scholarly journals Cotyledon Age and Somatic Embryogenesis of Cowpea

HortScience ◽  
1998 ◽  
Vol 33 (4) ◽  
pp. 594b-594
Author(s):  
Lurline Marsh

Somatic embryogenesis can be used to facilitate the improvement of traits in plants. The study was conducted to assess different ages of immature zygotic cowpea (Vigna unguiculata (L. Walp), “MN13” cotyledons for their ability to produce somatic embryos. Cotyledons were harvested weekly for the first 8 weeks following anthesis. After removal of their embryo axes, they were cultured in Murashige and Skoog (MS) medium containing 0.6% agar, B-5 vitamins, 3% sucrose, and 20 mg·L-1 2,4-dichlorophenoxy acetic acid (2-4D). Cotyledon explants of all the ages produced calli. The percentage of explants producing calli ranged from 32% to 92%. On transfer of the calli to similar medium containing 0.2% gellan gum instead of 0.6% agar, all ages except those from the 1-week cotyledons produced white globular somatic embryos. The largest of these embryos were 9 mm in length. The highest frequency of globular embryos was produced with the 3- to 5-week-old cotyledons.

HortScience ◽  
2002 ◽  
Vol 37 (7) ◽  
pp. 1122-1123 ◽  
Author(s):  
Mohammed A.M. Aly ◽  
Bala Rathinasabapathi ◽  
Sheevani Bhalsod

Many members of the Plumbaginaceae are important flower crops wherein propagation is hindered by poor seed germination. Micropropagation via organogenesis is commercially practiced for certain Limonium species. However, somatic embryogenesis was not reported for members of the Plumbaginaceae until recently for L. bellidifolium Durmort. The induction of somatic embryogenesis from cotyledon explants in a modified Murashige and Skoog (MS) medium was examined in four other members of this family, Limonium aureum O. Kuntze, L. latifolium O. Kuntze, L. sinuatum Mill., and Armeria maritima Willd. Induction of embryogenic callus was achieved in all the species examined on MS medium supplemented with 4.5 μm 2,4-D and 88 or 118 mm sucrose. Species of the genus Limonium responded better than A. maritima Willd. in somatic embryo induction and maturation. Somatic embryos of L. aureum O. Kuntze matured readily on MS medium supplemented with 0.93 μm kinetin and 88 mm mannitol. Chemical name used: 2,4-Dichlorophenoxy acetic acid (2,4-D).


1990 ◽  
Vol 115 (4) ◽  
pp. 691-696 ◽  
Author(s):  
Rebecca M. Cade ◽  
Todd C. Wehner ◽  
Frank A. Blazich

Two studies were conducted to test the effects of various tissue culture media on somatic embryogenesis from cotyledon tissue of cucumber (Cucumis sativus L.). The two best media for embryo initiation were Murashige and Skoog (MS) salts and vitamins containing either 1 or 2 mg 2,4-D/liter and 0.5 mg kinetin/liter. In the second study, embryos developed more normally. More plantlets developed when tissue was removed from the initiation medium after 3 weeks and transferred to MS containing 1 mg NAA/liter and 0.5 mg kinetin/liter for 3 weeks, rather than leaving the embryos on a medium containing 2,4-D. Histological evidence indicated that the embryos were multicellular in origin. Charcoal in the maturation medium inhibited embryo development. Chemical names used: (2,4-dichlorophenoxy) -acetic acid (2,4-D); N-(2-furanylmethyl)-lH-purine-6-amine (kinetin); 1-naphthaleneacetic acid (NAA).


Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 3 ◽  
Author(s):  
Xiuxia Ren ◽  
Ya Liu ◽  
Byoung Ryong Jeong

