基于SRAP标记的甜瓜耐盐种质资源遗传多样性分析
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摘要
研究不同耐盐程度甜瓜种质资源的亲缘关系和遗传多样性,为甜瓜种质资源有效利用提供理论依据。该试验以不同耐盐程度的27个甜瓜品种为材料,利用SRAP标记分析甜瓜种质资源的遗传多样性,从364对引物组合中筛选出24对扩增清楚且多态性丰富的引物,共扩增出415个清晰的位点,其中多态性位点241个,多态比率达58%。平均观测等位基因数(Na)为1.588 0,平均有效等位基因数(Ne)为1.347 0,平均Nei's基因多样性指数(H)为0.201 3,平均Shannon's信息指数(I)为0.301 0,表明供试甜瓜具有丰富的遗传多样性。UPGMA聚类结果显示:遗传相似系数在0.36～1.00之间,遗传相似性系数在0.72时,可将27个甜瓜品种划分为4个类群,耐盐型甜瓜在其中3个类群有分布,中等耐盐型甜瓜在4大类群中均有分布,盐敏感型均集中在第Ⅰ类群,表明耐盐型和中等耐盐型甜瓜种质的遗传多样性比盐敏感型种质丰富。遗传相似系数与UPGMA聚类结果的Cophentic矩阵相关系数为0.749,说明聚类分析结果较准确。研究认为,耐盐型甜瓜的遗传多样性较丰富,要得到耐盐的杂种优势品种,需要融入更多遗传关系较远的新种质。
To provide theoretical basis for effective use of melon germplasm resources,we studied the genetic relationship and diversity of melon with different degree of salt tolerance.In this study,the genetic diversity of melon germplasm resources was analyzed by Sequence-Related Amplified Polymorphism(SRAP)using 27 melon varieties with different salt tolerance levels.24 primer pairs with clear amplification and rich polymorphism were selected from 364 pairs,and 415 clear sites were amplified totally,of which 241 were polymorphic sites and the polymorphic ratio was 58%.Average observed number of alleles(Na)was 1.588 0,average effective number of alleles(Ne)was 1.347 0,average Neis gene diversity(H)was 0.201 3,average Shannons information index(I)was 0.301 0,and the results showed that melon germplasms had abundant genetic diversity.Unweighted pair-group method with arithmetic means(UPGMA)clustering results showed that the genetic similarity coefficients were between 0.36 and 1.00.27 melon varieties were divided into four classes at the genetic similarity coefficient of 0.72.Salt tolerant melon was distributed among three categories,medium salt tolerant melon was distributed through all the classes,and while salt sensitive melon was concentrated in the first group,the results demonstrated that genetic diversity of salt tolerant and medium salt tolerant melon were more abundant than that of salt sensitive melon.Cophenetic matrix correlation coefficient of genetic similarity coefficient and UPGMA clustering result was 0.749,which indicated that the cluster analysis was accurate.The study indicated that the genetic diversity of salt tolerant melon was rich,and in order to get salt tolerant heterosis varieties,more far genetic relationship germplasms need to be introduced.
To provide theoretical basis for effective use of melon germplasm resources,we studied the genetic relationship and diversity of melon with different degree of salt tolerance.In this study,the genetic diversity of melon germplasm resources was analyzed by Sequence-Related Amplified Polymorphism(SRAP)using 27 melon varieties with different salt tolerance levels.24 primer pairs with clear amplification and rich polymorphism were selected from 364 pairs,and 415 clear sites were amplified totally,of which 241 were polymorphic sites and the polymorphic ratio was 58%.Average observed number of alleles(Na)was 1.588 0,average effective number of alleles(Ne)was 1.347 0,average Neis gene diversity(H)was 0.201 3,average Shannons information index(I)was 0.301 0,and the results showed that melon germplasms had abundant genetic diversity.Unweighted pair-group method with arithmetic means(UPGMA)clustering results showed that the genetic similarity coefficients were between 0.36 and 1.00.27 melon varieties were divided into four classes at the genetic similarity coefficient of 0.72.Salt tolerant melon was distributed among three categories,medium salt tolerant melon was distributed through all the classes,and while salt sensitive melon was concentrated in the first group,the results demonstrated that genetic diversity of salt tolerant and medium salt tolerant melon were more abundant than that of salt sensitive melon.Cophenetic matrix correlation coefficient of genetic similarity coefficient and UPGMA clustering result was 0.749,which indicated that the cluster analysis was accurate.The study indicated that the genetic diversity of salt tolerant melon was rich,and in order to get salt tolerant heterosis varieties,more far genetic relationship germplasms need to be introduced.
