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.
GREEN GRAM (MUNG BEAN)
vigna radiata (2n = 22)
It is esteemed as the most wholesome among the pulses, free from the heaviness and tendency to cause flatulence, which is associated with other pulses.
Place of origin : India
Wild relative : Vigna radiata var. sublobata
BLACK GRAM (URAD, ULUNDU)
Vigna mungo (2n = 22, 24)
Origin : India
Putative parents
V. trinerivus / V. sublobata or V.mungo var. sylvestris.
Breeding objectives
1. Evolving medium duration high yielding varieties for dry land cultivation.
Co5 black gram. Suitable for dry land cultivation.
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.
GREEN GRAM (MUNG BEAN)
vigna radiata (2n = 22)
It is esteemed as the most wholesome among the pulses, free from the heaviness and tendency to cause flatulence, which is associated with other pulses.
Place of origin : India
Wild relative : Vigna radiata var. sublobata
BLACK GRAM (URAD, ULUNDU)
Vigna mungo (2n = 22, 24)
Origin : India
Putative parents
V. trinerivus / V. sublobata or V.mungo var. sylvestris.
Breeding objectives
1. Evolving medium duration high yielding varieties for dry land cultivation.
Co5 black gram. Suitable for dry land cultivation.
Tissue culture is the growth of tissue or cells separate from the organism
Strawberry (Fragaria x ananassa Duch.) is a natural hybrid of Fragaria chiloensis .
It is a perinnial, stoloniferous herb belongs to the Rosaceae family.
Strawberries have traditionally been a popular delicious fruit for its flavour, taste, fresh use, freezing and processing.
Tissue culture is the growth of tissue or cells separate from the organism
Strawberry (Fragaria x ananassa Duch.) is a natural hybrid of Fragaria chiloensis .
It is a perinnial, stoloniferous herb belongs to the Rosaceae family.
Strawberries have traditionally been a popular delicious fruit for its flavour, taste, fresh use, freezing and processing.
Production of Haploids Plants from Anther Culture of Musa Paradisiaca cv. ‘Pu...RSIS International
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.
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.
Effect of Plant Growth Promoting Rhizobacteria of the Growth of Cicer arietinumOpen Access Research Paper
This research was conducted to isolate the rhizospheric bacteria of chickpea plant and to check their effect on its growth. Out of ten bacterial strains isolated, six were checked. They included two strains of Pseudomonas sp., three strains of Bacillus sp. and one strain of Brevibacterium sp. Out of all strains, one Bacillus strain showed good results. The 16s rRNA sequencing showed it Bacillus velezensis MN611255. Early germination, enhanced number of leaves, shoots, roots, and increase in their weight were notable features of B. velezensis as a PGPR. Furthermore, its effect on the flavonoids, total flavonoids, phenols, carbohydrates and chlorophyll content of chickpea plant was more pronounced as compared to the control. PGPR did not show siderophore production but were positive to indole acetic acid and phosphate solubilization. It can be concluded from the observations that indigenous isolated B. velezensis showed promising results as a PGPR. Field trials can help in further elaborating its role as a biofertilizer.
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.
This study investigated the effect of two different concentrations (0.05% and 0.1%) of chitosan nano-particles (CsNPs) as priming solutions (for 6 h) of Vicia faba seeds cv. Sakha 1, followed by germination and subsequent growth of seedlings for seven days. Chitosan nanoparticles were prepared using methacrylic acid and showed a mean size of 20 ± 2 nm. Both concentrations of chitosan nanoparticles caused deleterious effects on germination and seedling growth criteria. Germination was greatly reduced in both concentrations as compared to control (distilled water). The magnitude of decrease was much pronounced with the higher concentration of chitosan nanoparticles (0.1%). On the other hand, the lower concentration of CsNPs (0.05%) increased the content of total phenols and the activities of antioxidant enzymes (catalase, ascorbate peroxidase, peroxidase and polyphenol oxidase) as compared with those of the control seedlings. This might indicate that the relatively low concentration of chitosan nanoparticles enhanced the defense system of seeds by increasing total phenols and antioxidant enzyme activities.
THE EFFECTS OF HELPING BACTERIA (PSEUDOMONAS SPP.) IN NITROGEN GREEN BEANS F...IJSIT Editor
Some- bacteria settle in the rhizosphere of legume plants and enhance the performance of ribosome
bacteria to nitrogen fixation and nodulation. In this paper, we used four isolated from two species of
Pseudomonas containing P.putida, P.fluorescens Chao, P.Flouresence Tabriz, P.flouresence B119 and Rhizobium
leguminosarumbv.phaseoli. In a factorial experiment with complete randomized blocks were used 5 levels of
helping bacteria(Pseudomonas spp.) and two rhizobium levels, four replicates were employed. Jamaran418
green bean was utilized as host plant. At the end, nodulation, growth and plant’s nitrogen indexes were
measured. The results showed that all above mentioned helping bacteria enhance the growth and nodulation
performance of green bean. It should be said that P.putida had the highest effect on the green bean
nodulation increase along with rhizobium (130%) followed by P.fluorescens Tabriz, P. fluorescens Chao and
P.fluorescens B119, ( 83, 63 and 17%, respectively). Also, we observed 45, 33, 22 and 8% performance
increase under the effect of P.putida, P. fluorescens Chao, P. fluorescens Tabriz and P. fluorescens B119,
respectively.
