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作物学报 ›› 2010, Vol. 36 ›› Issue (07): 1114-1125.doi: 10.3724/SP.J.1006.2010.01114

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦区试品系DUS测试的分子标记

王立新,常利芳**,李宏博,季伟,刘丽华,赵昌平*   

  1. 北京市农林科学院/北京杂交小麦工程技术研究中心,北京100097
  • 收稿日期:2010-02-03 修回日期:2010-04-12 出版日期:2010-07-12 网络出版日期:2010-04-28
  • 通讯作者: 赵昌平, E-mail: bjhwc2003@yahoo.com.cn, Tel: 010-51503968
  • 基金资助:
    本研究由北京市农业育种基础研究创新平台项目II(D08070500690801)和国家科技支撑计划项目(2006BAD01A02-4)资助。

Molecular Markers for Estimating Distinctness, Uniformity, and Stability of Wheat Lines in Regional Trials

WANG Li-Xin,CHANG Li-Fang**,LI Hong-Bo,JI Wei,LIU Li-Hua,ZHAO Chang-Ping*   

  1. Beijing Engineering and Technique Research Center for Hybrid Wheat / Beijign Academy of Agricultural and Forest Sciences, Beijing 100097, China
  • Received:2010-02-03 Revised:2010-04-12 Published:2010-07-12 Published online:2010-04-28
  • Contact: ZHAO Chang-Ping,E-mail: bjhwc2003@yahoo.com.cn, Tel: 010-51503968

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1437

被引次数

40

摘要：

为了确定测试小麦(Triticum aestivum L.)区试品系特异性、一致性、稳定性(DUS)的分子标记，采用156个来自我国不同麦区的品种对SSR、EST-SSR和AFLP-SCAR标记的1334对引物进行筛选，根据在染色体上分布均匀、多态性信息指数较高、带型清晰、不同等位变异的带型易于区分及PCR产物稳定的原则，筛选出105对小麦品种DUS测试的分子标记引物，包括63对SSR引物、21对EST-SSR引物和21对AFLP-SCAR引物，可以检测122个位点的754个等位变异，平均每条染色体上被检测位点5.8个，平均每个位点包含7.2个等位变异。根据DUS测试的需求、引物的染色体分布、PIC值大小和带型特点，将105对引物分为21对核心引物、29对一级备用引物和55对二级备用引物。核心引物分辨力较高，可以完成约80%品系的特异性检测，约95%品系的种子纯度检测和约60%品系的一致性、稳定性检测；备用引物用于确定品系DNA位点纯合率和相似品种(品系)之间的遗传相似系数，以判断DNA指纹相同或相似的品种(品系)之间的相似性和特异性，评价核心标记中具有非纯位点的品系的DNA位点纯合度，同时完成核心引物未能完成的少数品系的种子纯度检测。通过在2006—2007、2007—2008、2008—2009年度对464个冬小麦区试品系DUS测试中的应用，证明105对引物具有很好的代表性和实用性，可以完成90%以上参试品系的DUS检测。

关键词: 小麦, DUS测试, 分子标记, DNA指纹

Abstract:

