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作物学报 ›› 2015, Vol. 41 ›› Issue (07): 1039-1046.doi: 10.3724/SP.J.1006.2015.01039

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

青海大黄油菜Brsc1基因的精细定位及图谱整合

李新**,肖麓**,杜德志*   

  1. 青海大学农林科学院春油菜研究所 / 青海省春油菜遗传改良重点实验室 / 青海省高原作物种质资源创新与利用国家重点实验室培育基地，青海西宁，810016
  • 收稿日期:2014-12-04 修回日期:2015-04-02 出版日期:2015-07-12 网络出版日期:2015-04-14
  • 通讯作者: 杜德志, E-mail: qhurape@126.com, Tel: 0971-5366520
  • 基金资助:

    本研究由国家高技术研究发展计划(863计划)项目(2011AA10A104)，国家重点基础研究发展计划(973计划)项目(2012CB723007)，国家现代农业产业技术体系建设专项(CARS-13)和国家自然科学基金项目(31060196)资助。

Fine Mapping and Map Integration of Brsc1 Gene in Dahuang Rape (Brassica rapa L.)

LI Xin**,XIAO Lu**,DU De-Zhi*   

  1. Province for Spring Rapeseed Genetic Improvement / National Key Laboratory Breeding Base of Qinghai Province for Innovation and Utilization of Plateau Crop Germplasm, Xining 810016, China
  • Received:2014-12-04 Revised:2015-04-02 Published:2015-07-12 Published online:2015-04-14
  • Contact: 杜德志, E-mail: qhurape@126.com, Tel: 0971-5366520

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摘要：

大黄油菜是源于青海湟源的地方品种，种皮颜色鲜黄。研究表明，大黄油菜的黄籽性状受1对隐性基因(Brsc1)控制，该基因被定位于白菜A9染色体上一段1.7 Mb的区间内。为了更好地利用这一黄籽资源，我们对Brsc1基因进一步精细定位。利用青海大黄油菜和褐籽白菜型油菜09A-126构建BC4及F2分离群体。利用白菜同源区段内已公布的SSR标记，同时利用该区段序列信息开发新的SSR引物，共获得6个与目标基因紧密连锁的标记(BrID10711、BrA5-BrA9)，其中BrA5与目标基因共分离，BrA9为一侧最近标记，它与目标基因之间的遗传图距为0.69 cM。至此，Brsc1基因进一步被限定于标记Y06和BrA9之间约1.2 Mb的区间内。利用本研究中获得的标记检测F2群体中3种类型单株，鉴定出一个共显性标记BrA8。将本研究中获得的SSR标记与前人研究结果进行整合，加密了Brsc1基因所在区间的标记密度。同时，特异片段与拟南芥基因组进行序列比对的结果表明，共有5个标记与拟南芥的第1染色体有较好的共线性关系，暗示Brsc1基因的同源基因可能位于拟南芥的第一染色体上。本研究中获得的标记将为Brsc1基因的克隆及利用Brsc1基因进行黄籽油菜的分子辅助育种提供有利条件。

关键词: Brassica rapa L., Yellow-seeded, SSR marker, Genetic map, Physical map

Abstract:

Dahuang rape, a landrace originated from Qinghai Huangyuan, has bright yellow seed coat. Previous studies indicated that the yellow-seeded trait in Dahuang was controlled by a recessive gene (Brsc1), which was located in a 1.7 Mb interval on chromosome A9 of Brassica rapa (B. rapa). In order to better use the yellow-seeded resource, we further fine mapped the Brsc1 gene. BC4 and F2 populations, constructed from the cross of Dahuang and 09A-126 (brown-seeded, B. rapa), were used for fine mapping. New PCR markers were developed based on the information of the homologous region in B. rapa and the published SSR markers were used in polymorphism survey. A total of six markers (BrID10711, BrA5-BrA9) tightly linked to the target gene were obtained, wherein BrA5 co-segregated with Brsc1, and BrA9 was 0.69 cM away from the Brsc1 gene. So Brsc1 was further limited in a 1.2 Mb interval, approximately. The markers identified in this study were used to detect three types of individual plants in F2 population. BrA8 was identified as a co-dominant marker. Marker density of the region encompassing the Brsc1 gene was increased by integrating the markers from previous researches and this study. Sequence alignment with the genome of Arabidopsis thaliana was performed and a good homologous linear relationship between five markers and chromosome 1 of Arabidopsis thaliana was detected, which implied that homologous gene of the Brsc1 gene maybe located in the chromosome 1 of Arabidopsis thaliana. Markers obtained from this study would provide favorable conditions for map-based cloning of the Brsc1 and molecular marker-assisted selection (MAS) breeding of the yellow-seeded rape.

