红菜薹种质资源遗传多样性ISSR分析
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摘要
对47个红菜薹(Brassica campestris ssp. chinensis var. purpurea)种质进行了基于ISSR分子标记的遗传多样性分析。从20条随机引物中筛选出10条重复性好,条带清晰的引物,通过ISSR-PCR扩增后,共检测到83个位点,平均每引物扩增出近8个位点,其中多态性位点有80个(96.39%)。扩增产物片段大小都在100～2 000 bp,相似系数在0.47～0.90。当相似系数在0.72时,可将47份红菜薹资源分为7类。等位基因数平均多样性指数(I)为0.481 0;基因多样性0.322 2;每个位点的等位基因数1.975 9;有效等位基因数(Ne)为1.559 9。红菜薹具有丰富的遗传多样性。研究为净化红菜薹同名异种、同种异名的市场乱象,确定红菜薹种质资源相互之间的亲缘关系提供了理论支持,也为红菜薹资源的利用和育种提供了分子生物学依据。
With the help of ISSR molecular marker technique,the experiment screened out 10 primers from20 random primers that have clear and repeating bands through the extracted DNA from 47 kinds of purple flow?ering stalk(Brassica campestris ssp. chinensis var. purpurea),and 83 bands were amplified out totally,including80 polymorphism bands with polymorphism rate of 96.39%.The sizes of the amplified DNA fragments were 100-2 000 bp,and the similarity coefficients were between 0.47-0.90.47 accessions were grouped into 7 clusters at coefficient of 0.72.The average number of the allele diversity index(I)was 0.481 0;the genetic diversity was0.322 2;the number of alleles per locus was 1.975 9;the effective number of the alleles(Ne)was 1.559 9.The results show there is abundant genetic diversity in the purple flowering stalk germplasm of China.This provides the basis for molecular biology research for the utilization and breeding of purple flowering stalk resources.
With the help of ISSR molecular marker technique,the experiment screened out 10 primers from20 random primers that have clear and repeating bands through the extracted DNA from 47 kinds of purple flow?ering stalk(Brassica campestris ssp. chinensis var. purpurea),and 83 bands were amplified out totally,including80 polymorphism bands with polymorphism rate of 96.39%.The sizes of the amplified DNA fragments were 100-2 000 bp,and the similarity coefficients were between 0.47-0.90.47 accessions were grouped into 7 clusters at coefficient of 0.72.The average number of the allele diversity index(I)was 0.481 0;the genetic diversity was0.322 2;the number of alleles per locus was 1.975 9;the effective number of the alleles(Ne)was 1.559 9.The results show there is abundant genetic diversity in the purple flowering stalk germplasm of China.This provides the basis for molecular biology research for the utilization and breeding of purple flowering stalk resources.
引文
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[13]聂启军,邱正明,焦忠久,等.一种红菜薹新病害的病原鉴定及生物学特性研究[J].湖北农业科学,2016,55(24):6442-6444.Nie Q J,Qiu Z M,Jiao Z J,et al.Pathogen identification and biological characteristic of marginal scorch-a new Brassicacampestris ssp. chinensis var. utilis Disease[J].Hubei Agricultural Sciences,2016,55(24):6442-6444.
[14]魏林,梁志怀,唐炎英.红菜薹黑斑病发生规律及其综合防治[J].长江蔬菜,2018(11):53-54.Wei L,Liang Z H,Tang Y H.Occurrence regularity and integrated control of nigrities artis on flowering Chinese cabbage[J].Journal of Changjiang Vegetables,2018(11):53-54.
[15]Hosseini K S,Masoumiasl A,Dehdari M.A comparative analysis of RAPD and ISSR markers for assessing genetic diversityin Iranian populations of Nigellasativa L.[J].Cellular and Molecular Biolog(Noisy-le-Grand,France),2018,64(1):52-59.
[16]苏晓娜,王惠梅,黄雪雯,等.利用SSR标记分析东北野生菰遗传多样性和遗传结构[J].江西农业大学学报,2018,40(3):579-589.Su X N,Wang H M,Huang X W,et al.Genetic diversity and genetic structure of Zizania latifolia Griseb(Turcz)from north?east China based on SSR analysis[J].Acta Agriculturae Universitatis Jiangxiensis,2018,40(3):579-589.
