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 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 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 Saussurea heteromalla (D. Don.) Hand-Mazz Using Auxins and Current Status in Galiyat Abbottabad

2020 ◽  
Vol 10 (5) ◽  
pp. 1-7
Author(s):  
H. Mehreen ◽  
J. Zafar ◽  
G. Zishan
Keyword(s):  
In Vitro Regeneration ◽  
In Vitro Rooting ◽  
Current Status ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Indole Butyric Acid ◽  
Half Strength ◽  
Emergence Percentage

Current Status of Saussurea heteromalla was investigated in Galiyat areas of District Abbottabad, viz., Jahaffar, Seri, Beeran Gali, Banj, Haryala, Daryala Gali, Sarbhanna, Barriyan, Akhreela and Broangiala. Saussurea heteromalla was found common in Seri, Sarbhanna and Barriyan; endangered in Haryala, Jahaffar, Banj, Daryala Gali and Beeran Gali and absent in Akhreela and Broangiala. In vitro regeneration of Saussurea heteromalla (D. Don.) Hand-Mazz on MS media was conducted and shoots were developed on full strength MS medium supplemented with 1 mgL-1 GA3. The developed shoots were transferred for root induction to half strength MS medium fortified with various concentrations of Indole butyric acid (IBA) and α-Naphthalene acetic acid (NAA) i.e. T1 (control), T2 (2 mgL-1 IBA), T3 (3 mgL-1 IBA), T4 (4 mgL-1 IBA), T5 (1 mgL-1 NAA), T6 (2 mgL-1 NAA) and T7 (3 mgL-1 NAA). Maximum mean shoot length (6.3 cm), mean number of leaves (7), mean number of nodes (5.25); highest root emergence percentage (71%), means root length (1.5 cm), mean number of roots (3) and highest survival rate (100%) was recorded in treatment T4. However, treatments T5 and T6 also seem to be effective for in vitro rooting of Saussurea heteromalla. Treatments T2, T3 and T7 showed minimum root growth. It was concluded that IBA at higher concentration is more effective for in vitro rooting and better shoot growth of Saussurea heteromalla whereas NAA also initiate rooting but at lower concentration.

An effective protocol for improved regeneration capacity of Kabuli chickpeas

2012 ◽  
Vol 92 (6) ◽  
pp. 1057-1064 ◽  
Author(s):  
I. S. Yadav ◽  
N. P. Singh
Keyword(s):  
Shoot Regeneration ◽  
Culture Medium ◽  
Genetic Manipulation ◽  
In Vitro Regeneration ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Regeneration System ◽  
Embryonic Axes

Yadav, I. S. and Singh, N. P. 2012. An effective protocol for improved regeneration capacity of Kabuli chickpeas. Can. J. Plant Sci. 92: 1057–1064. An efficient protocol for in vitro regeneration is essential for genetic manipulation and micro-propagation of important plant species. A direct shoot regeneration system has been optimized for Desi chickpeas, but an effective regeneration protocol is still needed for Kabuli chickpeas. An efficient regeneration protocol for Kabuli chickpeas was developed, using whole embryonic axes, an embryonic axes slice and cotyledonary node explants from two genotypes L550 and JGK-1. Depending upon chickpea genotype, type of explant and culture medium, percentage of shoot producing explants (frequency) and the number of shoots per explant (efficiency) varied from 10 to 83% and from 1 to 58, respectively. The shoot regeneration capacity (SRC=frequency×efficiency), which is an indicator of the effectiveness of the protocol, varied from 47 to 2508 shoots per 100 explants cultured. On average, SRC of L550 was 1.8 times higher than JGK-1. Murashige and Skoog's (MS) medium+B5 vitamins supplemented with 8.0 µM benzyl amino purine (BAP)+0.5 µM α- naphthalene acetic acid (NAA) and 0.1 M sucrose plus embryonic axes was found to be the most effective culture medium and type of explants, respectively. Half strength MS medium+2% sucrose supplemented with 4 µM NAA, 3µ M IAA or 4µM IAA produced a high rooting percentage in both chickpea genotypes. The regeneration process starting from explant preparation to establishment of a complete plant in soil took 105–110 d. This optimized regeneration method holds promise for facilitating the insertion of interested genes through genetic transformation for improvement of Kabuli chickpeas.

scholarly journals Optimization of Callogenesis/Caulogenesis Induction Protocol in Saffron Plant (Crocus sativus L.) Using Response Surface Methodology

2021 ◽  
Vol 12 (4) ◽  
pp. 4731-4746
Keyword(s):  
Callus Formation ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Crocus Sativus ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Regeneration System ◽  
Indole Butyric Acid ◽  
2,4 Dichlorophenoxyacetic Acid