引文
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[19]陈芸,李冠,王贤磊.甜瓜种质资源遗传多样性的SRAP分析[J].遗传,2010,32(7):744-751.CHEN Y,LI G,WANG X L.Genetic diversity of a germplasm collection of Cucumis melo L.using SRAP markers[J].Hereditas,2010,32(7):744-751.
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[21]张羽,张晓娟,陈进,等.43份油菜菌核病抗性资源SCoT,SSR与SRAP标记分析[J].西北农林科技大学学报,2017,45(1):45-52.ZHANG Y,ZHANG X J,CHEN J,et al.Germplasm analysis of 43 Sclerotinia sclerotiorum resistant Brassica napus by SCoT,SSR and SRAP markers[J].Journal of Northwest A and F University,2017,45(1):45-52.
[22]张全芳,张荦嘉,梁水美,等.适用于多重荧光SSR检测的玉米基因组DNA提取方法[J].玉米科学,2016,24(2):29-34.ZHANG Q F,ZHANG L J,LIANG S M,et al.Apply to multiple fluorescent SSR detection of maize genome DNA extraction method[J].Journal of Maize Sciences,2016,24(2):29-34.
[23]黄春琼,刘国道,白昌军.SRAP标记在落花生属种质资源遗传多样性上的利用[J].基因组学与应用生物学,2015,34(3):622-627.HUANG C Q,LIU G D,BAI C J.Application of SRAPmarkers in the genetic diversity of Arachis Linn.germplasm resources[J].Genomics and Applied Biology,2015,34(3):622-627.
[24]梁宝萍,原玉香,朴凤植,等.大白菜非变性聚丙烯酰胺凝胶电泳技术的优化[J].河南农业科学,2012,41(5):129-132.LIANG B P,YUAN Y X,PIAO F Z,et al.The optimization of technique of native polyacrylamide gel electrophoresis(Native-PAGE)[J],Journal of Henan Agricultural Sciences,2012,41(5):129-132.
[25]李桂花,陈汉才,张艳,等.小白菜种质遗传多样性与亲缘关系的SRAP和SSR分析[J].广东农业科学,2017,44(5):37-45.LI G H,CHEN H C,ZHANG Y,et al.Genetic diversity and phylogenetic relationships analysis of Chinese cabbage germplasm resources by SRAP and SSR[J].Guangdong Agricultural Sciences,2017,44(5):37-45.
[26]常艳,金青,凌磊,等.16种平菇遗传关系的SRAP分析[J].安徽农业大学学报,2017,44(5):834-838.CHANG Y,JIN Q,LING L,et al.Phylogenetic relationship analysis of 16 Pleurotus ostreatus species by SRAPmarkers[J].Journal of Anhui Agricultural University,2017,44(5):834-838.
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[2]MUNNS R,TESTER M.Mechanisms of salinity tolerance[J].Annu.Rev.Plant Biol.,2008,59:651-681.
[3]张恒,郑宝江,宋保华,等.植物盐胁迫应答蛋白质组学分析[J].生态学报,2011,31(22):6 936-6 946.ZHANG H,ZHENG B J,SONG B H,et al.Salt-responsive proteomics in plants[J].Acta Ecologica Sinica,2011,31(22):6 936-6 946.
