水稻多酚氧化酶基因功能标记的开发与应用

Development and Application of Functional Markers for Polyphenol Oxidase (PPO) Alleles in Rice

水稻酚反应是水稻分类的重要依据。控制水稻酚反应的多酚氧化酶基因(PPO)已经克隆,根据其等位基因的DNA序列差异设计了InDel功能标记FMppo-18和FMppo-29。利用这两个标记对45份材料进行了标记基因型分析和酚反应鉴定。结果发现3份普通野生稻为非缺失带型;8份籼稻材料中6份为非缺失带型,而Dular为缺失29 bp带型,龙晴为缺失18bp带型,表明Dular和龙晴分别携带29 bp和18 bp缺失的PPO等位基因;11份粳稻材料中9份携带18 bp缺失的PPO等位基因,2份携带29 bp缺失的PPO等位基因。对23份杂草稻鉴定的结果表明,7份早年发现的杂草稻,即3份江苏省连云港穞稻和4份安徽省怀远、来安、全椒、肥东的塘稻为非缺失带型;而16份近年来发现的杂草稻中14份不缺失,2份表现为18 bp缺失的PPO等位基因,与江苏省大面积推广的栽培粳稻品种携带相同的PPO基因。酚反应结果表明带29 bp或18bp缺失的PPO等位基因的材料酚反应呈阴性,而不缺失的材料酚反应呈阳性。标记基因型与酚反应表现型完全对应。表明这两个功能标记可用于水稻种质资源鉴定、进化研究以及多酚氧化酶基因的分子标记辅助选择育种。

The phenol reaction（PHR） phenotype is one of the criteria of rice classification and the gene controlling PHR has been identified.The positive PHR phenotype is conditioned by functional polyphenol oxidase（PPO） gene,and negative PHR phenotype is conditioned by non-functional PPO gene.To seek the markers for rice classification,two InDel markers,FMppo18 and FMppo29,were developed flanking on the 18 bp or 29 bp deletion in exon 3 of PPO alleles.Forty-five rice materials were genotyped with the two markers and their PHR phenotypes were also investigated.Three accessions of wild rice and six indica rice varieties carried non-deletion functional PPO alleles.Among the 11 japonica rice materials,nine carried 18 bp deletion allele,and two carried 29 bp deletion alleles.However,the indica rice variety Dular carried 29 bp deletion allele and Longqing carried 18 bp deletion allele.Among the 23 weedy rice accessions,three accessions of Ludao collected from Lianyungang of Jiangsu Province,China,and four accessions of Tangdao collected from Huaiyuan,Laian,Quanjiao,and Feidong of Anhui Province,China in 1950s carried non-deletion PPO alleles.Among the 16 weedy rice accessions collected in recent years,14 carried normal PPO alleles,and two carried 18 bp deletion PPO allele which was consistent with the cultivated rice in Jiangsu Province.These results showed that the positive PHR materials carried normal PPO alleles,and the negative PHR materials carried deletion PPO alleles,indicating that the phenotype perfectly matched with the genotype.Therefore,the two markers can be used in germplasm screening,marker-assisted selection and evolution research in rice.