Factors affecting initiation stage
Conclusions
References
In vitro Shoot Micro Propagation of Medicinal Applications and Ornamental Val...Shafkat Shamim Rahman
Cestrum nocturnum L. a night Blooming jasmine belongs to Solanaceae family widely circulated all over tropical as well as subtropical areas of the World. It is mainly popular for ornamental fragrant flowering and hedge plant but also sometimes for traditional medicinal purpose. Due to strongest smelling characteristics of the plants, it is used in many industries for making Perfumes, essential Oils, Soaps, Candles, Body Oils, etc. The existence of natural plants of economic importance are threatened due to rapid urban development, including industrialization, residential development, educational, commercial etc., reduce the land for cultivation. Hence plant tissue culture protocol may be adapted for production and utilization of economically popular plants, including C. nocturnum involving limited space and short period of time. Shoot tip explants of naturally grown C. nocturnum were excised sterilized and endued on ‘Murashige and Skoog’ (MS) medium enriched changed concentration of BA, NAA, as well as GA3 singly or in combination. Excised micro shoots were examined for root development on 0.5 MS using IBA, NAA as well as IAA separately. The highest amount of multiple bud were observed in low concentration of BA (01.50 milligram × l-1), resulted no. of shoot 4.40 as well as 4.20/explant, no. of leaves 15.40 as well as 4.20/explant as well as size of different shoot 5.360 as well as 4.860 cm. The concentration of IBA and IAA were found to be best for root formation in micro shoots (13.20, 6.80 roots/micro shoots) as well as root size (8.39, 5.73 cm) individually. There are many opportunity of plant tissue culture which offer marvelous chances in plant propagation, plant development as well as creation of plants with necessary agronomical features. Finally often hardening plantlets were gradually adjusted to natural condition and acclimatized with 90% success. This established protocol could help plant cell biotechnology, horticulture, medical and industrial sector of the country.
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.
Micro-propagation of Alstroemeria Hybrida Cv. PlutoIJEAB
The experiment entitled micropropagation of Alstroemeria hybrida cv. Pluto was conducted to standardize protocol for aseptic establishment, callus induction, proliferation, and rooting from rhizome tips, rhizome sections, shoot tips, shoot nodal segments and inflorescence buds. Highest culture asepsis of 79.20 per cent at 2 weeks of culture and 68.08 per cent at 4 weeks of culture was recorded in rhizome tips following sterilization treatment with Carbendazim 200 ppm for 30 minutes + HgCl2 (0.1 %) dip for 10 minutes and final treatment with ethyl alcohol (70 %) for 1 minute. Rhizome tips and rhizome section explants survived sterilant treatment better than other explants. MS-liquid medium supplemented with BAP + IBA: 1.5 + 0.2 mg l-l proved best for culture establishment (89.42 %) in case of rhizome tips and (56.13 %) in case of rhizome sections. MS-solid medium with plant growth regulator combinations BAP + IBA: 1.0 + 0.2 mg l-1 fortified with activated charcoal resulted in an establishment of (78.25 %) in rhizome tips and (40.24 %) in case of rhizome sections. Callus induction was highest in MS-solid medium fortified with BAP + NAA: 0.5 + 4.5 mg l-l. Rhizome tips cultured on MS-medium BAP + IBA + GA3 + Activated charcoal: 2.0 + 0.4 + 0.5 + 1000 mg l-l gave highest proliferation (88.85 %) along with highest number of erect shoots (5.75) , number of new rhizome buds ( 3.75), rhizome fresh weight/shoot complex (6.05), and multiplication index (2.76). Highest Rooting (54.81 %) along with lowest days to appearance of root (10.87), highest number of roots (3.12) and highest root length (16.42 mm) was recorded in MS-liquid medium fortified with NAA 1.5 mg l-1. Abbreviations used— AC; Activated charcoal, BAP; 6-Benzyl amino purine, BA; 6-Benzyladenine, 2, 4-D; 2, 4dichloro-phenoxyacetic acid,GA3; Gibberelic acid, IAA; Indole-3-acetic acid, IBA; Indole-3-butyric acid, MS; Murashige and Skoog’s (1962) medium, NAA; Naphthalene acetic acid and µm; Micro molar.
