Improvement of plant regeneration and Agrobacterium-mediated genetic transformation of Stylosanthes guianensis
文献类型： 外文期刊
作者： Guo, Pengfei 1 ; Liu, Pandao 2 ; Lei, Jian 1 ; Chen, Caihong 1 ; Qiu, Hong 2 ; Liu, Guodao 2 ; Chen, Zhijian 2 ; Luo, Lij 1 ;
作者机构： 1.Hainan Univ, Coll Trop Crops, Hainan Key Lab Sustainable Utilizat Trop Bioresou, Haikou 570110, Hainan, Peoples R China
2.CATAS, Trop Crops Genet Resources Inst, Haikou 571101, Hainan, Peoples R China
关键词： Genetic engineering; GUS staining; plant hormones; tissue culture; tropical legumes
期刊名称：TROPICAL GRASSLANDS-FORRAJES TROPICALES （ 影响因子：0.611； 五年影响因子：0.897 ）
ISSN： 2346-3775
年卷期： 2019 年 7 卷 5 期
页码：
收录情况： SCI
摘要： As a pioneer tropical pasture legume, stylo (Stylosanthes guianensis) is well adapted to growth-limiting factors in acid soils. Considering the importance of stylo, there is a need to improve Agrobacterium-mediated genetic transformation to enable development of elite cultivars. In this study, S. guianensis cv. RY5 was used to systematically optimize Agrobacterium-mediated transformation based on its plant regeneration. Results showed that Murashige and Skoog (MS) medium containing 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 2 mg/L 6-benzylaminopurine (6-BA) was the optimal callus induction medium. MS medium supplemented with 2 mg/L 6-BA was suitable for shoot regeneration from cotyledon-derived calluses, and 0.5 mg/L indole-3-acetic acid (IAA) and 0.5 mg/L indole-3-butyric acid (IBA) applications were beneficial for rooting The highest transformation efficiency (67%) was obtained at an Agrobacterium concentration of optical density = 0.6 combined with an infection time of 15 min and 3 days of co-cultivation. Furthermore, 200 mg/mL carbenicillin (Carb) and 0.6 mg/L Basta (R) supplements were effective in eliminating excess bacterial growth and selecting transgenic plants, respectively. Subsequent polymerase chain reaction (PCR) analysis confirmed that the beta-glucuronidase (GUS) and BAR genes were successfully integrated into the stylo genome. Wider testing of this improved protocol as a means of enhancing genetic improvement and gene function analysis of stylo seems warranted.
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