Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

HortScience ◽  
2004 ◽  
Vol 39 (6) ◽  
pp. 1378-1380 ◽  
Author(s):  
C.K. Kim ◽  
J.Y. Oh ◽  
J.D. Chung ◽  
A.M. Burrell ◽  
D.H. Byrne

Somatic embryogenesis was initiated from in vitro-grown leaf explants of rose using an induction period of 4 weeks on MS basal medium supplemented with auxin followed by several subcultures on MS basal medium with cytokinin. `4th of July' showed the highest regeneration frequency (24.4%) on 5.3 μm NAA followed by culture on medium containing 18.2 μm zeatin. `Tournament of Roses' produced somatic embryos when cultured for 4 weeks on medium containing dicamba, 2.3 μm followed by three subcultures on medium containing 18.2 μm zeatin. Embryogenic callus matured on MS media containing 0.5 μm NAA, 6.8 μm zeatin, and 2.9 μm GA3. Long-term cultures were established for both cultivars. Somatic embryos germinated on MS medium containing IBA and BA. Silver nitrate (58.8 μm) enhanced shoot formation and germination of somatic embryos. Plants derived from somatic embryos were acclimatized and successfully established in the greenhouse.


2018 ◽  
Vol 41 (4) ◽  
Author(s):  
Marlúcia Souza Pádua ◽  
Raíssa Silveira Santos ◽  
Luciano Vilela Paiva ◽  
Vanessa Cristina Stein ◽  
Luciano Coutinho Silva

ABSTRACT Oil palm is a woody monocot of economic importance due to high oil production from its fruits. Currently, the conventional method most used to propagate oil palm is seed germination, but success is limited by long time requirements and low germination percentage. An alternative for large-scale propagation of oil palm is the biotechnological technique of somatic embryogenesis. The rooting of plants germinated from somatic embryos is a difficult step, yet it is of great importance for later acclimatization and success in propagation. The aim of this study was to evaluate the effect of the auxins indole acetic acid (IAA) and indole butyric acid (IBA) on the rooting of somatic embryos of Tenera hybrid oil palm. Plants obtained by somatic embryogenesis were inoculated in modified MS medium with 10% sucrose and 0.6% agar and supplemented with IAA or IBA at concentrations of 5 µM, 10 µM, and 15 µM, and the absence of growth regulators. After 120 days, the presence of roots, root type, length of the longest root, number of roots, number of leaves, and shoot length were analyzed. Growth regulators were favorable to rooting; plants cultivated with IBA growth regulator at 15 µM showed higher rooting percentage (87%) and better results for the parameters of number of roots (1.33) and shoot length (9.83).


2006 ◽  
Vol 1 (3) ◽  
pp. 1934578X0600100
Author(s):  
Bishnu P. Chapagain ◽  
Vinod Saharan ◽  
Dan Pelah ◽  
Ram C. Yadav ◽  
Zeev Wiesman

This study describes the effects of plant growth regulators, explants, and somatic embryogenesis on in vitro production of the steroidal sapogenin, diosgenin, in callus cultures of the Balanites aegyptiaca (L.) Del.(desert date). Root, shoot, hypocotyl, and epicotyl callus culture of B. aegyptiaca, were raised on MS basal media supplemented with various combinations of either 2,4-D and NAA alone, or with BAP. The diosgenin content (on a dry weight basis) was found to be highest when calli were cultured in MS basal medium supplemented with 1.0 mg l−1 2,4-D alone and/or in combination with 0.5 mg l−1 BAP. However, the callus growth was highest in media supplemented with 2.5 or 3.0 mg l−1 2,4-D. MS basal media supplemented with 2,4-D 2.5 mg l−1 alone and in combination with 0.5 mg l−1 BAP induced pre-embryogenic callus formation on root cultures. When these pre-embryogenic callus cultures were used to establish cell suspension cultures, two growth densities were obtained in embryogenic suspension cultures, inducing clusters of somatic embryos at various stages of development. The maximum number of somatic embryos were obtained at the fifth week on the medium supplemented with 1.0 mg l−1 2,4-D. However, the diosgenin content in these somatic cells was found to be lower compared to the explant calluses. This study revealed that production of diosgenin in callus cultures of B. aegyptiaca is possible, but the amount is significantly affected by the growth regulators, type of explants, and somatic embryogenesis.


