安调过 李立会. Application of molecular marker techniques in rye research

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1 Chinese Journal of Eco-Agriculture, March 2011, 19(2): DOI: /SP.J * 尹冬冬 1,3 安调过 1** 李立会 2 许红星 1 ( ; ; ) 黑麦 (Secale cereal L.) 作为小麦的近缘植物, 是改良小麦抗病性 产量和品质等性状的重要基因源 分子标记技术作为分子生物学研究中极具价值的一种研究工具已被广泛用于黑麦研究 本文论述了目前分子标记技术在黑麦遗传连锁图谱构建 有益基因定位和黑麦特异性分子标记开发应用等方面的研究进展, 并分析了该技术在黑麦研究中的应用前景 黑麦分子标记技术简单重复序列分子标记辅助选择 : S512.5 : A : (2011) Application of molecular marker techniques in rye research YIN Dong-Dong 1,3, AN Diao-Guo 1, LI Li-Hui 2, XU Hong-Xing 1 (1. Center for Agricultural Resources Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Shijiazhuang , China; 2. Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing , China; 3. Graduate University of Chinese Academy of Sciences, Beijing , China) Abstract Rye (Secale cereal L.) is an important alien gene resources in wheat (Triticum aestivum L.) genetic research as it is a related species of Triticinae. It has been used in improving disease resistance, yield and grain quality of wheat. Molecular marker technique (a valuable tool in molecular biological research) has been used extensively in rye research. This review summarized the progress in research regarding the applications of molecular marker techniques in rye. Such techniques included genetic linkage map construction, valuable gene identification and mapping, and specific rye genome marker development and application. The application prospects of molecular marker techniques in rye research were also discussed. Key words Secale cereal L., Molecular marker technique, Simple sequence repeat, Marker-assisted selection (Received May 4, 2010; accepted July 26, 2010) (Secale cereal L.) (Gramineae) (Triticeae) (Triticinae), (Tertiary gene pool) [1],,,, (Blumeria graminis f sp tritici) (Puccinia graminis f sp tritici) (Puccinia triticina Eriks.) (Puccinia graminis Pers. f. sp. tritici) (Tilletia controversa Kuhn, TCK) (Barley yellow dwarf virus, BYDV), [2 7], DNA 20 70,,, : (1) ; (2) ; (3) ; (4) ; (5) ; (6) * (2006BAD13B02) (863 ) (2011AA100103) ** : 安调过 (1965~), 女, 博士, 研究员, 研究方向为作物种质创新 优异基因的发掘和聚合育种 andiaoguo@163.com 尹冬冬 (1985~), 男, 硕士研究生, 研究方向为小麦遗传改良与种质创新 yindongdong2003@163.com : :

2 (Marker-assisted selection, MAS) 1 DNA 分子标记技术的类别与发展 DNA : (1)DNA, (Restriction fragment length polymorphisms, RFLPs); (2)PCR(Polymerase chain reaction), DNA(Random amplified polymorphic DNAs, RAPDs) (Amplified fragment length polymorphisms, AFLPs) (Microsatellites or simple sequence repeats, SSRs); (3) (Plant retrotransposons); (4) (Single nucleotide polymorphisms, SNPs) [8 9], 4 (Sequence tagged sites, STSs) (Inter simple sequence repeats, ISSRs) (Selective amplification of microsatellite polymorphic loci, SAMPL) (Sequencetagged microsatellites, STMs) EST-SSRs (Expressed sequence tags-simple sequence repeats) EST-SNPs (Expressed sequence tags-single nucleotide polymorphisms) [10] SSR 1.1 SSR (Repetitive sequences), : (1) (Simple sequence repeats, SSR) (Microsatellite), (CAG)n; (2) (Complex sequence repeats, CSR), (Transposable elements, TE); (3) (Mathematically defined repeats, MDR) SSR (Short tandem repeat, STR) SSR, CA/GT DNA,,,, SSR PCR, SSR : (1), ; (2), ; (3) ; (4), PCR, DNA SSR, RFLP, RAPD SSR DNA, DNA, SSR, [11 13] 1.2 EST-SSR (Expressed sequence tag, EST) EST-SSR 150~500 bp (cdna) Adams [14] 1991 (EST ) EST 1991 EST 2 000, (National Center for Biotechnology Information, NCBI)EST EST EST EST EST mrna,, EST [15] SSR(Genome-SSR, G-SSR), EST ; EST-SSR EST DNA,, EST-SSR G-SSR [16] SSR,, EST-SSR, EST-SSR Varshney [16] 165 EST-SSR 78.2% 75.2% 42.4%