[17]徐重益,李锡香,王海平,等.菜薹种质内不同大小群体间遗传多样性的RAPD鉴定和比较[J].植物遗传资源学报,2004,5(1):43-46.Xu C Y,Li X X,Wang H P,et al.RAPD identification and comparison of the genetic diversity among different populationssampled from same accession of Flowering Chinese Cabbage(Brassica campestris L. ssp. chinensis var. utilis Tsen et.Lee)[J].Journal of Plant Genetic Resources,2004,5(1):43-46.
[18]朱波华,金渭,刘昆,等.珍稀药材三叶青种质资源遗传多样性的ISSR分析[J].江西农业大学学报,2015,37(5):914-919.Zhu B H,Jin W,Liu K,et al.ISSR analysis of genetic diversity of Tetrastigma hemsleyauum,a rare Chinese medicinal herb[J].Acta Agriculturae Universitatis Jiangxiensis,2015,37(5):914-919.
[19]Mujeeb F,Bajpai P,Pathak N,et al.Genetic diversity analysis of medicinally important horticultural crop Aegle marmelosby ISSR markers[J].Methods in Molecular Biology,2017,1620:195-211.
[20]Hajiyeva S V,Akparov Z I,Hasanov N A,et al.ISSR analysis of variability of cultivated form and varieties of pomegranate(Punica granatum L.)from Azerbaijan[J].Russian Journal of Genetics,2018,54(2):188-197.
[21]王关林,方宏筠.植物基因工程(第2版)[M].北京:科学出版社,2002:742-744.Wang G L,Fang H J.Plant genetic engineering(2nd Edition)[M].Beijing:Science Press,2002:742-744.
[22]张凤银,戴希刚,潘磊.藜豆种质资源遗传多样性的ISSR分子标记分析[J].江汉大学学报(自然科学版),2013,41(6):100-104.Zhang F Y,Dai X G,Pan L.Genetic diversity analysis of velvet bean germplasm resources with ISSR markers[J].Journal ofJianghan University(Natural Science Edition),2013,41(6):100-104.
[23]戴希刚,包满珠.羽衣甘蓝DH群体遗传多样性分析.江西农业大学学报,2013,35(1):49-53.Dai X G,Bao M Z.Genetic diversity analysis of doubled haploid population in ornamental kale(Brassica oleracea var.aceph?ala)[J].Acta Agriculturae Universitatis Jiangxiensis,2013,35(1):49-53.
[24]谭雪,郭爽,郑少薇,等.菜薹种质资源亲缘关系的RAPD分析[J].华北农学报,2011,26(s1):18-22.Tan X,Guo S,Zheng S W,et al.Genetic relationship analysis of flowering Chinese cabbage germplasm resources by RAPD[J].Acta Agriculturae Boreali-Sinica,2011,26(s1):18-22.
[25]任梦云,陈彦君,张盾,等.ISSR标记技术在药用植物资源中的研究进展及应用[J].生物技术通报,2017,33(4):63-69Ren M Y,Chen Y J,Zhang D,et al.Application and research progress of inter-simple sequence repeat(ISSR)marker in Me?dicinal[J].Plants Biotechnology Bulletin,2017,33(4):63-69.
[26]Ding Z Y,Bai Z B,Wu Y F,et al.Study on genetic relationship of purple Tsai-tai germplasms with SSR Markers[J].Agricul?tural Science&Technology,2012,13(8):1664-1669.
[2]周晓波,白占兵,丁茁荑,等.利用SSR分析红菜苔的遗传多样性[J].植物遗传学报,2012,13(6):1088-1092.Zhou X B,Bai Z B,Ding Z Y,et al.Genetic diversity of Tsai-tai germplasm revealed by SSR markers[J].Journal of Plant Ge?netic Resources,2012,13(6):1088-1092.
[3]吴朝林.紫菜薹新品种五彩紫薹二号的选育[J].长江蔬菜,2006(6):48-49.Wu C L.Breeding of a new purple Cai-tai variety Wucaizitai No.2[J].Journal of Changjiang Vegetables,2006(6):48-49.
