scholarly journals In vitro Regeneration of Recalcitrant Green Gram (Vigna radiata L. Wilczek) from Immature Cotyledons for Genetic Improvement

2018 ◽  
Vol 7 (1) ◽  
pp. 3072-3080
Author(s):  
Aparna Priyadarshini Patra ◽  
Kailash Chandra Samal ◽  
Gyana Ranjan Rout ◽  
Simachal Sahu ◽  
Prem Narayan Jagadev
Keyword(s):  
Vigna Radiata ◽  
Genetic Improvement ◽  
In Vitro Regeneration ◽  
Green Gram ◽  
Immature Cotyledons

scholarly journals PHYTOCHEMICAL SCREENING AND GAS CHROMATOGRAPHY-MASS SPECTROSCOPY ANALYSIS OF BIOACTIVE COMPOUNDS AND BIOSYNTHESIS OF SILVER NANOPARTICLES USING SPROUT EXTRACTS OF VIGNA RADIATA L. AND THEIR ANTIOXIDANT AND ANTIBACTERIAL ACTIVITY

2019 ◽  
pp. 180-184
Author(s):  
SIVAKUMAR T
Keyword(s):  
Gas Chromatography ◽  
Antioxidant Activity ◽  
Silver Nanoparticles ◽  
Antibacterial Activity ◽  
Mass Spectroscopy ◽  
Vigna Radiata ◽  
Green Gram ◽  
Hexadecanoic Acid ◽  
Gas Chromatography Mass Spectroscopy

Objectives: The present study was aimed to investigate the facile synthesis of silver nanoparticles (AgNPs) using the green gram sprout extract (GGSE) of Vigna radiata L. and also in vitro studies of antioxidant and antimicrobial activities. Methods: Gas chromatography-mass spectroscopy techniques have been used for the qualitative and quantitative evaluation of the phytochemicals present in the green gram seedlings. The antioxidant activity of AgNPs and GGSE was analyzed using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay. In vitro antibacterial activity was performed using the agar well diffusion method. Results: The presence of various secondary metabolites such as flavonoids, steroids, terpenoids, alkaloids, amino acids, polyphenol, glycoside, and protein was found in samples. The major chemical compounds of V. radiata were n-hexadecanoic acid, stigmasterol, caffeine, hexadecanoic acid, cholest-5-en-3-ol (3.beta.)-, and cyclopentane. The percentage of DPPH activity was enhanced on increasing the concentration of AgNPs. In vitro antibacterial effect of the diverse concentrations of AgNPs was investigated against each Gram-negative (Klebsiella aerogenes, and Escherichia coli) and Gram-positive (Bacillus substilis and Staphylococcus aureus) bacterial strains. Conclusion: The result suggests that biosynthesized AgNPs have good antibacterial and antioxidant activity and might be a potential for the bioactive components.

scholarly journals In Planta Genetic Transformation of Mungbean (Vigna radiata (L.) Wilczek) with Marker Gene

2019 ◽  
Vol 29 (2) ◽  
pp. 245-255
Author(s):  
Mohammad Mahmood Hasan ◽  
Sujay Kumar Bhajan ◽  
M. Imdadul Hoque ◽  
R.H. Sarker ◽  
Mohammad Nurul Islam
Keyword(s):  
Genetic Transformation ◽  
Genetic Improvement ◽  
In Vitro Regeneration ◽  
Marker Gene ◽  
Gus Expression ◽  
In Planta ◽  
Leaf Tissues ◽  
Gus Gene Expression ◽  
Screenable Marker

In genetic improvement of mungbean much success has not been achieved due to its recalcitrant nature towards in vitro regeneration. An attempt was made to develop an Agrobacterium-mediated in planta genetic transformation protocol for a locally grown mungbean variety BARI Mung-3 using a screenable marker gene. Two minutes of vacuum infiltration followed by 60 minutes of incubation period in Agrobacterium suspension of Winans’ AB medium containing wounded tobacco leaf extract was found most suitable towards genetic transformation in pricked de-coated half seed explants. An optical density (OD600) of 0.7 was found most effective for transient gus gene expression. Chimeric GUS expression was observed in the root and leaf tissues from the successfully transformed plantlets obtained through in planta transformation. This methodology of genetic transformation was found more suitable, easier and less time consuming than tissue culture based genetic transformation, which may be used for the genetic improvement of mungbean.

