粉枝莓ISSR遗传多样性和果实香气成分多样性分析
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摘要
为明确西藏粉枝莓的多样性和亲缘关系,利用ISSR分子标记技术和HS-SPME/GC-MS技术,对粉枝莓7个居群的遗传多样性和果实香气成分多样性进行研究。结果表明:11条引物共得到102个清晰的扩增条带,其中多态性条带99个,PPB达到97.06%,香农信息指数I为0.490 9,表明粉枝莓具有较高的遗传多样性水平。Nei’s遗传结构(GST,0.287 2)和分子变异分析(AMOVA)的结果表明大部分的遗传变异分布在居群内(71.28%)。通过遗传分化系数GST估计的基因流为1.241 1。45个粉枝莓果实共检测出28种挥发性成分,基于果实香气成分的欧式距离为0.028 6~0.144 6。利用Mantel检验结果表明粉枝莓ISSR遗传距离和果实香气成分欧式距离无显著相关性(r=-0.153 2,P=0.676 0)。采用28种香气成分对45个样本进行主成分分析,前4个主成分的累积贡献率达到74.20%,较好的反映了粉枝莓果实香气成分的组成情况。研究表明,粉枝莓存在较丰富的遗传多样性,为西藏粉枝莓资源的进一步研究和利用提供参考。
To clarify the diversity and relationship of Rubus biflorus Buch,the genetic and phytochemical diversities of 7 populations of Rubus biflorus Buch were investigated using ISSR and HS-SPME/GC-MS.The results showed that:Among102 clear amplified bands obtained by 11 pairs primers,99 were polymorphic.The number of polymorphic bands accounted for 96.07% of total bands,the Shannon index of diversity I was 0.490 9,indicating that Rubus biflorus Buch had abundant genetic diversity.The coefficient of gene differentiation(GST,0.365 2)and AMOVA analysis revealed that most genetic diversities were distributed within populations(71.28%).The estimate of gene flow based on GST was 1.241 1.28 aromatic compounds were detected among 45 Rubus biflorus Buch in total.The Euclidean distance among the seven populations varied from 0.028 6-0.144 6.The Mantel test showed that there was no significant correlation between the genetic distance and Euclidean distance(r=-0.153 2,P=0.676 0).Principal component analysis was used to evaluate the 28 aromatic components.The results indicated that the former 4 principal components accounted for 74.20% of the cumulative contribution value.In conclusion,PCA could simplify the complicate original data to principal components,which reflected the aroma constitution preferably.This study showed that there existed a rich genetic diversity providing references for further research and utilization of Rubus biflorus Buch resources.
To clarify the diversity and relationship of Rubus biflorus Buch,the genetic and phytochemical diversities of 7 populations of Rubus biflorus Buch were investigated using ISSR and HS-SPME/GC-MS.The results showed that:Among102 clear amplified bands obtained by 11 pairs primers,99 were polymorphic.The number of polymorphic bands accounted for 96.07% of total bands,the Shannon index of diversity I was 0.490 9,indicating that Rubus biflorus Buch had abundant genetic diversity.The coefficient of gene differentiation(GST,0.365 2)and AMOVA analysis revealed that most genetic diversities were distributed within populations(71.28%).The estimate of gene flow based on GST was 1.241 1.28 aromatic compounds were detected among 45 Rubus biflorus Buch in total.The Euclidean distance among the seven populations varied from 0.028 6-0.144 6.The Mantel test showed that there was no significant correlation between the genetic distance and Euclidean distance(r=-0.153 2,P=0.676 0).Principal component analysis was used to evaluate the 28 aromatic components.The results indicated that the former 4 principal components accounted for 74.20% of the cumulative contribution value.In conclusion,PCA could simplify the complicate original data to principal components,which reflected the aroma constitution preferably.This study showed that there existed a rich genetic diversity providing references for further research and utilization of Rubus biflorus Buch resources.