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.
Improving the germination of somatic embryos in date palm Berhi cultivar in v...INNS PUBNET
Embryogenic suspension cultures of Date palm (Phoenix dactylifera L.) allow mass propagation of somatic embryos; Partial desiccation (0, 1, 2, 3 and 4 hours) and low temperature (0°C for 2 hr, 0°C for 4 hr, 4°C for 24 hr and 4°C for 48 hr) treatments were applied to improve germination of somatic embryos in vitro of date palm cultivar Berhi with or without AC. The highest germination percentage was achieved when embryos were desiccated for three hours as well as treatment of low temperature in 4°C for 24 hr. Also, the results proved that found activated charcoal) AC) in liquid media produced the highest somatic embryos number and weight and improving percentage of germination. Further, Partial desiccation and low temperature increased embryos proline content. The improvement of the germination of somatic embryos via low temperature and especially via partial desiccation embryos somatic is successful can be used for the purpose of commercial propagation especially for Berhi cultivar.
Production of Genetically Modified Grape (Vitis vinifera L.) PlantsAI Publications
Grape (Vitis vinifera L.) is one of the most economically important fruits in the world. High salinity stress adversely affects plant growth and limits agricultural production worldwide. This study describes a successful method of somatic embryogenesis using in vitro-derived leaf explants and introduction of a vacuolar-type Na+/H+ antiporter gene from a halophytic plant, Atriplex gmelini (AgNHX1) confers salt tolerance to grape cv. Superior Seedless using the Agrobacterium-mediated transformation. Callus embryogenic was induced on NN medium 2.0 mgL-1 2,4-D, 0.5 mgL-1 BAP and 0.5 mgL-1 NAA. Subsequent subculture of callus on NN medium containing 1.5 mgL-1 BAP, 0.5 mgL-1kinetin and 0.5 mgL-1NAA induced shoot organogenesis after eight weeks of culture. The leaf explants were co-cultivated with Agrobacterium strain LBA4404 harbouring the binary vector pBI121 which contained the AgNHX1 and nptII genes and putative transgenic plants were produced. The presence and stable integration of AgNHX1 gene in transgenic plants was confirmed by PCR and northern blot hybridization. The transgenic grape plants overexpressing the AgNHX1 gene showed a strong tolerance to salt stress under 250 mM NaCl, whereas non-transgenic plants died under the same conditions. Salt tolerance assays followed by salt treatments showed that the transgenic plants overexpressing AgNHX1 could survive under conditions of 250 mM NaCl for 4 weeks while the non-transgenic plants died under the same conditions. These results indicate that overexpression of the Na+/H+ antiporter gene in grape plants significantly improves their salt tolerance.
Welcome to TechSoup New Member Orientation and Q&A (May 2024).pdfTechSoup
In this webinar you will learn how your organization can access TechSoup's wide variety of product discount and donation programs. From hardware to software, we'll give you a tour of the tools available to help your nonprofit with productivity, collaboration, financial management, donor tracking, security, and more.
June 3, 2024 Anti-Semitism Letter Sent to MIT President Kornbluth and MIT Cor...Levi Shapiro
Letter from the Congress of the United States regarding Anti-Semitism sent June 3rd to MIT President Sally Kornbluth, MIT Corp Chair, Mark Gorenberg
Dear Dr. Kornbluth and Mr. Gorenberg,
The US House of Representatives is deeply concerned by ongoing and pervasive acts of antisemitic
harassment and intimidation at the Massachusetts Institute of Technology (MIT). Failing to act decisively to ensure a safe learning environment for all students would be a grave dereliction of your responsibilities as President of MIT and Chair of the MIT Corporation.
This Congress will not stand idly by and allow an environment hostile to Jewish students to persist. The House believes that your institution is in violation of Title VI of the Civil Rights Act, and the inability or
unwillingness to rectify this violation through action requires accountability.
Postsecondary education is a unique opportunity for students to learn and have their ideas and beliefs challenged. However, universities receiving hundreds of millions of federal funds annually have denied
students that opportunity and have been hijacked to become venues for the promotion of terrorism, antisemitic harassment and intimidation, unlawful encampments, and in some cases, assaults and riots.
