scholarly journals An efficient in vitro regeneration protocol to generate stable transgenic lines of Black gram (Vigna mungo)

2018 ◽  
Vol 78 (3) ◽  
Author(s):  
Bidyut Kumar Sarmah ◽  
Trishna Konwar ◽  
Borsha Borah ◽  
Arun Kumar Handique ◽  
Sumita Acharjee
Keyword(s):  
Transformation Efficiency ◽  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Vigna Mungo ◽  
Ms Medium ◽  
Black Gram ◽  
Transgenic Lines ◽  
Half Strength ◽  
The Individual

An efficient and quick in vitro regeneration protocol was developed for black gram (Vigna mungo) using wounded embryonic axis with cotyledon as explant. Murashige and Skoog (MS) medium supplemented with 4.44 μM BAP and 2.32 μM Kinetin was found to be effective in producing maximum number (mean 7.80) of multiple shoots. The individual shoots elongated to 4.5 cm when MS medium was supplemented with 2.89 μM GA3 along with 0.44 μM BAP and 0.46 μM KIN. A novel in vitro rooting technique was also optimized for black gram using half-strength liquid MS medium supplemented with 1.34 μM NAA. The shoots in this medium produced the highest number (mean 7.50) of roots with root length of 6.02 cm. The plantlets were transferred to soil mixture and placed in greenhouse where more than 80% successfully grew to maturity. The same protocol was successfully used to generate transgenic black gram lines carrying Bt-Cry2Aa gene through Agrobacteriummediated transformation with a transformation efficiency of 0.42%. The rooted T0 plants grew to maturity and produced T1 seeds with the presence and expression of transgene in T1 plants. Thus, we have standardized an in vitro regeneration protocol suitable for generation of stable transgenic black gram plants.

scholarly journals In vitro regeneration of two high-yielding eggplant (Solanum melongena L.) varieties of Bangladesh

1970 ◽  
pp. 08-12
Author(s):  
Sabina Yesmin, Mst Muslima Khatun, Tanzena Tanny ◽  
Anica Tasnim Protity ◽  
Md Salimullah ◽  
Iftekhar Alam
Keyword(s):  
In Vitro Regeneration ◽  
Solanum Melongena ◽  
Multiple Shoots ◽  
Multiple Shoot ◽  
Healthy Plant ◽  
Ms Medium ◽  
Best Response ◽  
Half Strength ◽  
Multiple Shoot Regeneration

An in vitro regeneration protocol was developed for two high-yielding eggplant varieties (Solanum melongena L.) namely BARI begun-4 and BARI begun-6. Multiple shoots were regenerated from cotyledonary explants through organogenesis with growth regulators of different combinations and concentrations.  The best response towards multiple shoot regeneration was achieved from cotyledon explants on MS media complemented with 1 mg/l BAP + 0.2 mg/l IAA in both the two varieties of eggplant. Elongation of shoots was achieved on hormone free MS medium. Regenerated shoots of both the varieties produced   active in vitro root system on half strength of MS medium supplemented with 0.2 mg/l IBA.  The in vitro grown plantlets were acclimatized in soil, grew up to maturity, flowered, fruited and produced seeds as normal healthy plant like the control.

scholarly journals Development of an effective in vitro Regeneration protocol for BARI Mash 2 (Vigna mungo L.) an important legume crop in Bangladesh

2017 ◽  
Vol 6 (1) ◽  
pp. 23-33
Author(s):  
P Saha ◽  
M Afrin ◽  
AKM Mohiuddin ◽  
AM Shohael
Keyword(s):  
Callus Formation ◽  
In Vitro Regeneration ◽  
Vigna Mungo ◽  
Young Leaf ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Black Gram ◽  
Regeneration System ◽  
Indole Butyric Acid

Black Gram (Vigna mungo L.), widely known as Mashkalai in Bangladesh is an important protein source used as human food as well as fodder. BARI Mash 2 is a popular black gram variety released by Bangladesh Agriculture Research Institute (BARI) which is cultivated throughout the country and very popular especially in the char areas. Establishment of a reliable regeneration system for BARI Mash 2 has been tried for further genetic improvement. A rapid, reproducible and efficient in vitro regeneration method was developed using hypocotyl and young leaf explants through callus formation. The frequency of callus formation was highest (75%) on Murashige and Skoog (MS) medium supplemented with a high concentration (31.66 ?M) of 2,4-Dichlorophenoxyacetic Acid (2,4-D) using the young leaf as explants’ source. Callus induction rate was less in hypocotyls in the same medium. No further progress was observed from those calluses. MS medium containing 16.11?M of ?- Naphthalene acetic acid (NAA) showed the 70% calli induction from hypocotyls segment. These calli were amenable to produce multiple shoots (5-6 shoot) in the medium containing 17.75 ?M of 6 Benzyl aminopurine (BAP) alone and the combination of BAP (17.75 ?M ) and NAA (2.68 ?M). Shoots were rooted most effectively (55%) in half strength MS basal medium containing 7.38 ?M of Indole-butyric Acid (IBA). Well rooted plantlets were successfully acclimatized, transferred to the soil and found to produce flowers and fruits. The efficient and reproducible regeneration protocol described here allows for successful in vitro regeneration of BARI Mash 2 that is vital for future genetic manipulation.Jahangirnagar University J. Biol. Sci. 6(1): 23-33, 2017 (June)

scholarly journals An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501
Author(s):  
Sanjog T. Thul ◽  
Arun K. Kukreja
Keyword(s):  
Shoot Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Ms Medium ◽  
Mentha X Piperita ◽  
Half Strength ◽  
Multiple Shoot Regeneration

