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Expression of a hevein-like gene in transgenic Agave hybrid No. 11648 enhances tolerance against zebra stripe disease

文献类型： 外文期刊

作者： Gao, Jianming 1 ; Yang, Feng 1 ; Zhang, Shiqing 1 ; Li, Jinzhi 2 ; Chen, Helong 1 ; Liu, Qiaolian 1 ; Zheng, Jinlong 3 ;

作者机构： 1.Chinese Acad Trop Agr Sci, Inst Trop Biosci & Biotechnol, Key Lab Trop Crop Biotechnol, Minist Agr, Haikou 571101, Peoples R China

2.Taizhou Univ, Sch Life Sci, Linhai 317000, Zhejiang, Peoples R China

3.Chinese Acad Trop Agr Sci, Environm & Plant Protect Res Inst, Haikou 571101, Peoples R China

关键词： Hevein;Transgenic;H. 11648;Tolerance;Zebra disease

期刊名称：PLANT CELL TISSUE AND ORGAN CULTURE （ 影响因子：2.711； 五年影响因子：2.73 ）

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收录情况： SCI

摘要： Zebra disease, which is caused by Phytophthora nicotianae, is a serious global threat to the main cultivar Agave hybrid No. 11648 (H.11648). This disease requires a long period to obtain anti-P. nicotianae variants by crossbreeding, given that 10 years are needed for this plant to bloom. Therefore, transgenic technology could be useful for the production of new agave lines that might develop significant tolerance to P. nicotianae within a relatively short period. In this study, a tissue culture system was tested on H.11648. Results showed that the optimum culture medium for callus induction was SH + 3 mg L-1 6-BA + 0.5 mg L-1 NAA + 0.1 mg L-1 2,4-D + 30 g L-1 sucrose + 6.5 g L-1 carrageenan, with pH of 5.8. The induction medium for budding was SH + 1.5 mg L-1 6-BA + 0.5 mg L-1 NAA + 30 g L-1 sucrose + 6.5 g L-1 carrageenan at pH 5.8. The induction medium for rooting was SH + 0.1 mg L-1 IAA + 30 g L-1 sucrose + 6.5 g L-1 carrageenan. The primary factors influencing plant transformation efficiency were also investigated. Results showed that the effective time for infection was 10 min, the optimum concentration of acetosyringone was 200 mu M, the optimum time for pre-culture of sisal callus was 3 days, and the optimum co-culture time for calli and Agrobacterium was 4 days. Sisal calli will die when phosphinothricin concentration reaches 2.0 mg L-1. Furthermore, a hevein-like protein gene from Pharbitis nil was introduced to H.11648. The integration stability and expression level of the transgene in the H.11648 genome were analyzed by using Southern blot and reverse transcription polymerase chain reaction. The mycelial growth rate of P. nicotianae was significantly inhibited by the crude leaf extracts from the transgenic H.11648 plants. After further study in vivo, the resistance of transgenic H.11648 plants to P. nicotianae infection was enhanced.

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