The House of Representatives will not countenance the use of federal funds to indoctrinate students into hateful, antisemitic, anti-American supporters of terrorism. Investigations into campus antisemitism by the Committee on Education and the Workforce and the Committee on Ways and Means have been expanded into a Congress-wide probe across all relevant jurisdictions to address this national crisis. The undersigned Committees will conduct oversight into the use of federal funds at MIT and its learning environment under authorities granted to each Committee.
• The Committee on Education and the Workforce has been investigating your institution since December 7, 2023. The Committee has broad jurisdiction over postsecondary education, including its compliance with Title VI of the Civil Rights Act, campus safety concerns over disruptions to the learning environment, and the awarding of federal student aid under the Higher Education Act.
• The Committee on Oversight and Accountability is investigating the sources of funding and other support flowing to groups espousing pro-Hamas propaganda and engaged in antisemitic harassment and intimidation of students. The Committee on Oversight and Accountability is the principal oversight committee of the US House of Representatives and has broad authority to investigate “any matter” at “any time” under House Rule X.
• The Committee on Ways and Means has been investigating several universities since November 15, 2023, when the Committee held a hearing entitled From Ivory Towers to Dark Corners: Investigating the Nexus Between Antisemitism, Tax-Exempt Universities, and Terror Financing. The Committee followed the hearing with letters to those institutions on January 10, 202
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3. A Presentation On
In vitro Micropropagation of Caladium bicolor L.
BY
Batule Prashant Dnyandev
(VSBT42/2018)
Course Title: Project Formulation, Execution and Presentation
Course No- (READY-482)
UNDER THE DEPARTMENT
PLANT BIOTECHNOLOGY
Vidya Pratishtan College of Agricultural Biotechnology Baramati.
(Affilated to Mahatma Phule Krishi Vidyapeeth, Rahuri, Dist: Ahmadnagar, 413722
Year 2021-22)
Project Co-Guide:
Prof. Sayli Badekar
Assistant Professor,
Department of Plant
Biotechnology, Baramati.
Project Guide:
Dr. Sharad Bhange
Sai Samruddhi Biotech
& Nursery.
A/p, Khadaka,
Ahamadnagar.
5. 1. INTRODUCTION
• Caladium was discovered in the 19th century growing wild in Amazon Basin South
America. (
• Caladium are of great importance due to their indoor and outdoor decorative value.
• Caladium are mostly used in vertical gardening.
• Caladium is large genus of ornamental plant in the Araceae family.
• Caladium also cultivated as House plant.
• Conventional propagation of Caladium bicolor through tuber cutting and
seed germination is slow and inconsistent with the demand
• The pollen and the inflorescence both have short lives, which means a large
collection of Caladium bicolor is necessary if cross breeding is to be done
successfully.
6. Classification of Caladium:
Kingdom Plantae
Division Angiosperms
Class Monocots
Order Alismatales
Family Araceae
Subfamily Aroideae
Genus Caladium vent
Species Caladium
Plate 1: Caladium bicolor plant
7. 2. OBJECTIVE
1. To identify a suitable hormonal combination for in vitro
regeneration.
8. • The qualitative and quantitative development
of plants.
• Conventional propagation of Caladium with
tuber and seed is slow inconsistent with
demand.
• Micro propagation is the best option to solve
this problem by providing consistent quality
plant.
3. PROJECT SIGNIFICANCE
9. Yazawa et al., (2003): Leaf explants of Caladium ‘Pink Cloud’ were cultured in vitro on MS medium
containing various auxins (NAA, IBA, IAA, 2,4,5-T and 2,4-D) in combination with cytokinin (BA).
NAA gave the most vigorous in vitro propagation of this plant, and only 15% of the plants were leaf-
colour variants on the medium containing 0.5mol NAA. Leaf colour variation was observed in all plants
regenerated on the medium containing 2,4-D at 0.5 4.5mol. In hormone-free medium, only a few leaf-
colour variants (6%) occurred, but the rate of plant regeneration was very low. Application of 0.5mol
NAA together with 4.5mol BA seemed to be the most appropriate for in vitro propagation of
Caladium‘Pink Cloud’ with only a few leaf-colour variants.
Ali et al., (2007): Rapid propagation of Caladium bicolor was achieved in vitro. Excellent results for shoot
induction from apical meristem were obtained when it was cultured on MS medium containing 1 mg L-1BAP
within 8 days of inoculation. For shoot multiplication maximum number of shoots was obtained when 0.25 mg
L-1NAA was added to shoot induction medium i.e., MS medium supplemented with 1 mg L-1BAP + 0.25 mg L-
1NAA. Cent percent rooting was achieved by transferring an individual microshoot to modified MS medium
containing combination of 2 mg L-1IBA + 1 mg L-1NAA.
Benega et al., (2007): established this protocol, in vitro shoots developed from petioles cultured on a semi-solid
medium were used as starting materials. When using temporary immersion the multiplication rate was more than
12 times higher than under a conventional propagation system after 45 days. The highest multiplication rate was
found when explants were cultured in sprouting medium (MS + 2.0 mg/l 6-BAP) in the temporary immersion
system for four weeks. The highest number of competent (i.e. ready for acclimatization) and uniform plants was
achieved when bud clusters were subcultured for four weeks on MS medium without plant growth regulators.
