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 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 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 Influence of growth regulators and explants on shoot regeneration in carnation

2009 ◽  
Vol 36 (No. 4) ◽  
pp. 140-146 ◽  
Author(s):  
J.K. Kanwar ◽  
S. Kumar
Keyword(s):  
Acetic Acid ◽  
Shoot Regeneration ◽  
Growth Regulators ◽  
Dianthus Caryophyllus ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Shoot Bud ◽  
Dichlorophenoxy Acetic Acid ◽  
Number Of Shoots

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in <I>Dianthus caryophyllus</I> L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.

scholarly journals GROWTH OF ‘PRATA-ANA’ BANANA’S MICROSHOOTS CLONE GORUTUBA FROM SYNTHETIC SEEDS:SUBSTRATES AND BAP CONCENTRATION

2017 ◽  
Vol 39 (5) ◽  
Author(s):  
JÚLIO CÉSAR GOMES PEREIRA ◽  
SELMA SILVA ROCHA ◽  
LUCIANA CARDOSO NOGUEIRA LONDE ◽  
MARCELA CAROLINE BATISTA DA MOTA ◽  
PABLO FERNANDO SANTOS ALVES ◽  
...  
Keyword(s):  
Culture Medium ◽  
Economic Importance ◽  
Leaf Length ◽  
Naphthalene Acetic Acid ◽  
Synthetic Seed ◽  
Ms Medium ◽  
World Production ◽  
Number Of Leaves ◽  
Alginate Matrix

ABSTRACT The banana crop stands out as an activity of great social and economic importance in Brazil, which occupies the fifth place in world production. Synthetic seed production is becoming promising for a micropropagation and in vitro conservation. The aim of the study was to analyze the conversion and growth of ‘Prata-anã’ banana’s microshoots clone Gorutuba from synthetic seed in MS medium and vermiculite, different substrates and concentrations of BAP (6-benzylaminopurine) associated with ANA (acetic naphthalene acid) in the constitution of its capsule were tested. The microshoots were immersed in the sodium alginate matrix (3%) and dripped in a solution of CaCl2.2H2O (100 mM) for complexation and then in KNO3 solution (100 mM) to decomplex. The experimental design was completely randomized in a 2 x 5 factorial design (substrate x BAP concentrations), containing different substrates (MS culture medium and vermiculite) and BAP concentrations (2.22, 4.44, 6.66, 8.88 and 13.32 µmol L-1) associated with NAA (naphthalene acetic acid) 0.54 µmol L-1, totaling 10 treatments, with 4 replicates, and that each replicate containing 5 seeds. The evaluations of conversion, number of leaves, leaf length, leaf height, number of roots, root length and oxidation were performed at 30 and 60 days.The use of the MS medium provided better growth results in relation to vermiculite as substrate, in which the different BAP concentrations did not differ from each other. It was found that, in MS culture medium, BAP concentrations above 8.88 µmol L-1 in the capsule composition are not indicated for microshoots growth.

scholarly journals In vitro Regeneration and Rapid Multiplication of Dendrobium bensoniae, an Indigenous Ornamental Orchid

2017 ◽  
Vol 14 (2) ◽  
pp. 24-31 ◽  
Author(s):  
S S Riva ◽  
A Islam ◽  
M E Hoque
Keyword(s):  
Shoot Regeneration ◽  
In Vitro Regeneration ◽  
Ms Medium ◽  
Shoot Induction ◽  
Rapid Multiplication ◽  
Number Of Shoots

An experiment was conducted on in vitro regeneration and multiplication of Dendrobium bensoniae. Different concentrations of BA and IBA alone or combination of both hormones were used as treatment for regeneration.  It was revealed that shoot regeneration from node was the best at 2.0 mg/l BA supplemented to MS medium. It gave better responses than all other concentrations and combinations of BA and BA+IBA, used in the present study. The highest number of shoots and leaves were found when 1.0 mg/l BA with 1.5 mg/l IBA was supplemented into MS medium.  For rooting, 0.5 mg/l BA with 1.0 mg/l IBA was found to be the most effective. The well-rooted plantlets were successfully acclimatized under 70-80% humidity and planted in pots and transferred to the shade house for establishment. Around 85% of plantlets survived in the field. From the present result, it may be recommended that MS medium supplemented with 2.0 mg/l BA may be used for rapid shoot induction and regeneration of D. bensoniae.The Agriculturists 2016; 14(2) 24-31

