大白菜-结球甘蓝易位系外源染色体区段基因标记的开发与应用

doi:10.7668/hbnxb.201751153

摘要/Abstract

摘要： 为了利用基因标记对大白菜-结球甘蓝易位系进行鉴定，针对大白菜和结球甘蓝共线性基因序列差异位点设计引物，以大白菜85-1和结球甘蓝11-1为试材，进行了多态性标记的筛选，并在大白菜-结球甘蓝易位系、易位系自交后代及大白菜和结球甘蓝不同商品种间进行了可利用性鉴定。结果表明，根据大白菜和结球甘蓝263个共线性基因共设计特异引物171对，66对引物在大白菜和结球甘蓝间表现出多态性扩增，所占比例为38.60%。66个基因标记中，基于大白菜基因序列获得的2个和基于结球甘蓝基因序列获得的48个，可作为结球甘蓝相对大白菜特异的基因标记，并成功用于大白菜-结球甘蓝易位系AT1-41和AT1-47及AT1-47自交后代的鉴定。进一步利用这66个基因标记在4个大白菜和4个结球甘蓝商品种中进行了PCR扩增，有52个标记能将供试的大白菜品种和结球甘蓝品种完全鉴定出来，有11个标记在部分品种中表现出多态性。结果为深入研究大白菜-结球甘蓝易位系外源基因的表达、调控、互作和标记辅助育种奠定了基础。

关键词: 大白菜, 结球甘蓝, 易位系, 基因标记

Abstract: In order to identify Chinese cabbage-cabbage translocation line by gene markers, the polymorphism primers were designed according to the nucleotide difference of collinear genes between Chinese cabbage and cabbage, and were screened with Chinese cabbage 85-1 and cabbage 11-1 as test materials. The applicability of the primers was detected using Chinese cabbage-cabbage translocation lines and their self-crossing progenies, Chinese cabbage varieties and cabbage varieties. The results showed that among 171 pairs of primers designed based on 263 collinear-gene sequences of Chinese cabbage and cabbage, 66 pairs of polymorphic primers were confirmed, the ratio being 38.60%. Among the 66 gene markers, two markers from Chinese cabbage gene sequences and 48 markers from cabbage gene sequences could be used as the specific gene markers to distinguish Chinese cabbage and cabbage, which were successfully applied to identify Chinese cabbage-cabbage translocation lines AT1-41 and AT1-47, and the self-crossing progenies of AT1-47. Furthermore, PCR amplification of the 66 gene markers was carried out in four Chinese cabbage varieties and four cabbage varieties, among which 52 markers could distinguished completely the Chinese cabbage varieties and the cabbage varieties, while other 11 markers presented the polymorphism in parts of the varieties. The results established the foundation for further studying the expression, regulation and interaction of exogenous genes in Chinese cabbage-cabbage translocation lines and for marker-assisted breeding.

Key words: Chinese cabbage, Cabbage, Translocation line, Gene marker

中图分类号:

苏建辉, 张乐强, 王汝颖, 王彦华, 张巍巍, 马聪, 轩淑欣, 申书兴. 大白菜-结球甘蓝易位系外源染色体区段基因标记的开发与应用[J]. 华北农学报, 2019, 34(3): 89-95. doi: 10.7668/hbnxb.201751153.

SU Jianhui, ZHANG Leqiang, WANG Ruying, WANG Yanhua, ZHANG Weiwei, MA Cong, XUAN Shuxin, SHEN Shuxing. Development and Application of Gene Markers in the Exogenous Chromosome Segment of Chinese Cabbage-Cabbage Translation Lines[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(3): 89-95. doi: 10.7668/hbnxb.201751153.

