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园艺学报 ›› 2018, Vol. 45 ›› Issue (1): 139-148.doi: 10.16420/j.issn.0513-353x.2017-0281

• 研究报告 • 上一篇    下一篇

平欧杂交榛转录组中SSR信息分析和引物筛选

程云清,张丽娜,赵永斌,刘剑锋*   

  1. 吉林师范大学植物资源科学与绿色生产重点实验室,吉林四平 136000
  • 出版日期:2018-01-25 发布日期:2018-01-25

Analysis of SSR Markers Information and Primer Selection from Transcriptome Sequence of Hybrid Hazelnut Corylus heterophylla × C. avellana

CHENG Yunqing,ZHANG Lina,ZHAO Yongbin,and LIU Jianfeng*   

  1. Jilin Provincial Key Laboratory of Plant Resource Science and Green Production,Jilin Normal University,Siping,Jilin 136000,China
  • Online:2018-01-25 Published:2018-01-25

摘要: 对平欧杂交榛的转录组数据进行了深入挖掘,以期开发其SSRs标记。在25 649条含SSR序列的Unigene中,共找到35 604条SSR序列;其中,单、二、三核苷酸重复为高频类型,其数量及所占百分比分别为:10 102(28.37%)、16 543(46.46%)、7 035(19.76%);在单核苷酸重复中,A/T重复最多(28.13%);二核苷酸重复中,AG/CT(35.79%)、AT/AT(7.47%)、AC/GT(3.17%)最为丰富;三、四、五、六核苷酸重复中,比例最高的重复序列分别为:AAG/CTT(7.24%)、AAAG/CTTT(0.33%)、AAAAG/CTTTT(0.38%)、AAAAAG/CTTTTT(0.15%)。依据转录组中的SSRs序列信息,随机合成了60对引物,并对含平榛(Corylus heterophylla Fisch)、平欧杂交榛(C. heterophylla × C. avellana)、欧洲榛(C. avellana)及毛榛(C. mandshurica Maxim.)的25份材料进行了PCR扩增和毛细管电泳,找到7对多态性良好的引物,其PIC(Polymorphism Information Content)值为0.35 ~ 0.70,获得多态性条带52条;UPGMA(Unweighted Pair Group Method Analysis)聚类结果显示,上述引物可将这些种质进行准确区分。以上研究初步明确了平欧杂交榛转录组中SSR的分布特征,为其遗传连锁图谱构建等研究提供了科学依据。

关键词: 榛子, 转录组, 简单序列重复, 种质鉴定, 引物

Abstract: Deep bio-information analysis was carried out using transcriptome data in order to develop SSR marks in hybrid hazelnut Corylus heterophylla × C. avellana. In total,35 604 SSR sequences were found in 25 649 SSR containing Unigenes. In all the SSR sequences,mono-nucleotide,dinucleotide and trinucleotide repeats belonged to high frequent type,and their number and percent were 10 102(28.37%),16 543(46.46%)and 7 035(19.76%)respectively. In mono-nucleotide repeats,A/T was most abundant,accounting for 28.13% of the total SSR amount. In dinucleotide repeats,AG/CT,AT/AT,AC/GT were more abundant,accounting for 35.79%,7.47% and 3.17% of the total. In repeats of trinucleotide,tetranucleotide,pentanucleotide and hexanucleotide,the highest percent sequence of each nucleotide repeat were AAG/CTT,AAAG/CTTT,AAAAG/CTTTT,AAAAAG/CTTTTT,and they accounted for 7.24%,0.33%,0.38% and 0.15% of the total SSR amount respectively. Sixty pairs of randomly chosen SSR primers were synthesized and amplified from 25 Corylus samples,including Corylus heterophylla Fisch,C. heterophylla × C. avellana,C. avellana and C. mandshurica Maxim. Polymorphism information content(PIC)values of seven polymorphic pairs of primers varied from 0.35 to 0.70,and 52 polymorphic bands were obtained. UPGMA(Unweighted Pair Group Method Analysis)clustering results indicated that these primers could precisely separate germplasms mentioned above into different groups. This study clarifies SSR distribution characteristics of hybrid hazelnut C. heterophylla × C. avellana and provides a scientific basis for further studies of genetic linkage map construction in these species.

Key words: hazelnut, transcriptome, simple sequence repeat, germplasm identification, primer