Plantlets could be effectively acclimatized (92%) on a 1:1 zeolite : sugarcane filter (i.e. derived from the sugar
milling process) substrate.
4. REVIEW OF LITERATURE
10. Shah et al., (2007): Different planting media had enormous effect on various growth parameters.
Maximum number of sprouts (5.7), number of leaves (17.8), leaf length (36.0 cm), lamina area (521.3
cm2), plant height (57.8 cm), number of tubers (4.8), tuber size/volume (55.2 cm3) and tuber weight
(53.8g) were noted in SMC. Control (soil + silt + FYM at 1:1:1 ratio), produced minimum values for the
mentioned parameters. In case of cultivars, cv. Calypso performed better in number of sprouts (5.7),
numbers of leaves (19.4), leaf length (37.4 cm), lamina area (595.7 cm2), plant height (64.4 cm), while
minimum number of sprouts (2.0), number of leaves (8.8), leaf length (22.1 cm), lamina area (196.9
cm2), number of tubers (1.6), tuber size (29.2 ml) and minimum tuber weight (32.0 g) was recorded for
cv. Gypsy Rose. Maximum tuber size (57.6 cm3) and tuber weight (56.7 g) was recorded in cv. White
Christmas, while maximum number of tubers (4.2) was noted in cv. Fannie Monsoon. The interaction
between media and cultivars was found non significant for all the parameters.
Maqsood et al., (2015): An efficient somatic embryo encapsulation and in vitro plant regeneration technique
were established with Caladium bicolor, an important ornamental plant. Tuber derived embryogenic callus
(95.50%) was obtained on Murashige and Skoog (MS) medium amended with 0.5 mg L-1 α-Naphthalene acetic
acid (NAA) + 0.5 mg L-1 6-Benzyladenine (BA). The embryogenic callus later differentiated into somatic
embryos in the same plant growth regulators (PGRs) added medium (NAA and BA). The induced embryos
matured and developed into plantlets in NAA and BA added media; maximum plantlets development was
observed at 1.0 mg L-1 NAA + 1.0 mg L-1 BA supplemented medium. Synthetic seeds were produced by
encapsulating embryos in gel containing 3.0% sucrose + 3.0% sodium alginate and 100 mM of calcium chloride.
The highest synthetic seed germination (97.6%) was observed on medium supplemented with 1.0 mg L-1 NAA
+ 1.0 mg L-1 BA.
Seydi et al., (2016): Leaf explants Caladium bicolor (Aiton) Vent., were cultured in vitro on MS media
supplemented with 25 different concentrations of BAP and NAA in order to determine the appropriate con-
centrations for micropropagation. All combinations induced callus formation on explants. Callus production on
leaves explants grown on control medium was very low. The medium enriched with 4 mg l-1 BAP + 0.5 mg l-1
NAA was the most effective for callus formation. The highest number of shoots (6.43 per explant) and roots
(5.56 per explant) were regenerated on media containing 1 mg l-1 BAP + 0.5 mg l-1 NAA and 3 mg l-1 BAP +
0.5 mg l-1NAA, respectively. The regenerated plantlets were grown in a greenhouse and acclimatized
successfully.
11. 5.Material and Methods
• Two to three month old mother plants of Caladium bicolor were
collected. Tuber segments of Caladium bicolor were used as
explants for in vitro micropropagation studies.
• Plants of caladium bicolor were collected from Samridhhi
Biotech, Newasa.
Plate 2. Mother Plant
12. • Chemicals (For surface
sterilization):-
1) 0.1% Bavistin
2) 0.1% Tween 20
3) 0.1% Mercuric chloride
• Equipments:-
1) Electronic weighing
balance
2) pH meter
3) Autoclave
4) Refrigerator
5) Laminar Air flow
Plate 3: Facilities available at work place.
13. • Equipments used in the inoculation room:-
1) Forceps
2) Scalpels
3) Cotton
4) Thermometer
14. • Glasswares: -
1) Beakers
2) Conical flasks
3) Measuring cylinders
4) Pipettes
5) Volumetric flasks
6) Glass rods
7) Culture bottles
• Chemicals:-
1) MS medium
2) Agar powder
3) Growth regulators (BAP and IBA)
Plate 4: Chemicals and instruments.
15. Table 1 . Medium preparation table :-
N
o.
