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作 者 ：王春梅;冯祎高;庄丽芳;曹亚萍;亓增军;别同德;曹爱忠;陈佩度

期 刊 ：作物学报 2007年 33卷 11期 页码：1741-1747

关键词：染色体特异分子标记；表达序列标签；位点标签序列；黑麦(Secalecereale)；簇毛麦(Haynaldiav；

Keywords:Chromosomespecificmarker, Expressedsequencetag(EST), Sitetaggedsequence(STS), Secalecereale, Haynaldiavillosa, Roegneriakamoji, TriticumaestivumL.,

摘 要 ：为筛选普通小麦近缘物种黑麦1R、簇毛麦1V及鹅观草1Rk^#1染色体特异分子标记，根据已定位于普通小麦第一部分同源群的EST序列设计104对STS引物，对中国春、鹅观草、黑麦及簇毛麦进行多态性分析。在104对STS引物中，有53对在对照普通小麦中国春与鹅观草、黑麦及簇毛麦之间存在多态性。利用普通小麦-黑麦1R~7R二体异附加系筛选出5对黑麦1R染色体特异标记，分别是CINAU 19-₅₀₀、CINAU20-₉₅₀、CINAU21-₁₅₀₀、CINAU22-₃₁₀和CINAU23-₂₀₀₀；利用普通小麦-簇毛麦1V~7V二体异附加系筛选出5对簇毛麦1V染色体特异标记，分别是CINAU23-₁₇₀₀、CINAU24-₁₀₅₀、CINAU25-₁₆₅₀、CINAU26-₅₀₀和CINAU27-₆₂₀；利用鹅观草二体异附加系DA1Rk^#1、异代换系DS1Rk^#1(1A)、端体系DA1Rk^#1L、易位系T1Rk^#1S·W、长臂缺失系Del1Rk#1L筛选出5对鹅观草1Rk^#1特异标记，分别是CINAU27-₉₆₀、CINAU28-₁₃₆₀、CINAU29-₄₈₀、CINAU30-₅₆₀和CINAU31-₅₂₀。研究表明，可以利用普通小麦的EST序列设计PCR引物，转化成STS标记，筛选普通小麦近缘物种黑麦、簇毛麦及鹅观草等染色体特异的分子标记，快速检测和追踪导入普通小麦背景中的黑麦1R、簇毛麦1V及鹅观草1Rk^#1染色体或染色体片段，为深入研究普通小麦远缘杂种材料提供新的工具。

Abstract:

Many useful genes of wheat-relative species have been transferred to common wheat by chromosome addition, substation and translation and need to be identified. The comparative genomics shows obvious colinearity between wheat and its relative species. A lot of molecular makers have been developed and used to identify alien chromosomes, chromosome segments and genes. Wheat EST sequences are especially valuable as molecular markers because they are derived from gene transcripts and are more likely to be conserved among species. In the present study, in order to develop chromosome-specific markers for chromosome 1R of Secale cereale, 1V of Haynaldia villosa and 1Rk^#1 of Roegneria kamoji, 104 STS primers were designed based on EST sequences downloaded from Graingenes distributed on chromosome 1A, 1B, and 1D of Triticum aestivum using software primer5.0. Out of them, fifty-three pairs of primers amplified polymorphic bands between T. aestivum “Chinese Spring (CS)” and S. cereale, H. villosa, and R. kamoji. Five STS markers, CINAU 19-₅₀₀, CINAU20-₉₅₀, CINAU21-₁₅₀₀, CINAU22-₃₁₀ and CINAU23-₂₀₀₀ amplified specific fragments from S. cereal, T. aestivum-S. cereale amphiploid and 1R addition line, but not from CS and 2R-7R addition lines, were specific to 1R of S. cereale. Five STS markers, CINAU23-₁₇₀₀, CINAU24-₁₀₅₀, CINAU25-₁₆₅₀, CINAU26-₅₀₀ and CINAU27-₆₂₀ were specific to 1V of H. villosa because they amplified corresponding patterns from H. villosa, T. durum-H. villosa amphiploid, and 1V addition, not from CS and 2V-7V addition lines. And five STS markers, CINAU27-₉₆₀, CINAU28-₁₃₆₀, CINAU29-₄₈₀, CINAU30-₅₆₀ and CINAU31-₅₂₀ specific to 1Rk^#1 of R. kamoji were screened by amplifying specific fragments from R. kamoji, Wheat-R. kamoji DA1Rk^#1 and DS1Rk^#1 (1A). Furthermore, markers CINAU27-₉₆₀ and CINAU29-₄₈₀ were located on the short arm, CINAU28-₁₃₆₀ and CINAU31-₅₂₀ on the long arm and CINAU30-₅₆₀ close to the region of centremere of chromosome 1Rk^#1, respectively. This result indicated that STS markers derived from the EST of common wheat are useful for identifying corresponding specific chromosomes of wheat relatives.

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