芥菜型油菜多室基因Bjmc2的精细定位

doi:10.3724/SP.J.1006.2016.01735

摘要/Abstract

摘要：

芥菜型多室油菜的产量比普通两室油菜更高，定位乃至克隆多室基因可为油菜遗传改良及解释多室角果形成机制创造条件。本研究通过验证JD11-2家系衍生群体仅在BjMc2位点上存在差异，可用于BjMc2的定位。采用AFLP结合BSA法分析BC₅和BC₆群体，筛选到1个与BjMc2连锁的AFLP标记并转化为SCAR标记SC1。基于该AFLP标记序列信息，利用白菜同源序列设计SSR引物和SCAR引物，获得11对SSR标记和1对SCAR标记。通过在芥菜型油菜BAC文库中的挑选，获得2个覆盖目标区域的单克隆，由此开发1个SSR标记。将获得的SCAR和SSR标记扫描BC₇群体，构建了两室性状基因BjMc2的遗传连锁图，两侧最近标记ZX17和BACsr96与目标基因之间的遗传距离分别为0.048 cM和0.340 cM，并定位到白菜A7 scaffold000019的946~1014 kb之间，约68 kb物理距离。

关键词: 芥菜型油菜, 多室角果, BjMc2, 分子标记

Abstract:

Multilocular plants have a higher seed yield than bilocular plants in B. juncea, mapping and cloning the multilocular gene(s) might be helpful for rapeseed genetic improvement and explanation of the multilocular trait development mechanism. This study verified backcross populations derived from JD11-2 family different at BjMc2 locus only can be used for BjMc2 mapping. Using AFLP assay and BSA method in BC₅ and BC₆ generations, one AFLP marker linked to the target gene was obtained and converted to SCAR marker (SC1). On the basic of homologous sequences of the AFLP maker in B. rapa, 11 SSR markers and one SCAR marker were identified. Through the screening in theZBjH BAC library of Brassica juncea, two BACs covered the target area were selected and one SSR marker was developed. All the developed SCAR and SSR markers were then used to detect the BC₇ population, and a linkage map for the bilocular gene BjMc2 was built. ZX17 and BACsr96, the closest flanking markers, were mapped at 0.048 cM and 0.340 cM distant from the BjMc2 gene, respectively. Bjmc2 is positioned of 68 kb between 946 kb and 1014 kb in the scaffold000019 physical map of A7 in B. rapa. This result would lay a foundation for cloning polycyetic gene Bjmc2 and selectingpolycyetic lines by marker-assisted selection.

Key words: Brassica juncea, Polycyetic silique, BjMc2, Molecular marker

王刚，张向向，徐平，吕泽文，文静，易斌，马朝芝，涂金星，傅廷栋，沈金雄*. 芥菜型油菜多室基因Bjmc2的精细定位[J]. 作物学报, 2016, 42(12): 1735-1742.

WANG Gang,ZHANG Xiang-Xiang,XU Ping,LYU Ze-Wen,WEN Jing,YI Bin,MA Chao-Zhi,TU Jin-Xing, FU Ting-Dong,SHEN Jin-Xiong*. Fine Mapping of Polycyetic Gene (Bjmc2) in Brassica juncea L.[J]. Acta Agron Sin, 2016, 42(12): 1735-1742.