Components MS Medium
(1 L)
Initiation
Medium (4 L)
Multiplicati
on Medium
(2 L)
Rooting
Medium (2
L)
1 Macronutrient 50 ml 200 ml 100 ml 100 ml
2 Micronutrient 5 ml 20 ml 10 ml 10 ml
3 Iron ( EDTA, FeSo4) 5 ml 20 ml 10 ml 10 ml
4 Vitamin 1 ml 4 ml 2 ml 2 ml
5 Glycine 1 ml 4 ml 2 ml 2 ml
6 KI 1 ml 4 ml 2 ml 2 ml
7 Inositol 100 mg 400 mg 200 mg 200 mg
8 Sucrose 30 gm 120 gm 60 gm 60 gm
9 Hormones - BAP (0.5, 1,
1.5, 2 mg/l)
BAP
(0.5,1.0,1.5
mg/l)
IBA
(0.5,1.0,1.5
mg/l)
10 Agar 6 gm 24 gm 12 gm 12gm
11 pH 5.7 5.7 5.7 5.7
16. MEDIA PREPARATION
The medium was
prepared by adding
required amount of
stock solution and final
volume was made up
with distilled water.
The pH was adjusted to 5.7
with the help of 1N
NaOHand 0.1N Hcl&
Agar is added to solidify
the medium.
The medium was poured in
washed culture bottles and
autoclaved at 121C for 20
min at 15 lbs. pressure.
17. Surface Sterilization :-
Selection of explant (Tuber segment).
The explants were cut into small pieces
about 1 to 2 cm
The explants were washed under
running tap water for 10 min
Explant treated with Bavistin 0.1% For
20 min.
Explant washed with distilled water for
20 min.
Then Explant treated with 5 drops of
Tween-20 for 10 min.
Plate5: Bavistin Treatment
Plate 6: Tween-20 Treatment
18. Explant treated with 0.1% HgCl2 for 5
min.
Explant were throughly washed with
sterile distilled water for 4 times.
Explant washed with sterile distilled
water.
Then treated with streptocyclin 40 ppm
for 10 min.
Explant washed with distilled water for
20 min.
Plate8: HgCl2 Treatment
Plate7: Streptocycline Treatment
19. Inoculation of Explant
All inoculation and aseptic manipulation
were carried out in laminar air flow cabinet.
Initiation medium were divided into 5 parts
given below.
Explants were cut into very small pieces
with sterile scalpel, and inoculate in the
medium.
1) MS Medium (control)
2) MS Medium + 0.5 mg/l BAP
3) MS Medium + 1.0 mg/l BAP
4) MS Medium + 1.5 mg/l BAP
5) MS Medium + 2.0 mg/l BAP
20. SR. NO. Treatments Medium + Hormones
1. T0 MS Medium
2. T1 MS + 0.5 mg/l BAP
3. T2 MS + 1.0 mg/l BAP
4. T3 MS + 1.5 mg/l BAP
5. T4 MS + 2.0 mg/l BAP
Table No. 2: Shoot Initiation hormone combinations
21. S.R. NO. Treatments Medium + Hormone
1. T0 MS Medium (Control)
2. T1 MS + 0.5 mg/l BAP
3. T2 MS + 1.0 mg/1BAP
4. T3 MS + 1.5 mg1/BAP
Table No. 3 Shoot Multiplication hormones
22. Rooting:
Tuber segments were cutted into 1.5 to 2 cm
size and cultured on rooting medium.
Rooting was carried out using 4-5 cm long and
vigorous shoots from the shoot multiplication
stage.
23. S.R. NO. Treatments Medium + Hormone
1. T0 MS Medium (Control)
2. T1 MS + 0.5 mg/l BAP
3. T2 MS + 1.0 mg/1BAP
4. T3 MS + 1.5 mg1/BAP
Table No. 4: Root Initiation Hormones
24. Acclimatization
• Newly formed adventitious roots washed with
distilled water to remove the traces of agar.
• For acclimatization, plantlet will be transferred
into pots containing sterile mixture of sand,
soil and compost in ratio 2:1:3
• After that these pots will be shifted to green
house condition for acclimatization after that
plantlet will be shifted to field condition for
Hardening.
26. Shoot Multiplication
Plate 11: Shoot Multiplication
MS medium + 0.5,1.0,1.5 mg/l BAP were
used for shoot multiplication medium.
Tuber segments were cutted into 1.5 -2 cm
size and cultured on shoot multiplication
medium.
Shoot multiplication was carried out using
the 5-6 cm long and most vigorous shoots
from the initiation stage.
27. Table 5: Establishment of Caladium bicolor tuber explant using different
combinations of BAP.
