scholarly journals In VitroPlant Regeneration from Commercial Cultivars of Soybean

2017 ◽  
Vol 2017 ◽  
pp. 1-9 ◽  
Author(s):  
Ghulam Raza ◽  
Mohan B. Singh ◽  
Prem L. Bhalla
Keyword(s):  
Genetic Transformation ◽  
Adventitious Shoots ◽  
Cotyledonary Node ◽  
Regeneration Capacity ◽  
Shoot Induction ◽  
Regeneration Rate ◽  
Soybean Cultivars ◽  
Legume Crop ◽  
High Regeneration ◽  
Regeneration Response

Soybean, a major legume crop, is the source of vegetable oil and protein. There is a need for transgenic approaches to breeding superior soybean varieties to meet future climate challenges. Efficient plant regeneration is a prerequisite for successful application of genetic transformation technology. Soybean cultivars are classified into different maturity groups based on photoperiod requirements. In this study, nine soybean varieties belonging to different maturity group were regenerated successfully from three different explants: half split hypocotyl, complete hypocotyl, and cotyledonary node. All the genotypes and explant types responded by producing adventitious shoots. Shoot induction potential ranged within 60–87%, 50–100%, and 75–100%, and regeneration rate ranged within 4.2–10, 2.7–4.2, and 2.6–10.5 shoots per explant using half split hypocotyl, complete hypocotyl, and cotyledonary explants, respectively, among all the tested genotypes. Bunya variety showed the best regeneration response using half split and complete hypocotyl explants and the PNR791 with cotyledonary node. The regenerated shoots were successfully rooted and acclimatized to glasshouse conditions. This study shows that commercial varieties of soybean are amenable to shoot regeneration with high regeneration frequencies and could be exploited for genetic transformation. Further, our results show no correlation between shoots regeneration capacity with the maturity grouping of the soybean cultivars tested.

scholarly journals Genetic Diversity and Heritability of In Vitro Leaf Regeneration Ability in Malus Species

HortScience ◽  
2017 ◽  
Vol 52 (10) ◽  
pp. 1396-1400
Author(s):  
Yi Tan ◽  
Baisha Li ◽  
Yi Wang ◽  
Ting Wu ◽  
Zhenhai Han ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Genetic Transformation ◽  
Callus Formation ◽  
Formation Rate ◽  
Regeneration Ability ◽  
Regeneration Rate ◽  
Malus Sieversii ◽  
High Regeneration ◽  
Germplasm Accessions

Agrobacterium-mediated genetic transformation is commonly used in dicotyledon plants such as apples. The regeneration ability of the recipient is an important factor in transformation efficiency. Here, the variations in bud regeneration rate (BRR) and the number of adventitious buds (NAB) formed per explant in Malus germplasm accessions with phenological stage were estimated. Both BRR and NAB of explants at the dormancy broken and spring sprouting stages were significantly higher than those at the autumn sprouting stage. The genetic diversity and inheritance of BRR and NAB were evaluated using 153 Malus germplasm accessions and 78 hybrid trees of Jonathan × Golden Delicious. Malus sieversii 31, Liberty, and Smoothee exhibited significantly high BRR (98.33%, 98.33%, and 93.33%, respectively) and a large NAB without vitrification. BRR and NAB linearly correlated with each other but not with callus formation rate. The broad sense heritability of the regeneration rate was 92.16%. The three Malus accessions that had high regeneration ability, and some of their sexual descendants, might be outstanding genetic resources for future genetic transformation.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaves of Formosan Sweetgum (Liquidambar formosana L.)