[4]王春梅,辛复林,朱大社,等.红菜薹新品种佳红1号的选育及应用[J].长江蔬菜,2009(8):20-21.Wang C M,Xin F L,Zhu D S,et al.Development and application of a new Purple Cai-tai cultivar,Jiahong No.1[J].Journalof Changjiang Vegetables,2009(8):20-21.
[5]聂启军,邱正明,朱凤娟,等.红菜薹新品种鄂红5号的选育[J].湖北农业科学,2017,56(24):4828-4829,4928.Nie Q J,Qiu Z M,Zhu F J,et al.Breeding of a new purple-caitai variety Ehong No.5[J].Hubei Agricultural Sciences,2017,56(24):4828-4829.
[6]许明,白明义,魏毓棠.不同发育期紫菜苔细胞质雄性不育系及保持系植株体内物质代谢的差异[J].沈阳农业大学学报,2008,39(2):152-155.Xu M,Bai M Y,Wei Y T.Changes in substance metabolism between B.campestris ssp.Chinensis var. purpurea Hort CMSlines and their maintainer line at different development stages[J].Journal of Shenyang Agricultural University,2008,39(2):152-155.
[7]邵明珠,徐跃进,万正杰.新型红菜薹细胞质雄性不育系的花药发育细胞学观察[J].华中农业大学学报,2012,31(4):436-439.Shao M Z,Xu Y J,Wan Z J,et al.Cytological study of anther abortion in a novel cytoplasmic male-sterile line of purple Cai-tai[J].Journal of Huazhong Agricultural University,2012,31(4):436-439.
[8]邵明珠,徐跃进,万正杰,等.红菜薹细胞质雄性不育系的农艺性状调查及POD同工酶分析[J].中国蔬菜,2012(2):30-35.Shao M Z,Xu Y J,Wan Z J,et al.Agronomic traits investigation and POD isozyme analysis of cytoplasmic male sterility inHongcaitai[Brassica campestris L. ssp. chinensis(L.)Makino var. utilis Tsen et Lee][J].China Vegetables,2012(2):30-35.
[9]吴艺飞,丁茁荑,戴雄泽,等.红菜薹小孢子培养技术研究[J].作物研究,2016,30(1):73-77.Wu Y F,Ding Z Y,Dai X Z,et al.Study on microspore culture techniques in purple flowering stalk(Brassica campestris)[J].Crop Research,2016,30(1):73-77.
[10]邓耀华,周国林,黄新学,等.气候条件对红菜薹游离小孢子培养的影响[J].长江蔬菜,2010(24):15-16.Deng Y H,Zhou G L,Huang X X,et al.Effect of climatic conditions on Hong Caitai isolated microspore training[J].Journalof Changjiang Vegetables,2010(24):15-16.
[11]聂启军,邱正明.高山红菜薹栽培关键技术[J].中国蔬菜,2013(21):42.Nie Q J,Qiu Z M.Key techniques for cultivation of purple Cai-tai on high mountain[J].China Vegetables,2013(21):42.
[12]陈祥金,张颖芳,熊迪,等.露地丝瓜-红菜薹高效栽培模式[J].长江蔬菜,2016(10):71-72.Chen X J,Zhang Y F,Xiong D,et al.High efficiency cultivation mode of loofah and purple Cai-tai in open field[J].Journalof Changjiang Vegetables,2016(10):71-72.
[13]聂启军,邱正明,焦忠久,等.一种红菜薹新病害的病原鉴定及生物学特性研究[J].湖北农业科学,2016,55(24):6442-6444.Nie Q J,Qiu Z M,Jiao Z J,et al.Pathogen identification and biological characteristic of marginal scorch-a new Brassicacampestris ssp. chinensis var. utilis Disease[J].Hubei Agricultural Sciences,2016,55(24):6442-6444.
[14]魏林,梁志怀,唐炎英.红菜薹黑斑病发生规律及其综合防治[J].长江蔬菜,2018(11):53-54.Wei L,Liang Z H,Tang Y H.Occurrence regularity and integrated control of nigrities artis on flowering Chinese cabbage[J].Journal of Changjiang Vegetables,2018(11):53-54.