SR. NO. Medium +
Hormones
Average number
of shoots per
explants
Average
length of shoot
(cm)
Survival %
1. MS Medium 5 ± 0.47 3.3 ± 0.3 50
2. MS + 0.5 mg/l BAP 7.75 ± 0.47 6.1 ± 0.2 90
3. MS + 1.0 mg/l BAP 5.5 ± 0.28 3.7 ± 0.3 80
4. MS + 1.5 mg/l BAP 2.25 ± 0.47 1.6 ± 0.3 70
5. MS + 2.0 mg/l BAP 2.25 ± 0.25 1.5 ± 0.2 60
28. Fig 4.1 Effect of BAP on Caladium Bicolor
3.3
6.1
3.7 1.6 1.5
50
90
80
70
60
5 7.75 5.5
2.25 2.25
0
10
20
30
40
50
60
70
80
90
100
Control BAP 0.5 mg/l BAP 1.0 mg/l BAP 1.5 mg/l BAP 2.0 mg/l
Average length of
shoot (cm)
Survival %
Average no of shoots
29. Table 6: Shoot multiplication of using Caladium bicolor 0.5 mg/l BAP.
S.R. NO. Medium + Hormone Average number
of shoots per
explants
Average length
of shoot (cm)
Survival%
1. MS Medium
(Control)
8.50± 0.17 3.0±.027 50%
2. MS + 0.5 mg/l
BAP
11.66 ± 0.37 6.0 ± 0.47 90%
3. MS + 1.0
mg/1BAP
9.50± 0.13 4.0± 0.17 80%
4. MS + 1.5
mg1/BAP
7.40 ± 0.12 4.7 ± 0.15 70%
31. Fig4.2 Effect of BAP on shoot multiplication
3
6
4 4.7
50
90
80
70
8.5
11.66
9.5
7.4
0
10
20
30
40
50
60
70
80
90
100
Control BAP 0.5 mg/l BAP 1.0 mg/l BAP 1.5 mg/l
Average number
of shoot per
explant (cm)
Survival %
Average length
of shoot (cm)
32. Table No. 7: Rooting of Caladium bicolor using IBA
SR. NO. Medium +
Hormones
Average
number of roots
per explants
Average
length of root
(cm)
Survival %
1. MS Medium 5.5 ± 0.40 1.2 ± 0.1 50
2. MS + 0.5 mg / l IBA 6.6 ± 0.50 1.7 ± 0.2 90
3. MS + 1.0 mg/l IBA 4.2 ± 0.15 1.1 ± 0.1 80
4. MS + 1.5 mg /1 IBA 3.5 ± 0.10 1.0 ± 0.2 70
34. Plate13: Tuber segment inoculate
for rooting
Medium used for rooting:
MS Medium+ 0.5,1.0,1.5mg/l
IBA
Tuber segments cultured on
rooting medium.
35. Fig. 4.3 Effect of IBA and NAA on rooting
5.5 6.6
4.2 3.5
50
90
80
70
1.2 1.7 1.1 1
0
10
20
30
40
50
60
70
80
90
100
Control IBA 0.5 mg/l IBA 1.0 mg/l IBA 1.5 mg/l
Chart Title
Average number
of roots per
explant
Survival %
Average length of
root (cm)
36. Primary Hardening
• The well rooted caladium plantlets
were removed gently from the culture
bottles after four weeks of rooting.
• Then washed thoroughly with
running tap water to remove all the
traces of the adhering agar
• After that the plantlets were
transplanted in 4 cm diameter small
polythene cup tray filled with coco
peat and soil (1:1).
37. Initiation
i. Initiation of explants tuber segments were used on MS medium with
different concentration of BAP (0.5, 1.0, 1.5, 2.0 mg/l) for shoot
initiation.
ii. Out of all these concentrations BAP (0.5 mg/l) gave best shoot
initiation. Ahmed et al., (2002) .
iii. MS medium containing BAP (0.5 mg/l) provided best result from
tuber segment explants .
iv. The highest survival percentage 90% and average number of shoots
per explants (7.75 ± 0.47) and average length of shoots (6.1 ± 0.2 cm)
was obtained on MS medium supplemented with 0.5 mg/l BAP .
Outcomes
38. Multiplication
i. MS medium with combination of BAP (0.5 mg/l) were used for shoot
multiplication.
ii. In this treatment BAP showed excellent length for shoot
multiplication and highest survival percentage (90%).
iii. The average number of shoots per explants (11.66 ± 0.37) and best
average shoot length (6.0 ± 0.47 cm) was observed on MS Medium
supplemented with 0.5 mg/l BAP .
iv. Ahamd et al., (2002) also reported that multiple shoots from tuber
segments was the highest in MS medium supplemented with 0.5 mg/l
BAP.
39. Rooting
i. The best rooting response was observed on medium
containing 0.5 mg/l IBA showed highest average root
length (1.7 ± 0.2 cm).
ii. It showed highest average number of roots per explant
(6.6 ± 0.50) as compared to medium supplemented with
0.5 mg/l IBA.
iii. The highest survival percentage 90% was observed when
medium was supplemented with 0.5 mg/l IBA (Table 4.3).
iv. Ali et al.,(2007) also reported that the maximum root
induction (100%) was observed on medium
supplemented with 0.5 mg/l IBA.
40. Primary hardening
• The well rooted plantlets were transferred to
seedling tray containing cocopeat and soil (1:1)
mixture.