HortScience ◽  
2007 ◽  
Vol 42 (3) ◽  
pp. 721-723 ◽  
Author(s):  
L. Xu ◽  
G.F. Liu ◽  
M.Z. Bao
Keyword(s):  
Shoot Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Naphthalene Acetic Acid ◽  
Regeneration Rate ◽  
High Regeneration

Plantlets were regenerated from in vitro-grown leaf explants of five genotypes of Liquidambar formosana on WPM basal medium supplemented with different concentrations of TDZ and NAA. With the addition of 0.27 μm NAA, regeneration efficiency was increased by 2- to 4-fold over that with TDZ alone. Lower concentrations of TDZ (0.45–2.27 μm) were beneficial for regenerating shoot clusters. Four genotypes (P2, P6, P9, and P11) showed high regeneration rates (up to 90%), whereas genotype P13 showed a low capability for shoot regeneration on all media tested (<35%). For all five genotypes, the optimum medium for inducing adventitious shoots was WPM supplemented with 1.14 μm TDZ and 0.27 μm NAA, on which regeneration rate ranged from 72.6% to 89.5% and adventitious shoot clusters per regenerating leaf explant ranged from 2.63 to 3.11 in four genotypes (P2, P6, P9, and P11), while for P13, the regeneration rate and number of shoot clusters per regenerating explant were 23% and 1.39, respectively. Transfer of shoot clusters to WPM basal medium containing 0.54 μm NAA, 2.22 μm BA, and 1.44 μm GA3, resulted in shoot elongation. All the elongated shoots were rooted on WPM supplemented with 9.84 μm IBA, and plantlets were transplanted to soil successfully. Chemical names used: 6-benzyladenine (BA), gibberellic acid (GA3), indole-3-butyric acid (IBA), 1-naphthalene acetic acid (NAA), plant growth regulator (PGR), thidiazuron (TDZ), woody plant medium (WPM).

scholarly journals Establishment of a Genetic Transformation System in Guanophilic Fungus Amphichorda guana

2021 ◽  
Vol 7 (2) ◽  
pp. 138
Author(s):  
Min Liang ◽  
Wei Li ◽  
Landa Qi ◽  
Guocan Chen ◽  
Lei Cai ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Genetic Manipulation ◽  
Major Facilitator Superfamily ◽  
Protoplast Regeneration ◽  
Transformation System ◽  
Regeneration Rate ◽  
Bioactive Natural Products ◽  
Genetics Research ◽  
Genetic Transformation System ◽  
Major Facilitator

Fungi from unique environments exhibit special physiological characters and plenty of bioactive natural products. However, the recalcitrant genetics or poor transformation efficiencies prevent scientists from systematically studying molecular biological mechanisms and exploiting their metabolites. In this study, we targeted a guanophilic fungus Amphichorda guana LC5815 and developed a genetic transformation system. We firstly established an efficient protoplast preparing method by conditional optimization of sporulation and protoplast regeneration. The regeneration rate of the protoplast is up to about 34.6% with 0.8 M sucrose as the osmotic pressure stabilizer. To develop the genetic transformation, we used the polyethylene glycol-mediated protoplast transformation, and the testing gene AG04914 encoding a major facilitator superfamily transporter was deleted in strain LC5815, which proves the feasibility of this genetic manipulation system. Furthermore, a uridine/uracil auxotrophic strain was created by using a positive screening protocol with 5-fluoroorotic acid as a selective reagent. Finally, the genetic transformation system was successfully established in the guanophilic fungus strain LC5815, which lays the foundation for the molecular genetics research and will facilitate the exploitation of bioactive secondary metabolites in fungi.

Potential SNPs related to microspore culture in Raphanus sativus based on a single-marker analysis

2018 ◽  
Vol 98 (5) ◽  
pp. 1072-1083 ◽  
Author(s):  
Yong Suk Chung ◽  
Yun Gyeong Lee ◽  
Renato Rodrigues Silva ◽  
Suhyoung Park ◽  
Min Young Park ◽  
...  
Keyword(s):  
Raphanus Sativus ◽  
Microspore Culture ◽  
P Value ◽  
Nucleotide Polymorphisms ◽  
Single Marker Analysis ◽  
Regeneration Rate ◽  
Marker Analysis ◽  
Wide Range ◽  
Single Marker ◽  
High Regeneration

Radish (Raphanus sativus) is an economically important crop grown for its edible roots and leaves. It is a self-incompatible, outcrossing species, making the production of homozygous lines and the development of breeding populations difficult. However, this can be overcome with haploids production techniques using isolated microspores, providing the rapid production of homozygous lines for breeding. Thus, it would be useful to identify radishes with a high regeneration rate from microspore culture. In the current study, 96 radish cultivars or germplasms were evaluated for high regeneration rates. Also, a single-marker analysis (SMA) was applied to detect single nucleotide polymorphisms (SNPs) potentially associated with this trait using genotype-by-sequencing (GBS) technology. The regeneration rate from microspore culture of 96 lines showed a wide range, from 0% to 269.5%. From the SMA, 52 markers were detected at a p value of 0.001 and a total of 11 physically nearby genes with high levels of similarity in various species were identified as candidates for high regeneration rates. This result could be used for clarifying the genetic basis underlying these traits and developing molecular markers associated with regeneration rates and would be beneficial for generating homozygous inbred lines.