Key words: Brassica rapa L., Yellow-seeded, SSR marker, Genetic map, Physical map

[1]刘后利. 油菜遗传育种学. 北京: 中国农业大学出版社, 2000. pp 215–225



Liu H L. Rapeseed Genetics and Breeding. Beijing: Chinese Agricultural University Press, 2000. pp 215–225 (in Chinese)



[2]高永同. 黄籽油菜的遗传和育种研究进展. 中国油料, 1984, (3): 89–93



Gao Y T. Research progress of genetic and breeding on yellow-seeded rape. ZHONGGUOYOULIAO, 1984, (3): 89–93 (in Chinese)



[3]陈功华, 田森林, 刘淑艳. 黄籽油菜的育种及推广. 作物研究, 2005, 19(2): 126–128



Chen G H, Tian S L, Liu S Y. Breeding and promotion of yellow-seeded rape. Crop Res, 2005, 19(2): 126–128 (in Chinese)



[4]刘显军, 袁谋志, 官春云, 陈社员, 刘淑艳, 刘忠松. 芥菜型油菜黄籽性状的遗传、基因定位和起源探讨. 作物学报, 2009, 35: 839–847



Liu X J, Yuan M Z, Guan C Y, Chen S Y, Liu S Y, Liu Z S. Inheritance, mapping, and origin of yellow-seeded trait in Brassica juncea. Acta Agron Sin, 2009, 35: 839–847 (in Chinese with English abstract)



[5]刘后利. 油菜的遗传和育种. 上海: 科学技术出版社, 1985. pp 9–23



Liu H L. Genetic and Breeding of Rapeseed. Shanghai: Shanghai Scientific and Technical Publishers, 1985. pp 9–23 (in Chinese)



[6]何余堂, 涂金星, 傅廷栋, 李殿荣, 陈宝元. 中国白菜型油菜种质资源的遗传多样性研究. 作物学报, 2002, 28: 697–703



He Y T, Tu J X, Fu T D, Li D R, Chen B Y. genetic diversity of germplasm resources of Brassica campestris L. in China by RAPD markers. Acta Agron Sin, 2002, 28: 697–703 (in Chinese with English abstract)



[7]何余堂, 龙卫华, 胡进平, 傅廷栋, 李殿荣, 陈宝元, 涂金星. 白菜型油菜角果多室性状的遗传及解剖学研究. 中国油料作物学报, 2003, 25(1): 1–4



He Y T, Long W H, Hu J P, Fu T D, Li D R, Chen B Y, Tu J X. Anatomic and genetic studies on multicapsular character in Brassica campestris L. Chin J Oil Crop Sci, 2003, 25(1): 1–4 (in Chinese with English abstract)



[8]李霞. 人工合成甘蓝型黄籽油菜粒色基因的精细定位. 华中农业大学硕士学位论文, 湖北武汉, 2009. p 21



Li X. Fine mapping of seed color gene in the resynthesized yellow-seed Barssica napus. MS Thesis of Huazhong Agricultural University, Wuhan, China, 2009. p 21 (in Chinese with English abstract)



[9]Li X, Chen L, Hong M Y, Zhang Y, Zu F, Wen J, Yi B, Ma C Z, Shen J X, Tu J X, Fu T D. A Large Insertion in bHLH Transcription Factor BrTT8 Resulting in Yellow Seed Coat in Brassica rapa. PloS One, 2012, 7: e44145



[10]Mohammad A, Irka S M, Aziz M A. Inheritance of seed color in some Brassica oleiferous. Ind J Genet Breed, 1942, 2: 112–127



[11]Rahman M, McVetty P B, Li G. Development of SRAP, SNP and multiplexed SCAR molecular markers for the major seed coat color gene in Brassica rapa L. Theor Appl Genet, 2007, 115: 1101–1107



[12]Schwetka A. Inheritance of seed colour in turnip rape (Brassica campestris L.). Theor Appl Genet, 1982, 62: 161–169



[13]李正强. 白菜型油菜黄籽遗传研究. 种子, 1992, (6): 13–16



Li Z Q. A Study on Inheritance of Yellow Seed in B. campestris. Seed, 1992, (6): 13–16 (in Chinese with English abstract)



[14]Chen B Y, Heneen W K, Jonsson R. Resynthesis of Brassica napus L. through interspecific hybridization between B. alboglabra bailey and B. campestris L. with special emphasis on seed color. Plant Breed, 1988, 101: 52–59



[15]Kebede B, Cheema K, Greenshields D L, Li C, Selvaraj G, Rahman H. Construction of genetic linkage map and mapping of QTL for seed color in Brassica rapa. Genome, 2012, 12: 813–823



[16]Xiao L, Zhao Z, Du D Z, Yao Y M, Zhao L. Genetic characterization and fine mapping of a yellow-seeded gene in Dahuang (Brassica rapa landrace). Theor Appl Genet, 2012, 124: 903–909



[17]Zhang J F, Ying L, Yuan Y X, Zhang X W, Geng J F, Chen Y, Cloutier S, McVetty P B E, Li G Y. Map-based cloning and characterization of a gene controlling hairiness and seed coat color traits in Brassica rapa. Plant Mol Biol, 2009, 69: 53–63



[18]罗玉秀, 杜德志. 青海大黄油菜主要农艺性状研究. 西北农业学报, 2007, 16(1): 136–139



Luo Y X, Du D Z. Research on desirable traits of Qinghai Dahuang (Brassica rapa L.). J Northwest Agric, 2007, 16(1): 136–139 (in Chinese with English abstract)