[15]Hosseini K S,Masoumiasl A,Dehdari M.A comparative analysis of RAPD and ISSR markers for assessing genetic diversityin Iranian populations of Nigellasativa L.[J].Cellular and Molecular Biolog(Noisy-le-Grand,France),2018,64(1):52-59.
[16]苏晓娜,王惠梅,黄雪雯,等.利用SSR标记分析东北野生菰遗传多样性和遗传结构[J].江西农业大学学报,2018,40(3):579-589.Su X N,Wang H M,Huang X W,et al.Genetic diversity and genetic structure of Zizania latifolia Griseb(Turcz)from north?east China based on SSR analysis[J].Acta Agriculturae Universitatis Jiangxiensis,2018,40(3):579-589.
[17]徐重益,李锡香,王海平,等.菜薹种质内不同大小群体间遗传多样性的RAPD鉴定和比较[J].植物遗传资源学报,2004,5(1):43-46.Xu C Y,Li X X,Wang H P,et al.RAPD identification and comparison of the genetic diversity among different populationssampled from same accession of Flowering Chinese Cabbage(Brassica campestris L. ssp. chinensis var. utilis Tsen et.Lee)[J].Journal of Plant Genetic Resources,2004,5(1):43-46.
[18]朱波华,金渭,刘昆,等.珍稀药材三叶青种质资源遗传多样性的ISSR分析[J].江西农业大学学报,2015,37(5):914-919.Zhu B H,Jin W,Liu K,et al.ISSR analysis of genetic diversity of Tetrastigma hemsleyauum,a rare Chinese medicinal herb[J].Acta Agriculturae Universitatis Jiangxiensis,2015,37(5):914-919.
[19]Mujeeb F,Bajpai P,Pathak N,et al.Genetic diversity analysis of medicinally important horticultural crop Aegle marmelosby ISSR markers[J].Methods in Molecular Biology,2017,1620:195-211.
[20]Hajiyeva S V,Akparov Z I,Hasanov N A,et al.ISSR analysis of variability of cultivated form and varieties of pomegranate(Punica granatum L.)from Azerbaijan[J].Russian Journal of Genetics,2018,54(2):188-197.
[21]王关林,方宏筠.植物基因工程(第2版)[M].北京:科学出版社,2002:742-744.Wang G L,Fang H J.Plant genetic engineering(2nd Edition)[M].Beijing:Science Press,2002:742-744.
[22]张凤银,戴希刚,潘磊.藜豆种质资源遗传多样性的ISSR分子标记分析[J].江汉大学学报(自然科学版),2013,41(6):100-104.Zhang F Y,Dai X G,Pan L.Genetic diversity analysis of velvet bean germplasm resources with ISSR markers[J].Journal ofJianghan University(Natural Science Edition),2013,41(6):100-104.
[23]戴希刚,包满珠.羽衣甘蓝DH群体遗传多样性分析.江西农业大学学报,2013,35(1):49-53.Dai X G,Bao M Z.Genetic diversity analysis of doubled haploid population in ornamental kale(Brassica oleracea var.aceph?ala)[J].Acta Agriculturae Universitatis Jiangxiensis,2013,35(1):49-53.
[24]谭雪,郭爽,郑少薇,等.菜薹种质资源亲缘关系的RAPD分析[J].华北农学报,2011,26(s1):18-22.Tan X,Guo S,Zheng S W,et al.Genetic relationship analysis of flowering Chinese cabbage germplasm resources by RAPD[J].Acta Agriculturae Boreali-Sinica,2011,26(s1):18-22.
[25]任梦云,陈彦君,张盾,等.ISSR标记技术在药用植物资源中的研究进展及应用[J].生物技术通报,2017,33(4):63-69Ren M Y,Chen Y J,Zhang D,et al.Application and research progress of inter-simple sequence repeat(ISSR)marker in Me?dicinal[J].Plants Biotechnology Bulletin,2017,33(4):63-69.
[26]Ding Z Y,Bai Z B,Wu Y F,et al.Study on genetic relationship of purple Tsai-tai germplasms with SSR Markers[J].Agricul?tural Science&Technology,2012,13(8):1664-1669.