Influence of Plant Growth Regulators on Somatic Embryogenesis Induction in Se...IJEABJ
Seriphidium herba-album (syn. Artemisia herba-alba) is a medicinal, aromatic, greenish-silver herb. It is used widely in folk medicine for treatment of diarrhea, abdominal cramps and in the healing of external wounds. It's also used for the treatment of diabetes mellitus, neurological disorders as epilepsy, Alzheimer’s disease, depression and jaundice. In this study we assessed the protocol for callus induction, maturation of somatic embryogenesis, frequency of germination and conversion into plantlets for leaf explants of Seriphidium herba-album using different concentrations of PGRs. Highest induction frequencies of embryogenic calli occurred after 35 days on MS medium supplemented with 1.5 mg L-1 2,4-D and 0.5 mg L-1 BAP. Optimum MS medium for higher frequency of matured somatic embryos was recorded using 5.0 mg L-1 BAP and 0.5 mg L-1 NAA and somatic embryos also induced young in vitro grown plantlets when cultured in the medium containing GA3 and kinetin. Hence, attempts to induce direct somatic embryogenesis have been achieved up to embryo regeneration and maturation.
The production of haploid plants exploiting the totipotency of microspore.
Androgenesis is the in vitro development of haploid plants originating from totipotent pollen grains through a series of cell division and differentiation.
PREFERENTIAL ASSOCIATION OF ENDOPHYTIC BRADYRHIZOBIA WITH DIFFERENT RICE CULT...Anamika Rana
All of rice oligotrophic endophytic Bradyrhizobia in this study were obtained except SUT-R74.
6 Bradyrhizobial strains were obtained from 98 bacterial strains.
Bradyrhizobium is found only in rice root, with 10% relative abundance of total Alphaproteobacteria.
Endophytic Bradyrhizobia could not be obtained from the monoculture system.
Thai rice cultivars, the Thai Bradyrhizobial strains could promote rice growth better than Japanese strains.
Three rice cultivars (Pathum Thani 1, Kasalath, and Nipponbare), cultivar Pathum Thani 1 responded only to putative Thai rice endophytic Bradyrhizobia.
This phenomenon was not found in Japanese rice cultivars.
Non-PB strains are also capable of forming a natural endophytic association with rice.
Strains SUT-PR9, WD16, RP5, and RP7 displayed non-PB phenotypes but were genotypically close to PB strains.
The Studies of Effect of Bio Fertilizers Rhizobium, Phosphobacteria, and Root...IIJSRJournal
A pot culture experiment was carried out and to the find out the effect of Rhizobium, Phosphobacteria and Root Nodule extract on the vegetative growth and biochemical changes in Black gram and Maize. The Black gram and Maize is vegetative parameters such as seed germination, shoot and root length, leaf fresh weight, leafs dry weight, shoot and root dry weight had analyzed and biochemical studies of both Black gram and Maize was carried such as total chlorophyll content, leave soluble protein and in vivo nitrate reductase activity. Among microbial inoculants, the Rhizobium + PB mixer was found most effective in terms of seed germination, shoot and root length, leaf fresh weight, leafs dry weight, shoot and root dry weight and also showed increase total chlorophyll content, soluble protein and nitrate activities in both Black gram and Maize.