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals In vitro Propagation of Vanda testacea (Lindl.) Reichb.f. – A Rare Orchid of High Medicinal Value

2010 ◽  
Vol 19 (1) ◽  
pp. 1-7 ◽  
Author(s):  
Saranjeet Kaur ◽  
K.K. Bhutani
Keyword(s):  
Leaf Explants ◽  
Subsequent Development ◽  
Individual Treatment ◽  
Chemical Stimulus ◽  
Medicinal Value ◽  
Best Response ◽  
High Regeneration ◽  
Regeneration Response ◽  
High Regeneration Frequency

Foliar explants of Vanda testacea (Lindl.) Reichb. f. were cultured on Mitra (M) medium with 1.0 mg/l BAP, Kn  each and 1.0 mg/l NAA individually and in combination for initiation of regeneration response, proliferation of regenerants and subsequent development of plantlets. Juvenility of the tissues and chemical stimulus were important factors in initiating the regeneration response in the explants. The relatively older leaf explants (>1cm in length) remained recalcitrant to regeneration the representing younger ones (<1cm in length) responded to certain chemical regimes. BAP, Kn individually in the medium should direct PLB regeneration whereas when used with NAA, the explants showed callus proliferation and further differentiated into PLBs. An individual treatment with NAA (1.0 mg/l) impaired the response frequency and delayed further morphogenetic processes leading to plantlet development. The best response in the explants (in terms of high regeneration frequency, early initiation, PLB proliferations, and plantlet development) was observed in 1.0 mg/l BAP alone/with 1.0 mg/l NAA + activated charcoal. Plantlets were transferred to pots containing epiphytic compost (1 charcoal : 1 brick pices : 1 bats). Nearly 75% of plantlets survival was recorded.  Key words: In vitro, Orchid, Vanda testacea, Micropropagation, Protocorm-like bodies, callus D.O.I. 10.3329/ptcb.v19i1.4077 Plant Tissue Cult. & Biotech. 19(1): 1-7, 2009 (June) 

EFFECT OF SINGLE-WALL CARBON NANOTUBE ON SOYBEAN (GLYCINE MAX) REGENERATION FROM MATURE COTYLEDONARY NODE EXPLANTS

Nano LIFE ◽  
2012 ◽  
Vol 02 (04) ◽  
pp. 1250014 ◽  
Author(s):  
X. H. WANG ◽  
Q. M. HUANG ◽  
L. WANG ◽  
L. Z. WANG
Keyword(s):  
Carbon Nanotube ◽  
Adventitious Shoots ◽  
Cotyledonary Node ◽  
Shoot Elongation ◽  
Regeneration Frequency ◽  
Single Wall Carbon Nanotube ◽  
Single Wall Carbon ◽  
Plantlet Formation ◽  
Shoot Initiation ◽  
Cotyledonary Node Explants

One of the major limitations in producing transgenic soybeans using the agrobacterium-mediated cotyledonary-node method is low regeneration frequency. An improved highly efficient regeneration system of soybean was established herein. Cotyledonary node explants were placed in shoot initiation medium with single-wall carbon nanotube (SWNT) for adventitious shoots regeneration, and adventitious shoots were subcultured in shoot elongation medium with SWNT for shoot elongation and rooting. 40 mg/L SWNT supplemented in shoot initiation medium was found to be the optimal concentration with shoots regeneration frequency significantly increased by 21.5% compared with the control treatment, while for 4 mg/L and 400 mg/L, the increase was 4.6% and 6.5%, respectively. Faster elongation and rooting of adventitious shoots was observed in shoot elongation medium with 40 mg/L SWNT. Soybean plantlet formation frequency within the limited four weeks supplemented with 40 mg/L SWNT reached 48.4% while in the other three treatments: 4 mg/L, 400 mg/L and control, 0% were observed. These results indicate that supplement of SWNT in the soybean medium can efficiently promote adventitious shoots formation frequency, increase plantlet formation frequency and shorten the regeneration period.

