38个欧美草莓栽培品种SSR指纹图谱构建.docVIP

38个欧美草莓栽培品种SSR指纹图谱的构建 　　摘 要：为了建立草莓品种指纹图谱数据库，从而对不同草莓品种进行分子鉴定，以早光、因都卡、达赛莱克特、森加拉等38个欧美草莓栽培品种为试材，利用8％非变性聚丙烯酰胺凝胶电泳进行扩增产物检测。通过对各种影响因素不同浓度梯度的比较，得出优化的SSR扩增反应体系，25 μL PCR的反应体系中各成分为：1×Buffer；2.5 mmol?L-1 MgCl2；0.4 μmol?L-1 引物；1.5 U Taq酶；0.2 mmol?L-1 dNTP；60 ng DNA。从44对引物中筛选出带型清晰、多态性丰富的10条SSR引物。最终确定了4对SSR引物作为核心引物对38个草莓品种进行了分子图谱的构建，29个草莓品种可以完全与其他品种区分开。 　　关键词： 草莓； 指纹图谱； 分子标记； SSR 　　中图分类号：Ｓ６６8．4 文献标识码：Ａ 文章编号：１００９－９９８０?穴２０11?雪０6－1032－０6 　　 　　Establishment of SSR fingerprinting database for 38 cultivars of strawberry (Fragaria ananassa) native to Europe and America 　　WANG Zhuang-wei，ZHAO Mi-zhen， YUAN Ji，WU Wei-min， QIAN Ya-ming 　　（Institute of Horticulture， Jiangsu Academy of Agricultural Sciences， Nanjing， Jiangsu 210014 China） 　　Abstract: The total DNA of 38 strawberry cultivars originating in Europe and America ，such as Earliglow， Induka， Darselect ，Senga， Litessa， were amplified by 44 pairs of SSR primers. The PCR products were separated in 8% polyacrylamide sequencing gels. Some factors which affected amplified products，such as dNTPs，Taq DNA polymerase，primer and so on were also studied. Optimum condition was as follows：25 μL PCR reaction system consisted of 1×PCR Buffer， 2.5 mmol?L-1 MgCl2， 0.4 μmol primer， 0.2 mmol dNTP， 1.5 U Taq polymerase and 60 ng DNA. The amplification conditions were 94 ℃ for 1 min，35 cycles of 94 ℃ for 30 s，annealing temperature for 30 s ，72 ℃ for 30 s and a final extension step at 72 ℃ for 7 min． Ten pairs of primers which could amplify stable and distinct band were selected. The electrophoresis results of the 38 strawberry DNA samples were evaluated and analyzed. Eventually， 29 cultivars could be distinguished by the fingerprint map constructed by only 4 pairs of SSR primers． 　　Key words: Strawberry； Fingerprinting； Molecular marker； SSR 　　草莓在世界上广泛种植，是重要的经济果树。全世界草莓属约20个种，2 000多个品种。草莓通过匍匐茎营养繁殖，容易造成种质资源圃或育苗地品种混杂。长期以来，人们主要根据形态特征来鉴定区分草莓品种。草莓育种往往集中利用少数优良亲本，遗传基础狭窄，栽培草莓多为八倍体，众多农艺性状为数量性状，且草莓的植物学性状易受环境、栽培、气候等影响，传统的形态学方法常常难以区分相近的草莓品种[1-2]，品种的纯度及品种