Definition of hairy root culture ,multiple shoot culture ,Production of hairy root and multiple shoot , advantages an disadvantages of hairy root and multiple shoot culture, Sterilization and sterilizing agents wit concentration and exposure time
The experiment was conducted at the experimental farm and laboratory of Institute of
Sustainable Agrotechnology, University Malaysia Perlis, Padang Besar, Perlis, Malaysia, with the objective
toinvestigate the inhibitory and stimulatory effects of aqueous extract of mungbean on seed germination and
seedling growth of three crop species, mungbean, sweet corn and okra. Different treatments of mungbean
aqueousextracts (vegetative fresh, vegetative after 2 weeks drying, vegetative after 4 weeks drying, flowering
fresh, flowering after 2 weeks drying, flowering after 4 weeks drying, flowering fresh, flowering after 2 weeks
drying, flowering after 4 weeks drying, maturity fresh, maturity after 2 weeks drying, maturity after 4 weeks
drying and water as control) were used to test their effect on the test species. The experiment was randomly
distributed and according to Completely Randomized Design(CRD) with five replicates. The results showed the
fresh vegetative aqueous extract of mungbean had a significant effect (stimulatory) on germination percent and
growth parameters such as number of root, root length and shoot height, of the three crop species. The study
revealed that the aqueous extract of mungbean have different effects (inhibitory and stimulatory) on the
seedlings and the mode of action depends on the associated plant species.Our results suggest that the aqueous
extract of mungbean from the different growth stages and drying periods have an allelopathic effect.
The ability of an explant to regenerate into a whole plant under in vitro asceptic conditions by providing a proper artificial nutrient medium is called as Plant Tissue Culture.
Banana is the fourth largest produced food crop of the world and its demand is increasing day by day. It is available throw out the year and its cost is very less in comparison to other fruits. With the development in science new tissue culture protocols are standardized for mass propagation of Musa (Banana) on the basis of effects of plant growth regulators. BAP (6-Benzyl Amino Purine), KN (Kinetin) are most widely used cytokinins for shoot proliferation and IAA (Indole -3-acetic acid), NAA (Naphathalene acetic acid) are widely used auxins for root induction.
How to Build a Module in Odoo 17 Using the Scaffold MethodCeline George
Odoo provides an option for creating a module by using a single line command. By using this command the user can make a whole structure of a module. It is very easy for a beginner to make a module. There is no need to make each file manually. This slide will show how to create a module using the scaffold method.
Safalta Digital marketing institute in Noida, provide complete applications that encompass a huge range of virtual advertising and marketing additives, which includes search engine optimization, virtual communication advertising, pay-per-click on marketing, content material advertising, internet analytics, and greater. These university courses are designed for students who possess a comprehensive understanding of virtual marketing strategies and attributes.Safalta Digital Marketing Institute in Noida is a first choice for young individuals or students who are looking to start their careers in the field of digital advertising. The institute gives specialized courses designed and certification.
for beginners, providing thorough training in areas such as SEO, digital communication marketing, and PPC training in Noida. After finishing the program, students receive the certifications recognised by top different universitie, setting a strong foundation for a successful career in digital marketing.
Acetabularia Information For Class 9 .docxvaibhavrinwa19
Acetabularia acetabulum is a single-celled green alga that in its vegetative state is morphologically differentiated into a basal rhizoid and an axially elongated stalk, which bears whorls of branching hairs. The single diploid nucleus resides in the rhizoid.
Executive Directors Chat Leveraging AI for Diversity, Equity, and InclusionTechSoup
Let’s explore the intersection of technology and equity in the final session of our DEI series. Discover how AI tools, like ChatGPT, can be used to support and enhance your nonprofit's DEI initiatives. Participants will gain insights into practical AI applications and get tips for leveraging technology to advance their DEI goals.
A workshop hosted by the South African Journal of Science aimed at postgraduate students and early career researchers with little or no experience in writing and publishing journal articles.
Thinking of getting a dog? Be aware that breeds like Pit Bulls, Rottweilers, and German Shepherds can be loyal and dangerous. Proper training and socialization are crucial to preventing aggressive behaviors. Ensure safety by understanding their needs and always supervising interactions. Stay safe, and enjoy your furry friends!
Macroeconomics- Movie Location
This will be used as part of your Personal Professional Portfolio once graded.
Objective:
Prepare a presentation or a paper using research, basic comparative analysis, data organization and application of economic information. You will make an informed assessment of an economic climate outside of the United States to accomplish an entertainment industry objective.
