小麦2B染色体新SSR标记开发

doi:10.7668/hbnxb.201751761

摘要/Abstract

摘要： 为开发用于小麦育种的新SSR分子标记，利用普通小麦中国春2B染色体25 Mb的DNA序列进行了SSR筛选，在检测到的2 852个SSR中80.40%是二核苷酸重复。二、三、四、五核苷酸重复序列分别有6，30，17，1种类型。二核苷酸中GA/CT、AG/TC、AT/TA数量最多，分别占SSR总数的20.44%，19.00%，17.15%；三核苷酸中CTT/GAA的数量最多，占SSR总数的2.56%；四、五核苷酸重复出现的频率都较低。进一步分析SSR的重复次数，发现二核苷酸重复次数总体较高，重复次数≥10的SSR数量有327个，占所有二核苷酸序列的14.26%，其中，AG/TC、AT/TA的重复次数≥10的SSR数量分别有98，97个。三核苷酸的重复次数总体偏低，重复次数≥8的SSR数量仅有62个，占所有三核苷酸序列的11.55%，其中TTC/AAG的重复次数≥8的SSR数量最多，有12个。四核苷酸中只有ACAT/TGTA、TAGA/ATCT的重复次数分别达到了16，14次，其他重复基元的重复次数主要为5~7次。根据筛选到的SSR位点共设计合成了135对SSR引物，发现有101对（74.81%）引物在小麦品系Cf5019-21和Cf5240-41扩增出清晰的DNA条带，17对引物在二者间表现出差异扩增。

关键词: 小麦, SSR, 引物, 2B染色体

Abstract: In order to develop novel SSR markers for wheat breeding, simple sequence repeat(SSR) was identified from a 25 Mb region according to the Chinese Spring 2B chromosome reference sequence. In 2 852 SSR loci identified, 80.40% belonged to dinucleotides repeat.Six, thirty, seventeen, one kinds of SSR repeats were found in dinucleotide, trinucleotide, tetranucleotide, pentanucleotide repeats, respectively. For dinucleotides, GA/CT, AG/TC, AT/TA had the highest frequencies, occupying 20.44%, 19.00%, 17.15% of all SSR loci identified, respectively; for trinucleotides, CTT/GAA had the highest frequency, occupying 2.56% of all SSR loci identified; whereas the frequencies of tetranucleotide and pentanucleotide repeats were relatively low. Furthermore, the replication numbers were higher in dinucleotide than others, and 327 SSR loci which occupied 14.26% of all dinucleotides loci identified were with ≥ 10 replications. The numbers of SSR loci with ≥ 10 replications for AG/TC and AT/TA reached 98 and 97, respectively. The replication numbers in trinucleotide were relatively low, and 62 SSR loci which occupied 11.55% of all trinucleotides loci identified were with ≥ 8 replications. The number of TTC/AAG with ≥ 8 replications was the biggest, reached 12. For tetranucleotide, the numbers in ACAT/TGTA and TAGA/ATCT reached 16 and 14, respectively, and the replication numbers in other repeats of tetranucleotide varied between 5 and 7. Based on the found SSR loci, 135 SSR primer pairs were designed and synthesized. Of these, 101(74.81%) SSR primer pairs amplified clear products in wheat lines Cf5019-21 and Cf5240-41, and 17 SSR primer pairs showed polymorphism between them.

Key words: Wheat, SSR, Primer, 2B chromosome

中图分类号:

S512.1

李小军, 徐鑫, 张自阳, 刘明久, 茹振钢. 小麦2B染色体新SSR标记开发[J]. 华北农学报, 2019, 34(3): 68-74. doi: 10.7668/hbnxb.201751761.

LI Xiaojun, XU Xin, ZHANG Ziyang, LIU Mingjiu, RU Zhengang. Development of Novel SSR Markers on Wheat Chromosome 2B[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 68-74. doi: 10.7668/hbnxb.201751761.

http://www.hbnxb.net/CN/Y2019/V34/I3/68

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