大麦TILLING体系在抗病基因研究中的应用

TILLING-BASED ANALYSIS OF DISEASE RESISTANCE GENES IN BARLEY

基因组靶向定位诱导损伤技术(Targeting Induced Local Lesions In Genomes,TILLING)是在化学诱变和PCR定向筛选基础上发展起来的检测点突变的反向遗传学研究方法,在多种重要农作物上都有应用。本研究使用化学诱变剂甲基磺酸乙酯(Ethyl methane sulfonate,EMS)处理大麦品种‘Tamalpais’,获得了2154个M2株系,同时开发了基于芹菜内切酶CEL I(celery juice extract)的酶切筛选体系。针对植物水杨酸抗病途径相关的两个重要基因EDR1和NPR1,检测到5个M2突变株系。序列分析表明,2个突变发生在内含子、1个NPR1基因的同义突变、2个EDR1基因的错义突变(His351Tyr和Pro556Ser)。本项研究为麦类作物反向遗传学研究奠定了基础。

TILLING（Targeting Induced Local Lesions In Genomes） is a powerful reverse genetic tool widely used in many important crops.In this study,2154 M2 mutant lines were developed from barley genotype ＇Tamalpais＇ which was treated by ethyl methane sulfonate（EMS）.A CEL Ⅰ（celery juice extract） based screening system was developed.In plants,EDR1 and NPR1 genes regulate plant defense responses through the Salicylic acid（SA） signaling pathway.Mutants of both EDR1 and NPR1 genes were screened in the current M2 population.Five mutants were obtained.Two mutations occurred in the intron region;the rest three mutations included one silent mutation of the NPR1 gene and two missense mutations（His351Tyr and Pro556Ser） in the EDR1 gene.This work provides preliminary data to develop reverse genetics tool in Triticeae Crops.