scholarly journals Tissue Culture and Plant Regeneration of Cowpea (Vigna unguiculata)

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 627f-627
Author(s):  
Lurline Marsh

Explants (cotyledon, cotyledonary node, second node, hypocotyl, epicotyl, and leaf) of cowpea (Vigna unguiculata) genotypes MN13 and Pinkeye Purple Hull were cultured on Murashige and Skoog basal nutrient medium. The medium was supplemented with 1 mg·L–1 benzyladenine (BA) or 1 mg·L–1 benzyladenine plus naphthalene acetic acid (BA + NAA) or 2 mg·L–1 2,4-dichlorophenoxy acetic acid (2,4-D). Cultures were maintained at 22°C for 1 month, after which they were transferred to 1 mg·L–1 BA + NAA. Cotyledons, hypocotyl, epicotyl, and leaf segments produced only calli after subculturing in BA + NAA. The second node and cotyledonary node explants cultured on the BA or BA + NAA followed by subculture on BA + NAA produced calli, shoots, and roots. The plants were then transplanted to promix but later died.

HortScience ◽  
1993 ◽  
Vol 28 (4) ◽  
pp. 276E-276
Author(s):  
Lurline Marsh ◽  
Moshen Dkhill

Tissue culture of four cowpea (Vigna unguiculata) and two pigeonpea (Cajanus cajan) genotypes was tested on Blaydes' medium supplemented with different hormone concentrations. Explants of cotyledonary nodes, cotyledons and leaves of the cowpea genotype IT82E-16, IT64E-124, Pinkeye Purple Hull and MN13 produced callus after 4 weeks in Blaydes' medium. The hormone combinations in the medium were 2-l dichlorophenoxy acetic acid (2.4-D) (2 mg/liter) and kinetin at 0.05, 0.1, 0.5 mg/liter, or 2,4-D and thidiazuron (TDZ) at 2.2, 4.4 and 6.6 mg/liter or benzylaminopurine (BA) at 2.25, 4.5 or 6.75 mg/liter. Shoots occured on cotyledonary nodes of Pinkeye Purple Hull In the TDZ (6.6 mg/liter). Roots were produced from the leaf and cotyledonary nodes of Pinkeye Purple Hull and on cotyledons of IT-64E-124 cultured In media with kinetin (0.5 mg/liter). Leaf and cotyledon explants of pigeonpea genotype; ICPL 146 1965HK and ICPL 65024 produced callus and some shoots in BA (2.25 mg/liter) after 4 weeks. The callus when subcultured on BA (0.5 mg/liter) and NAA (0.1 mg/liter) produced shoots. Regenerated shoots rooted in the Blaydes' medium with kinetin (0.01 mg/liter) and NAA (0.6 mg/liter).


HortScience ◽  
1990 ◽  
Vol 25 (5) ◽  
pp. 569-571 ◽  
Author(s):  
A. Raymond Miller ◽  
Craig K. Chandler

A protocol was developed for excising and culturing cotyledon explants from mature achenes of strawberry (Fragaria × ananassa Duch.). Cotyledon explants formed callus with multiple shoot buds on agar-solidified Murashige and Skoog media containing several combinations of hormones (1 μm 2,4-D; 10 μm 2,4-D; 1 μm BA + 1 μm 2,4-D; 1 μm BA + 10 μm 2,4-D; 5 μm BA; 5 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μ m 2,4-D; 5 μ m BA + 5 μm NAA; 5 μ m BA + 15 μ m NAA). After three subcultures, only tissues maintained on the medium containing 5 μm BA + 5 μm NAA continued to form shoots. Tissues transferred to other media eventually died (1 μm 2,4-D; 1 μ m BA + 10 μ m 2,4-D; 5 μ m BA; 5 μ m BA + 1 μ m 2,4-D), became unorganized (1 μm BA + 1 μm 2,4-D; 5 μm BA + 10 μm 2,4-D; 5 μm BA + 15 μm NAA), or formed roots (10 μm 2,4-D). Whole plantlets were produced by transferring callus with buds to medium lacking hormones. The rapid regeneration of clonal plantlets from cotyledon explants may be useful for reducing variability in future developmental studies. Chemical names used: N-(phenylmethyl)-1H-purin-6-amine (BA); (2,4-dichlorophenoxy) acetic acid (2,4-D); and 1-naphthaleneacetic acid (NAA).