2. Introduction
• Micropropagation is a proven means of producing millions
of identical plants under a controlled and aseptic condition,
independent of seasonal constraints.
• It not only provides economy of time and space but also
gives greater output and allows further augmentation of elite
disease free propagules.
• India is homeland of many important fruit crops such as
Indian gooseberry (Emblica officinalis Gaertn), bael (Aegle
marmelos Corr.), Guava (, Psidium guajava), jamun or
black plum (Syzygium cuminii L. Skeels.), Mango
(Mangifera indica) and Papaya (Carica papaya).
3. • Most of these crops have medicinal value and are suitable
for growing under marginal situations.
• The commercial production of these crops is restricted due
to the shortage of desirable planting material.
• Micropropagation can play an important role in rapidly
increasing new cultivars of these fruit crops.
5. Introduction
• Aegle marmelos Corr., belongs to family Rutaceae, is more
prized for its pharmacological virtues than its edible quality.
• Because of pharmacological importance, it’s become potential
candidate for developing transgenics to enhance its medicinal
properties.
• Maximum mortality of micropropagated plants occur during
acclimatization phase because plantlets undergo rapid and
extreme changes in physiological functioning, histological
and biochemical changes.
• In order to investigate the actual reason of this limitation, test
samples were collected at different micropropagation stages
(In vitro, and acclimation).
6. Findings
• Rapid clonal micropropagation protocol of Aegle marmelos Corr.
cv. CISH-B1 and CISH-B2 was achieved by nodal stem segment
of mature bearing tree.
• Three centimeter long shoots having one axillary bud excised
from 10-15th nodal region of shoots during September gave quick
in vitro bud burst (5.33 days) when cultured on MS medium
supplemented with BAP, 8.84 μM + IAA 5.7 μM.
• The maximum number of proliferated shoots (9.0/explant) were
obtained on same medium supplemented with BAP 8.84 μM +
IAA 5.7 μM.
• The micro shoots were rooted (100 %) on ½ strength MS medium
supplemented with IBA 49.0 + IAA 5.7 μM. .
7. • In vitro rooted plants were acclimatized on coconut husk
containing ½ strength MS plant salt mixture and under shade
net house (50 % shade 70-80 % RH).
• The plants were established in the field after acclimatization.
• The micropropagated plants were tested for its genetic fidelity
using 13 RAPD, 3 ISSR and 2 DAMD primers.
• Profile obtained by all the three Single Primer Amplification
Reaction (SPAR) technique from mother tree and
micropropagated plants revealed genetic uniformity of
micropropagated plants with that of mother tree.
8. A
C
B
D
E
G
F
Fig: Micropropagation of Aegle marmelos (bael). (A) In vitro shoot bud
induction,
(B) proliferation, (C) Rooting, (D) Acclimatization, (E)
Acclimatized plants (F) 3 month old acclimatized plants in polyhouse (G)
Plant growing in field
10. Histological and biochemical changes
• Maximum mortality of micropropagated plants occur during
acclimatization phase, because plantlets undergo rapid and
extreme changes in physiological functioning, histological
and biochemical changes.
• In order to investigate the actual reason of this limitation, test
samples (Leaf, stem and root) were collected at different
stages of micropropagated plants (In vitro and acclimation).
• The biochemical result showed that micropropagated plantlets
produced significantly low total chlorophyll (0.042 mg/g fresh
weight), reducing sugar (3.227%), NR activity (1.353
NO2/h/g fresh weight) and but higher protein (0.048 μg/g)
during in vitro phase.
11. • The in vitro raised plants showed abnormal histological features
like altered leaf mesophyll, absence of thick cuticle, sunken
stomata, poorly developed stem and root histology.
• Photoautrophic mode of nutrition during in vitro phase increased
the survival rate during acclimatization compared to
photoheterotrophic mode of nutrition
• Photoautotrophism phenemoneon has substantial influence on
the physiology and development of in vitro regenerated Aegle
marmelos Corr. plantlets.