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals In Vitro Regeneration of Phyllanthus amarus Schum. and Thonn.: An Important Medicinal Plant

Our Nature ◽  
1970 ◽  
Vol 7 (1) ◽  
pp. 110-115 ◽  
Author(s):  
A. Sen ◽  
M.M. Sharma ◽  
D. Grover ◽  
A. Batra
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Phyllanthus Amarus ◽  
Regenerated Plants ◽  
Half Strength ◽  
Important Medicinal Plant

An efficient in vitro plant regeneration protocol was developed for the medicinally potent plant species Phyllanthus amarus Schum. and Thonn. (Euphorbiaceae) using nodal segment as explant. Maximum multiplication of shoots (15.275±0.96) was achieved on Murashige and Skoog’s medium supplemented with BAP (0.5 mg/l) after 3-4 weeks of inoculation. The shoots were separated from cluster and subcultured for their elongation on the same medium. In vitro flowering was also observed on the elongated shoots after 3–4 weeks of sub culturing on the shoot elongation medium. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l).  Regenerated plants were successfully hardened and acclimatized, 80 % of plantlets survived well under natural conditions after transplantation.Key words: In vitro regeneration, multiple shoots, nodal segments, Phyllanthus amarusDOI: 10.3126/on.v7i1.2557Our Nature (2009) 7:110-115

scholarly journals Establishment of in vitro regeneration protocol for Adhatoda vasica Nees

2016 ◽  
Vol 51 (1) ◽  
pp. 75-80 ◽  
Author(s):  
S Khan ◽  
S Akter ◽  
A Habib ◽  
TA Banu ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Garden Soil ◽  
Root Induction ◽  
Ms Medium ◽  
Adhatoda Vasica

An in vitro regeneration protocol of Adhatoda vasica has been developed using excised nodal segments and juvenile leaves for multiple shoots regeneration directly or through callus induction. Explants were cultured on MS medium with different concentrations of IAA, NAA, BAP, GA3 and Kn singly or in combinations. MS medium supplemented with BAP (10.0 mg/l) was found best for multiple shoot formation, in which 93.33% explants produced multiple shoots. After two months, maximum number of multiple shoots were 10.6 ± 1.82, highest length of plantlets was 5.2 ± 2.20 cm. 100% calli formation were observed on MS medium supplemented with IAA (0.05 mg/l) + NAA (0.05 mg/l) + BAP (1.0 mg/l). Callus initiation started after 14 days and gave light green colored callus. Best callus mediated shoot regeneration was found on MS+10.0 mg/l BAP medium. Root induction of in vitro raised shoots was best on ½ MS + IBA (1.0 mg/l). Well rooted plantlets were transferred to plastic pots containing garden soil and compost in a ratio of 2:1 for hardening. The ultimate survival rate under natural condition was about 80%.Bangladesh J. Sci. Ind. Res. 51(1), 75-80, 2016

scholarly journals In vitro Regeneration of Grass Pea (Lathyrus sativus L.)

2016 ◽  
Vol 4 (2) ◽  
pp. 1-8 ◽  
Author(s):  
P Saha ◽  
M Afrin ◽  
AKM Mohiuddin ◽  
AM Shohael
Keyword(s):  
In Vitro Regeneration ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Nodal Explants ◽  
Lathyrus Sativus ◽  
Naphthalene Acetic Acid ◽  
Grass Pea ◽  
Half Strength ◽  
Lathyrus Sativus L

In vitro regeneration protocol for grass pea (Lathyrus sativus L.) was optimized using different concentrations and combinations of growth regulators. Direct shoot regeneration obtained through shoot organogenesis from different explants of grass pea cultured on MS medium supplemented with Gamborg B5 vitamin containing 6-benzylaminopurine (BAP), Thidiazuron (TDZ) and ?-naphthalene acetic acid (NAA). Highest percentage of shoots were obtained at 4.0 mg/l of BAP on nodal explants. Stunted multiple shoots were developed from nodal explants while 1.5 mg/l TDZ was used. About 56% of direct shoots were also obtained, while the combination of BAP (4.0 mg/l) and NAA (0.5 mg/l) were used. Regenerated plantlets were rooted most effectively (40%) in rooting medium containing half strength of MS basal medium containing 1.0 mg/l NAA. Well rooted plantlets were further successfully acclimatized to ambient humidity level and grown in controlled environment until hardening.Jahangirnagar University J. Biol. Sci. 4(2): 1-8, 2015 (December)

scholarly journals An efficient protocol for rapid plant regeneration from deembryonated cotyledons of black gram [Vigna mungo (L.) Hepper]