• The same mixture also used by Cooper et al, (1983)
for primary hardening.
41. • After four weeks of initiation, maximum number of shoots
were obtained in medium supplemented with 0.5 mg/l BAP.
and the highest survival percentage 95% was obtained when
the MS medium was supplemented with 0.5 mg/l BAP.
• The highest number of roots per explant as well as the
highest root length was recorded on medium containing 0.5
mg/l IBA
• The effect of hormone like BAP gave better response in
shoot multiplication and IBA hormone show highest root
number in rooting growth.
7. Summary
42. Conclusion:-
• Caladium is sweet herb and an important ornamental plant, but is
avilability less due to its infertile and small size seed.
• The method of vegetative propagation are not efficient to save this rare
plant.
• The production of large number of caladium plants were possible through
in vitro propagation techniques.
• In caladium bicolor MS medium containing 0.5 mg/l BAP was the best
for culture initiation.
• In caladium bicolor MS medium containing 0.5 mg/l BAP was suitable
for shoot multiplication and it shows best shooting response.
• IBA has been widely used as root induction hormone and showed
positive role during in vitro rooting. In caladium 0.5 mg/l IBA showed to
be the best for in vitro rooting.
43. 9.References
• Ali, AAMIR., Munawar, ASIFA., &Naz, SHAGUFTA. (2007). An in vitro
study on micropropagation of Caladium bicolor. Int J Agri Biol, 5, 731-735.
• Atta-Alla, H., McAlister, B. G., & Van Staden, J. (1998). In vitro culture
and establishment of Anthurium parvispathum. South African journal of
botany, 64(5), 296-298.
• Ahmed, E. U., HaSrujani, T., &Yazawa, S. (2004). Auxins increase the
occurrence of leaf-colour variants in Caladium regenerated from leaf
explants. Scientia horticulture, 100(1-4), 153-159.
• Allan, P. (1981). Plant propagation through tissue culture. South African
Avocado Growers Association Yearbook, 4, 22-26.
44. • Deng, Z., & Harbaugh, B. K. (2006). Garden White'—A Large White Fancy-
leaved Caladium for Sunny Landscapes and Large Containers. Hort Science,
41(3), 840-842.
• Hartman, R.D. (1974). Dasheen mosaic virus and other phytopathogens
eliminated from Caladium, taro, and cocoyam by culture of shoot tips.
Phytopath.64: 237-240.
• Shah, M., Khattak, A. M., & Amin, N. U. (2007). Effect of Various Amended
Organic Media on the Tuberization of Caladium cultivars. SARHAD JOURNAL
OF AGRICULTURE, 23(4), 899.
• von Aderkas, P., & Bonga, J. M. (2000). Influencing Micropropagation and
somatic embryogenesis in mature trees by manipulation of phase change, stress
and culture environment. Tree Physiology, 20(14), 921-928.
46. A
Presentation
on
Entrepreneurship Development in Micropropagation of Caladium bicolor L.
and study of their cost
AT
(Samridhhi Biotech, Newasa)
By
Batule Prashant Dnyandev
[VSBT-42/2018]
Course Title:- Entrepreneurial Development in Biotechnology
Course No.:- READY-483
Project Guide: Prof. Sayali Badekar
VIDYA PRATISHTHAN’S
COLLEGE OF AGRICULTURAL BIOTECHNOLOGY, BARAMATI,413133.
(AFFILIATED TO MAHATMA PHULE KRISHI VIDYAPEETH, RAHURI, DIST-
AHMEDNAGAR,413722.)
(2021-2022)
46
47. 47
CONTENTS
1. Introduction
2. Profile of the Industry
3. Objectives taken for entrepreneur skill development
4. Scientific Information Gathered
5. Cost Analysis
6. SWOT Analysis
7. Institute Related Photographs
8. Experiance Gained
48. 48
1. Introduction
Plant tissue culture technology has been globally accepted as one of the important
tools for Direct application in agriculture. It has a strong and positive influence on
the agricultural sector worldwide.
The main advantage of tissue culture technology lies in the production of high
quality and Uniform planting material.
The demand for micropropagated plants in agriculture, horticulture and in social
forestry is growing now a day. There is a large gap between the demand and
supply. There is a need for boosting the youths in setting up additional units and
training manpower and supply plants with more competitive prices for improving
productivity.
49. 49
2. Profile of the Industry
Samridhhi Biotech, Newasa.
Fig.No.1- Logo of Samridhhi Biotech
50. SAMRIDHHI BIOTECH PVT. LTD TISSUE CULTURE LAB & NURSERY
A renowned name of the market, an expert tissue culture manufacturer and supplier & a customer
friendly private limited company are the identifying factors of our company.
People trust them because of our positive image in the market. Currently, we are engaged in
helping thousands of clients in finding the best 1 Year Growth Caladium Plants, Uniform Banana
Bunches and many other variety of plants.
