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【目的】建立简便高效的水稻突变体筛选体系,为克隆参与调控水稻xa13/Xa13表达以及xa13/Xa13介导的抗/感病和花粉发育信号途径的基因奠定基础。【方法】利用EMS化学诱变携带显性Xa13启动子驱动绿色荧光蛋白(GFP)基因的水稻转基因种质IRBB13(PXa13:GFP),构建突变群体,分3个时期进行突变体筛选:幼苗期筛选根和芽中绿色荧光减弱或消失的突变体、成株期筛选回复感病表型的突变体和孕穗期筛选育性下降的突变体。【结果】室内检测代M2突变体株系3 000个,筛到幼根和幼芽荧光消失的突变体14个,根部荧光正常而芽无荧光的突变体9个。田间试验种植突变体株系3 000个,成株期接种PXO99初步筛到回复感病表型的突变体株系25个。孕穗期筛选到育性下降且花药中绿色荧光消失的突变体株系7个,其中,包含1个回复感病的突变体株系。【结论】建立了简便高效的筛选水稻突变体筛选体系,结合GFP表达模式异常的突变体、回复感病突变体以及育性下降的突变体分析可判断xa13/Xa13参与的水稻抗/感病与花粉发育这2条信号网络途径并不是完全独立的,而是存在一定程度的交叉。
【Objective】 To establish a simple and efficient screening system for rice mutant and lay a foundation for cloning the genes involved in the regulation of xa13 / Xa13 expression and xa13 / Xa13-mediated resistance / susceptibility and pollen development pathway in rice. 【Method】 Mutant groups were constructed by EMS chemical mutagenesis of rice transgenic rice IRBB13 (PXa13: GFP) carrying the dominant Xa13 promoter and green fluorescent protein (GFP) gene. The mutants were screened in three stages: seedling stage Mutants with diminished or disappeared green fluorescence in roots and shoots, mutants with susceptible phenotypes recovered in adult stage, and mutants with reduced fertility at booting stage. 【Result】 The results showed that there were 3 000 M2 mutant lines detected in laboratory, 14 mutants with fluorescence disappeared in young and young shoots, 9 with normal fluorescence in roots and no fluorescence in shoots. In the field experiment, 3 000 mutant lines were planted and inoculation of PXO99 in adult stage was initially screened to 25 mutant lines which responded to the susceptible phenotype. At the booting stage, 7 mutant lines with reduced fertility and disappearance of green fluorescence in anthers were screened, including 1 mutant line which responded to the disease. 【Conclusion】 A simple and efficient screening system for rice mutant screening was established. Mutants with abnormal expression pattern of GFP, response to susceptible mutants and mutants with reduced fertility could be used to determine the resistance / susceptibility of rice to xa13 / Xa13 Pollen development of these two signaling networks is not completely independent of the way, but there is a certain degree of cross-over.