参考文献

[1]宋稀, 刘凤兰, 郑普英, 张学昆, 陆光远, 付桂萍, 程勇. 高密度种植专用油菜重要农艺性状与产量的关系分析. 中国农业科学, 2010, 43: 1800–1806
Song X, Liu F L, Zheng P Y, Zhang X K, Lu G Y, Fu G P, Cheng Y. Correlation analysis between agronomic traits and yield of rapeseed (Brassica napus L.) for high-density planting. Sci Agric Sin, 2010, 43: 1800–1806 (in Chinese with English abstract)
[2]Zhu L X, Zhang D X, Fu T D, Shen J X, Wen J. Analysis of yield and disease resistance traits of new winter rapeseed varieties over the past twenty years in China. Agric Sci Tech, 2011, 12: 842–846
[3]Katiyar R K, Chamola R, Chopra V L. Tetralocular mustard, Brassica juncea: new promising variability through interspecific hybridization. Plant Breed, 1998, 117: 398–399
[4]赵洪朝, 杜德志, 刘青元, 李秀萍, 余青兰, 付忠. 芥菜型多室油菜的主要性状表现. 西北农业学报, 2003, 12(3): 62–64
Zhao H C, Du D Z, Liu Q Y, Li X P, Yu Q L, Fu Z. Performances in main characteristics of multilocular Brassica juncea L. Acta Agric Boreali-Occident Sin, 2003, 12(3): 62–64 (in Chinese with English abstract)
[5]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)
[6]朱彦涛. 甘蓝型三棱角油菜新材料的遗传与选育研究初报. 西北农业学报, 2005, 14(1): 23–26
Zhu Y T. Preliminary report on inheritance and breeding of a new triquetrum-pod rapeseed material in Brassica napus L. Acta Agric Boreali-Occident Sin, 2005, 14(1): 23–26 (in Chinese with English abstract)
[7]吕泽文, 徐平, 张向向, 文静, 易斌, 马朝芝, 涂金星, 傅廷栋, 沈金雄. 芥菜型油菜多室角果的解剖特征及遗传分析. 中国油料作物学报, 2012, 34: 461–466
Lü Z W, Xu P, Zhang X X, Wen J, Yi B, Ma C Z, Tu J X, Fu T D, Shen J X. Primary study on anatomic and genetic characteristics of multi-loculus in Brassica juncea L. Chin J Oil Crop Sci, 2012, 34: 461–466 (in Chinese with English abstract)
[8]Paritosh K, Yadava S K, Gupta V, Panjabi M P, Sodhi Y S, Pradhan A K, Pental D. RNA-seq based SNPs in some agronomically important oleiferous lines of Brassica rapa and their use for genome-wide linkage mapping and specific-region fine mapping. BMC Genom, 2013, 14(1): 1–13
[9]Fan C C, Wu Y D, Yang Q Y, Yang Y, Meng Q W, Zhang K Q, Li J G, Wang J F, Zhou Y M. A novel single-nucleotide mutation in a CLAVATA3 gene homolog controls a multilocular silique trait in Brassica rapa L. Mol Plant, 2014, 7: 1788–1792
[10]Xiao L, Zhao H Y, Zhao Z, Du D Z, Xu L, Yao Y M, Zhao Z G, Xing X R, Shang G X, Zhao H C. Genetic and physical fine mapping of a multilocular gene Bjln1 in Brassica juncea to a 208-kb region. Mol Breed, 2013, 32: 373–383
[11]吕泽文. 芥菜型油菜多室性状的遗传分析及其基因的分子标记. 华中农业大学硕士学位论文, 湖北武汉, 2011
Lü Z W. Study on Inheritance and Molecular Markers for Multilocular Trait in Brassica juncea L. MS Thesis of Huazhong Agricultural University, Wuhan, China, 2011 (in Chinese with English abstract)
[12]Xu P, Lv Z W, Zhang X X, Wang X H, Pu Y Y, Wang H M, Yi B, Wen J, Ma C Z, Tu J X, Fu T D. Identification of molecular markers linked to trilocular gene (mc1) in Brassica juncea L. Mol Breed, 2014, 33: 425–434
[13]Doyle J J, Doyle J L. Isolation of plant DNA from fresh tissue. Focus, 1990, 12: 13–15
[14]Michelmore R W, Paran I, Kesseli R V. Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proc Natl Acad Sci USA, 1991, 88: 9828–9832
