西瓜种子大小相关性状分子标记与验证
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摘要
以种子大小差异显著的黏籽西瓜(Citrullus lanatus ssp.mucosospermus)品系PI186490和普通栽培西瓜品系(Citrullus lanatus ssp.vulgaris)W1-1为亲本材料,通过杂交及自交获得的F2群体,对西瓜种子大小相关性状进行精细定位,并构建遗传连锁图谱。结果表明:(1)获得与种子长度及宽度相关的QTL位点各一个,贡献率分别为24.57%和24.79%。(2)利用与种子长度及宽度紧密连锁的CAPS标记(CAPS13,CAPS14)对72份不同种子大小的自然群体材料上进行标记与表型的符合度的验证:标记CAPS13在大粒种子中检出率为36.11%,小粒检出率为80.56%;标记CAPS14在大粒中检出率为30.56%,小粒检出率为83.33%。
In this research,the genetic linkage map and size related traits of watermelon seeds were constructed and accurately located by using the F2 population derived from the hybridization and selfing of Egusi watermelon(Citrullus lanatus ssp.mucosospermus) PI186490 and cultivated watermelon(Citrullus lanatus ssp.vulgaris) W1-1.The results showed that(1) Two QTL locis associated with seed length and width were obtained with the contribution rates of 24.57% and 24.79%,respectively.(2) The CAPS markers(CAPS13,CAPS14) which were closely linked with the length and width of the seeds were used to verify the conformance of 72 different natural populations with different seed sizes.The detection rate of CAPS13 in large seed group was 36.11%,and in small seed group was80.56%; And the counterpart of CAPS14 in large group was 30.56%,and in small group was 83.33%.
In this research,the genetic linkage map and size related traits of watermelon seeds were constructed and accurately located by using the F2 population derived from the hybridization and selfing of Egusi watermelon(Citrullus lanatus ssp.mucosospermus) PI186490 and cultivated watermelon(Citrullus lanatus ssp.vulgaris) W1-1.The results showed that(1) Two QTL locis associated with seed length and width were obtained with the contribution rates of 24.57% and 24.79%,respectively.(2) The CAPS markers(CAPS13,CAPS14) which were closely linked with the length and width of the seeds were used to verify the conformance of 72 different natural populations with different seed sizes.The detection rate of CAPS13 in large seed group was 36.11%,and in small seed group was80.56%; And the counterpart of CAPS14 in large group was 30.56%,and in small group was 83.33%.
引文
Che n L.L.,Qin Z.W.,Zhou X.Y.,Xin M.,and Wu T.,2012,Molecular mark er and genetic analysis of cucumber seed length,Zhongguo Nongxue Tongbao(Chinese Agricultural Science Bulletin),28(16):165-170(陈璐璐,秦智伟,周秀艳,辛明,武涛,2012,黄瓜种子长度的遗传分析及分子标记,中国农学通报,28(16):165-170)
Chi Y.Y.,Gao P.,Zhu Z.C.,Luan F.S.,Li G.Y.,and Yu P.,2017,The QTL analysis of fruit and seed associated traits in watermelon based on CAPS markers,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),50(7):1282-1293(迟莹莹,高鹏,朱子成,栾非时,李桂英,于鹏,2017,基于CAPS标记的西瓜果实与种子相关性状QTL分析,中国农业科学,50(7):1282-1293)
Hu S.,2015,Molecular mechanism of Arabidopsis Lec RK-8Ainvolved in seed size determination,Thesis for M.S.,Biology College,Hunan University,Supervisor:Guo X.H.,pp.22-34(胡帅,2015,拟南芥LecRK-8A基因调控种子大小分子机理研究,硕士学位论文,湖南大学生物学院,导师:郭新红,pp.22-34)