Somatic embryogenesis is a preferred method for vegetative propagation due to its high propagation efficiency. In this study, zygotic embryos, cotyledons, and hypocotyls of Paeonia ostii ‘Fengdan’ were used as the explant to induce somatic embryogenesis. The results showed that a combination of 0.5 mg·L−1 thidiazuron (TDZ) and 0.5 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D) was effective in inducing somatic embryos from the zygotic embryo and cotyledon explants. Hypocotyls only formed somatic embryos on Murashige and Skoog (MS) medium supplemented with both 0.5 mg·L−1 TDZ and 0.5 mg·L−1 1-naphthylacetic acid (NAA). Moreover, the compact callus was effectively produced from zygotic embryo, cotyledon, and hypocotyl explants in medium supplemented with a combination of 3.0 mg·L−1 6-benzylaminopurine (BA) and 1.0 mg·L−1 NAA, and then converted into somatic embryos in the same medium, and the ratio of the explants with embryo induction and number of embryos induced per explant were much higher than those induced by 0.5 mg·L−1 TDZ and either 0.5 mg·L−1 2,4-D or 0.5 mg·L−1 NAA. The MS medium was better than the woody plant medium (WPM) for inducing somatic embryos from zygotic embryo and hypocotyl explants, whereas the WPM was better than the MS medium for somatic embryogenesis induction from cotyledon explants. All of the somatic embryos developed well into mature embryos on their respective media supplemented with both 3.0 mg·L−1 BA and 1.0 mg·L−1 NAA. Overall, the protocols for indirect somatic embryogenesis from zygotic embryo, cotyledon, and hypocotyl of P. ostii ‘Fengdan’ were successfully established, which can greatly facilitate their propagation and breeding processes.


1995 ◽  
Vol 43 (4) ◽  
pp. 385-390 ◽  
Author(s):  
S. Kulothungan ◽  
A. Ganapathi ◽  
A. Shajahan ◽  
K. Kathiravan

Embryogenic callus was induced from seedling leaf explants of cowpea (Vigna unguiculata (L.) Walp. cv. C152 on Murashige and Skoog (MS) medium containing 2.0 mg 1−1 2,4-dichlorophenoxyacetic acid (2,4-D). The maximum frequency of somatic embryogenesis was noticed when this callus was transferred to MS liquid medium supplemented with 2 mg 1−1 2,4-D. Further studies on ontogeny of somatic embryos showed that the cells destined to become somatic embryos divided into spherical or filamentous proembryos. Subsequent divisions in the proembryo led to globular, heart, torpedo-shaped, and cotyledonary-stage somatic embryos. Tiny plantlets were obtained by transferring the cotyledonary-stage somatic embryos to MS liquid medium containing 0.5 mg 1−1 2,4-D.


HortScience ◽  
1993 ◽  
Vol 28 (6) ◽  
pp. 672-673 ◽  
Author(s):  
S.A. Merkle ◽  
B.A. Watson-Pauley

Bigleaf magnolia (Magnolia macrophylla Michx.) cultures were initiated from immature seeds on an induction medium containing 9.0 μm 2,4-D, 1.1μm BA, and 1 g casein hydrolysate/liter. After 2 months on induction medium, one culture produced adventive embryos. Clumps of embryos transferred to liquid induction medium proliferated as nodules, which grew in diameter, but failed to produce embryos while maintained in induction medium. Nodules transferred to basal medium produced clumps of somatic embryos, which continued to produce repetitive embryos with monthly transfer to fresh basal medium. Individual embryos transferred to basal medium lacking casein hydrolysate germinated and leaves expanded. Plantlets derived from these embryos were transferred to potting mix and acclimatized to greenhouse conditions. Chemical names used: (2,4-dichlorophenoxy)acetic acid (2,4-D); N -(phenylmethyl)-lH-purin-6-amine (BA).


1990 ◽  
Vol 115 (5) ◽  
pp. 858-860 ◽  
Author(s):  
S.A. Merkle ◽  
A.T. Wiecko

Cultures were initiated from immature seeds of three species of magnolia: sweetbay magnolia (Magnolia virginiana L.), fraser magnolia (M. fraseri Walt.) and yellow cucumbertree [M. acuminata var. cordata (Michx.) Sarg.]. Immature seeds were bisected longitudinally and cultured on a solidified conditioning medium containing 2 mg 2,4-D/liter, 0.25 mg BA/liter, 40 g sucrose/liter, and 1 g casein hydrolysate/liter. Cultures were maintained in the dark at 22C and transferred to fresh medium at monthly intervals. Within 2 months of culture, somatic embryos or proembryogenic masses proliferated from one end of the endosperm mass. Somatic embryos and proembryogenic masses of each species were cultured on a hormone-free version of the conditioning medium to complete maturation and then transferred to the same hormone-free medium, minus casein hydrolysate, to initiate germination. Germinants were transferred to a hormone-free plantlet development medium for conversion. Plantlets of all three species survived transfer to soil mix and continued to grow. Chemical names used: (2,4 -dichlorophenoxy) acetic acid (2,4-D), N- (phenylmethyl)-1H-purin-6-amine (BA).


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 23 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Dinesh Giri ◽  
Sushma Tamta

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013  (December)


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