scholarly journals Somatic Embryogenesis and Plant Regeneration from Commercial Soybean Cultivars

Plants ◽  
2019 ◽  
Vol 9 (1) ◽  
pp. 38 ◽  
Author(s):  
Ghulam Raza ◽  
Mohan B. Singh ◽  
Prem L. Bhalla

The efficient regeneration of plants from commercial genotypes is a pre-requisite for successful genetic transformation, to apply modern crop improvement techniques such as CRISPR-based genome editing. Plant regeneration through the somatic embryogenesis pathway offers an advantage over the organogenesis approach, avoiding the risk of developing chimeras. Plant genotype, explant type, and media compositions play an essential role in the in-vitro regeneration of plants. This study aimed to characterize the commercially grown Australian soybean genotypes for their potential to induce somatic embryos, embryo proliferation, maturation, germination, and plant regeneration. Overall, nine soybean cultivars belonging to different maturity groups were evaluated. Immature cotyledon ranging from 2–4 and 4–6 mm in size were used as explants for somatic embryogenesis induction. Maximum somatic embryo induction frequency (86%) was observed from 4–6 mm immature cotyledons of the cv. Jack (MG III), followed by 66%, 26%, 21%, and 6% in cultivars Williams (MG III), Snowy (MG III), MoonB1 (MG V), and PNR791 (MG V), respectively. On the other hand, cv. Snowy showed maximum somatic-embryo-inducing potential (67%) in 2–4 mm immature cotyledons followed by Williams, Jack, MoonB1, and PNR791. Somatic embryos from Jack, Williams, and Snowy cultivars were further tested for embryo proliferation, maturation, and germination. Maximum proliferation and maturation were observed in cv. Jack, followed by Snowy and Williams. However, cv. Snowy showed a significantly higher conversion of cotyledonary stage embryos to plantlets (85%), than both Jack and Williams cultivars (53% each). In conclusion, this study outlined a protocol for somatic embryogenesis and plant regeneration from three soybean cultivars. Our findings suggest commercial cv. Snowy could be a good candidate for developing transgenic plants through somatic embryogenesis.

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 694c-694
Author(s):  
Sung-Do Oh ◽  
Won-Seob Song ◽  
Man-Sang Lee

From one week through 7 weeks after artificial pollination, immature ovules of yooza(Citrus junos Sieb. et Tanaka) were excised and cultured in vitro on MT media. Even though there was only a little difference in percentage of somatic embryo formation depending upon the time of excision, immature ovules of 4-week-old showed the highest ratio of somatic embryo formation without callus outgrowth. Various growth regulators or other stimulators were added to the MT media to increase the somatic embryogenesis, In general, BAP was more effective than 2,4-D for somatic embryo formation and the combinations of 0.01mg/l 2,4-D and 0,01 or 0.1mg/l BAP were particularly effective in stimulating somatic embryo formation. When 500mg/l malt extract was added to the medium, the percentage of somatic embryo formation increased reaching as high as 86.7%. Plant regeneration from somatic embryos reached to 66.7% on the medium containing 1.0mg/l zeatin. Isozyme banding patterns were also analyzed to confirm the variations of characteristics of the plantlets derived from direct somatic embryos.


1970 ◽  
Vol 14 ◽  
pp. 31-38 ◽  
Author(s):  
M Rahman ◽  
M Asaduzzaman ◽  
N Nahar ◽  
MA Bari

Somatic embryos were obtained from cotyledon and midrib explants of Solanum melongena L., cultivar Loda. For callus induction, medium was supplemented with different concentrations of auxin singly or in combination with BAP. The best callusing 83-85% was obtained from both of the explants cultured on MS medium containing 2.0 mgl-1NAA + 0.05 mgl-1BAP. Somatic embryogenesis and shoot regeneration was achieved after transferring the calli to MS medium supplemented with BAP, GA3, NAA and Zeatin. Cotyledon derived calli showed better performance (87%) for regeneration than that of midrib (82%) when sub cultured on MS medium having 2.0 mgl-1 Zeatin + 1.0 mgl-1 BAP. For root induction, MS + 3.0 mgl-1 IBA was proved to be better treatment for average number (14-15) and mean length (12 cm) of roots than those of other treatments. Key words: Eggplant; cotyledon; midrib; callus induction; somatic embryo J. bio-sci. 14: 1-9, 2006


2015 ◽  
Vol 24 (2) ◽  
pp. 247-262 ◽  
Author(s):  
El Sawy A Mohamed ◽  
Amina Gomaa ◽  
Nancy Danial

Better results were obtained when stigma explants of variegated lemon and citron were used. After ten months, somatic embryos developed into plantlets at a frequency ranged from 13.3 for lime to 66.7% for lemon. Virus presence was tested by ELISA and RT?PCR. The results indicated that the plantlets regenerated through somatic embryogenesis are CTV?free. RAPD analysis was used to asses the genetic stability of plantlets as compared to the mother plants. The results indicated that most plantlets belong to the respective mother plants and the polymorphism percentage was genotype and explant?dependant.Plant Tissue Cult. & Biotech. 24(2): 247-262, 2014 (December


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%.


