蓝莓优良杂交品系父本鉴定及DNA指纹图谱构建

Male Parent Identification of Blueberry Excellent Hybridization Strains and Construction of DNA Fingerprint

自然杂交实生苗选育是获得蓝莓优良品系或新品种的重要手段之一,但是父本未知是该种选育方法的最大缺点,所以利用分子生物学技术从授粉群体中鉴定其父本来源并建立相应的DNA指纹图谱,对于蓝莓优良品系或新品种的利用与保护具有重要价值。本研究利用EST-SSR分子标记技术对优良品系的父本进行了鉴定,并构建了相应的DNA指纹图谱。结果表明,从40对EST-SSR引物中筛选出10对条带清晰、多态性较高的引物组合,其中检测出等位基因23个,且每个位点的等位基因数在2~3之间,平均每个位点包含2.3个等位基因;另外,平均Shannon多样性指数为0.700 3。基于遗传相似系数,利用(unweighted pair-group method with arithmetic means,UPGMA)非加权类平均法对8个自然杂交蓝莓优良品系与其待选父本的亲缘关系进行了分析,明确了父本来源并获得杂交系谱:即SMN-72(爱国者×斯巴坦);SMN-86(日出×艾克塔);SMN-234(蓝丰×迪克西);SMN-291(爱国者×大粒蓝金);SMN-339(爱国者×斯巴坦);SMN-350(陶柔×塞拉);SMN-460(柳叶×陶柔);SMN-461(晚蓝×蓝鸟)。另外,还利用CA94和CA231两对引物构建了8个自然杂交蓝莓优良品系的DNA指纹图谱,为保护该优良品系提供科学依据。

Breeding of natural hybrid seedlings is one of the important ways to obtain excellent blueberry strains or new varieties, but the unknown male is the biggest drawback of breeding method, so the use of molecular biology techniques to identify the paternal origin from pollination groups and the establishment of DNA fingerprint have the great value for the utilization and protection of blueberry clones or new varieties. In this study, the male parent of fine strain was identified by EST-SSR molecular marker, and the corresponding DNA fingerprinting was constructed. The results showed that 10 pairs of primer combinations with clear bands and high polymorphism were screened out from 40 pairs of EST-SSR primers, the amount of each primer alleles ranged from 2 to 3, and 10 primers were detected 2.3 alleles in total. In addition, Shannon diversity index was 0.700 3 on average. Based on the genetic similarity coefficient, UPGMA was used to identify the genetic relationship of 8 blueberry excellent natural hybridization strains and their male parents. SMN-72 （PatriotxSpartan）, SMN-86 （SunrisexEchota）, SMN-339 （Patriot×Spartan）, SMN-350 （Toro×Sierra）, The hybridization pedigree results showed as follows： SMN-234 （BluecropxDixi）, SMN-291 （Patriot×Bluegold）, SMN-460 （Willow×Yoro）, SMN-461 （Lateblue×Bluejay）. Furthermore, DNA fingerprinting of 8 natural hybrid blueberry strains was constructed by using two primers of CA94 and CA231, which provided a scientific basis for the protection of the excellent strains.