[4]王迎儿,高旭,王毓洪,等.南瓜耐盐种质的筛选鉴定及耐盐基因的标记[J].浙江农业学报,2015,27(3):372-379.WANG Y E,GAO X,WANG Y H,et al.Screening of salt tolerance germplasms and tagging of the tolerance gene(s)using RAPD and SSR markers in pumpkin[J].Acta Agriculturae Zhejiangensis,2015,27(3):372-379.
[5]ABEL G H,Mackenzie A J.Salt tolerance of soybean varieties during germination and later growth[J].Crop Sci,1964,(4):157-160.
[6]赵丽娜,张芙蓉,莫霏,等.甜瓜盐碱逆境生理响应及相关基因研究进展[J].上海农业学报,2016,32(6):176-180.ZHAO L N,ZHANG F R,MONG F,et al.Research progress on physiological responses of melon under saline stress and their related genes[J].Acta Agriculture Shanghai,2016,32(6):176-180.
[7]SAJID H,ZHANG J H,ZHONG C,et al.Effects of salt stress on rice growth,development characteristics,and the regulating ways:A review[J].Journal of Integrative Agriculture,2017:16(11):2 357-2 374.
[8]REDDY I N B L,KIM B K,YOON I S,et al.Salt tolerance in rice:Focus on mechanisms and approaches[J].Rice Science,2017,24(3):123-144.
[9]田艺心,高凤菊,曹鹏鹏,等.大豆耐盐基因研究进展[J].大豆科学,2018,37(4):629-636.TIAN Y X,GAO F J,CAO P P,et al.Recent developments of salt tolerance gene in soybean[J].Soybean Science,2018,37(4):629-636.
[10]郝德荣,程玉静,徐辰武,等.玉米耐盐种质筛选及群体遗传结构分析[J].植物遗传资源学报,2013,14(6):1 153-1 160.HAO D R,CHENG Y J,XU C W,et al.Screening of maize germplasms for salt-tolerance and evaluation of population genetic structure[J].Journal of Plant Genetic Resources,2013,14(6):1 153-1 160.
[11]刘雅辉,王秀萍,鲁雪林,等.棉花耐盐相关序列扩增多态性(SRAP)分子标记筛选[J].江苏农业学报,2015,31(3):484-488.LIU Y H,WANG X P,LU X L,et al.Selection of sequence-related amplified polymorphism molecular marker associated with salt tolerance of cotton[J].Jiangsu J.of Agr.Sci.,2015,31(3):484-488.
[12]BEHROOZ S,SAHEBALI B,NASSER G,et al.Genotypic differences in physiological and biochemical responses to salinity stress in melon(Cucumis melo L.)plants:prospects for selection of salt tolerant landraces[J].Plant Physiology and Biochemistry,2017,119(2017):294-311.
[13]王澍,朱春燕,俞晨玺,等.NaCl对不同甜瓜品种植株中K+和Na+含量的影响[J].上海交通大学学报,2011,29(3):41-46.WANG S,ZHU C Y,YU C X,et al.K+and Na+contents in plants of muskmelon varieties under NaCl stress[J].Journal of Shanghai Jiaotong University,2011,29(3):41-46.
[14]赵卫星,常高正,高宁宁,等.甜瓜种质资源芽苗期和幼苗期耐NO3-性评价及鉴定指标筛选[J].河南农业科学,2018,47(1):84-89.ZHAO W X,CHANG G Z,GAO N N,et al.Evaluation and selection of identification Indices for NO3-tolerance at sprout and seedling stages of melon germplasms[J].Journal of Henan Agricultural Sciences,2018,47(1):84-89.
[15]陈嘉贝,张芙蓉,黄丹枫,等.盐胁迫下两个甜瓜品种转录因子的转录组分析[J].植物生理学报,2014,50(2):150-158.CHEN J B,ZHANG F R,HUANG D F,et al.Transcriptome analysis of transcription factors in two melon(Cucumis melo L.)cultivars under salt stress[J].Plant Physiology Journal,2014,50(2):150-158.
[16]刘东明,杨丽梅,方智远,等.甘蓝类蔬菜作物分子育种研究进展[J].中国农业科技导报,2015,17(1):15-22.LIU D M,YANG L M,FANG Z Y,et al.Progress of brassica crops molecular breeding[J].Journal of Agricultural Science and Technology,2015,17(1):15-22.