2009 ◽  
Vol 36 (No. 4) ◽  
pp. 140-146 ◽  
Author(s):  
J.K. Kanwar ◽  
S. Kumar

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.


2014 ◽  
Vol 9 (12) ◽  
pp. 1182-1195
Author(s):  
Samar Naseer ◽  
Tariq Mahmood

AbstractThe present study was designed to analyze genetically somaclonal variants using biochemical and molecular markers. Efficient tissue culture protocol for Solanum melongena L. cv. Nirrala was developed. Maximum callus induction (100%) was observed for Murashige and Skoog (MS) media supplemented with 2.0 mg L−1 naphthalene acetic acid +0.5 mg L−1 6-benzylaminopurine; and nodal explants gave best callusing response (88.8%) as compared to internodes (88.3%) and leaves (87.7%). The best shooting was induced on nodal and internodal callus in the presence of 2.0 mg L−1 6-benzylaminopurine. Total soluble protein content of callus and regenerated variant plants was estimated for biochemical analysis, and largest amount of soluble protein was found in callus (6.54 mg g−1 fresh tissue) followed by variant plant grown on 2.0 mg L−1 6-benzylaminopurine (5.96 mg g−1 fresh tissue). Random amplification of polymorphic DNA technique was done with five decamer primers (OPC1-OPC5) and maximum polymorphism was detected by OPC 2 (26.99%) among all samples, whereas nodal callus on media containing 1.0 mg L−1 naphthalene acetic acid +1.0 mg L−1 6-benzylaminopurine showed highest polymorphism producing 22 bands, out of which 8 bands were polymorphic. The study shows that this marker system can provide better evaluation of genetic variation induced by tissue culture.


2015 ◽  
Vol 44 ◽  
pp. 38-44 ◽  
Author(s):  
H. Sandhya ◽  
Rao Srinath

Suitable protocol for induction of callus and regeneration was developed from different explants viz., node, stem and leaves in Physalis minima. MS basal medium supplemented with various concentrations (1.0-4.0mg/l) of auxins like 2,4-Dichlorophenoxy acetic acid (2,4-D), α-naphthalene acetic acid (NAA) and Indole-3-acetic acid (IAA) and cytokinins (0.5-1.5mg/l) like BAP or Kn were used. All the three explants responded for induction of callus, however stem explants were found superior, followed by node and leaf. Callus induction was observed in all the auxins and combination of growth regulators used with varied mass (2010±1.10) and highest percentage of callus induction was observed from stem at 2.0mg/l 2,4-D (90%) followed by NAA (70%) and IAA (50%). Organogenesis was induced when nodal explants were transferred on MS medium supplemented with 2,4-D and Kn at various concentrations, maximum being on 2.0mg/l 2,4-D + 1.0mg/l Kn (90%). Regenerated shoots were elongated on 0.5mg/l GA3. The shoots were subsequently rooted on MS + 1.0mg/l IBA (95%) medium. Rooted shoots were hardened and acclimatized, later they were transferred to polycups containing soil, cocopeat and sand in the ratio 1:2:1.Keywords:Physalis minima, Node, Stem, Leaf, callus and growth regulators.


1998 ◽  
Vol 20 (20) ◽  
pp. 131
Author(s):  
Elcí Terezinha Henz Franco ◽  
Cinara Echart Almeida

Petiole explants of Begonia rex were cultured on basal medium (MURASHIGE & SKOOG, 1962). The medium was suplemented with naphthalene acetic acid (0.01; 0.1 and 0.5 mg/I) and kinetin ( 0.1; 0.2; 0.5 and 1.0 mg/I). In these experiments, were as also used coconut water (15% and 20%) or potato extract ( 15% and 20%). Buds were formed in several treatments, but the best combination was coconut water 0.01 mg/L NAA and 0.1 mg/I KIN. Whole plants (40% of the explants) were obtained when was added coconut water . The optimal combination for plant regeneration (100%) was 0.01 mg/I NAA plus 0.1 mg/I KIN.


