水稻白叶枯病菌室内链霉素抗性菌株的诱导及其抗性分子机制和风险评价

徐颖; 祝晓芬; 周明国

水稻白叶枯病菌室内链霉素抗性菌株的诱导及其抗性分子机制和风险评价

1.
南京农业大学植物保护学院, 南京 210095
2.
上海市园林科学研究所, 上海 200232

基金项目:

国家高技术研究发展计划(863计划)(2012AA101502);国家自然科学基金(31100100);公益性行业(农业)科研专项(201303023).

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出版历程
- 收稿日期: 2012-06-24
- 修回日期: 2012-10-06

Molecular mechanism and resistance risk assessment of laboratory-induced streptomycin-resistant mutants of Xanthomonas oryzae pv.oryzae

1.
College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, China
2.
Shanghai Landscape Gardening Research Institute, Shanghai 200232, China

摘要

摘要: 通过紫外光照射诱导获得了6株水稻白叶枯病菌Xanthomonas oryzae pv.oryzae抗链霉素突变体。测得链霉素对水稻白叶枯病菌敏感菌株ZJ173及其抗性突变体的最低抑制浓度(MIC)分别为0.10和600 μg/mL;对敏感菌株的有效抑制中浓度(EC50)为0.03 μg/mL,对抗性菌株的平均EC50值为11.64 μg/mL,平均抗性倍数为388。通过PCR扩增了敏感菌株ZJ173及5株抗性菌株的rpsL基因(编码S12核糖体蛋白)和rrs基因(编码16S rRNA),并检测了strA基因是否存在。序列分析表明,5株被测抗性菌株的rpsL基因均发生了突变,其中4株在氨基酸43位、1株在88位,均由赖氨酸突变为精氨酸,而rrs基因未发生突变,strA基因未被检测到。表明实验室诱导获得的水稻白叶枯病菌抗性菌株对链霉素的抗药性是由rpsL基因突变引起的。抗性风险研究表明,抗性突变体的抗药性在无药剂压力下可稳定保持,其致病性、生长速率与敏感菌株相比无明显差异,竞争性低于或略低于敏感菌株,抗性自发突变率较高,且抗性突变为单一位点突变,病害循环为多循环,因此由rpsL基因突变引起的水稻白叶枯病菌对链霉素的抗性风险较高。
- 水稻白叶枯病菌 /
- 链霉素 /
- 诱导抗性 /
- rpsL基因 /
- 抗性风险
Abstract: Six streptomycin-resistant mutants of Xanthomonas oryzae pv.oryzae were obtained by ultraviolet light induction in the laboratory.The minimal inhibition concentrations(MICs) of streptomycin inhabiting wild-type strain ZJ173 and resistant mutants were 0.10 and 600 μg/mL,respectively.The medium effective concentration(EC50) of wild-type strain ZJ173 was 0.03 μg/mL and the mean EC50 value of six mutants was 11.64 μg/mL. The resistance factor(RF) of laboratory mutants relative to their parent strain ZJ173 was 388.Mutations in the rpsL(encoding S12 protein) and rrs gene(encoding 16S rRNA) and the presence of strA gene were examined in sensitive and resistant strains of X.oryzae pv.oryzae by PCR amplification and sequencing.Four of five tested laboratory mutants had mutation at codon 43(Lys→Arg) and the other one at 88(Lys→Arg) in the rpsL gene.Neither the strA gene nor mutation in the rrs has been found.The resistant mutants maintained their resistance through 10 times of transfers on NA plates.The pathogenicity and growth rate showed no difference between resistant mutants and sensitive strain,while the competitiveness of resistant mutants is lower or slightly lower than that of sensitive strain.Considering the high spontaneous mutation frequency,single-gene mutation pattern and a typical multi-cycle disease,the risk of streptomycin-resistance caused by rpsL mutation in X.oryzae pv.oryzae would be high.
- Xanthomonas oryzae pv.oryzae /
- streptomycin /
- induced resistance /
- rpsL gene /
- resistance risk

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