Jiang L.L.,Zhang G.Y.,and Zhang J.N.,2016,Purity identification of watermelon hybrid combination T-1 seeds using SSRmolecular marker,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University Natural Science Edition),44(6):93-98,110(蒋莉莉,张高原,张建农,2016,西瓜杂交组合T-1种子纯度鉴定的SSR标记研究,西北农林科技大学学报(自然科学版),44(6):93-98,110)
Li N.,Wang J.M.,Shang J.L.,Li N.N.,Xu Y.Y.,and Ma S.W.,2017,Fine-mapping of QTL and development of in del markers for Fusarium oxysporum race 1 resistance in watermelon,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),50(1):131-141(李娜,王吉明,尚建立,李楠楠,徐永阳,马双武,2017,西瓜枯萎病生理小种1抗性QTL精细定位与InDel标记开发,中国农业科学,50(1):131-141)
Li Z.D.,2015,QTL analysis for pod and seed traits in peanut,Thesis for M.S.,Biotechnology Research Institute CAAS,Supervisor:Jiang H.F.,pp.9-20(李振动,2015,花生荚果及种子大小相关性状的QTL分析,硕士学位论文,中国农业科学院研究生院,导师:姜慧芳,pp.9-20)
Liu C.Q.,Gao P.,and Luan F.S.,2014,Construction of a genetic linkage map and QTL analysis of fruit-associated traits in watermelon,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(14):2814-2829(刘传奇,高鹏,栾非时,2014,西瓜遗传图谱构建及果实相关性状QTL分析,中国农业科学,47(14):2814-2829)
Liu S.,Wang X.Z.,Luan F.S.,and Zhu Z.C.,2013,QTL analysis for total sugar content in watermelon,Guoshu Xuebao(Jour nal of Fruit Science),30(1):75-80(刘识,王学征,栾非时,朱子成,2013,西瓜果实总糖含量QTL分析,果树学报,30(1):75-80)
Long Y.L.,Zhu B.B.,Xu Z.J.,Zhang W.,and Jiang X.F.,2016,Sequence analysis of APX gene in watermelon and its expression characteristic a fter treated with MeJA,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(8):2125-2132(龙娅丽,朱白婢,徐子健,张文,江雪飞,2016,西瓜APX基因的序列分析及其茉莉酸甲酯诱导表达特性,基因组学与应用生物学,35(8):2125-2132)
Lu B.Y.,Zhou H.W.,Chen X.,Luan F.S.,Wang X.Z.,and Jiang Y.,2016,QTL analysis of fruit traits in watermelon,Guoshu Xuebao(Journal of Fruit Science),33(10):1206-1218(卢丙洋,周慧文,陈欣,栾非时,王学征,姜羽,2016,西瓜果实几个性状的QTL分析,果树学报,33(10):1206-1218)
Luan F.S.,Delannay I.,and Staub J.E.,2008,Chinese melon(Cucumis melo L.)diversity analyses provide strategies for germplasm curation,genetic improvement,and evidentiary support of domestication patterns,Euphytica,164(2):445-461
Luan F.S.,Lu B.Y.,Zhou H.W.,and Wang X.Z.,2016,Construction of genetic linkage map and QTL location for fruit-related,Dongbei Nongye Daxue Xuebao(Journal of Northeast Agricultural University),47(8):9-20(栾非时,卢丙洋,周慧文,王学征,2016,瓜遗传连锁图谱构建及果实相关性状QTL定位,东北农业大学学报,47(8):9-20)
Ma S.W.,and Liu J.P.,eds.,2005,Description specifi cation of waterlemon germplasm resources and data criteria,China Agricultural Press,Beijing,China,pp.79(马双武,刘君璞,编著,2005,西瓜种质资源描述规范和数据标准,中国农业出版社,中国,北京,pp.79)
Miao H.,Gu X.F.,Zhang S.S.,Zhang Z.H.,Huang S.W.,Wang Y.,Cheng Z.C.,Zhang R.W.,Mu S.Q.,Li M.,Zhang Z.X.,and Fang Z.Y.,2011,Mapping QTLs for fruit-associated traitsin Cucumis sativus L.,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(24):5031-5040(苗晗,顾兴芳,张圣平,张忠华,黄三文,王烨,程周超,张若纬,穆生奇,李曼,张振贤,方智远,2011,黄瓜果实相关性状QTL定位分析,中国农业科学,44(24):5031-5040)