12. Anatomy of leaf
A
B
(A) Whole mount of in vitro leaf showed open type stomata with fully turgid
guard cells, (B) whole mount of acclimatized leaf showed partially
closed stomata.
13. Anatomy of leaf
A
B
(A) T.S. of in vitro leaf, showed single layered epidermis with almost no cuticle and
single layered and poorly developed palisade parenchyma with poorly
developed vascular tissue.
(B) T.S. of acclimatized leaf, showed single layered epidermis with thick cuticle and
well developed double layered palisade mesophyll while the lower side had
spongy parenchyma with air spaces and sunken type stomata.
14. Anatomy of stem
A
B
(A) T.S. of in vitro stem, had a polystelic structures, vascular bundles were arranged
in a ring and they were conjoint collateral and open no corck cambium, showed
primary medullary rays, uniseriate epidermis, no cuticle, distinct endoderm was
absent while cortex was parenchymatous.
(B) T.S. of acclimatized stem, well developed cork cambium, parenchymatous pith
but the pith was mucilaginous, distinct secondary growth with well developed
wood and large woody vessels, epidermis was uniseriate and covered with cuticle,
and cortex was collenchymatous.
15. Anatomy of root
A
B
(A) T.S. of in vitro root, showed poorly developed pith, undifferentiated cortex with
very less amount of storage tissue and no secondary growth at all.
(B) T.S. of acclimatized Root, had a well developed parenchymatous cortex with tannin
cells, pericycle is made of stone cells vascular cylinder consisted of secondary
xylem towards inner side and secondary phloem towards outer side, secondary
xylem showed wide vessels scattered among trachieds, medullary rays were present
while pith was negligible.
17. Introduction
• Guava (Psidium guajava Linn.) commonly known for its
food, nutraceutical and commercial values throughout
the world.
• The guava plant parts are used for the development of
various industrial and pharmaceutical products.
• Guava contains broad spectrum of phytochemicals
including polysaccharides, vitamins, essential oils,
minerals, enzymes, proteins, sesquiterpenoid alcohols
and triterpenoid acids , alkaloids, glycosides, steroids,
flavanoids, tannins, saponins .
• Guava is very rich in antioxidants and vitamins and also
high in lutein, zeaxanthine and lycopene .
18. Findings
• Micropropagation through in vitro shoot bud culture has
been developed in guava cv. Allahabad Safeda.
• 3 cm long nodal segments were surface sterilized using
Carbendazime 0.1% and PVP 100 mg/l.
• Explants were further treated with HgCl2 0.1% for 5
minutes aseptically, followed by six washing in sterile
distilled water.
• Quick shoot bud induction was achieved within five days in
MS medium supplemented with BAP 3.0 mg/l.
• The proliferation of microshoots (2.67 shoots/explant) was
achieved in same medium (MS+BAP 3.0 mg/l).
19. • Rooting was achieved in MS medium containing IBA 10
mg/l within 17.6 days.
• In vitro rooted plants were acclimatized on coconut husk
containing ½ strength MS plant salt mixture and under
shade net house (50 % shade 70-80 % RH).
• Around 88.48%
acclimatization.
plants
were
survived
during
• Acclimatized plantlets of guava cv. Allahabad Safeda were
planted in field during rainy season.
20. A
B
D
E
C
F
Fig: Micropropagation of Psidium guajava (Guava). (A) In vitro shoot bud
induction, (B) proliferation, (C) Rooting, (D) Acclimatization, (E) Acclimatized
plants in polyhouse (F) Plant growing in field
22. Introduction
• Indian gooseberry or aonla (Emblica officinalis Gaertn.)
belongs to the family euphorbiaceae and is well known for
its medicinal and therapeutic properties.
• It is a rich source of Vit C and used in fruit processing
industries.
• The use of micropropagation approach for accelerating the
production of clonal stock of commercial cultivars in Indian.