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 Callus Induction and Shoot Formation for Mexican Red Bean (Phaseolus vulgaris L.) Pinto Cultivar in Vitro

2020 ◽  
pp. 1887-1893
Author(s):  
Rasha K. Mohammed Al-Saedi ◽  
Ansam G. Abdulhalem
Keyword(s):  
Growth Regulators ◽  
Culture Medium ◽  
Shoot Formation ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Phaseolus Vulgaris L ◽  
Different Types ◽  
Red Bean ◽  
Internode Explants

     The current study aimed to adopt a method for inducing callus cells and regenerating the important common red bean using different types of growth regulators such as N6-benzylaminopurine (BAP), Naphthalene acetic acid (NAA), and Thidiazuron (TDZ). Different types of common bean pinto cultivar explants, such as  internodes, cotyledons and roots,  were inoculated on Murashige and Skoog medium (MS) provided with different combinations of plant growth regulators, including 1- BAP (5 mg/l) 2-BAP (4.5 mg/l) NAA (0.5 mg/l), 3- BAP (4.5 mg/l), and TDZ (0.1mg/l). Callus was initiated on MS culture medium supplied with 5 mg/l BAP for all explants (internodes, cotyledons, and roots) at 50, 20, and 10% respectively, while adding NAA with 0.5mg/l showed a low percentage of callus (30%) only in the internode explants. Optimum results were obtained by growing the internodes on MS medium with 4.5 mg/l BA and either 0.5 mg/l NAA or 0.1 mg/l TDZ, transplanting the derived shoots into internodes and cotyledons with 70 and 10% respectively. This study concludes that the internodes as explants have the best growth results.

scholarly journals Indirect in vitro Regeneration of the Medicinal Plant, Aspilia africana, and Histological Assessment at Different Developmental Stages

2021 ◽  
Vol 12 ◽  
Author(s):  
Denis Okello ◽  
Sungyu Yang ◽  
Richard Komakech ◽  
Yuseong Chung ◽  
Endang Rahmat ◽  
...  
Keyword(s):  
Medicinal Plant ◽  
Shoot Regeneration ◽  
Callus Induction ◽  
Developmental Stages ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Regeneration Capacity ◽  
Root Explants ◽  
Indirect Regeneration

The medicinal plant, Aspilia africana, has been traditionally used in several African countries to treat many diseases such as tuberculosis, cough, inflammation, malaria, osteoporosis, and diabetes. In this study, we developed a protocol for in vitro propagation of A. africana using indirect shoot organogenesis from leaf and root explants of in vitro-grown seedlings and assessed the tissues at different developmental stages. The highest callus induction (91.9 ± 2.96%) from leaf explants was in the Murashige and Skoog (MS) medium augmented with 1.0 mg/L 6-Benzylaminopurine (BAP) and 1.0 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) while from root explants, the highest callus induction (92.6 ± 2.80%) was in the same plant tissue culture medium augmented with 0.5 mg/L BAP and 1.0 mg/L 2,4-D. The best shoot regeneration capacity from leaf-derived calli (i.e., 80.0 ± 6.23% regeneration percentage and 12.0 ± 6.23 shoots per callus) was obtained in medium augmented with 1.0 mg/L BAP and 0.05 mg/L α-Naphthaleneacetic acid (NAA); the best regeneration capacity for root-derived calli (i.e., 86.7 ± 6.24% shoot regeneration percentage and 14.7 ± 1.11 shoots per callus) was obtained in the MS medium augmented with 1.0 mg/L BAP, 0.05 mg/L NAA, and 0.1 mg/L Thidiazuron (TDZ). Regenerated plantlets developed a robust root system in 1/2 MS medium augmented with 0.1 mg/L NAA and had a survival rate of 93.6% at acclimatization. The in vitro regenerated stem tissue was fully differentiated, while the young leaf tissue consisted of largely unorganized and poorly differentiated cells with large intercellular airspaces typical of in vitro leaf tissues. Our study established a protocol for the indirect regeneration of A. africana and offers a basis for its domestication, large-scale multiplication, and germplasm preservation. To the best of our knowledge, this is the first study to develop an indirect regeneration protocol for A. africana and conduct anatomical assessment through the different stages of development from callus to a fully developed plantlet.