[15]Negi M S, Devic M, Delseny M, Lakshmikumaran M. Identification of AFLP fragments linked to seed coat colour in Brassica juncea and conversion to a SCAR marker for rapid selection. Theor Appl Genet, 2000, 101: 146–152
[16]Yi B, Chen Y N, Lei S L, Tu J X, Fu T D. Fine mapping of the recessive genic male-sterile gene (Bnms1) in Brassica napus L. Theor Appl Genet, 2006, 113: 643–650
[17]刘显军. 芥菜型油菜黄籽基因克隆和黄籽形成机制分析. 湖南农业大学博士学位论文, 湖南长沙, 2013
Liu X J. Positional Cloning of The Gene for Seed Color and Molecular Mechanism of Yellow Seed Formation in Brassica juncea L. PhD Dissertation of Hunan Agricultural University, Changsha, China, 2013 (in Chinese with English abstract)
[18]Liu R H, Meng J L. MapDraw: a microsoft excel macro for drawing genetic linkage maps based on given genetic linkage data. Hereditas, 2003, 25: 317–321
[19]朱彦涛, 田建华. 甘蓝型油菜三棱角自交群体的角果性状. 中国油料作物学报, 2015, 37: 730–734
Zhu Y T, Tian J H. Investigation on triquetrum-pod selfing populations in Brassica napus L. Chin J Oil Crop Sci, 2015, 37: 730–734 (in Chinese with English abstract)
[20]赵洪朝, 杜德志, 刘青元, 余青兰, 王瑞生. 芥菜型油菜多室性状的遗传研究. 西北农林科技大学学报(自然科学版), 2003, 31(6): 90–92
Zhao H C, Du D Z, Liu Q Y, Yu Q L, Wang Y S. Study on multilocular heredity of Brassica juncea L. J Northwest Sci Tech Univ Agric For (Nat Sci Ed), 2003, 31(6): 90–92 (in Chinese with English abstract)
[21]Truco M J, Hu J, Sadowski J, Quiros C F. Inter- and intra-genomic homology of the Brassica genomes: implications for their origin and evolution. Theor Appl Genet, 1996, 93: 1225–1233
[22]Chalhoub B, Denoeud F, Liu S Y, Parkin I A P, Tang H B, Wang X Y, Chiquet J, Belcram H, Tong C B, Samans B Correa M, Da Silva C, Just J, Falentin C, Koh C S, Le Clainche I, Bernard M, Bento P, Noel B, Labadie K, Alberti A, Charles M, Arnaud D, Guo H, Daviaud C, Alamery S, Jabbari K, Zhao M, Edger P P, Chelaifa H, Tack D, Lassalle G, Mestiri I, Schnel N, Le Paslier M C, Fan G Y, Renault V, Bayer P E, Golicz A A, Manoli S, Lee T H, Thi V H D, Chalabi S, Hu Q, Fan C C, Tollenaere R, Lu Y, Battail C, Shen J S, Sidebottom C H D, Wang X F, Canaguier A, Chauveau A, Berard A, Deniot G, Guan M, Liu Z S, Sun F M, Lim Y P, Lyons E, Town C D, Bancroft I, Wang X W, Meng J L, Ma J X, Pires J C, King G J, Brunel D, Delourme R, Renard M, Aury J M, Adams K L, Batley J, Snowdon R J, Tost J, Edwards D, Zhou Y M, Hua W, Sharpe A G, Paterson A H, Guan C Y, Wincker P. Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome. Science, 2014, 345: 950–953
[23]Clark S E, Running M P, Meyerowitz E M. CLAVATA1, a regulator of meristem and flower development in Arabidopsis. Development, 1993, 119: 397–418
[24]Clark S E, Williams R W, Meyerowitz E M. The CLAVATA1 gene encodes a putative receptor kinase that controls shoot and floral meristem size in Arabidopsis. Cell, 1997, 89: 575–585
[25]Dievart A, Dalal M, Tax F E, Lacey A D. Huttly A, Li J M, Clark S E. CLAVATA1 dominant-negative alleles reveal functional overlap between multiple receptor kinases that regulate meristem and organ development. Plant Cell, 2003, 15: 1198–1211

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