[17]SOLMAZ I6),KACAR A Y,SARI N,et al.Genetic diversity within Turkish watermelon(Citrullus lanatus(Thunb.)Matsumura&Nakai)accessions revealed by SSR and SRAPmarkers[J].Turk.J.Agric.For.,2016,40:407-419.
[18]杨永,王豪杰,张学军,等.新疆甜瓜地方种质资源遗传多样性的SRAP分析[J].植物遗传资源学报,2017,18(3):436-448.YANG Y,WANG H J,ZHANG X J,et al.Genetic diversity analysis of melon landraces(Cucumis melo)in Xinjiang based on SRAP markers[J].Journal of Plant Genetic Resources,2017,18(3):436-448.
[19]陈芸,李冠,王贤磊.甜瓜种质资源遗传多样性的SRAP分析[J].遗传,2010,32(7):744-751.CHEN Y,LI G,WANG X L.Genetic diversity of a germplasm collection of Cucumis melo L.using SRAP markers[J].Hereditas,2010,32(7):744-751.
[20]刘雅辉,王秀萍,鲁雪林,等.棉花耐盐相关序列扩增多态性(SRAP)分子标记筛选[J].江苏农业学报,2015,31(3):484-488.LIU Y H,WANG X P,LU X L,et al.Selection of sequence-related amplified polymorphism molecular marker associated with salt tolerance of cotton[J].Jiangsu Journal of Agriculture Science,2015,31(3):484-488.
[21]张羽,张晓娟,陈进,等.43份油菜菌核病抗性资源SCoT,SSR与SRAP标记分析[J].西北农林科技大学学报,2017,45(1):45-52.ZHANG Y,ZHANG X J,CHEN J,et al.Germplasm analysis of 43 Sclerotinia sclerotiorum resistant Brassica napus by SCoT,SSR and SRAP markers[J].Journal of Northwest A and F University,2017,45(1):45-52.
[22]张全芳,张荦嘉,梁水美,等.适用于多重荧光SSR检测的玉米基因组DNA提取方法[J].玉米科学,2016,24(2):29-34.ZHANG Q F,ZHANG L J,LIANG S M,et al.Apply to multiple fluorescent SSR detection of maize genome DNA extraction method[J].Journal of Maize Sciences,2016,24(2):29-34.
[23]黄春琼,刘国道,白昌军.SRAP标记在落花生属种质资源遗传多样性上的利用[J].基因组学与应用生物学,2015,34(3):622-627.HUANG C Q,LIU G D,BAI C J.Application of SRAPmarkers in the genetic diversity of Arachis Linn.germplasm resources[J].Genomics and Applied Biology,2015,34(3):622-627.
[24]梁宝萍,原玉香,朴凤植,等.大白菜非变性聚丙烯酰胺凝胶电泳技术的优化[J].河南农业科学,2012,41(5):129-132.LIANG B P,YUAN Y X,PIAO F Z,et al.The optimization of technique of native polyacrylamide gel electrophoresis(Native-PAGE)[J],Journal of Henan Agricultural Sciences,2012,41(5):129-132.
[25]李桂花,陈汉才,张艳,等.小白菜种质遗传多样性与亲缘关系的SRAP和SSR分析[J].广东农业科学,2017,44(5):37-45.LI G H,CHEN H C,ZHANG Y,et al.Genetic diversity and phylogenetic relationships analysis of Chinese cabbage germplasm resources by SRAP and SSR[J].Guangdong Agricultural Sciences,2017,44(5):37-45.
[26]常艳,金青,凌磊,等.16种平菇遗传关系的SRAP分析[J].安徽农业大学学报,2017,44(5):834-838.CHANG Y,JIN Q,LING L,et al.Phylogenetic relationship analysis of 16 Pleurotus ostreatus species by SRAPmarkers[J].Journal of Anhui Agricultural University,2017,44(5):834-838.
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