To determine the molecular markers for testing the distinctness, uniformity，and stability (DUS) of wheat (Triticum aestivum L.) lines in regional trials, 145 wheat cultivars were used to screen 1 334 pairs of primers of SSR, EST-SSR, and AFLP-SCAR markers. According to the even distribution of molecular markers on 21 wheat chromosomes, the higher polymorphism information content (PIC), the clear PCR bands, the easy discrimination of different alleles, and the stable PCR amplification, 105 pairs of primers, including 63 pairs of SSR primers, 21 pairs of EST-SSR primers, and 21 pairs of AFLP-SCAR primers, were selected for wheat cultivar (lines) DUS testing. A total of 754 alleles at 122 loci can be identified with 105 pairs of primers, with 5.8 loci per chromosome and 7.2 alleles per locus on average. The three kinds of molecular markers can amplify the DNA fragments that are inside/outside of genes and repeative/non-repetitive sequences. The 105 pairs of primers were classified into 21 pairs of core primers and 84 pairs of standby primers based on their pattern definition, distribution on 21 chromosoms, and PIC value. The 21 core primers distributing on 21 chromosoms could determine the distinctness test for about 80% of total cultivars (lines), the seed purity test for 95%, and the uniformity and stability test for 60%. The 84 standby primers were used as the first grade (29 pairs) and the second grade (55 pairs) to the homozygous DNA locus ratio of lines with no-homozygous loci of coer markers and the genetic similarities or the distinctness of similar cultivars (lines) pairs revealed by the core primers. At the same time, we determined the seed purity for the other 5% of lines. These 105 pairs of primers could finish DUS test for more than 90% of lines in the winter wheat regional trials in 3 cropping seasons from 2006 to 2009. Although less than 10% of lines should be evaluated in combination with phenotypic identification in the field, the effectiveness and efficiency of the 105 pairs of primers are satisfactory in bulk screening for wheat DUS test.

Key words: Wheat, DUS, Molecular markers, DNA fingerprint


[1] Dai J(戴剑), Li H-Y(李华勇), Ding K-M(丁奎敏), Hong D-L(洪德林). Current status and prospects on DUS testing technique systerm for new plant variety protection. Seed (种子), 2007, 26(9): 44-47 (in Chinese)

[2] Liu P-W(刘平武), Zhou G-L(周国岭), Yang G-S(杨光圣), Fu T-D(傅廷栋). Fingerprints construction of hybrid parentsin brassicanapus and its utilization in hybrid purity test. Acta Agron Sin (作物学报), 2005, 31(5): 640-646 (in Chinese with English abstract)

[3] Wang J-F(王俊芳) , Yang W-H(杨伟华), Kuang M(匡猛), Xu H-X(许红霞), Wang Y-Q(王延琴), Zhou D-Y(周大云), Feng X-A(冯新爱). Fingerprints construction of cotton varieties and seed identification. China Cotton (中国棉花), 2009, 36(3): 6-24 (in Chinese)

[4] Shi H-B(石海波), Wang L-X(王立新), Li H-B(李宏博), Zhang F-T(张风廷), Ma Q(马庆), Zhao C-P(赵昌平). Division of seeds motley and SSR loci impurity in wheat cultivar using SSR markers. Mol Plant Breed (分子植物育种), 2006, 4(6): 513-519 (in Chinese with English abstract)

[5] Wang L-X(王立新), Chang L-F(常利芳), Li H-B(李宏博), Ge L-L(葛玲玲), Xin A-H(信爱华), Gao S-Q(高世庆), Ji W(季伟), Sun H(孙辉), Zhao C-P(赵昌平). Method of testing wheat seeds purity by molecular markers.J Triticeae Crops (麦类作物学报), 2009, 29(1): 1-8 (in Chinese with English abstract)

[6] Ji W(季伟), Wang L-X(王立新), Sun H(孙辉), Wang M-Y(王茅雁), Zhao C-P(赵昌平). Predigestion of wheat SSR analysis protocol. J Agric Biotechnol (农业生物技术学报), 2007, 15(5): 907-908 (in Chinese with English abstract)

[7] Wang L-X(王立新), Li H-B(李宏博), Ge L-L(葛玲玲), Sun H(孙辉), Yao J(姚骥), Qiu J(邱军), Zhao C-P(赵昌平). A new labeling method for wheat SSR parerns. J Triticeae Crops (麦类作物学报), 2006, 26(4): 164-168 (in Chinese with English abstract)

[8] Wang L-X(王立新), Li Y-F(李云伏), Chang L-F(常利芳), Huang L(黄岚), Li H-B (李宏博), Ge L-L(葛玲玲), Liu L-H(刘丽华), Yao J(姚骥), Zhao C-P(赵昌平) . Method of ID constitution for wheat cultivars. Acta Agron Sin (作物学报), 2007, 33(10): 1738-1740 (in Chinese with English abstract)