http://www.hbnxb.net/CN/Y2019/V34/I3/89

参考文献

[1] Qi W L,Tang Y,Zhu W,Li D Y,Diao C D,Xu L L,Zeng J,Wang Y,Fan X,Sha L A,Zhang H Q,Zheng Y L,Zhou Y H,Kang H Y. Molecular cytogenetic characterization of a new wheat-rye 1BLaEuro cent 1RS translocation line expressing superior stripe rust resistance and enhanced grain yield[J].Planta,2016,244(2):405-416.doi:10.1007/s00425-016-2517-3.
[2] Wang L,Yuan J H,Bie T D,Zhou B,Chen P D. Cytogenetic and molecular identification of three Triticum aestivum-Leymus racemosus translocation addition lines[J]. Journal of Genetics and Genomics,2009,36(6):379-385.doi:10.1016/S1673-8527(08) 60127-7.
[3] 陈雅玲,罗向东,张帆涛,戴亮芳,胡标林,谢建坤. 东乡野生稻基因渐渗系中逆转座子逆转录酶序列的克隆及表达分析[J]. 植物学报,2013,48(2):138-144.doi:10.3724/SP.J.1259.2013.00138.
Chen Y L,Luo X D,Zhang F T,Dai L F,Hu B L,Xie J K. Cloning and expression analysis of retrotransposon reverse transcriptase in introgression lines from Dongxiang wild rice[J]. Bulletin of Botany,2013,48(2):138-144.
[4] 王虹,白杨,李集临,张延明. 小麦-黑麦小片段易位系的分子细胞遗传学分析[J]. 植物遗传资源学报,2014,15(2):383-388.doi:10.13430/j.cnki.jpgr.2014.02.023.
Wang H,Bai Y,Li J L,Zhang Y M. Molecular cytogenetic analysis of wheat-rye small fragment translocation lines[J]. Journal of Plant Genetic Resources,2014,15(2):383-388.
[5] 曹立超,樊双虎,张春义,陈茹梅. 大刍草渐渗系雌穗表型统计及SNP分子标记开发[J]. 种子,2014,33(4):42-47,68.doi:10.3969/j.issn.1001-4705.2014.04.013.
Cao L C,Fan S H,Zhang C Y,Chen R M. Female spike phenotypic statistics of teosinte introgression and development of SNP marker[J]. Seed,2014,33(4):42-47,68.
[6] 孟雅宁,王彦华,顾爱侠,李晓峰,申书兴. 大白菜-结球甘蓝5号二体异附加系的选育及鉴定[J]. 中国农业科学,2010,43(14):2937-2943.doi:10.3864/j.issn.0578-1752.2010.14.012.
Meng Y N,Wang Y H,Gu A X,Li X F,Shen S X. Generation and identification of 5 Chinese cabbage-cabbage disomic alien addition lines[J]. Scientia Agricultura Sinica,2010,43(14):2937-2943.
[7] 顾爱侠,王彦华,轩淑欣,陈雪平,申书兴.结球甘蓝相对于大白菜的连锁群特异SSR标记建立[J]. 园艺学报,2009,36(8):1221-1226.doi:10.3864/j.issn.0578-1752.2010.14.012.
Gu A X,Wang Y H,Xuan S X,Chen X P,Shen S X. Establishment of specific SSR from different linkage groups of cabbage compared with Chinese cabbage[J]. Acta Horticulturae Sinica,2009,36(8):1221-1226.
[8] 耿倩倩,王彦华,轩淑欣,毛清云,赵建军,申书兴.添加甘蓝2号染色体片段的大白菜易位系的获得及其遗传稳定性分析[J]. 园艺学报,2016,43(2):261-270.doi:10.16420/j.issn.0513-353x.2015-0858.
Geng Q Q,Wang Y H,Xuan S X,Mao Q Y,Zhao J J,Shen S X. Obtaining and genetic stability of Chinese cabbage-cabbage translocation lines with fragment of cabbage chromosome 2[J]. Acta Horticulturae Sinica,2016,43(2):261-270.
[9] 闫珍臣,王彦华,轩淑欣,赵建军,申书兴.添加甘蓝8号染色体片段的大白菜易位系的获得及其遗传稳定性分析[J]. 园艺学报,2015,42(6):1085-1092.doi:10.16420/j.issn.0513-353x.2015-0127.
Yan Z C,Wang Y H,Xuan S X,Zhao J J,Shen S X. Obtaining and genetic stability of Chinese cabbage-cabbage translocation lines with fragment of cabbage chromosome 8[J]. Acta Horticulturae Sinica,2015,42(6):1085-1092.
[10] 李雪姣,王彦华,轩淑欣,赵建军,顾爱侠.大白菜相对于结球甘蓝连锁群特异SSR标记的筛选[J]. 植物遗传资源学报,2013,14(4):694-698.
Li X J,Wang Y H,Xuan S X,Zhao J J,Gu A X. Screening of specific SSR markers on different linkage groups of Chinese cabbage compared with cabbage[J]. Journal of Plant Genetic Resources,2013,14(4):694-698.
[11] 朱东旭,王彦华,赵建军,刘博,李晓峰. 轩淑欣,申书兴.结球甘蓝相对于大白菜连锁群特异InDel标记的建立及应用[J]. 园艺学报,2014,41(8):1699-1706.
Zhu D X,Wang Y H,Zhao J J,Liu B,Li X F,Xuan S X,Shen S X. Establishment and application of specific InDel markers on different linkage groups of cabbage compared with Chinese cabbage[J]. Acta Horticulturae Sinica,2014,41(8):1699-1706.