Plants ◽  
2020 ◽  
Vol 9 (3) ◽  
pp. 318
Author(s):  
Mehtab Muhammad Aslam ◽  
Joseph K. Karanja ◽  
Qian Zhang ◽  
Huifeng Lin ◽  
Tianyu Xia ◽  
...  

The tissue culture regeneration system of Lupinus albus has always been considered as recalcitrant material due to its genotype-dependent response and low regeneration efficiency that hamper the use of genetic engineering. Establishment of repeatable plant regeneration protocol is a prerequisite tool for successful application of genetic engineering. This aim of this study was to develop standardized, efficient protocol for successful shoot induction from cotyledonary node of white lupin. In this study, 5 day old aseptically cultured seedlings were used to prepare three explants (half cotyledonary node, HCN; whole cotyledonary node, WCN; and traditional cotyledonary node, TCN), cultured on four concentrations of M519 medium (M519, ½ M519, 1/3 M519, and ¼ M519), containing four carbohydrate sources (sucrose, fructose, maltose, and glucose), and stimulated with various combinations of KT (kinetin), and NAA (naphthalene acetic acid) for direct shoot regeneration. High frequency of 80% shoot regeneration was obtained on ½ M519 medium (KT 4.0 mg L−1 + NAA 0.1 mg L−1) by using HCN as an explant. Interestingly, combinations of (KT 4.0 mg L−1 + NAA 0.1 mg L−1 + BAP 1.67 mg L−1), and (KT 2.0 mg L−1 + NAA 0.1 mg L−1) showed similar shoot regeneration frequency of 60%. Augmentation of 0.25 g L−1 activated charcoal (AC) not only reduced browning effect but also improved shoot elongation. Among the all carbohydrate sources, sucrose showed the highest regeneration frequency with HCN. Additionally, 80% rooting frequency was recorded on ½ M519 containing IAA 1.0 mg L−1 + KT 0.1 mg L−1 (indole acetic acid) after 28 days of culturing. The present study describes establishment of an efficient and successful protocol for direct plant regeneration of white lupin from different cotyledonary nodes.


2016 ◽  
Vol 41 (3) ◽  
pp. 521-528
Author(s):  
MR Kabir ◽  
S Ahmed ◽  
MAY Akhond

Seedling-derived cotyledonary nodes and hypocotyl explants of BARI Dherosh- 1 were cultured in vitro on MS medium supplemented with varying concentrations of 2, 4-Dichlorophenoxy acetic acid (2, 4-D), 6- Benzylaminopurine (BAP), Thidiazuron (TDZ), BAP with 1-Nepthaleneacetic acid (NAA), BAP with Indole 3-butyric acid (IAA) and Zeatin with IAA along with a control. Shooting response (100%) with callus was only observed from cotyledonary nodes on thidiazuron (TDZ) where hypocotyls produced only callus or callus with roots on different concentrations of plant growth regulators. Considering the shooting response, the cotyledonary nodes of BARI Dherosh-1 were cultured on various concentrations of TDZ for regeneration. The highest percentage (64.0) with maximum number (6.8) of shoots per explant were observed in 0.044 ?M TDZ in 8.4 days. The regenerated shoots were rooted on ½ strength MS, MS supplemented with 2.46 ?M IBA and 0.53 ?M NAA. The highest percentage (83.3) and minimum days (9.7) required for root induction were recorded in 2.46 ?M IBA. The rooted plantlets were transferred to soil and hardened in the plastic pots under green house conditions. The rooted shoots grew normally under natural conditions following acclimatization.Bangladesh J. Agril. Res. 41(3): 521-528, September 2016


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