An Efficient in vitro Regeneration System for Australian-Grown Chickpea (Cicer arietinum) Cultivars

1995 ◽  
Vol 43 (5) ◽  
pp. 491 ◽  
Author(s):  
AL Adkins ◽  
ID Godwin ◽  
SW Adkins
Keyword(s):  
Cicer Arietinum ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Immature Cotyledon ◽  
Efficient Regeneration ◽  
Immature Cotyledons ◽  
Dim Light ◽  
Indole 3 Acetic Acid

A comparison of methods for efficient in vitro regeneration of Australian-grown chickpea (Cicer arietinum L.) cultivars was undertaken. The most efficient regeneration system was one where immature cotyledon and embryonic axis explants, 14-21 days post-pollination, were cultivated on Murashige and Skoog's salts with Gamborg's vitamins, 1.0, 3.0 or 5.2 mg L-1 zeatin, 0 or 35 μg L-1 indole-3-acetic acid, 30 g L-1 sucrose and 8 g L-1 Phytagar. The first embryoid structures appeared after 2 weeks of culture at 25 ± 1°C in dim light (150 μmol m-2 s-1) and formed directly on the edges of the immature cotyledons or petiole stumps. Between 10 and 20 structures were produced on each cotyledon explant in two cultivars, however, the embryogenic structures which developed on cv. Narayen were more efficiently transformed into shoots than far cv. Amethyst. An efficient regeneration medium (2 mg L-1 naphthaleneacetic acid, 1/2 Murashige and Skoog's salts with Gamborg's vitamins, and 0.5 g L-1 activated charcoal) was used to develop a portion of the shoots into morphologically normal plants growing in a vermiculite and soil potting mix in a growth room. Less efficient in vitro regeneration was observed when hypocotyl and shoot sections, and shoot apices were induced to form callus and plants by organogenesis. These plants could not be established in a potting mix. The amount and type of callus produced varied between explant type and cultivar.

Agrobacterium tumefaciens-Mediated Genetic Transformation of Green Gram [Vigna radiata (L.) Wilczek] - A Recalcitrant Grain Legume

2021 ◽  
Author(s):  
Aparna Priyadarshini Patra ◽  
Kailash Chandra Samal ◽  
Gyana Ranjan Rout ◽  
Simachal Sahu ◽  
Prem Narayan Jagadev
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Vigna Radiata ◽  
In Vitro Regeneration ◽  
Cotyledonary Node ◽  
Green Gram ◽  
Grain Legume ◽  
Marker Genes ◽  
Soil Productivity ◽  
Ms Medium

Background: Green gram is grown in many parts of India as a source of dietary protein (21-25%). It is an important nitrogen fixing crop which fixes atmospheric nitrogen (119-140 kg/ha) to soil and enhance the soil productivity. In the present investigation, efficient Agrobacterium-mediated genetic transformation of Vigna radiata L. (Wilczek) has been achieved with VrTIP1 gene for abiotic stress resistance i.e. moisture and salinity stress. Methods: Four days old shoot tip and cotyledonary node were used for in vitro regeneration with MS medium supplemented with BAP 2.0 mg/l, kinetin 0.5 mg/l and 50 mg/l kanamycin for co-cultivation with Agrobacterium tumefaciens strains, LBA 4404. The modified binary vector pCXSN, EHA105 containing hygromycin phosphotransferase II (hpt II) marker genes and a synthetic TIP1 gene under a constitutive CaMV35S promoter were used for transformation of Vigna radiata L. cotyledonary node explants. Putative transformants selected from hygromycin resistant shoots were subsequently rooted on MS medium supplemented with 1.0 mg/l NAA and later transferred to sterile vermiculite followed by transfer to the transgenic green house. Result: The T1 plants were produced from PCR positive T0 plants and analysed for presence and integration of transgenes in putative T1 plants were confirmed by polymerase chain reaction (PCR) amplification of 752 bp of hpt II fragment. This protocol can be effectively used for transferring new traits in greengram and other legumes for their quantitative and qualitative improvements.

scholarly journals In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Local Varieties of Mungbean (Vigna radiata (L). Wilczek)

2019 ◽  
Vol 29 (1) ◽  
pp. 81-97
Author(s):  
Sujay Kumar Bhajan ◽  
Setara Begum ◽  
Mohammad Nurul Islam ◽  
M Imdadul Hoque ◽  
Rakha Hari Sarker
Keyword(s):  
Genetic Transformation ◽  
Vigna Radiata ◽  
Zygotic Embryo ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Direct Organogenesis ◽  
Pcr Analysis ◽  
Root Induction ◽  
Half Strength

An efficient Agrobacterium-mediated transformation compatible in vitro regeneration protocol was developed for two important varieties of mungbean (Vigna radiata (L.) Wilczek) cultivated in Bangladesh, namely Binamoog-5 and BARI Mung-6. Two different zygotic embryo derived explants, such as cotyledonary node (CN) and cotyledon attached decapitated embryo (CADE) were used for direct organogenesis of shoot. MS supplemented with 4.0 μM BAP was found to be the best for the development of highest number of multiple shoots from CADE in both the varieties of mungbean. While in case CN the best shoot formation was achieved on MS containing 4.0 μM BAP and 0.5 μM NAA in both varieties. Half strength of MS with 2.0 μM IBA was found to be most effective for producing healthy root from regenerated shoots. Following root induction, the in vitro raised plantlets were successfully transplanted to soil for their establishment. Considering overall responses, genetic transformation efficiency was found to be better with CADE explant using Agrobacterium tumefaciens strain LBA4404 harboring the binary plasmid pBI121 conferring GUS and nptII genes. Different factors influencing transformation was optimized during this study. Selection of transformed shoots was carried out by gradually increasing the concentration of kanamycin and such transformed shoots were eventually selected using 200 mg/l kanamycin. Stable expression of the GUS gene was detected in various parts of regenerated transformed plantlets. Transformed shoots were rooted on half strength MS containing 2.0 μM IBA and 100 mg/l ticarcillin. Rooted transformed plantlets were successfully transferred to soil. Stable integration of GUS and nptII genes in the putative transformed shoots was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 81-97, 2019 (June)