引文
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[6]李钰莹,范希峰,侯新村,朱毅,赵春桥,武菊英.利用ISSR和SSR标记分析芒荻类植物资源遗传多样性[J].中国农业大学学报,2014,19(2):21-27Li Y Y,Fan X F,Hou X C,Zhu Y,Zhao C Q,Wu J Y.Genetic diversity of Miscanthus&Triarrhena based on ISSR and SSR analysis[J].Journal of China Agricultural University,2014,19(2):21-27(in Chinese)
[7]戴智慧,龙骏,俞雷民,倪穗.宁波地区野生换锦花遗传多样性的ISSR分析[J].核农学报,2016,30(10):1898-1905Dai Z H,Long J,Yu L M,Ni S.Genetic diversity of wide Lycorissprengeri populations from different habitats of Ningbo based on ISSR analysis[J].Journal of Nuclear Agricultural Sciences,2016,30(10):1898-1905(in Chinese)
[8]Nudin N F H,Ali A M,Ngah N,Mazlan N Z,Mat N,Ghani M N A,Alias N,Zakaria A J,Jahan M S.ISSR marker-assisted genetic diversity analysis of Dioscorea hispida and selection of the best variety for sustainable production[J].Comptes Rendus Biologies,2017,340(8):359-366
[9]Oliveira L A R,Machado C A,Cardoso M N,Oliveira A C A,Amaral A L,Rabbani A R C,Silva A V C,Ledo A S.Genetic diversity of Saccharum complex using ISSR markers[J].Genetics and Molecular Research,2017,16(3):gmr 16039788
[10]孙淑霞,李靖,陈栋,谢红江,涂美艳,江国良.ISSR分子标记技术在桃品种鉴定中的应用[J].中国农学通报,2011,27(4):173-177Sun S X,Li J,Chen D,Xie H J,Tu M Y,Jiang G L.Molecular identification of peach germplasm by ISSR Markers[J].Chinese Agricultural Science Bulletin,2011,27(4):173-177(in Chinese)
[11]戴正,陈力耕,童品璋.香榧品种遗传变异与品种鉴定的ISSR分析[J].园艺学报,2008(8):1125-1130Dai Z,Chen L G,Tong P Z.Genetic variation and fingerprinting of Torreyagrandis cultivars detected by ISSR markers[J].Acta Horticuhurae Sinic,2008(8):1125-1130(in Chinese)
[12]刘崇琪,陈学森,王金政,陈晓流,王海波,田长平,吴传金.新疆野生樱桃李果实部分表型性状的遗传多样性分析[J].园艺学报,2008,35(9):1261-1268Liu C Q,Chen X S,Wang J Z,Chen X L,Wang H B,Tian C P,Wu C J.Studies on genetic diversity of phenotypic traits in wild M yrobalan Plum(Prunus cerasifera Ehrh)[J].Acta Horticuhurae Sinic,2008,35(9):1261-1268(in Chinese)
[13]周莉,刘莉,刘翔,徐伟欣,张平,李志文.不同变种甜瓜果实成熟性状及其香气成分的多样性分析[J].华北农学报,2013,28(3):102-108Zhou L,Liu L,Liu X,Xu W X,Zhang P,Li Z W.Diversity analysis of aroma and other mature fruit characters among different varieties of Cucumis melo L[J].Acta Agriculturae Boreali-Sinica,2013,28(3):102-108(in Chinese)
[14]Korbin M,Kuras A,Straczynska K,Orzel A,Danek J.Biotechnologicaldirections in polish breeding of Rubus[C].In:ISHS Acta Horticulturae 585:9th International Rubus and Ribes Symposium.Pucón:International Society for Horticultural Science,2008(1):133-139
[15]Hong Y P,Kin M J,Hong K N.Genetic diversity in natural populations of two geographic isolates of Korean black raspberry[J].Journal of Horticultural Science and Biotechnology,2003,78(3):350-354
[16]李金璐,王硕,于婧,王玲,周世良.一种改良的植物DNA提取方法[J].植物学报,2013,48(1):72-78Li J L,Wang S,Yu J,Wang L,Zhou S L.A modified CTAB protocol for plant DNA extraction[J].Chinese Bulletin of Botany,2013,48(1):72-78(in Chinese)
[17]尹燕雷,苑兆和,冯立娟,招雪晴,王金政,王超.不同栽培条件下凯特杏果实发育过程中香气成分的GC/MS分析[J].林业科学,2010,46(7):92-98Yin Y L,Yuan Z H,Feng L J,Zhao X Q,Wang J Z,Wang C.GC/MS analysis of aromatic components in Katy apricot fruit in various developmental periods under different cultivation condition[J].Scientia Silvae Sinicae,2010,46(7):92-98(in Chinese)
[18]孔祥琪,施瑞城,张彦军,谭乐和,皋香.气相色谱质谱联用技术分析热处理前后番木瓜汁挥发性香气[J].食品与发酵工业,2016,42(1):189-194Kong X Q,Shi R C,Zhang Y,Tan L H,Gao X.HS-SPME/GCMS analysis of the aromatic components of papayajuice before and after the heat treatment[J].Food and Fermentation Industries Editorial Staff,2016,42(1):189-194(in Chinese)