Ramamurthy R.K.,and Waters B.M.,2015,Identification of fruit quality and morphology QTLs in melon(Cucumis melon)using a population derived from flexuosus and cantalupensis botanical groups,Euphytica,204(1):1-15
Ren Y.,2016,QTL Mapping of sugar content and functional analysis of sugar transporter genes in watermelon fruit,Dissertation for Ph.D.,Biotechnology Research Institute CAAS,Supervisor:Xu Y.,pp.23-35(任毅,2016,西瓜果实含糖量QTL定位及糖转运蛋白基因功能初析,博士学位论文,中国农业科学院研究研究生院,导师:许勇,pp.23-35)
Sandlin K.,Prothro J.,Heesacker A.,Khalilian N.,Okashah R.,Xiang W.,Bachlava E.,Caldwell D.G.,Taylor C.A.,Seymour D.K.,White V.,Chan E.,Tolla G.,White C.,Safran D.,Graham E.,Knapp S.,and McGregor C.,2012,Compar ative mapping in watermelon(Citrullus lanatus(Thunb.)Matsum.et Nakai),Theor.Appl.Genet.,125(8):1603-1618
Tong J.P.,Han A.N.,Wang S.J.,Liu X.J.,and Su J.P.,2011,Research progress in DNA marker analysis of crop quantitative character,Shengwu Jishu Jinzhan(Current Biotechnology),1(2):98-104(童继平,韩傲男,王胜军,刘学军,苏京平,2011,作物数量性状研究进展,生物技术进展,1(2):98-104)
Wang X.J.,2012,Study on development and nutrient accumulation of seeds in edible seed watermelon,Thesis for M.S.,Agriculture College,Gansu Agricultural University,Supervisor:Zhang J.N.,pp.11-14(王晓娟,2012,籽瓜种子发育中组织结构及营养成分积累规律的研究,硕士学位论文,甘肃农业大学农学院,导师:张建农,pp.11-14)
Wang Y.P.,Wang G.F.,Wu Z.F.,Wang Z.Q.,Shi Q.H.,Pan X.H.,and Wu Z.,2016,Identification of QTLs for rice grain shape traits fora large grain material,Henongxue Bao(Journal of Nuclear Agricultural Sciences),30(12):2295-2303(汪欲鹏,王根发,武志峰,王智权,石庆华,潘晓华,吴自明,2016,大粒型水稻材料粒型性状的QTL定位,核农学报,30(12):2295-2303)
Wang Z.J.,2013,The genetic and QTL analysis of fruit shape trait of watermelon,Thesis for M.S.,Horticulture College,Northeast Agricultural University,Supervisor:Luan F.S.,pp.25-35(王兆吉,2013,西瓜果形相关性状遗传及QTL分析,硕士学位论文,东北农业大学园艺学院,导师:栾非时,pp.25-35)
Zhang Z.Z.,2014,Advances in genetic engineering for disease resistance in watermelon,Nongxuexue bao(Journal of Agriculture),4(1):85-89(张志忠,2014,西瓜抗病基因工程研究进展,农学学报,4(1):85-89)
Zhao P.,Liu R.X.,Li C.P.,Xing X.R.,Cao X.L.,Tao Y.S.,and Zhang Z.X.,2011,QTL mapping for grain yield associated traits using Ye478 introgression lines in maize,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(17):3508-3519(赵璞,刘瑞响,李成璞,邢向茹,曹晓良,陶勇生,张祖新,2011,基于掖478导入系的玉米产量性状QTL鉴定,中国农业科学,44(17):3508-3519)
Zhou H.W.,Lu B.Y.,Ma H.Y.,Gao P.,Luan F.S.,Gao Q.F.,and Qi G.A.,2016,QTL mapping of watermelon seed traits,Yuanyi Xuebao(Acta Horticulturae Sinica),43(4):715-723(周慧文,卢丙洋,马鸿艳,高鹏,栾非时,高启帆,齐国安,2016,西瓜种子大小形状相关QTL分析,园艺学报,43(4):715-723)
Chi Y.Y.,Gao P.,Zhu Z.C.,Luan F.S.,Li G.Y.,and Yu P.,2017,The QTL analysis of fruit and seed associated traits in watermelon based on CAPS markers,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),50(7):1282-1293(迟莹莹,高鹏,朱子成,栾非时,李桂英,于鹏,2017,基于CAPS标记的西瓜果实与种子相关性状QTL分析,中国农业科学,50(7):1282-1293)