[12] 李岩宾,轩淑欣,王彦华,冯大领,赵建军,申书兴.大白菜-结球甘蓝1号染色体二体异附加系自交后代InDel标记分析和染色体数鉴定[J]. 园艺学报,2015,42(7):1329-1337.doi:10.16420/j.issn.0513-353x.2015-0106.
Li Y B,Xuan S X,Wang Y H,Feng D L,Zhao J J,Shen S X. Analysis of InDel markers and identification of chromosome number in selfing progenies of Chinese cabbage head cabbage chromosome 1 disomic alien addition line[J]. Acta Horticulturae Sinica,2015,42(7):1329-1337.
[13] 岳晓鹏. 基于甘蓝性油菜基因组重测序开发InDel标记[D]. 武汉:华中农业大学,2014.
Yue X P. Developmeng of indel markers based on whole genome resequencing in Brassica napus[D].Wuhan:Huazhong Agricultural University,2014.
[14] 张涛,杨蛟,蒋开锋,曹应江,杨莉,杨乾华,万先齐,游书梅,罗婧,高磊,李昭祥,郑家奎. 利用产量功能基因标记分析三系杂交水稻亲本的遗传多样性[J]. 中国农业科学,2014,47(1):11-23.doi:10.3864/j.issn.0578-1752.2014.01.002.
Zhang T,Yang J,Jiang K F,Cao Y J,Yang L,Yang Q H,Wan X J,You S M,Luo J,Gao L,Li Z X,Zheng J K. Genetic diversity of the main Chinese three-line hybrid rice parents based on functional genetic markers related to yield[J]. Scientia Agricultura Sinica,2014,47(1):11-23.
[15] 王军,杨杰,陈志德,仲维功. 水稻香米基因标记的开发与应用[J]. 分子植物育种,2008,6(6):1209-1212.doi:10.3969/j.issn.1672-416X.2008.06.030.
Wang J,Yang J,Chen Z D,Zhong W G. Development and application of fragrance gene markers in rice[J]. Molecular Plant Breeding,2008,6(6):1209-1212.
[16] 田志喜,严长杰,钱前,严松,谢会兰,王芳,徐洁芬,刘贵富,王永红,刘巧泉,汤述翥,李家洋,顾铭洪. 水稻淀粉合成相关基因分子标记的建立[J]. 科学通报,2010,55(26):2591-2601.doi:10.1007/s11434.
Tian Z X,Yan C J,Qian Q,Yan S,Xie H L,Wang F,Xu J F,Liu G F,Wang Y H,Liu Q Q,Shang S Z,Li J Y,Gu M H. Development of gene-tagged molecular markers for starch synthesis-related genes in rice[J]. Chinese Science Bulletin,2010,55(26):2591-2601.
[17] Yonemaru J I,Choi S H,Sakai H,Ando T,Shomura A,Yano M,Wu J Z,Fukuoka S. Genome-wide indel markers shared by diverse Asian rice cultivars compared to Japanese rice cultivar Koshihikari[J].Breeding Science,2015,65(3):249-256.doi:10.1270/jsbbs.65.249.
[18] Liu L F,Dang P M,Chen C Y.Development and utilization of InDel markers to identify peanut (Arachis hypogaea) disease resistance[J].Frontiers in Plant Science,2015,6:988.doi:10.3389/fpls.2015.00988.
[19] 侯璐,宋晓贺,路亚明,胡茂林,贺苗苗,井金学,王保通. 小簇麦易位系V9128-3抗条锈病基因的遗传分析和SSR分子标记[J]. 植物病理学报,2009,39(1):67-75.doi:10.3321/j.issn:0412-0914.2009.01.010.
Hou L,Song X H,Lu Y M,Hu M L,He M M,Jing J X,Wang B T. Genetic analysis and SSR molecular mapping of translocation line V9128-3 drived from Haynaldia villosa resistance to stripe rust[J]. Acta Phytopathologica Sinica,2009,39(1):67-75.
[20] Hirani A H,Zelmer C D,Mcvetty P B,Li G Y. Homoeologous GSL-ELONG gene replacement for manipulation of aliphatic glucosinolates in Brassica rapa L. by marker assisted selection[J].Frontiers In Plant Science,2013,4:55.doi:10.3389/fpls.2013.00055.
[21] 赵振国. 黄瓜-酸黄瓜抗霜霉病渐渗系分子标记的筛选及细胞程序性死亡相关基因的表达分析[D]. 南京:南京农业大学,2011.
Zhao Z G. Screening of molecular for downy mildew resistance introgression line of Cucumis hystris-C.sativus and ananlysis of programmed cell death[D].Nanjing:Nanjing Agricultural University,2011.
[22] Geleta M,Heneen W K,Stoute A I,Scott R J,King G J,Kurup S,Bryngelsson T. Assigning brassica microsatellite markers to the nine c-genome chromosomes using Brassica rapa var. trilocularis-B. oleracea var. alboglabra monosomic alien addition lines[J].Theoretical and Applied Genetics,2012,125(3):455-466.doi:10.1007/s00122-012-1845-3.

选择文件类型/文献管理软件名称

选择包含的内容