1990 ◽  
Vol 68 (3) ◽  
pp. 487-491 ◽  
Author(s):  
N. Arumugam ◽  
Sant S. Bhojwani

In vitro multiplication of Podophyllum hexandrum Royle (Podophyllaceae) via somatic embryogenesis is reported. The callus derived from zygotic embryos on Murashige and Skoog medium containing 2 μM BA and 0.5μM IAA differentiated globular embryos. On this medium the globular embryos continued to multiply but failed to mature. Further development of the embryos occurred if the sucrose level in the basal medium was raised to 6% or the medium was supplemented with 1–10 μM NAA. Light and temperatures higher than 25 °C suppressed embryogenesis. Embryogenic potential of the callus has been maintained for over 20 months through subcultures. The somatic embryos developed into plantlets on the basal medium. Key words: endangered species, podophyllotoxin, Podophyllum, somatic embryogenesis.


HortScience ◽  
1994 ◽  
Vol 29 (4) ◽  
pp. 251f-251 ◽  
Author(s):  
Christopher S. Cramer ◽  
Mark P. Bridgen

Disinfected midrib sections of Mussaenda `Queen Sirikit' ≈3 to 4 mm in size were cultured on a basal medium of Murashige and Skoog salts and vitamins, 87.7 mm sucrose, and 5 g Sigma agar/liter supplemented with several concentrations of indole-3-acetic acid (IAA) (0, 5.0, 10.0, 20.0 μm) and 6-benzylaminopurine (BAP) (0, 0.5, 1.0, 2.5, 5.0, 10.0, 25.0, 50.0 μm). Cultures were subculture onto the same treatment after 5 weeks and observed weekly for 15 weeks for the presence of somatic embryos. As somatic embryos were produced, they were subculture onto basal medium supplemented with 0.5, 1.0, 2.5, or 25.0 μm BAP. Callus was first observed at 2 weeks in cultures grown on basal medium supplemented with 5.0–20.0 μm IAA and 0–50.0 μm BAP. Somatic embryos were observed at 8 weeks on basal medium supplemented with 5.0–10.0 μm IAA and 2.5–5.0 μm BAP. Callus cultured on 0–10 μm IAA and 5.0–10.0 μm BAP produced the greatest number of somatic embryos by 15 weeks. Somatic embryos subculture to basal medium supplemented with 25.0 μm BAP proliferated shoots, while eliminating BAP from the medium resulted in root and callus production. Shoots and entire plants were removed from in vitro conditions and successful] y acclimated to greenhouse conditions. Somatic embryo-derived plants flowered sporadically 25 to 35 weeks after removal from in vitro conditions. Variations in sepal number and leaf number per node were observed at 1% to 5%.


2012 ◽  
Vol 64 (2) ◽  
pp. 809-817 ◽  
Author(s):  
Irina Holobiuc ◽  
R. Catana

Our aim was to establish an efficient and reproducible system for producing synthetic seeds from recurrent somatic embryogenesis in long-term cultures of Gentiana lutea L. This species is a vulnerable medicinal plant, protected both at the national and international levels, and is included in different Red Lists and Books. In vitro culture, as an alternative to classical methods of preservation, allows for the cyclic multiplication of plant material and short-, medium- and long-term preservation of tissue collections. Biotechnological approaches allow for maintenance of the plant material in a confined space and protection against biotic and abiotic factors. Somatic embryogenesis (SE) is the most efficient way to regenerate plants, ensuring material for preservation and fundamental research. In our experiment, recurrent somatic embryogenesis was developed in long-term cultures in the presence of sugar alcohols (mannitol, sorbitol) and in the absence of growth factors. This process proceeded at a high rate, with adventive somatic embryos being generated in a continuous process, followed by maturation, germination and development into plants. To follow the somatic embryogenesis process, histological samples were made. We used these embryogenic cultures for synthetic seed production and medium-term conservation. The viability of somatic embryos after moderate osmotic stress treatment was tested using TTC. Our methodology relied on the induction of somatic embryogenesis in the presence of auxins in the first cycle of in vitro cultures, long-term high embryogenic culture maintenance in the presence of sugar alcohols and synthetic seed production.


2014 ◽  
Vol 17 (2) ◽  
pp. 100-107
Author(s):  
Nhung Thi Tuyet Tran ◽  
Hoang Ngo Phan ◽  
Sanh Du Nguyen

Pseuderanthemum palatiferum contains many compounds which have pharmaceutical and medicinal values. In this study, the in vitro somatic embryogenesis of callus formed from Pseuderanthemum palatiferum’s leaves was performed and analyzed. The calli were induced in three weeks in MS medium with 1 mg.l-1 or 2 mg.l-1 of 2,4-D. The somatic embryos were developed as follows: the globular embryos formed after 9 days, the heart embryos formed after 13 days, the torpedo embryos formed after 15 days and the cotyledonary embryos formed after 17 days. The mean number of obtained embryos was 28.5 embryos/cm2 of leaf tissue. The intensity of respiration and biological activity of endogenous plant growth regulators of leaf tissue during culture were recorded.


HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 452B-452
Author(s):  
Harrison Hughes ◽  
Leigh Towill

There are turfgrasses species that are clonally propagated; notably bermudagrass, buffalograss, and zoysiagrass. Some of the early cultivars of these species are no longer widely grown, and may eventually be lost if not preserved. In order to facilitate studies on the long-term cryopreservation of these species and specific lines of saltgrass, it is necessary to develop suitable micropropagation procedures. We have developed protocol for the isolation and establishment of clean cultures in vitro for all four species. A 1/2-strength MS basal medium with Nitsch & Nitsch vitamins, 5 mg/L of thiamine, 2 mg/L of glycine, 30 g of sucrose, 7 g of agar with varying growth regulators has been used. Explant materials are prewashed in the greenhouse prior to a 15- to 30-min soapy wash in the laboratory. After a 30- to 60-min rinse in running water, nodal sections are surface-disinfested in 10% bleach with Tween 20 for 15 min, followed by three sterile water rinses. This procedure, sometimes with PPM (a proprietary antimicrobial compound), results in 50% or greater clean cultures. Rapidly growing nodal sections work best and preferably those not established in soil. We have tested various growth regulator combinations and have found that 10 mg/L of BA results in proliferation of buffalograss and saltgrass. However, proliferation remains relatively slow, requiring 8 to 12 weeks to develop sufficiently for subculture. Although we have succeeded in obtaining clean cultures of bermudagrass and zoysiagrass, proliferation is minimal, Further research is ongoing to develop a proliferative system with these two species.


1991 ◽  
Vol 116 (4) ◽  
pp. 753-757 ◽  
Author(s):  
Ana M. Vieitez ◽  
Carmen San-José ◽  
F. Javier Vieitez ◽  
Antonio Ballester

Somatic embryos were induced on the roots of Camellia japonica L. plantlets regenerated from an in vitro clone of juvenile origin. The embryos appeared to differentiate from epidermic cells and to be connected with the root via a few parenchymatous cells. Somatic embryogenesis occurred on basal medium and with or without various combinations of zeatin, BA, and IBA. Secondary embryos were induced on cotyledons and/or hypocotyl regions of somatic embryos. Two morphological types of somatic embryos were developed, seed-like and bud-like types, and their formation was influenced by the presence of BA in the medium. Embryogenic capacity has been maintained for more than 24 months by subculturing secondary embryos at 7- to 8-week intervals. The best gibberellin/auxin combination for inducing the germination of isolated somatic embryos was GA at 5 mg·liter-1 G A3 and IAA at 1 mg·liter-1. P1antlets were successfully established in planting medium and have continued to grow in a greenhouse. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA); (1α, 2β, 4aα, 4bβ, 10β)-2,4a,7-trihydroxy-l-methyl-8-methylenegibb-3-ene-1,10-dicarboxylic acid l,4a-lactone (GA); 1 H -indole-3-acetic acid (IAA); 1 H- indole-3-butyric acid (IBA); 2-methyl-4-(1 H- purine-6-ylamino)-2-buten-l-ol (zeatin).


2021 ◽  
Author(s):  
Thiago Sanches Ornellas ◽  
Yohan Fritsche ◽  
Edison Cardona Medina ◽  
Miguel Pedro Guerra

Abstract Bamboos are an important worldwide non-timber forest product with current rising interest due to their environmentally friendly applications. Besides the consolidated uses of the sweet shoots and culms for structural uses, Dendrocalamus asper is an imposing ornamental bamboo for horticulture. The present work aimed to establish in vitro calli culture and plant regeneration through somatic embryogenesis starting from young inflorescences of the giant bamboo, D. asper. Pre-anthesis inflorescences were collected, disinfested, and subjected to callus induction on MS basal medium supplemented by 0 µM, 9 µM, 18 µM, 27 µM, and 36 µM of 2,4-D in combination with 9 µM of 2-iP or 9 µM Kin. The different obtained calli types were characterized and subcultured in 0 µM, 4.5 µM, 9 µM, and 18 µM of 2,4-D in combination with 9 µM of both cytokinins for multiplication and differentiation. Additionally, the explant incision and its inoculation orientation onto culture media were tested for callus induction improvement. The 2,4-D was essential for callus induction, and its combination with both cytokinins resulted in embryogenic callus induction and further somatic embryos regeneration. The subsequent reduction of this auxin to 4.5 µM resulted in somatic embryo maturation. Somatic embryos transferred to a plant growth regulator-free medium resulted in plantlet conversion. The present work showed the feasibility of using inflorescences as explants and the efficiency of using the 2-iP in combination with 2,4-D to callus induction and in vitro bamboo plant regeneration through somatic embryogenesis.


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