花药培养快速培育聚合抗3种水稻病害基因的新种质研究

Study of Quick Cultivation of New Seeds with Three Diseases Resistance by Anther Culture Method

为培育抗水稻白叶枯病、稻瘟病、条纹叶枯病3种病害的品种,保证水稻的稳产高产,利用花药培养与常规育种技术、分子标记辅助选择技术相结合,进行聚合抗3种病害的新种质研究。结果表明,用含有抗白叶枯病Xa23基因的BG152和含有抗稻瘟病Pi-1基因的R118分别与含抗条纹叶枯病Stvb-i基因的花育409进行有性杂交,共获得聚合双抗抗病基因的F0种子分别为180,105粒;鉴别真杂种后彼此再杂交,获得复交F0种子1 398粒。利用分子标记辅助选择技术,筛选出聚合抗3种病基因的杂合型单株52株,从中选择田间无病害的进行花药培养,经H0自然加倍,获得双倍体花培植株378株。经PCR检测,筛选出聚合3种抗病基因的花培H1株系14个;对其H2植株进行重复PCR与抗病鉴定,获得聚合3种抗病基因的花培材料所含的抗病基因能稳定遗传,抗病性鉴定有10份均表现为抗(R),且高效表达抗病。最后对水稻花药培养中存在的问题进行了分析,并提出了相应防治措施。

In order to breeding disease-resistant rice varieties to ensure stable and high-yields of rice,the breeding of disease-resistant germplasm was studied by methods of conventional,anther culture and molecular mark-er-assisted technology. The results showed that,after the gene Stvb-i resistant to stripe disease was hybridized by BG152 of bacterial leaf blight resistant gene Xa23 and R118 of rice blast resistant gene Pi-1 respectively,180 and 105 the seeds F0 with double resistant disease gene were obtained. Those seeds were re-crossed after identification of the true ones and 1 398 F0 hybrids were screened. 52 single plants with disease resistant to three genes were ob-tained by molecular marker-assisted technology. Furthermore,378 plants with double anther-culture were gotten based on the selection and culture of no disease samples in field after H0 natural doubling. 14 strains of anther cul-ture with three disease-resistant genes were obtained though the PCR detection. After multiple identification of PCR and disease resistance of their H2 plants,10 new seeds with stabilized hereditary and high level expression were ob-tained. At last,the analysis of the problem during the course of rice anther culture was carried out and the preven-tion and corresponding actions were proposed also.