Enhanced Stem Nematode Resistance of Transgenic Sweetpotato Plants Expressing Oryzacystatin-I Gene

• Published in: Agricultural sciences in China Vol. 10; no. 4; pp. 519 - 525
• Main Authors: GAO, Shang, YU, Bo, ZHAI, Hong, HE, Shao-zhen, LIU, Qing-chang
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.04.2011; Key Laboratory of Crop Genomics and Genetic Improvement, Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education/China Agricultural University, Beijing 100193, P.R.China
• Subjects: Agrobacterium radiobacter; Agrobacterium tumefaciens; beta-glucuronidase; cell aggregates; cysteine proteinase inhibitors; gene overexpression; genetic transformation; Ipomoea batatas; Ipomoea batatas (L.) Lam; I基因; Nematoda; Oryzacystatin-I gene; pest resistance; plant parasitic nematodes; quantitative polymerase chain reaction; reporter genes; somatic embryogenesis; Southern blotting; stem nematode resistance; sweet potatoes; transgenes; transgenic plant; transgenic plants; 体细胞胚胎发生; 农杆菌介导转化; 抗性基因; 植物表达; 甘薯茎线虫病; 转基因植株; 转基因植物; Agrobacterium tumefaciens; Oryzacystatin-I gene; stem nematode resistance; Ipomoea batatas (L.) Lam; transgenic plant; Ipomoea batatas (L.) Lain
• ISSN: 1671-2927
• DOI: 10.1016/s1671-2927(11)60032-1

Enhanced stem nematode resistance of transgenic sweetpotato （cv. Lizixiang） was achieved using Oryzacystatin-I （OCI） gene with Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harbors a binary vector pCAMBIA1301 with OCI gene, gusA gene and hptII gene. Selection culture was conducted using 25 mg L-1 hygromycin. A total of 1 715 plants were produced from the inoculated 1 450 cell aggregates of Lizixiang via somatic embryogenesis. GUS assay and PCR analysis of the putative transgenic plants randomly sampled showed that 90.54% of them were transgenic plants. Transgenic plants exhibited significantly enhanced resistance to stem nematodes compared to the untransformed control plants by the field evaluation with stem nematodes. Stable integration of the OCI gene into the genome of resistant transgenic plants was confirmed by Southern blot analysis, and the copy number of integrated OCI gene ranged from 1 to 4. Transgene overexpression in stem nematode-resistant plants was demonstrated by quantitative real-time PCR analysis. This study provides a way for improving stem nematode resistance in sweetpotato.