The Crocus sativus, an endangered medicinal and aromatic plant in Morocco, has a low propagation rate in natural conditions and, therefore, an efficient method for in vitro propagation is required. This study investigated the effects of various hormones on the induction of callogenesis and callogenesis in C. sativus corms using the Box-Behnken experimental design. The best shoot formation was obtained with Murashige and Skoog fortified with 3 mg/L 6-Benzylaminopurine. On the other hand, callus formation was obtained with 3 mg/L 1-Naphthaleneacetic Acid or 3 mg/L 2,4-Dichlorophenoxyacetic Acid. However, a combination of 3 mg/L 6-Benzylaminopurine, 1.056 mg/L Indole Butyric Acid, and 3 mg/L 2,4-Dichlorophenoxyacetic Acid allows 50% caulogenesis and 60% callogenesis. The in vitro regeneration system could be utilized for both conservation and largescale multiplication of Crocus sativus corms.

scholarly journals Effect of Different Media on in vitro Seed Germination and Seedling Development of Cymbidium aloifolium (L.) Sw.

2013 ◽  
Vol 14 (1) ◽  
pp. 51-56 ◽  
Author(s):  
Shreeti Pradhan ◽  
Tripti Regmi ◽  
Gaurav Parmar ◽  
Bijaya Pant
Keyword(s):  
Seed Germination ◽  
In Vitro Regeneration ◽  
Seedling Development ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Regeneration System ◽  
High Concentration ◽  
Plantlet Formation ◽  
Cymbidium Aloifolium

A comparative study on in vitro seed germination and seedling development of Cymbidium aloifolium (L.) Sw., an epiphytic medicinal orchid, was carried out on four different conditions of Murashige and Skoog (MS) and Knudson (KC) medium viz. full, ½. ¼ strength and medium supplemented with 0.5mg/l BAP (benzyl amino purine) and 0.5mg/l NAA (Naphthalene acetic acid). Varied response in terms of seed germination, protocorm formation and seedling development was observed on two different media. Medium supplemented with hormones favored optimum condition for the germination (approx. 90%) of seeds followed by full, ½ and ¼ strength on both MS & KC media. MS medium supplemented with 0.5mg/l BAP and 0.5mg/l NAA showed comparatively better response within 7 weeks of culture than other conditions of MS medium as well as KC medium. Based upon the results, it was found that MS medium was more effective than KC medium for germination, protocorm and plantlet formation. The present study has provided useful information that the high concentration of nutrient compounds supplemented with hormones are required for earlier in-vitro germination and plantlet development from immature seeds of C. aloifolium. It could be an important protocol to conserve this important orchid species by establishing an efficient in vitro regeneration system using immature seed culture. Nepal Journal of Science and Technology Vol. 14, No. 1 (2013) 51-56 DOI: http://dx.doi.org/10.3126/njst.v14i1.8878

scholarly journals Callus Induction in Baru (Dipteryx alata Vog.) Explants

2018 ◽  
Vol 47 (2) ◽  
pp. 538-543
Author(s):  
Rodrigo Kelson S. REZENDE ◽  
Ana Maria N. SCOTON ◽  
Maílson V. JESUS ◽  
Zeva V. PEREIRA ◽  
Fernanda PINTO
Keyword(s):  
Ascorbic Acid ◽  
Callus Induction ◽  
Callus Formation ◽  
Leaf Explants ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Alternative Technique ◽  
Propagation Methods ◽  
Dipteryx Alata

Baru (Dipteryx alata Vog.) is a species with great economic and environmental potential; it has popular acceptance, besides being a very productive species. Alternative propagation methods are important for species maintenance and exploration. Thus, micropropagation emerged as an alternative technique, providing genetic stability and the production of a large number of seedlings. The aim of the present investigation was to develop a callus induction protocol for in vitro baru explants. The tested explants were nodal, internodal and foliar segments. The explants were disinfected for 30 seconds in 70% alcohol (v/v) and 2 minutes in sodium hypochlorite (1.25% active chlorine). This was followed by triple washing. The inoculation was carried out in test tubes containing 15 mL MS medium (30 g L-1 sucrose, 6 g L-1 agar and 100 mg L-1 ascorbic acid) supplemented with 2.0 mg L-1 naphthalene acetic acid (NAA). The solution also contained 0.0, 2.5 or 5.0 mg L-1 of 6-benzylaminopurine (BAP) with the pH adjusted to 5.8. In the incubation phase, the explants were cultured for seven days in the dark and then subjected to a photoperiod of 16 hours (43 µmol m-2 s-1) at 25 ± 2 °C. The treatments were studied with 2.5, 5.0, 7.5 or 10.0 mg L-1 BAP additions to the MS. Callus formation, contamination and oxidation evaluations were undertaken. The results obtained when using 2.0 mg L-1 NAA concluded that such a treatment should be used to induce callogenesis from nodal explants, while for the tested baru leaf explants, the best results for callus formation were given by the combination of 2.0 mg L-1 NAA with 2.5 mg L-1 of BAP to.

scholarly journals In vitro regeneration protocol of Rauvolfia serpentina L.