Hu S.,2015,Molecular mechanism of Arabidopsis Lec RK-8Ainvolved in seed size determination,Thesis for M.S.,Biology College,Hunan University,Supervisor:Guo X.H.,pp.22-34(胡帅,2015,拟南芥LecRK-8A基因调控种子大小分子机理研究,硕士学位论文,湖南大学生物学院,导师:郭新红,pp.22-34)
Jiang L.L.,Zhang G.Y.,and Zhang J.N.,2016,Purity identification of watermelon hybrid combination T-1 seeds using SSRmolecular marker,Xibei Nonglin Keji Daxue Xuebao(Journal of Northwest A&F University Natural Science Edition),44(6):93-98,110(蒋莉莉,张高原,张建农,2016,西瓜杂交组合T-1种子纯度鉴定的SSR标记研究,西北农林科技大学学报(自然科学版),44(6):93-98,110)
Li N.,Wang J.M.,Shang J.L.,Li N.N.,Xu Y.Y.,and Ma S.W.,2017,Fine-mapping of QTL and development of in del markers for Fusarium oxysporum race 1 resistance in watermelon,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),50(1):131-141(李娜,王吉明,尚建立,李楠楠,徐永阳,马双武,2017,西瓜枯萎病生理小种1抗性QTL精细定位与InDel标记开发,中国农业科学,50(1):131-141)
Li Z.D.,2015,QTL analysis for pod and seed traits in peanut,Thesis for M.S.,Biotechnology Research Institute CAAS,Supervisor:Jiang H.F.,pp.9-20(李振动,2015,花生荚果及种子大小相关性状的QTL分析,硕士学位论文,中国农业科学院研究生院,导师:姜慧芳,pp.9-20)
Liu C.Q.,Gao P.,and Luan F.S.,2014,Construction of a genetic linkage map and QTL analysis of fruit-associated traits in watermelon,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),47(14):2814-2829(刘传奇,高鹏,栾非时,2014,西瓜遗传图谱构建及果实相关性状QTL分析,中国农业科学,47(14):2814-2829)
Liu S.,Wang X.Z.,Luan F.S.,and Zhu Z.C.,2013,QTL analysis for total sugar content in watermelon,Guoshu Xuebao(Jour nal of Fruit Science),30(1):75-80(刘识,王学征,栾非时,朱子成,2013,西瓜果实总糖含量QTL分析,果树学报,30(1):75-80)
Long Y.L.,Zhu B.B.,Xu Z.J.,Zhang W.,and Jiang X.F.,2016,Sequence analysis of APX gene in watermelon and its expression characteristic a fter treated with MeJA,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(8):2125-2132(龙娅丽,朱白婢,徐子健,张文,江雪飞,2016,西瓜APX基因的序列分析及其茉莉酸甲酯诱导表达特性,基因组学与应用生物学,35(8):2125-2132)
Lu B.Y.,Zhou H.W.,Chen X.,Luan F.S.,Wang X.Z.,and Jiang Y.,2016,QTL analysis of fruit traits in watermelon,Guoshu Xuebao(Journal of Fruit Science),33(10):1206-1218(卢丙洋,周慧文,陈欣,栾非时,王学征,姜羽,2016,西瓜果实几个性状的QTL分析,果树学报,33(10):1206-1218)
Luan F.S.,Delannay I.,and Staub J.E.,2008,Chinese melon(Cucumis melo L.)diversity analyses provide strategies for germplasm curation,genetic improvement,and evidentiary support of domestication patterns,Euphytica,164(2):445-461
Luan F.S.,Lu B.Y.,Zhou H.W.,and Wang X.Z.,2016,Construction of genetic linkage map and QTL location for fruit-related,Dongbei Nongye Daxue Xuebao(Journal of Northeast Agricultural University),47(8):9-20(栾非时,卢丙洋,周慧文,王学征,2016,瓜遗传连锁图谱构建及果实相关性状QTL定位,东北农业大学学报,47(8):9-20)
Ma S.W.,and Liu J.P.,eds.,2005,Description specifi cation of waterlemon germplasm resources and data criteria,China Agricultural Press,Beijing,China,pp.79(马双武,刘君璞,编著,2005,西瓜种质资源描述规范和数据标准,中国农业出版社,中国,北京,pp.79)