[9] Wang L-X(王立新), Ji W(季伟), Li H-B(李宏博), Ge L-L(葛玲玲), Xin H(信爱华), Wang L-X(王丽霞), Chang L-F(常利芳), Zhao C-P(赵昌平). Estimating the uniformity and stability of wheat cultivars based on the homozygous DNA Locus Ratio. Acta Agron Sin (作物学报), 2009, 35(12): 1-8 (in Chinese with English abstract)

[10] Cheng B-Y(程本义), Shi Y-F(施勇峰), Shen W-F(沈伟峰), Zhang J-Y(庄杰云), Yang S-H(杨仕华). Microsatellite marker- based analysis of rice varieties in national regional yield trial of southern china. Chin J Rice Sci (中国水稻科学), 2007, 21(1): 7-12 (in Chinese with English abstract)

[11] Zhang J-Y(张金渝), Zhang J-H(张建华), Yang X-H(杨晓洪), Jin H(金航), Mi Y-H(米艳华), Xiao Z-W(肖植文), Kong L-M(孔令明), Xiao Q(肖卿). Buliding of SSR fingerprinting map of standard varieties on maize in DUS testing. J Maize Sci (玉米科学), 2006, 14(4): 47-52 (in Chinese with English abstract)

[12] Lu G-Y(陆光远), Wu X-M(伍晓明), Zhang D-X(张冬晓), Liu F-L(刘凤兰), Chen B-Y(陈碧云), Gao G-Z(高桂珍), Xu K(许鲲). SSR-based evaluation of distinctness and uniformity of rapeseed (Brassica napus L.) varieties under Chinese national official field tests. Sci Agric Sin (中国农业科学), 2008, 41(1): 32-42 (in Chinese with English abstract)

[13] Wang F-G(王凤格), Zhao J-R(赵久然), Dai J-R(戴景瑞), Wang L(王璐), Yi H-M(易红梅), Guo J-L(郭景伦), Sun S-X(孙世贤), Liao Q(廖琴),Yang G-H(杨国航). Uniformity analysis of maize varieties by a set of SSR markers. Mol Plant Breed (分子植物育种), 2007, 5(1): 95-104 (in Chinese with English abstract)

[14] Li H-W(李宏伟), Gao L-F(高丽锋), Liu S-D(刘曙东), Jia J-Z(贾继增). Changes of gene diversity among Chinese wheat cuitivars at different decades revealed by EST-SSRs. Acta Bot Borea1-Occident Sin (西北植物学报), 2005, 25(1): 27-32 (in Chinese with English abstract)

[15] Wang L-X(王立新), Chang L-F(常利芳), Huang L(黄岚), Wang X-W(王晓维), Zhao C-P(赵昌平). Sequence polymorphism of wheat AFLP fragments and conversion of AFLP marker to SCAR marker. J Triticeae Crops (麦类作物学报), 2007, 27(6): 943-951 (in Chinese with English abstract)

[16] Wang X-W(王晓维), Wang L-X(王立新), Chang L-F(常利芳), Wei J-M(魏建民), Zhao C-P(赵昌平). Exploitation and application of AFLP-SCAR markers. J Triticeae Crops (麦类作物学报), 2008, 28(5): 738-744 (in Chinese with English abstract)

[17] Li G-Y(李根英), Dreisigacker S, Warburton M L, Xia X-C(夏先春), He Z-H(何中虎), Sun Q-X(孙其信). Development of a fingerprinting database and assembling an SSR reference kit for genetic diversity analysis of wheat. Acta Agron Sin (作物学报), 2006, 32(12): 1771-1778 (in Chinese with English abstract)