2017 ◽  
Vol 44 (3) ◽  
pp. 261
Author(s):  
Adam Saepudin ◽  
Nurul Khumaida ◽  
Didy Sopandie ◽  
Dan Sintho Wahyuning Ardie

ABSTRACT<br /><br />Somatic embryo induction medium was reported to be genotype dependent for soybean. This study was aimed to obtain the optimum medium for embryo somatic induction and proliferation, and to regenerate somatic embryo of five soybean genotypes. Five soybean genotypes (Tanggamus, Anjasmoro, Yellow Biloxi, CG-22-10, and SP-10-4) were used in this study. The research was divided into four steps: (1) embryogenic callus induction of  five soybean genotypes, (2) embryogenic callus proliferation of five soybean genotypes, (3) optimation of embryo somatic induction on five soybean genotypes and (4) embryo somatic regeneration of five soybean genotypes. The induction experiment showed that based on number of embryogenic callus, the best somatic embryo-induction medium was 3% sucrose+ NAA 5 mg L-1+2,4-D 5 mg L-1+ Vitamin B5. Embryogenic callus number for each genotype tested was increased on proliferation media of 3% sukrosa + 2,4-D 5 mg L-1 + NAA 5 mg L-1+ Vit B5, and Yellow Biloxi gave the highest number of proliferated somatic embryos compared to other genotypes. Increasing number of globular somatic embryo of all genotypes was obtained from the optimation of somatic embryo induction media being used, and Tanggamus genotype gave the highest number of globular somatic embryo which followed by Yellow Biloxi genotype. Tanggamus and Yellow Biloxi genotypes were also successfully formed the four steps of somatic embryos (globular, heart, torpedo, and cotyledonary stages), but in regeneration medium of MS0 and media MS + sukrosa 10 g L-1 + GA3 2 mg L-1 + BAP 4 mg L-1 + Vit B5 only Tanggamus genotype was regenerated into plantlet.  <br /><br />Keywords: 2,4-D, NAA, somatic embryos, induction, proliferation<br /><br />


2019 ◽  
Vol 49 (12) ◽  
pp. 1604-1612
Author(s):  
Tingyu Sun ◽  
Yanli Wang ◽  
Lihua Zhu ◽  
Xiaoqin Wu ◽  
Jianren Ye

Pine wilt disease (PWD) is a severe threat to pine forests in East Asia. Screening and breeding of resistant varieties is a very effective way to prevent and control PWD; however, no reliable somatic embryogenesis system has yet been developed for the elite nematode-resistant Pinus thunbergii Parl. line. In this study, we studied the plant regeneration via somatic embryogenesis of nematode-resistant P. thunbergii. Initiation of embryogenic tissue was significantly affected by seed family (p = 0.017), immature zygotic embryo stage (p = 0.032), and initiation medium (p = 0.004). Seed family 37 was the most favorable female parent for initiation of P. thunbergii. Furthermore, the initiation rate increased from the pre-embryonic stage to the cleavage polyembryonic stage. The optimal medium was I2, containing 2,4-dichlorophenoxyacetic acid (9 μmol·L−1) and 6-benzyladenine (4.4 μmol·L−1). A statistically significant interaction between cell line and subculture time (24 months) was observed in the influence on proliferation rate, somatic embryo production, and percentage germination (p < 0.001). In this study, the highest somatic embryo production was achieved using cell line 37-1 (1983 somatic embryos per gram fresh mass), with approximately 83.5% of somatic embryos germinating after transferring to germination medium, of which 77.6% converted into plantlets.


2006 ◽  
Vol 66 (2a) ◽  
pp. 455-462 ◽  
Author(s):  
E. T. H. Franco ◽  
L. B. Gavioli ◽  
A. G. Ferreira

The present study aimed at establishing a complete plant regeneration protocol for Didymopanax morototoni (matchwood), a native Brazilian forest species. Four types of explants (root, shoot, node, and cotyledonary leaves) were obtained from in vitro germinated seeds. In the first step, woody plant medium (WPM) with casein hydrolysate (250 mgL-1 ) and 2,4-D (1.0 and 5.0 mgL-1) were used combined with kinetin (0.1 and 1.0 mgL-1). Twenty days after inoculation, the material was evaluated. Embryogenic calli were split, transferred to expression medium with several combinations of NAA and KIN, and moved to fresh medium after 60 days. Light did not interfere in embryo expression. Somatic embryos were formed either from individual cells or cell clusters. Plantlets were obtained in WPM medium and 10 gL-1 of sucrose with no plant regulator, or using 0.1 mgL-1 BAP and 0.5 mgL-1GA. Plantlets from somatic embryos of D. morototoni developed in 33% of the cases.


2010 ◽  
Vol 45 (7) ◽  
pp. 715-720 ◽  
Author(s):  
Annette Droste ◽  
Anelise Machado da Silva ◽  
Iliane Freitas de Souza ◽  
Beatriz Wiebke-Strohm ◽  
Lauro Bücker-Neto ◽  
...  

The aim of this work was to identify Brazilian soybean (Glycine max) genotypes with potential to respond to in vitro culture stimuli for primary somatic embryo induction, secondary embryo proliferation and plant regeneration. Differences among eight tested cultivars were observed at each stage. Two cultivars, IAS-5 and BRSMG 68 Vencedora, were selected for the evaluation of the capacity for embryo differentiation and plant regeneration. These cultivars had high embryo induction frequencies, repetitive embryogenic proliferation, and low precocious embryo germination in the initial experiment. The effect of abscisic acid (ABA) and charcoal addition on plant regeneration was investigated. The addition of ABA to proliferation medium and of ABA and activated charcoal to maturation medium increased embryo differentiation rates, which resulted in a higher number of regenerated plants. The BRSMG 68 Vencedora cultivar was found to have a high potential for embryo induction, embryo proliferation and plant regeneration. The potential of this cultivar for somatic embryogenesis was similar to that observed for cultivar IAS-5, which is currently used for soybean transformation in Brazil. BRSMG 68 Vencedora may be a good alternative genotype for soybean genetic engineering via somatic embryogenesis protocols.


Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.


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