Total phenol content of guava fruit and development of an in vitro regeneration protocol amenable to genetic improvement

2011 ◽  
Vol 4 (2/3/4) ◽  
pp. 225 ◽  
Author(s):  
Viji Sitther ◽  
Donna L. Harris ◽  
Sadanand A. Dhekney ◽  
Jinhe Bai ◽  
Elizabeth A. Baldwin ◽  
...  
Keyword(s):  
Genetic Improvement ◽  
In Vitro Regeneration ◽  
Total Phenol ◽  
Total Phenol Content ◽  
Phenol Content ◽  
Guava Fruit

Advances in in vitro studies for genetic enhancement of blackgram: A review

2019 ◽  
Author(s):  
Paras Pandey ◽  
Meenal Rathore ◽  
N. P. Singh
Keyword(s):  
Genetic Transformation ◽  
Genetic Improvement ◽  
In Vitro Regeneration ◽  
Vigna Mungo ◽  
In Vitro Studies ◽  
Genetic Enhancement ◽  
Black Gram ◽  
Treasure Trove ◽  
Genetic Base

Black gram (Vigna mungo) is a treasure trove legume in terms of nutritional efficiency and medicinal values and an important pulse that is cultivated throughout India in all three seasons. However, its production and productivity is constrained due to abiotic and biotic stresses.Though technology has aided in genetic improvement of the crop, the narrow genetic base remains a potential constraint. In vitro regeneration and genetic transformation have aided to its genetic improvement, yet a lot remains to be explored and harnessed for efficient and effective genetic improvement of the pulse. The review highlights the progress and achievements in context of in vitro regeneration and genetic transformation in blackgram.

scholarly journals Somatic Embryogenesis and Plant Regeneration from Commercial Soybean Cultivars

Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 38 ◽  
Author(s):  
Ghulam Raza ◽  
Mohan B. Singh ◽  
Prem L. Bhalla
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
In Vitro Regeneration ◽  
Crop Improvement ◽  
Soybean Cultivars ◽  
Immature Cotyledons ◽  
Soybean Genotypes

The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration. Overall, nine soybean cultivars belonging to different maturity groups were evaluated. Immature cotyledon ranging from 2–4 and 4–6 mm in size were used as explants for somatic embryogenesis induction. Maximum somatic embryo induction frequency (86%) was observed from 4–6 mm immature cotyledons of the cv. Jack (MG III), followed by 66%, 26%, 21%, and 6% in cultivars Williams (MG III), Snowy (MG III), MoonB1 (MG V), and PNR791 (MG V), respectively. On the other hand, cv. Snowy showed maximum somatic-embryo-inducing potential (67%) in 2–4 mm immature cotyledons followed by Williams, Jack, MoonB1, and PNR791. Somatic embryos from Jack, Williams, and Snowy cultivars were further tested for embryo proliferation, maturation, and germination. Maximum proliferation and maturation were observed in cv. Jack, followed by Snowy and Williams. However, cv. Snowy showed a significantly higher conversion of cotyledonary stage embryos to plantlets (85%), than both Jack and Williams cultivars (53% each). In conclusion, this study outlined a protocol for somatic embryogenesis and plant regeneration from three soybean cultivars. Our findings suggest commercial cv. Snowy could be a good candidate for developing transgenic plants through somatic embryogenesis.

scholarly journals Plant Regeneration and Genetic Transformation in Buckwheat (Fagopyrum spp.), A Multipurpose Gluten Free Crop of High Nutraceutical Importance: A Critical Review

2018 ◽  
Vol 7 (1) ◽  
pp. 1954 ◽  
Author(s):  
Mithilesh Kumar
Keyword(s):  
Plant Regeneration ◽  
Genetic Transformation ◽  
High Frequency ◽  
Genetic Improvement ◽  
In Vitro Regeneration ◽  
Gluten Free ◽  
Success Rates ◽  
Plant Propagation ◽  
Cultivated Species

Buckwheat (Fagopyrum sp.) from family Polygonaceae, with its two main cultivated species (F. esculentumMoench. and F. tataricum) is mostly grown in hilly regions of Eurasia. It is a multipurpose gluten free crop having great nutraceutical value. For plant propagation several reports have been published and significant differences were noticed in culture protocols, genotypes and their success rates. A few studies report genetic transformation in buckwheat with varying degree of success. The present study reviews the in-vitro regeneration and genetic transformation reports in Fagopyrum spp. available in publically available literature and concludes on what needs to be done for the sustainable genetic improvement of buckwheat. A genotype independent high frequency in-vitro regeneration protocol compatible with high end stable genetic transformation is the need of the hour for sustained genetic improvement of this crop.

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