23. Findings
• 3 cm long nodal segments were surface sterilized using
Carbendazime 0.1% and PVP 100 mg/l.
• Explants were further treated with HgCl2 0.1% for 8
minutes aseptically, followed by six washing in sterile
distilled water.
• The maximum survival (66.49%) and minimum
contamination (33.51%) of shoot buds were observed
during surface sterilization.
• MS medium supplemented with Kinetin 13.9 3 +GA3 4.33
+Glutamine 342.11 mM gave the early bud break (7days)
and highest shoot proliferation (13.33 shoots/culture).
24. • 2-3 cm long microshoots were cultured on ½ MS+ IBA
49.20 mM+ NAA 10.74 mM and activated charcoal 0.1%,
in this treatment 4 roots/explant was achieved in 7.33 days.
• In vitro rooted plants were acclimatized on
Soil+Sand+FYM (1:1:1) containing ½ MS plant salt
mixture containing paclobuterazole 1.0 mg/l under shade
net house (50 % shade 70-80 % RH).
• 72.22% plants were survived during acclimatization.
• Acclimatized plants were successfully transferred to field
during rainy season.
25. A
B
C
D
E
F
Fig: Micropropagation of Emblica officinalis (Aonla). (A) In vitro shoot bud
induction,
(B‐C) proliferation, (D) Rooting, (E) Acclimatization, (F)
Acclimatized plants in polyhouse
27. Introduction
•
Jamun or black plum (family myrtaceae) is an important
indigenous underutilized fruits of commercial value.
•
It is good source of iron and anti-diabetic compounds.
•
The seeds are claimed to contain alkaloid, jambosine,
and glycoside jambolin or antimellin, which halts the
diastatic conversion of starch into sugar and seed
extract has lowered blood pressure.
•
Jamun orcharding can help in the management of
wastelands.
•
In order to produce large scale true-to-type planting
material, micropropagation can be gainfully employed.
28. Findings
• The shoot buds were taken from eight years old plant for
protocol establishment.
• 3 cm long nodal segments were surface sterilized using
Carbendazime 0.1% and PVP 100 mg/l followed by
treatment with HgCl2 0.1% for 6 minutes aseptically,
followed by six washing in sterile distilled water.
• Nodal segments were cultured on MS medium +BAP 2.0
mg/l. This treatment gave earlier shoot bud induction in just
5.6 days.
• Highest shoot proliferation was found in MS medium+BAP
2.0 mg/l+ GA3 0.5 mg/l+Casein hydrosylate 150 mg/l.
29. • Microshoots were rooted cent percent on ½ MS+ IBA 5
mg/l+Activated charcoal 100 mg/l in just 12 days.
• In vitro rooted plants were acclimatized on
Soil+Sand+FYM (1:1:1) containing ½ MS plant salt
mixture fortifird with paclobuterazole 1.0 mg/l under
shade net house (50 % shade 70-80 % RH).
• 80% plants were survived during acclimatization.
• After three month of acclimatization plants were transfer
to field for further analysis.
30. A
B
D
E
C
Fig: Micropropagation of Syzygium cuminii Skeels.(Jamun). (A) In vitro
shoot bud induction, (B) Culture establishment (C) Proliferation in
microshoots, (D) Rooting, (E) Acclimatization,
(F) Acclimatized plant
growing in field.
32. Introduction
• Mango ( Mangifera indica L.) is the most important fruit crop
because of its wide adaptability, high nutritive value, richness
in variety, delicious taste, excellent flavour, attractive
appearance and commercial utility in India as well as in many
part of the world.
• Conventional breeding in perennial crops is difficult and time
consuming.
• A standard uniform protocol of regeneration is the prime and
foremost prerequisite for not only improve the productivity, the
production from the existing area but also development of
transgenics in mango for various traits.
33. Findings
• Nucellar embryogenesis was induced in different
monoembryonic and polyembryonic cultivars of mango
(Mangifera indica L) viz., Dashehari, Amrapali, Bapakkai,
Kurukkan and Moovandan.