Miao H.,Gu X.F.,Zhang S.S.,Zhang Z.H.,Huang S.W.,Wang Y.,Cheng Z.C.,Zhang R.W.,Mu S.Q.,Li M.,Zhang Z.X.,and Fang Z.Y.,2011,Mapping QTLs for fruit-associated traitsin Cucumis sativus L.,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(24):5031-5040(苗晗,顾兴芳,张圣平,张忠华,黄三文,王烨,程周超,张若纬,穆生奇,李曼,张振贤,方智远,2011,黄瓜果实相关性状QTL定位分析,中国农业科学,44(24):5031-5040)
Ramamurthy R.K.,and Waters B.M.,2015,Identification of fruit quality and morphology QTLs in melon(Cucumis melon)using a population derived from flexuosus and cantalupensis botanical groups,Euphytica,204(1):1-15
Ren Y.,2016,QTL Mapping of sugar content and functional analysis of sugar transporter genes in watermelon fruit,Dissertation for Ph.D.,Biotechnology Research Institute CAAS,Supervisor:Xu Y.,pp.23-35(任毅,2016,西瓜果实含糖量QTL定位及糖转运蛋白基因功能初析,博士学位论文,中国农业科学院研究研究生院,导师:许勇,pp.23-35)
Sandlin K.,Prothro J.,Heesacker A.,Khalilian N.,Okashah R.,Xiang W.,Bachlava E.,Caldwell D.G.,Taylor C.A.,Seymour D.K.,White V.,Chan E.,Tolla G.,White C.,Safran D.,Graham E.,Knapp S.,and McGregor C.,2012,Compar ative mapping in watermelon(Citrullus lanatus(Thunb.)Matsum.et Nakai),Theor.Appl.Genet.,125(8):1603-1618
Tong J.P.,Han A.N.,Wang S.J.,Liu X.J.,and Su J.P.,2011,Research progress in DNA marker analysis of crop quantitative character,Shengwu Jishu Jinzhan(Current Biotechnology),1(2):98-104(童继平,韩傲男,王胜军,刘学军,苏京平,2011,作物数量性状研究进展,生物技术进展,1(2):98-104)
Wang X.J.,2012,Study on development and nutrient accumulation of seeds in edible seed watermelon,Thesis for M.S.,Agriculture College,Gansu Agricultural University,Supervisor:Zhang J.N.,pp.11-14(王晓娟,2012,籽瓜种子发育中组织结构及营养成分积累规律的研究,硕士学位论文,甘肃农业大学农学院,导师:张建农,pp.11-14)
Wang Y.P.,Wang G.F.,Wu Z.F.,Wang Z.Q.,Shi Q.H.,Pan X.H.,and Wu Z.,2016,Identification of QTLs for rice grain shape traits fora large grain material,Henongxue Bao(Journal of Nuclear Agricultural Sciences),30(12):2295-2303(汪欲鹏,王根发,武志峰,王智权,石庆华,潘晓华,吴自明,2016,大粒型水稻材料粒型性状的QTL定位,核农学报,30(12):2295-2303)
Wang Z.J.,2013,The genetic and QTL analysis of fruit shape trait of watermelon,Thesis for M.S.,Horticulture College,Northeast Agricultural University,Supervisor:Luan F.S.,pp.25-35(王兆吉,2013,西瓜果形相关性状遗传及QTL分析,硕士学位论文,东北农业大学园艺学院,导师:栾非时,pp.25-35)
Zhang Z.Z.,2014,Advances in genetic engineering for disease resistance in watermelon,Nongxuexue bao(Journal of Agriculture),4(1):85-89(张志忠,2014,西瓜抗病基因工程研究进展,农学学报,4(1):85-89)
Zhao P.,Liu R.X.,Li C.P.,Xing X.R.,Cao X.L.,Tao Y.S.,and Zhang Z.X.,2011,QTL mapping for grain yield associated traits using Ye478 introgression lines in maize,Zhongguo Nongye Kexue(Scientia Agricultura Sinica),44(17):3508-3519(赵璞,刘瑞响,李成璞,邢向茹,曹晓良,陶勇生,张祖新,2011,基于掖478导入系的玉米产量性状QTL鉴定,中国农业科学,44(17):3508-3519)
Zhou H.W.,Lu B.Y.,Ma H.Y.,Gao P.,Luan F.S.,Gao Q.F.,and Qi G.A.,2016,QTL mapping of watermelon seed traits,Yuanyi Xuebao(Acta Horticulturae Sinica),43(4):715-723(周慧文,卢丙洋,马鸿艳,高鹏,栾非时,高启帆,齐国安,2016,西瓜种子大小形状相关QTL分析,园艺学报,43(4):715-723)