scholarly journals Physiological Response of In Vitro Cultured MAGNOLIASP. to Nutrient Medium Composition

2014 ◽  
Vol 22 (1) ◽  
pp. 49-61 ◽  
Author(s):  
Rossen S. Sokolov ◽  
Bistra Y. Atanassova ◽  
Elena T. Iakimova
Keyword(s):  
Nutrient Medium ◽  
Culture Conditions ◽  
Medium Composition ◽  
Naphthaleneacetic Acid ◽  
Axillary Shoots ◽  
Regeneration Rate ◽  
Primary Explants ◽  
High Regeneration ◽  
Basal Salts

AbstractThe objective of this study was to assess the regeneration response of in vitro cultured Magnolia × soulangeana ‘Alexandrina’ and Magnolia liliiflora ‘Nigra’ to nutrient medium composition. In the primary culture (initiated from dormant axillary buds) combinations of Murashige and Skoog (MS) basal salts with 6-benzylaminopurine and α-naphthaleneacetic acid were tested. The primary explants of cv. ‘Alexandrina’ expressed higher regeneration rate than cv. ‘Nigra’. For both species, the regen eration was most strongly potentiated at addition of 0.25 mg dm−3 of the cytokinin alone. The auxin exerted undesir–able effects. Several basal salts media were applied in proliferation stage and their physiological effects were evaluated in reference to traditionally used MS. At culturing on Chée & Pool C2d Vitis Medium (VM) that is for the first time introduced to magnolia and on MS, M. liliiflora formed more but less elongated shoots than M. soulangeana. However, on VM, substantial increase (25-30%) of the number of axillary shoots and leaves, shoot length and fresh and dry weights over MS was established for both species. This suggested VM as promising composition of nutrients in multiplication stage. Microshoots obtained on MS, VM, Rugini Olive Medium and DKW Juglans Medium were successfully rooted in vitro and subsequently established ex vitro. The findings expand the information on magnolia response to culture conditions and contribute to elaboration of innovative elements of protocols for establishing tissue cultures with high regeneration capacity.

Establishment of regeneration system and transformation of Zm401 gene in Lilium longiflorum×L. formosanum

2008 ◽  
Vol 5 (2) ◽  
pp. 113-119 ◽  
Author(s):  
Li Qiu-Hua ◽  
Hong Bo ◽  
Tong Zheng ◽  
Ma Chao ◽  
Guan Ai-Nong ◽  
...  
Keyword(s):  
Genetic Transformation ◽  
Lilium Longiflorum ◽  
Bacterial Suspension ◽  
Specific Gene ◽  
Naphthaleneacetic Acid ◽  
Transformation System ◽  
Regeneration System ◽  
Regeneration Rate ◽  
Maize Pollen ◽  
Genetic Transformation System

AbstractIn vitro bulb scales of Lilium longiflorum×L. formosanum were used as explants to develop a highly efficient regeneration system. A high regeneration rate (100%) was reached through organogenesis on basal Murashige and Skoog (MS) medium supplemented with 1.0 mg/l 6-benzylaminopurine (6-BA) and 1.0 mg/l naphthaleneacetic acid (NAA). A genetic transformation system for the lily was developed using an Agrobacterium tumefaciens-mediated method. An improved genetic transformation rate (12‰) was obtained when the explants were pre-cultured for 3 days, immersed in bacterial suspension (OD600≈0.8) for 5 min, and co-cultivated for 5 days. The binary vector pBI121 containing Zm401, a maize pollen-specific gene, was introduced into the Agrobacterium strain LBA4404 and transformed into the explants using the genetic transformation system. Gene integration into the lily genome was confirmed by polymerase chain reaction (PCR) and PCR–Southern analysis. These results could lead to the production of new pollenless lily plants.

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