scholarly journals Long-term in Vitro Regeneration of Hamelia patens

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873G-874
Author(s):  
D. Sankhla ◽  
T.D. Davis ◽  
N. Sankhla ◽  
A. Upadhyaya
Keyword(s):  
Culture Medium ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Prolific Shoot ◽  
Ornamental Shrub

This report describes an efficient in vitro regeneration protocol for H. patens (firebush), a heat-tolerant ornamental shrub native to tropical and subtropical America. Shoot cultures were initially established using shoot tips placed on MS-revised medium containing 2.3 μM 2,4-D, 2.3 μM kinetin, and 0.25% polyvinylpyrrolidone. Other types of explants (nodal and internodal segments, leaf pieces, floral buds) did not regenerate shoots when placed on this medium. Two-month-old plantlets derived from the shoot tips were subcultured on MS medium supplemented with 0.5 μM thidiazuron (TDZ), and within 3 to 4 weeks, some callus was produced at the root–shoot junction. When this callus, with a small portion of the root and shoots, was placed on MS medium with 0.05 μM TDZ and 0.01 μM ABA, prolific shoot formation occurred within 3 to 4 weeks followed by root formation. By regular subculturing every 5 to 6 weeks, hundreds of plantlets have been obtained over the past 3 years with no apparent decline in regeneration potential. Addition of activated charcoal (0.5%) to the culture medium has greatly improved growth of the plantlets.

scholarly journals Micropropagation and Genetic Fidelity of the Regenerants of Aglaonema ‘Valentine’ Using Randomly Amplified Polymorphic DNA

HortScience ◽  
2016 ◽  
Vol 51 (4) ◽  
pp. 398-402 ◽  
Author(s):  
Mohammed Elsayed El-Mahrouk ◽  
Yaser Hassan Dewir ◽  
Yougasphree Naidoo
Keyword(s):  
In Vitro Regeneration ◽  
Shoot Proliferation ◽  
Genetic Fidelity ◽  
Naphthalene Acetic Acid ◽  
Axillary Shoot ◽  
Ms Medium ◽  
Randomly Amplified Polymorphic Dna ◽  
Simple Protocol ◽  
Regenerated Plantlets

The present study reports a simple protocol for in vitro regeneration of Aglaonema ‘Valentine’ using axillary shoot explants for rapid multiplication and production of true-to-type plants. Different concentrations of benzyladenine (BA; 0, 1, 3, 5, and 7 mg·L−1), kinetin (Kin; 0, 1, 3, 5, and 7 mg·L−1), thidiazuron (TDZ; 0, 0.5, 1.0, 1.5, and 2.0 mg·L−1), naphthalene acetic acid (NAA; 0, 0.5, and 1.0 mg·L−1), and indole-3-butyric acid (IBA; 0, 0.5, and 1.0 mg·L−1) were used for shoot regeneration. The highest shoot proliferation (5.0) was obtained on Murashige and Skoog (MS) medium supplemented with 1.5 mg·L−1 TDZ and 1 mg·L−1 NAA. In vitro rooting was easily achieved with 100% at all concentrations of NAA and IBA supplemented to half- or full-strength MS medium. Regenerated plantlets were acclimatized in greenhouse with 100% survival rate. Randomly amplified polymorphic DNA (RAPD) analysis confirmed the genetic fidelity of the regenerated plantlets and mother plant.

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