• Nucellus tissue excised from 3.5 cm long fruits of these
cultivars developed pro-embryonic calli on modified MS
medium supplemented with 2,4-D 4.52μM + malt extract
0.05% and spermidine 13.78μM.
• Among all the cultivars, polyembryonic cultivars gave higher
level of somatic embryogenesis in comparison to
monoembryonic.
34. • Among all polyembryonic cultivars. Bappakai produced 187.33
embryos per explant followed by Kurukkan 158.33 embryos per
expalnt and Movandan 146.45 embryos per explant, where as
monoembryonic cultivars shows comparatively lower level of
somatic embryogenesis.
• Dashehari gave rise to 97.25 embryos per explant followed by
Amrapali 82.33 embryos in 100 days under dark culture
conditions.
• However, all the differentiated embryos proliferated on medium
having low level of sucrose (4% w/v) and auxin (2, 4-D 2.26μM).
• Most of the proembryonic calli converted into heart shaped and
cotyledonary embryos by reducing temperature to 15oC.
35. • Somatic embryos were matured on modified MS medium
fortified with ABA 0.38μM+ IAA 0.57μM and PEG
30.30μM.
• Matured somatic embryos germinated on MS medium
supplemented with NAA 2.68μM+ kinetin 11.60μM and
glutamine 2736.9μM.
• Among all cultivars, Bappakai showed higher germination
(39.33) followed by Kurukkan (29.97 %), Movandan
(28.25%), Deshahari (26.45%) and Amrapali (25.25%).
36. Monoembryonic cultivars of mango
Dashehari
Amrapali
Polyembryonic cultivars of mango
Bapakkai
Kurukkan
Moovandan
37. Different stages of somatic embryogenesis in mango
A
B
C
D
E
F
Different stages of somatic embryogenesis in mango (Mangifera indica L).
A-B Somatic embryo induction from nucellar tissues, C-Proliferation and development of
globular embryos, D- conversion of SE into heart shaped, E- early cotyledonary stage, Flate cotyledonary shaped embryo.
Cont…
38. Different stages of somatic embryogenesis in mango
H
G
I
Different stages of somatic embryogenesis in mango (Mangifera indica L). Grooting and conversion into early stage of plants and H-rooted plants growing
vigorously in liquid culture medium and I- plant in polyhouse during acclimatization.
40. Introduction
• Papaya (Carica papaya L.) is a popular and economically
important and medicinal fruit tree of tropical and
subtropical countries.
• It has varied uses in the beverage, food and pharmaceutical
industries, in chill‐proofing beer, tenderizing meat, drug
preparations for digestive ailments and treatment of
gangrenous wounds.
• Papaya is host to various species of pests and pathogens and
affected by various viral, fungal and nematode diseases.
• Here we are reporting, Development of transgenic papaya
resistant to Papaya Leaf Curl Virus (PLCuV) and Papaya
Ringspot Virus (PRSV) using A. tumifaciens.
41. Findings
• In vitro seedling plants were raised from immature zygotic
embryos excised from 90-100 days old white, plump
immature seeds of Pusa Delicious in ½ MS medium
containing BAP 0.2 mg/l+ MS Vitamins 1.0 ml/l (extra),
agar 0.8% and sucrose 3%.
•
A. tumifaciens was grown in LB medium containing 50
mg/l Kanamycin in the dark at 28oC in incubator shaker at
120 rpm for 24 hrs (1.0 OD at OD600).
•
One week old Shoot tips (0.5cm), Carborandum
wounding, 30 min infection period, 72 hrs co-cultivation
period, Cefotaxime 500 mg/l, Acetosyringone 100 µM
and Spermidine 1.0 mM gave higher (8.8%) putative
transformats.
42. • Plants were rooted on ½ MS medium containing IBA 3.0
mg/l.
• Transformed plants were diagnosed using forward and
reverse primers of cp (410 bp), rep (479 bp) and npt II
(480 bp) PCR reactions.