[19]罗玉秀. 青海白菜型大黄油菜优良特性的研究. 青海大学硕士学位论文, 青海西宁, 2007. p 2



Luo Y X. Study on Good Properties of Qinghai Dahuang B. campestris. MS Thesis of Qinghai University, Xining, China, 2007. p 2 (in Chinese with English abstract)



[20]赵会彦, 肖麓, 赵志, 杜德志. 青海大黄油菜粒色性状分子标记的开发和图谱整合. 作物学报, 2014, 40: 965–972



Zhao H Y, Xiao L, Zhao Z, Du D Z. Development of molecular markers and map integration for seed color traits in Dahuang rape (Brassica rapa L.). Acta Agron Sin, 2014, 40: 965–972 (in Chinese with English abstract)



[21]严明理, 刘忠松, 官春云, 陈社员. 芥菜型油菜种皮颜色的遗传和SCAR标记. 云南农业大学学报, 2006, 21: 100–106



Yan M L, Liu Z S, Guan C Y, Chen S Y. Inheritance and SCAR Marker for the Seed Coat Color in Brassica juncea. J Yunnan Agric Univ, 2006, 21: 100–106 (in Chinese with English abstract)



[22]Padmaja K L, Arumugam N, Gupta V, Mukhopadhyay A, Sodhi Y S, Pental D, Pradhan A K. Mapping and tagging of seed coat color and the identification of microsatellite markers for marker-assisted manipulation of the trait in Brassica juncea. Theor Appl Genet, 2005, 111: 8–14



[23]Doyle J J. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochem Bull, 1987, 19: 11–15



[24]陆光远, 杨光圣, 傅廷栋. 一个简便的适合于分析油菜中SSR位点的检测体系. 中国油料作物学报, 2003, 25(3): 79–81



Lu G Y, Yang G S, Fu T D. An effective SSR detection system in rapeseed. Chin J Oil Crop Sci, 2003, 25(3): 79–81 (in Chinese with English abstract)



[25]梁宏伟, 王长忠, 李忠, 罗相忠, 邹桂伟. 聚丙烯酰胺凝胶快速、高效银染方法的建立. 遗传, 2008, 10: 1379–1382



Liang H W, Wang C Z, Li Z, Luo X Z, Zou G W. Improvement of the silver-stained technique of polyacrylamide gel electrophoresis. Hereditas (Beijing), 2008, 10: 1379–1382 (in Chinese with English abstract)



[26]Lincoln S, Daly M, Lander E. Mapping genetic mapping with MAPMAKER/EXP3.0. Cambridge: Whitehead Institute Technical Report, 1992



[27]Kosambi D D. The estimation of map distances from recombination values. Ann Human Genet, 1943, 12: 172–175



[28]刘仁虎, 孟金陵. MapDraw, 在Excel中绘制遗传连锁图的宏. 遗传, 2003, 25: 317–321



Liu R H, Meng J L. MapDraw: A microsoft excel macro for drawing genetic linkage maps based on given genetic linkage data. Hereditas (Beijing), 2003, 25: 317–321 (in Chinese with English abstract)



[29]路小春. 甘蓝型油菜编码WIP-锌指蛋白的TT1基因家族的克隆及在黄、黑籽之间的差异表达. 云南农业大学硕士学位论文, 云南昆明, 2006. pp 33–37



Lu X C. Molecular Cloning of Transparent Testa1 (BnTT1) Gene Family Eoconding WIP-Zinc Finger Proteins and Differential Expression between Yellow- and Black-Seeded Lines of Brassica napus. MS Thesis of Yunnan Agricultural University, Kunming, China, 2006. pp 33–37 (in Chinese with English abstract)



[30]Wei Y L, Li J N, Lu J, Tang Z L, Pu D C, Chai Y R. Molecular cloning of Brassica napus TRANSPARENT TESTA 2 gene family encoding potential MYB regulatory proteins of proanthocyanidin biosynthesis. Mol Biol Rep, 2006, 34: 105–120



[31]Fu F Y, Liu L Z, Chai Y R, Chen L, Yang T, Jin M Y, Ma A F, Yan X Y, Zhang Z S, Li J N. Localization of QTLs for seed color using recombinant inbred lines of Brassica napus in different environments. Genome, 2007, 50: 840–854



[32]Chai Y R, Lei B, Huang H L, Li J N, Yin J M, Tang Z L, Chen L. TRANSPARENT TESTA 12 genes from Brassica napus and parental species: cloning, evolution, and differential involvement in yellow seed trait. Mol Genet Genom, 2008, 281: 109–123



[33]李运涛, 李加纳, 柴友荣, 杨成军, 雷波. 白菜型油菜DFR基因的克隆与序列分析. 分子植物育种, 2005, 3: 485–492



Li Y T, Li J N, Chai Y R, Yang C J, Lei B. Cloning and sequence analysis of a DFR gene from Brassica campestris L. var. oleifera DC. Mol Plant Breed, 2005, 3: 485–492 (in English with English abstract)
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