2019 ◽  
Author(s):  
R. Anandan ◽  
T. Deenathayalan ◽  
R. Bhuvaneshwari ◽  
M. Merlin Monisha ◽  
M. Prakash
Keyword(s):  
Shoot Elongation ◽  
Multiple Shoot ◽  
Vigna Mungo ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Black Gram ◽  
Regeneration System ◽  
Direct Shoot Regeneration ◽  
Multiple Shoot Induction

Here an efficient protocol for micropropagation of black gram [Vigna mungo (L.) Hepper] cv. VBN 3 is reported. The deembryonated cotyledonary explants were cultured on MS medium containing different concentrations of plant growth regulators. The maximum frequency (72%) of direct shoot regeneration (devoid of callus phase), multiple shoot induction and shoot elongation was achieved from culturing the explants on MS medium containing 3.0 mg/l of 6-benzylaminopurine (BAP). Up to 65% of the regenerated shoots were rooted on MS medium containing 0.25 mg/l of á-naphthalene acetic acid (NAA) within 3 weeks after subculturing. The in vitro-raised plantlets were successfully hardened first under culture room conditions with 62% survival rate and then in greenhouse. The identified regeneration system could be efficiently used in various in vitro manipulation studies in black gram as well.

scholarly journals (175) In Vitro Regeneration of Cladrastis kentukea

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1052C-1052
Author(s):  
Denita Hadziabdic ◽  
Robert N. Trigiano ◽  
Stephen Garton ◽  
Mark T. Windham ◽  
William E. Klingeman
Keyword(s):  
Growth Regulators ◽  
Root Length ◽  
Woody Plant ◽  
In Vitro Regeneration ◽  
Cambial Activity ◽  
Ms Medium ◽  
Total Root Length ◽  
Woody Plant Medium ◽  
Half Strength

Axillary buds from a single Cladrastis kentukea tree were initially cultured on two media, woody plant medium (WPM) and Murashige and Skoog (MS) containing 0, 1, 2, or 4 μm 6–benzylaminopurine (BA). Cultures were transferred to fresh media every 4 weeks. Elongated shoots were harvested after 39 weeks and transferred to half-strength MS medium supplemented with the following concentrations of IBA: 0, 3, 30, 100, and 300 μm for 3 d, then returned to half-strength MS without growth regulators. Explants exposed to 300 μm of IBA produced significantly more roots (75%) compared to explants exposed to other treatments. Fifty-four and 45% of the microshoots rooted when exposed to 100 and 30 μm IBA, respectively. Only 4% of the microshoots rooted when exposed to 3 μm IBA and none of the control microshoots rooted. Although the 300 μm treatment yielded the most rooted plantlets, there was significantly higher terminal meristem abortion compared to other treatments. There were no statistical differences between the numbers of roots and total root length among all treatments. Additionally, all microshoots that rooted had lenticels, suggesting that presence of lenticel cambial activity can possibly improve rooting abilities of selected microshoots. Rooted microshoots were gradually acclimatized to nonsterile environment.

scholarly journals Effects of N-Benzyl -9-(2-tetrahydropyranyl and Indole –3-Acetic Acid In Vitro Culture of Bauhinia purpurea L.

2019 ◽  
pp. 238-242
Author(s):  
Belai Meeta Suwal Singh
Keyword(s):  
Analytical Procedure ◽  
Multiple Shoots ◽  
Nodal Explants ◽  
Leguminous Plant ◽  
Ms Medium ◽  
Half Strength ◽  
Bauhinia Purpurea ◽  
Mature Seeds ◽  
Indole 3 Acetic Acid

<p>Bauhinia purpurea L. is a leguminous plant moderate sized tree with multipurpose value. It is distributed in sub-Himalayan tracts. It has been cultivated in the plain region up to the elevation of 1350 m. Mature seeds of Bauhinia purpurea L. were cultured on half strength Murashige and Skoog (1962) (MS) medium. Nodal explants obtained from germinated seedlings were cultured on MS medium containing 0.5 M BAP produced multiple shoots which were used for experimental purposes. Nodal explants obtained from cultured were subcultured on different concentrations of N-Benzyl -9-(2-tetrahydropyranyl) (BPA) and Indole-3acetic acid (IAA). The best proliferation of nodes and shoots were observed on the MS medium supplemented with 0.5 M BPA and 0.1 M IAA. After 8 weeks of culture, the propagated plants were acclimatized and transferred to the sand box containing 1:1 soil and sand. Well rooted plants were then established in the field. All the data collected were worked out statistically with SPSS, a system of analytical procedure.</p>

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