转多基因1年生库安托杨对溃疡病的抗性分析

Resistance Evaluation of the One-year-old Multiple Transgenic Populus×euramericana 'Guariento' to Poplar Canker

[目的]本研究对转多基因库安托杨株系的溃疡病抗病性检测及抗病相关基因的表达分析,为通过基因工程手段培育抗病林木新树种提供有价值的参考。[方法]以转JERF36等多个外源基因的1年生库安托杨株系D5-9、D5-20、D5-21及非转基因受体D5-0为材料,对主干人工接种溃疡病菌的各株系病情指数进行了测定,同时对接种5 d后树皮组织中5个抗病基因的表达情况进行了实时定量PCR(qPCR)分析。[结果]在溃疡病菌胁迫下,3个转基因株系对溃疡病菌的抗性显著优于非转基因受体株系,转基因株系间抗病性也具有一定差异,D5-21的病情指数显著低于其它2个转基因株系;肉桂醇脱氢酶基因在3个转基因株系树皮中表达量均高于对照,类甜蛋白基因仅在D5-19树皮中的表达量高于对照,其他3个基因在转基因株系中的表达量或与对照相当或低于对照。[结论]转多基因库安托杨株系的溃疡病抗性与非转基因对照相比均有显著提高,且转基因株系间差异显著;同时转基因株系中不同抗病基因的表达模式也有很大差异,说明溃疡病菌胁迫下转多基因库安托杨抗病调控的复杂性,其分子调控机制还有待深入研究。

[Objective] To evaluate the resistance to poplar canker and mRNA expression of disease resistance-related genes of three transgenic Populus× euramericana ‘Guariento’ lines so as to provide reference for breeding disease-resistant trees through transgenic technology.[Method] Using 1-year-old stem of multiple transgenic P.× euramericana ‘Guariento’ clones (D5-9, D5-20, and D5-21) and the wild control (D5-0) as material, the disease index of canker was analyzed after inoculated with Botryosphaeria dothidea for 60 days, and the expression of five disease resistance related genes in bark of these clones after inoculated with B. dothidea for 5 days were detected by relative quantitative PCR (qPCR).[Result] The results showed that the resistance of the three transgenic clones to poplar canker was significantly higher than that of the non-transgenic clone and obvious difference in canker resistance was observed among the three transgenic poplar clones with D5-21 was higher than the other two clones. The expression of cinnamyl-alcohol dehydrogenase family protein gene in the bark of the 3 transgenic clones was higher than that of the control, and the expression of thaumatin-like protein gene was higher than control only in bark of D5-19, whereas the expressions of the other three genes were similar to or lower than the control.[Conclusion] The difference in the canker resistance of the three transgenic clones and various expression profiles of the five disease resistant genes indicates the regulation complexity in the disease resistance of the multiple transgenic poplars under stress of B. dothidea, suggesting further study is needed.