These are offered under our name with a guarantee of long life, healthiness and exceptional
quality of the plants.
Samruddhi biotech deals with the production of best quality tissue culture plants like Caladium,
Banana, Bamboo, Teak etc.
The disease free plants that we offer to clients are packed in individual corrugated boxes as well
as crates and is delivered to clients on time
Upto 5 lakh capacity of various varieties of plants in nurseries
Upto 20-30 workers and 300 working days within year.
51. Objectives of Samridhhi Biotech
1. To achieve huge demand of planting material for farmers.
2. To propagate disease free high quality planting material.
3. To make good quality material availabale to farmers.
51
52. 52
3. Objectives taken for
entrepreneur skill development
To study the commercialization of plant tissue culture raised plants.
To collect information of plant prices, marketing of plants
and company.
To understand the Marketing set up of company.
53. 53
4. Scientific information gathered
Marketing Strategies are as follows:
Marketing and Advertising are the important tools for the business. Marketing is important because
it helps you sell the products and Advertising is also important it helps in marketing of
product.Marketing plays a very essential role in the success of a company. It educates people on the
latest market trends, helps boost a company’s sales and profit, and develops company reputation.
• Small Nursery Collaboration
• Dealers
• Pamphlets
1. Farmers Meeting: Institute organizes farmers meeting for guidance.
2. Field/ Plot Visit: They organizes plot visit for farmers on field demonstration are also given.
3. Exhibition: Exhibition event provides a best marketing tool.
4. Bonding with dealers and retailers: To keep the good relation with dealers and retailers.
55. 55
6. Cost Analysis
Table No.1: Equipments used in Tissue Culture Lab
Equipments Quantity Approx. Rate (Rs.) Total (Rs.)
Bottles with caps 15,000 5 75000
White Trays + Crates 150 200 30000
Autoclave with pulley and double bucket 1 1,25,000 1,25,000
Measuring equipments- plastic and glass 1 5,000 5000
Refrigerator 1 25,000 25000
pH meter 1 15,000 15000
Distilled water unit 1 30,000 30,000
Balances 2 20,000 40,000
Mechanical stirrer 1 18,000 18,000
Table 2 5000 10,000
Sealing machine 1 2000 2,000
High Pressure Gas burner with burton 1 3000 3,000
Table and exhaust fan 3 1500 4,500
Air conditioner 1 45000 45,000
56. 56
Table No.2: Equipments used in Tissue Culture Lab
Equipments Quantity Approx. Rate
(Rs.)
Total (Rs.)
LAF (6×2) feet with sterilizer 3 80000 2,40,0000
Air Conditioner 2 45000 90000
Chairs 2 5000 10,000
SS trollies 3 5000 15,000
Cutting equipments 15 5000 75,000
Appron, cap, mask,slipper and dustbins 6 6000 36,000
Cupboard 1 5000 5,000
Vacuum cleaner 1 15000 15,000
Rack with LED lights 5 50000 25,000
Furniture 1 8000 8000
Ladder 1 4500 4500
Thermometer and lux meter 2 3000 6000
Bottle washing machine 1 25000 25000
Total - - 10,22,500
57. 57
Table No.3: Fixed Investments
Expenses Amount (Rs.)
Equipments cost 10,22,500
Platform preparation and Tyling work 2,00,000
Lab irrection cost (900 sq.ft.)- Puff Panel 5,00,000
Plumbing and electrification work 1,50,000
Hardening unit (Tunnel/ Fogger System) 2,50,000
Consultancy charges (Equipments, Chemicals,Tissues
procurement, design, production protocol and planning, lab
handling training for 1 person)
2,00,000
58. 58
Table No.5: Sale and Profit of One Year for Caladium
Sale and Profit per year Approximate Values
Number of plants produced 35,000
Sale cost per plant Rs. ,50 per plant
Number of Plants sale 35,,000
Total sale Rs. 17,50,000
Total expenses Rs. 10,22,500
Profit Rs. 7,27,500
Profit % 70.93%
59. Strengths
• Production of beneficial quality
products for farmer’s crop
• Marketing of products by social
media
• High quality products
• Environment sustainability.
Weakness
• Competitors can offer similar
Products
• Monsoon effect
Opportunity
• Increasing demand-supply gap in
different states of India
• Continued expansion for
online sales
• Increasing demand of organic
products
Threats
• Taxation
• Risk of contamination with other
microbes.
• High volatility in the prices of raw
materials & chemicals
SWOT analysis
61. 61
9. Experiance Gained
I gained a lots of experience and practical knowledge by working
with Samrudhhi Biotech.
This training give an exposure to new and interesting information
about manufacturing units of plant tissue culture and their
Marketing.
It helps me to improve entrepreneurial skills like time
management, risk taking,
communication skills, problem solving and financial adjustments.