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| 农杆菌介导的棕色棉胚珠离体培养遗传转化体系的建立 | |
| 引用本文： | 丁一,耿晓丽,何守朴,潘兆娥,贾银华,王立如,吴立强,杜雄明. 农杆菌介导的棕色棉胚珠离体培养遗传转化体系的建立[J]. 分子植物育种, 2021, 19(1): 218-223 |
| 作者姓名： | 丁一 耿晓丽 何守朴 潘兆娥 贾银华 王立如 吴立强 杜雄明 |
| 作者单位： | 河北农业大学农学院,保定,071000;中国农业科学院棉花研究所,棉花生物学国家重点实验室,安阳,455000;中国农业科学院棉花研究所,棉花生物学国家重点实验室,安阳,455000;河北农业大学农学院,保定,071000 |
| 基金项目： | 转基因生物新品种培育科技重大专项子课题(2016ZX08005-005)资助。 |
| 摘 要： | 本研究以棕色棉品种'棕1-61'为受体材料,以β-葡萄糖苷酸酶基因(GUS)为报告基因,探索农杆菌介导离体培养胚珠的遗传转化条件.结果表明最适合的遗传转化条件是:1 DPA胚珠预培养4d后,用针头进行扎孔处理形成伤口,在OD600为0.6～0.8的农杆菌菌液中侵染3 min,22 ℃黑暗条件下共培养20h后转移至含50... |
| 关 键 词： | 棕色棉 胚珠 农杆菌介导法 遗传转化 离体培养 |
Establishment of Agrobacterium-mediated Genetic Transformation System of In Vitro Culture Brown Cotton Ovules |
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| Ding Yi,Geng Xiaoli,He Shoupu,Pan Zhaoe,Jia Yinhua,Wang Liru,Wu Liqiang,Du Xiongming. Establishment of Agrobacterium-mediated Genetic Transformation System of In Vitro Culture Brown Cotton Ovules[J]. Molecular Plant Breeding, 2021, 19(1): 218-223 | |
| Authors: | Ding Yi Geng Xiaoli He Shoupu Pan Zhaoe Jia Yinhua Wang Liru Wu Liqiang Du Xiongming |
| Affiliation: | (College of Agronomy,Agriculture University of Hebei,Baoding,071000;State Key Laboratory of Cotton Biology,Institute of Cotton Research of the Chinese Academy of Agricultural Sciences,Anyang,455000) |
| Abstract: | In this study,we used the beta-glucuronidase(GUS)gene as the reporter gene and the in vitro cultured ovules of cotton cultivar’Brown 1-61’as the explants to investigate the optimum conditions for Agrobacteriummediated transformation.We found that the optimum transformation system was:4 day pre-cultured ovules stabbed by syringe needle were infection with Agrobacterium(OD600=0.6~0.8)for 3 min,and after cocultivation at 22℃in darkness for 20 h the ovules were then transferred to the BT medium containing 500 mg/L kanamycin for growth in darkness.The GUS stainning results found that 20 d cultured ovules and the cotton fibers on the 30 d cultured ovules could been stained and proved that the transformation was success.We further constructed the RNAi vector of Gh UFGT and introduced into pre-cultured ovules via Agrobacterium-mediated transformation.And the q PCR result demonstrated that the Gh UFGT showed significant decreased expression levels in positive ovules when compared to the control.Our study provides a fast and efficient transformation method for studying the function of genes which involved in fiber development and pigment deposition. |
| Keywords: | Brown cotton Ovule Agrobacterium-mediated method Genetic transformation In vitro culture |
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