Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro

Дубровна, О. В.; Slivka, L. V.

doi:10.7124/feeo.v26.1264

Cited by 4 publications

(3 citation statements)

References 13 publications

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“…Different concentrations of IBA, 6-BA, and Vc added to the differentiation medium had positive effects on callus differentiation [19]. Zhang et al [20] suggested that the most suitable 6-BA concentration for wild wheat was 0.5 mg/L, and a higher 6-BA concentration would increase the browning rate and decrease the callus differentiation rate, which could be because of the varying tolerance levels of crops to hormones. Sansar et al [21] found that the combination of IBA and 6-BA had the best effect on adventitious bud proliferation in Poplar xinjiangensis, and the bud proliferation ratio was the best when 6-BA and NAA were used in combination for bud proliferation in Scutellaria stems [22].…”

Section: Discussionmentioning

confidence: 99%

Optimization of Agrobacterium tumefaciens-Mediated Genetic Transformation of Maize

Liu¹,

Zhou²,

Liu³

et al. 2022

Phyton

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Immature embryos of inbred maize (Zea mays) lines (H8183, H8184, and H8185) were used for Agrobacterium infection. We used the β-glucuronidase gene (GUS) as the target gene and the glufosinate resistance gene (bar) as the selection marker. We conducted research on several aspects, such as different genotypes, coculture conditions, screening agent concentrations, and concentrations of indole-3-butytric acid (IBA), 6-benzylaminopurine (6-BA), and ascorbic acid (Vc) in the differentiation medium. We optimized the genetic transformation system, and the obtained results indicated that among the three lines studied, the induction rate of H8185 was the highest at 93.2%, followed by H8184, with H8183 having the lowest induction rate (80.1%). The best coculture method was that using the N6 coculture medium layered with a sterile filter paper. Using orthogonal analysis, we found that the optimal combination of the three factors in the differentiation medium was A 3 (1 mg mL −1 IBA), B 3 C 1 (1.6 mg mL −1 6-BA), and D 3 (1.5 mg mL −1 Vc). Through GUS staining analysis, Bar test-strip analysis, and polymerase chain reaction, five transgenic plants were finally obtained. This study established the optimal conditions for genetic transformation in maize.

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Section: Discussionmentioning

confidence: 99%

Optimization of Agrobacterium tumefaciens-Mediated Genetic Transformation of Maize

Liu¹,

Zhou²,

Liu³

et al. 2022

Phyton

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“…Agrobacterium-опосередковану трансформацію проводили згідно розроблено нами протоколу [14] Інтеграцію елементів векторної конструкції встановлювали ПЛР-методом за наявністю фрагментів екзона та інтрона гена ProDH1 арабідопсису та селективного гена неоміцинфосфотрансферази -nptII. Екстракцію ДНК із листків рослин проводили з використанням комплекту реагентів «ДНК-сорб-С» (ФБУН ЦНДІ, Росія).…”

Section: матеріали і методиunclassified

Obtaining genetically modified winter wheat plants with partial suppression of the prolin dehydrogenase gene

Dubrovna,

Slyvka,

Velikozhon

2023

Fakt. eksp. evol. org.

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Aim. Production of genetically modified plants of new promising winter wheat genotypes with partial suppression of the proline dehydrogenase gene in in vitro culture and determination of proline content in transgenic and control plants. Methods. Agrobacterium-mediated transformation in culture in vitro, molecular genetic analysis; biochemical determination of proline content; of mathematical statistics. Results. Using the method of Agrobacterium-mediated transformation of callus cultures of new promising genotypes of winter wheat, transgenic plants carrying a double-stranded RNA suppressor of the proline dehydrogenase gene were obtained. The frequency of transformation with the use of the LBA 4404 strain in the studied genotypes was 1.7-2.0 %, and with the use of the AGL0 strain, it was 2.0-2.3 %. It was established that plants with reduced activity of proline dehydrogenase are characterized by a significantly higher content of free L-proline compared to the control. Conclusions. The relatively greater efficiency of using the AGL0 strain for obtaining transgenic plants of winter wheat genotypes with partial suppression of the proline dehydrogenase gene in in vitro culture is shown. The presence of a double-stranded RNA suppressor of the ProDH gene in transgenic plants leads to an increase in the level of free L-proline accumulation.

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“…Нами шляхом Agrobacteriumопосередкованої трансформації в культурі in vitro отримано трансгенні рослини 4 нових перспективних генотипів озимої м'якої пшениці (Ук 065; Ук095/17; Ук 209h; Ук 322/17), які несуть дволанцюговий РНК-супресор гена проліндегідрогенази Arabidopsis thaliana та селективний ген неоміцинфосфотрансферази II (nptII) E.coli [13]. Із цих рослин шляхом самозапилення отримано рослини насіннєвого покоління Т 2 , трансгенна природа яких була підтверджена за допомогою ПЛР з праймерами, специфічними до генів ProDH та nptII.…”

Section: ключові словаunclassified

Physiological, biochemical and economic characteristics of transgenic winter wheat plants with RNA suppressor of the proline dehydrogenase gene

Slivka¹,

Дубровна²

2022

Fakt. eksp. evol. org.

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Aim. To analyze the physiological, biochemical and economic characteristics of genetically modified plants of new promising genotypes of winter bread wheat of seed generation T2 with a double-stranded RNA suppressor of the proline dehydrogenase gene. Methods. Agrobacterium-mediated transformation in vitro; biochemical determination of proline dehydrogenase enzyme activity and free proline content; morphometric indicators and elements of crop structure; mathematical statistics. Results. It is shown that transgenic plants, in contrast to control, grow on a medium with mannitol more intensely, retaining a green color. It was found that both under normal conditions and under conditions of water deficiency, plants of seed generation T2 have an increased level of free Proline in the leaves compared to control genotypes. It was found that transformants are characterized by reduced activity of the enzyme proline dehydrogenase, which is manifested by changes in normal – stress – normal conditions. Transgenic T2 plants had a higher tolerance to water deficiency compared to the original, which was reflected in the nature of their growth. In conditions of soil moisture deficiency, the yield of most transformed lines was higher compared to untransformed plants. Conclusions. The results suggest that the use of a vector construct with a double-stranded RNA suppressor of the ProDH gene is effective for creating transgenic winter bread wheat plants with increased tolerance to water deficiency.

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Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro

Cited by 4 publications

References 13 publications

Optimization of Agrobacterium tumefaciens-Mediated Genetic Transformation of Maize

Optimization of Agrobacterium tumefaciens-Mediated Genetic Transformation of Maize

Obtaining genetically modified winter wheat plants with partial suppression of the prolin dehydrogenase gene

Physiological, biochemical and economic characteristics of transgenic winter wheat plants with RNA suppressor of the proline dehydrogenase gene

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