43. Fig: Papaya shoot tip transformation using Agrobacterium tumifaciens. (A) 0.5 cm long shoot tip
as an explant, (B) Selection and regeneration of transformed shoot tip using Kanamycin 150 mg/l
after 12 weeks, (C) Rooting in transformed papaya shootlets using IBA 3.0 mg/l, (D)
Acclimatization of PCR positive plantlet, (E) Acclimatized plants are under evaluation in
Transgenic glass house, (F) Gel picture showing 480 bp npt II bands.
44. Publications
•
Pati R, Mishra M, Chandra R and Muthukumar M (2013). Histological and
biochemical changes in Aegle marmelos Corr. before and after acclimatization.
Tree Genetics and Molecular Breeding. 3(3): 12-18.
•
Pati R, Chandra R, Chauhan UK, Mishra M and Srivastiva N (2008). In vitro
clonal propagation of bael (Aegle marmelos Corr.) cv. CISHB1 through
enhanced axillary branching. Physiology and Molecular Biology of Plants.
14(4): 337-346.
•
Pati R, Chandra R, Chauhan UK and Mishra M (2008). In vitro plant
regeneration from mature explant of Aegle marmelos Corr.) CV. CISH-B2.
Science and Culture. 74(9-10): 359-367.
•
Mishra, M. Chandra, R., Pati, R. and Bajpai, A. (2007). Micropropagation of
Guava (Psidium guajava L.). Acta Horticulturae, 735: 155-158.
•
Mishra M, Pati R and Chandra R (2006). Clonal micropropagation of Indian
gooseberry (Emblica officinalis Gaertn). Indian Journal of Genetics and Plant
Breeding. 66(4): 361-361.
45. • Mishra M, Chandra R, Pati R (2008). In vitro regeneration and genetic fidelity
testing of Aegle marmelos (Corr.) plants. Indian Journal of Horticulture. 65(1):
6-11.
• Mishra M, Pati R, Chandra R. et al. R (2010). Micropropagation of Mangifera
indica L. cv. Kurakkan through somatic embryogenesis. Indian Journal of
Genetics and Plant Breeding. 70(1): 85-90.
• Mishra M, Chandra R, Pati R, Jain RK and Agarawal S (2010). Shoot tip
transformation of papaya (Carica papaya L.). Acta Horticulturae, 851: 219226.
• Mishra M, Chandra R, Tiwari RK, Pati R and Pathak RK (2005).
Micropropagation of certain under utilized fruit crops: A Review. Small fruits
Review. 4(4): 7-18.
• Pati R. and Muthukumar M. (2013). Genetic transformation in Aegle marmrlos
Corr. In: Biotechnology of neglected and underutilized crops, edited by S.M.
Jain and S. Dutta Gupta. Springer . p.343-365. [ISBN: 978-94-007-5500-0].
46. • Chandra R, Pati R and Mishra M (2010). Mango. In: Advances in Horticultural
Biotechnology Vol.-1 Regeneration Systems- Perennial Fruit Crops and Spices.
Eds., Singh HP, Parthasarathy VA and Nirmal Babu K. Westville Publishing
House, New Delhi, pp. 73-90. [ISBN-978-11-85873-65-7].
• Mishra M, Pati R and Chandra R (2009). “Clonal micropropagation of
subtropical fruit trees”. In: Forest Biotechnology in India, edited by Mandal
AK, Ansari SA and Narayanan C. Vedams eBooks (P) Ltd., New Delhi, India.
[ISBN-81-89304-56-9].
• Pati R, Gupta VK, Yadava LP, Srivastiva N, Gupta A and Modi DR (2008).
“Agrobacterium- As Natural Tool for Plant Genetic Engineering”: In: Potential
Microorganism for Sustainable Agriculture, edited by Prof. D.K. Maheshwari
and R.C. Dubey, I.K. International Pvt. Ltd., New Delhi, pp.436-459. [ISBN8190746205].