[18] Pestsova E, Ganal M W, Röder M S. Isolation and mapping of microsatellite markers specific for the D genome of bread wheat. Genome, 2000, 43: 689-697

[19] Röder M S, Korzun V, Wandehake K, Planschke J, Tixier M H, Leroy P, Ganal M W. A microsatellite map of wheat. Genetics, 1998, 149: 2007-2023

[20] Gao L-F(高丽锋). Molecular markers of SSRs and SNPs derived from ESTs and their application. Post-doctoral Work Report of Chinese Academy of Agricultural Sciences, 2003 (in Chinese)

[21] Chen H-M(陈海梅), Li L-Z(李林志), Wei X-Y(卫宪云), Li S-S(李斯深), Lei T-D(雷天东), Hu H-Z(胡海州), Wang H-G(王洪刚), Zhang X-S(张宪省). Exploitation, chromosomal location and genetic mapping of EST-SSRs in wheat. Chin Sci Bull (科学通报), 2005, 50(20): 2208-2216 (in Chinese with English abstract)

[22] Chen J-F(陈军方), Ren Z-L(任正隆), Gao L-F(高丽锋), Jia J-Z(贾继增). Developing new SSR markers from EST of wheat. Acta Agron Sin (作物学报), 2005, 31(2): 154-158 (in Chinese)

[23] Nei M, Li W, Mathematical Model for studying genetic variation in terms of restriction endonuclesses. Proc Natl Acad Sci USA, 1979, 76: 5269-5273

[24] Li Y-Q(李艳秋), Su Z-F(苏志芳), Wang L-X(王立新), Ji W(季伟), Yao J(姚骥), Zhao C-P(赵昌平). Increasing density of wheat genetic linkage map with molecular makers. Acta Agron Sin (作物学报), 2009, 35(5): 861-866 (in Chinese with English abstract)

[25] Yang X-Q(杨新泉), Liu P(刘鹏), Han Z-F(韩宗福), Ni Z-F(倪中福), Liu W-Q(刘旺清), Sun Q-X(孙其信). Comparative analysis of genetic diversity revealed by genomic-SSR, EST-SSR and pedigree in wheat (Triticum asetivum L.). Acta Geneti Sin (遗传学报), 2005, 32(4): 406-416 (in Chinese with English abstract)

[26] Chen X-H(陈先红), Xu L-Y(徐利远), Peng Z-S(彭正松), Du W-P (杜文平), Yu G-R(余桂蓉), Zhong C-S(钟昌松), Qu J-P(曲继鹏), Hu F-L(胡凤林). SSR analysis of genetic diversity for wheat cultivars (Lines) in the southwest of China. J Triticeae Crops (麦类作物学报), 2008, 28(1): 6-10 (in Chinese with English abstract)

[27] Hao C-Y(郝晨阳), Dong Y-C(董玉琛), Wang L-F(王兰芬), You G-X(游光霞), Zhang H-N(张洪娜), Ge H-M(盖红梅), Jia J-Z(贾继增), Zhang X-Y(张学勇). Genetic diversity and construction of core collection in Chinese wheat genetic resources. Chin Sci Bull (科学通报), 2008, 53(8): 1518-1526 (in Chinese)

[28] Liu L-H(刘丽华), Wang L-X(王立新), Zhao C-P(赵昌平), Yao J(姚骥), Zhang F-T(张凤廷), Zhang H(张华), Ye Z-J(叶志杰), Qin Z-L(秦志列), Zheng Y-L(郑用琏). Genetic diversity and alterations of population structure in restorers of dual cross-line hybrid wheat with thermo-photoperiod sensitive male sterile. Chin J Biochem Mol Biol (中国生物化学与分子生物学报), 2009, 25(9): 867-875 (in Chinese with English abstract)

[29] Yao J, Wang L X, Liu L H, Zhao C P, Zheng Y L. Association mapping of agronomic traits on chromosome 2A of wheat. Genetica, 2009, 137: 67-75
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