3 2 : 479 Thiel [17] 311 EST-SSR, 80%, 60%, 40% EST-SSR,, EST 2 分子标记技术在黑麦研究中的具体应用 2.1 RFLP RAPD SSR, [18 30] Masojć [24] 1 RFLP 282, cm, 139 RFLP 69 RAPD 13 SSR Milczarski [26] 99 RAPD 18 SSR 14 STS 9 SCAR 7 ISSR, cm ; Masojć [24], 611, 70~109, cm, 3.1 cm [25], Khlestkina [25] 39 EST-SSR 60 WMS, SSR 1R 2R 3R 4R 5R 6R 7R,, 44 Plaschke [27] RFLP 29, 129 cm 5R (Consensus linkage map) BÖrner [28] 12 RFLP, 374 RFLP 24 15, 413 ; 15 5, Stojałowski [29] 6R, 1.3 cm, cm Gustafson [30] JoinMap ( UC90 E-line, P87 P105, I0.1-line I0.1-line, E-line R-line Ds2 RxL10 ), cm 1.6 cm 5, 2.2, 1R 7R [31 32] ;, (Pm8 Pm17) (Yr9) (Lr26) (Sr31) (Ir) (Vr) 1RS Mohler [33] 1 RFLP STS, Pm8 Pm17 Mago [34 35] (Hordeum vulgare)1hs (Aegilop tauschii)1ds RFLP 1RS Lr26 Sr31 Yr9; 3, cm Wehling [36] SSR STS, 2 Pr1 Pr2 6RL 7RL Anderson [37] 1A 1B 1D 1H 1R 111 RFLP, Dn7 1R, Xbcd1434 XksuD cm 7.4 cm, Gallego [38 39] 22 RAPD 11, 13 3R 4R 6R ; 2 6RS Alt1 SCAR ScR ScB , 2 Alt1 2.1 cm 5.5 cm, MAS Miftahudin [40 41] RFLP Alt3 4R, 2 Alt3 0.4 cm 0.7 cm Alt cm, BÖrner [28] RFLP (Ct2 Sp1) Korzun [42]

4 RFLP Ddw1 Hp 5R, Xwg199 Ddw1 5.6 cm, Ddw1 5AL Rht12 ; Tenhola-Roininen [43] Ddw1 67.8% QTL WMS6 REMS1218com, REMS1218com 13 cm Stracke [44] RAPD AFLP Rfp1 Rfp2, SCAR, 2.9 cm 5.2 cm Malyshev [45] SSR 2 sy1 sy9; sy1 2RL, Xscm43 Xgwm132 ; sy9 7RL, 2 Xrems1188 Xrems cm 0.1 cm Dobrovolskaya [46] G-SSR EST-SSR 2 (Multi row spike, MRS) ; Mrs1 Xgwm988 Xgwm484; Mol Xrms5b Xcfe209, 15.7 cm 10.7 cm;, MAS 2.3,,,,,,,, [47 48], , Sharp [49] RFLP Katto [50] psc20h F3 R3 [51] AF BL/1RS, [52] 50 SSR 13 DNA, 3 gwm232, gwm260 gwm644, 2.3.2,,, Koebner [53] rrna, 1R NOR-R1, 1RS [54] NOR-R1 1R, NOR-R1 1R [55] RAPD, SCAR PSCM 1082, 6R [56 57] 1 EST 5 1R EST-STS ; 2 EST-STS 1RS, 1RS Kofler [58] BAC 74 1RS [59] 7 88 SSR 9 gwm232, 6R, 6R Zhuang [60] EST 81 EST-SSR, 30, 8 1R 4R 5R R7, 7 Lee [61] 2 EST 114, 2BS/2RL 6 2RL, RT-PCR(Real time PCR), 1 Wang [2] 1RS EST-STS,

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