【作者】 张霞；

【导师】 于肖夏；

【作者基本信息】 内蒙古农业大学 ， 作物遗传育种， 2022， 硕士

【摘要】 花青素含量是彩色马铃薯的重要品质性状。针对四倍体彩色马铃薯花青素含量相关分子标记的研究相对较少的问题,本研究通过MISA软件搜索马铃薯基因组序列中的SSR位点,设计SSR引物,以6个彩色马铃薯F₂代分离单株及亲本为材料筛选出多态性好的SSR适宜引物。同时结合NCBI上公布的SSR引物,利用BSA与SSR标记相结合的方法,开发与花青素含量紧密连锁的SSR标记,并通过彩色马铃薯F₂代分离群体对开发的标记进行验证。为今后深入开展彩色马铃薯花青素含量相关基因的挖掘和分子标记辅助育种等研究奠定了基础。主要研究结果如下:1.利用MISA软件对马铃薯基因组序列进行SSR位点搜索,共检索到218,997个SSR位点,平均密度为295.30 SSR·Mb^-1,对全基因组SSR序列特征分析得知,单核苷酸重复类型最多,达62.05%。基因组中含有215种基元类型,SSR基序长度主要分布在12～20 bp,呈现出中等多态性。2.基于检索到的SSR位点成功设计出100个引物对。利用彩色马铃薯双亲的基因组DNA进行SSR-PCR扩增,初步筛选出48对可以扩增出特异条带的引物,有效扩增率达48%。进一步以彩色马铃薯F₂代分离群体中随机选择的6个单株进行扩增,共筛选获得条带清晰稳定,多态性较高的SSR适宜引物26对,平均多态性比率达72.52%。3.根据自主设计和公开发表的共56对SSR引物,利用BSA与SSR标记相结合的方法,筛选到与花青素含量相关的3个SSR标记,经过F₂代分离群体的验证,3个SSR标记均与花青素含量极显著相关,可用于高花青素含量彩色马铃薯新品系的分子标记辅助选育。更多还原

【Abstract】 The anthocyanin content of colored potato is an important quality trait.Aiming at the relatively few number of molecular markers related to anthocyanin content of tetraploid colored potato,this study used MISA software to search for SSR loci in potato genome sequences,designed SSR primers,and screened out suitable SSR primers with good polymorphism by six colored potato F₂individuals and parents.At the same time,combined with the SSR primers published on the NCBI,the SSR markers closely linked to the anthocyanin content were developed by combining BSA with SSR markers,and which were then verified by the F₂population of colored potato.It laid the foundation for the study of the future research on the mining of genes related to anthocyanin content in colored potato and the marker-assisted selection.The main results were as follows:1.A total of 218,997 SSR loci with an average density of 295.30 SSR·Mb^-1were retrieved by MISA software,and the characteristics of the whole genome SSR sequence were analyzed.It showed that the mononucleotide was the most,up to 62.05%.The genome contained 215 motif types,and the length of SSR motifs ranged from 12 bp to 20bp,with medium polymorphism.2.Based on the retrieved SSR loci,a total of 100 primers were successfully designed.The SSR-PCR was used for the genomic DNA of parents of colored potato,and 48 pairs of primers that could amplify specific bands were preliminarily screened,with an effective amplification rate of 48%.Then six randomly selected F₂individuals of colored potato were amplified,and 26 suitable SSR primers with clear and stable bands and high polymorphism were obtained,with the average polymorphism ratio of 72.52%.3.Based on the self-designed and published 56 SSR primers,three SSR markers related to anthocyanin content were screened using the method of combining BSA and SSR markers.After verification by F₂isolated population,all three SSR markers were significantly correlated with anthocyanin content,which can be used for marker-assisted selection of new varieties of colored potato with high anthocyanin content.更多还原