2018 ◽  
Vol 53 (2) ◽  
pp. 133-138 ◽  
Author(s):  
S Khan ◽  
TA Banu ◽  
S Akter ◽  
B Goswami ◽  
M Islam ◽  
...  
Keyword(s):  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Nodal Segments ◽  
Root Induction ◽  
Ms Medium ◽  
Regeneration System ◽  
Rauvolfia Serpentina ◽  
Number Of Shoots

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13∙4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Bangladesh J. Sci. Ind. Res.53(2), 133-138, 2018

scholarly journals In vitro adventitious shoot regeneration from leaf explants of some apricot cultivars

2019 ◽  
Vol 43 ◽  
Author(s):  
Olga Vladimirovna Mitrofanova ◽  
Irina Vjacheslavovna Mitrofanova ◽  
Tatyana Nikolaevna Kuzmina ◽  
Nina Pavlovna Lesnikova-Sedoshenko ◽  
Sergey Vladimirovich Dolgov
Keyword(s):  
Callus Formation ◽  
System Development ◽  
Low Frequency ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Limiting Factor ◽  
Ms Medium ◽  
Regeneration System ◽  
Propagation Methods

ABSTRACT Apricot is one of the most valuable commercial fruits. In vitro propagation of apricot is very important for rapid multiplication of cultivars with desirable traits and production of cleaning up and virus-free plants. Low frequency of multiplication is the main limiting factor for traditional propagation methods. In this regard, the objective of our investigation was to study the morphogenetic capacity of apricot leaf explants of the promising cultivars ‘Iskorka Tavridy’, ‘Magister’ and ‘Bergeron’ for regeneration system development and solving some breeding questions. The source of explants was in vitro plants regenerated and cultured on QL medium. Leaves were maintained in the dark at 24±1 °C in thermostat for three-four weeks. Morphogenic callus and structures were mainly formed at the central and proximal parts of leaves on MS, QL and WPM media with 1.5 or 2.0 mg L-1 BAP and 1.5 or 2.0 mg L-1 IAA in different combinations, or TDZ (0.6 and 1.3 mg L-1). Callus with adventive buds was transferred to regeneration medium and placed into a growth chamber at 24±1 °C and 16-hour photoperiod with a light intensity of 37.5 μmol m-2 s-1. The best results were obtained when adaxial leaf surface was in contact with the culture medium. Frequency of leaf callus formation on MS medium with 1.5 mg L-1 BAP and 1.5 mg L-1 IAA was higher in the explants of ‘Iskorka Tavridy’ (80.0%) than in - ‘Bergeron’ (50.0%) and ‘Magister’ (36.7%). The best results of callogenesis for ‘Magister’ was obtained on MS medium with 1.3 mg L-1 TDZ (53.3%). Active microshoot regeneration in ‘Iskorka Tavridy’ cultivar was shown on MS medium with BAP and IAA and in ‘Magister’ cultivar - on MS medium with TDZ. Rhizogenesis was obtained on half strength MS medium with 2.0 mg L-1 IBA.

scholarly journals Phytohormones Influence on the In-Vitro Regeneration of Saccharum Officinarum L. from Apical Meristem Culture

2014 ◽  
Vol 5 (2) ◽  
pp. 85 ◽  
Author(s):  
Ejiroghene Felix Lawyer ◽  
Z. O. Jamaleddine ◽  
P. T. Lyam ◽  
I. T. Borokini ◽  
A. A. Adedeji ◽  
...  
Keyword(s):  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Apical Meristem ◽  
In Vitro Regeneration ◽  
Saccharum Officinarum ◽  
Naphthalene Acetic Acid ◽  
Root Induction ◽  
Indole Butyric Acid

Growth regulators especially auxins and cytokinins are critical for plant in-vitro regeneration. The effect of these plant growth regulators on in-vitro propagation of Saccharum officinarum L (Sugarcane) was investigated. In vitro response of two different varieties of sugarcane (NCS 005 and NCS 008) to Plant Growth Regulators was obtained in this study. Formation of buds was obtained on shoot apical meristem when cultured on MS (Murashige and Skoog) medium supplemented with 0.1mg/l BAP (6-Benzylaminopurine). After two weeks of initiation, regenerated meristem was inoculated into MS (Murashige and Skoog) fortified with different concentrations and combination of cytokinins. Shoot multiplication was optimal on 0.5mg/l BAP + 0.25 mg/l Kin(Kinetin) for NCS 005 variety while for NCS 008 variety, no significant (P≥0.05) difference was observed between 1.5mg/l BAP and 1.5mg/l BAP +0.5mg/l Kin. The best root induction for in vitro derived shoots was obtained on 1.0 mg/l NAA (Naphthalene acetic acid) and 2.0 mg/l IBA( Indole butyric acid) for both varieties of sugarcane within ten days of culture transfer. Successfully established plantlets showed excellent growth response when weaned under regulated green house conditions.

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