[19]Nybom H.Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants[J].Molecular Ecology,2004,13:1143-1155
[20]和志娇,和加卫,程在全,杨正松,杨洪涛,和文佳.三叶悬钩子自然居群遗传多样的ISSR分析[J].园艺学报,2012,39(11):2142-2150He Z J,He J W,Cheng Z Q,Yang Z S,Yang H T,He W J.Genetic diversity in the natural populations of Rubus delavayi demonstrated by inter-simple sequence repeats[J].Acta Horticuhurae Sinic,2012,39(11):2142-2150(in Chinese)
[21]和志娇,和加卫,杨正松,程在全,杨燕林,李燕,黄杏娥.红泡刺藤居群的遗传多样性研究[J].西北植物学报,2011,31(12):2406-2411He Z J,He J W,Yang Z S,Cheng Z Q,Yang Y L,Li Y,Huang X E.ISSR anlysis of genetic diversity of Rubus niveus Thunb populations[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(12):2406-2411(in Chinese)
[22]Wright S.The genetical structure of populations[J].Annals of Eugenices,1951,15(4):323-354
[2]康淑荷.粉枝莓根挥发油化学成分研究[J].西北民族大学学报:自然科学版,2007,28(65):27-29Kang S H.Study on chemical constituents of volatile oil from the roots of Rubus biflorus[J].Journal of Northwest University for Nationalities:Natural Science,2007,28(65):27-29(in Chinese)
[3]康淑荷,师永清,杨彩霞.粉枝莓根中的三萜及甾体化合物[J].中药材,2008,31(11):1669-1671Kang S H,Shi Y Q,Yang C X.Triter penoids and steroids of root of Rubus biflorus[J].Journal of Chinese Medicinal Materials,2008,31(11):1669-1671(in Chinese)
[4]康淑荷,郑尚珍.粉枝莓中的两种新黄酮成分[J].药学学报,2007,42(12):1288-1291Kang S H,Zheng S Z.Two new flavones from Rubus biflorus Buch[J].Acta Pharmaceutica Sinica,2007,42(12):1288-1291(in Chinese)
[5]李维林,蒋振军,蒋续银.粉枝莓资源开发利用研究[J].中国水土保持,1994(6):37-39Li W L,Jiang Z J,Jiang X Y.Study on exploitation and utilization of Rubus biflorus Buch resources[J].Soil and Water Conservation in China,1994(6):37-39(in Chinese)
[6]李钰莹,范希峰,侯新村,朱毅,赵春桥,武菊英.利用ISSR和SSR标记分析芒荻类植物资源遗传多样性[J].中国农业大学学报,2014,19(2):21-27Li Y Y,Fan X F,Hou X C,Zhu Y,Zhao C Q,Wu J Y.Genetic diversity of Miscanthus&Triarrhena based on ISSR and SSR analysis[J].Journal of China Agricultural University,2014,19(2):21-27(in Chinese)
[7]戴智慧,龙骏,俞雷民,倪穗.宁波地区野生换锦花遗传多样性的ISSR分析[J].核农学报,2016,30(10):1898-1905Dai Z H,Long J,Yu L M,Ni S.Genetic diversity of wide Lycorissprengeri populations from different habitats of Ningbo based on ISSR analysis[J].Journal of Nuclear Agricultural Sciences,2016,30(10):1898-1905(in Chinese)
[8]Nudin N F H,Ali A M,Ngah N,Mazlan N Z,Mat N,Ghani M N A,Alias N,Zakaria A J,Jahan M S.ISSR marker-assisted genetic diversity analysis of Dioscorea hispida and selection of the best variety for sustainable production[J].Comptes Rendus Biologies,2017,340(8):359-366
[9]Oliveira L A R,Machado C A,Cardoso M N,Oliveira A C A,Amaral A L,Rabbani A R C,Silva A V C,Ledo A S.Genetic diversity of Saccharum complex using ISSR markers[J].Genetics and Molecular Research,2017,16(3):gmr 16039788
[10]孙淑霞,李靖,陈栋,谢红江,涂美艳,江国良.ISSR分子标记技术在桃品种鉴定中的应用[J].中国农学通报,2011,27(4):173-177Sun S X,Li J,Chen D,Xie H J,Tu M Y,Jiang G L.Molecular identification of peach germplasm by ISSR Markers[J].Chinese Agricultural Science Bulletin,2011,27(4):173-177(in Chinese)
[11]戴正,陈力耕,童品璋.香榧品种遗传变异与品种鉴定的ISSR分析[J].园艺学报,2008(8):1125-1130Dai Z,Chen L G,Tong P Z.Genetic variation and fingerprinting of Torreyagrandis cultivars detected by ISSR markers[J].Acta Horticuhurae Sinic,2008(8):1125-1130(in Chinese)
[12]刘崇琪,陈学森,王金政,陈晓流,王海波,田长平,吴传金.新疆野生樱桃李果实部分表型性状的遗传多样性分析[J].园艺学报,2008,35(9):1261-1268Liu C Q,Chen X S,Wang J Z,Chen X L,Wang H B,Tian C P,Wu C J.Studies on genetic diversity of phenotypic traits in wild M yrobalan Plum(Prunus cerasifera Ehrh)[J].Acta Horticuhurae Sinic,2008,35(9):1261-1268(in Chinese)
[13]周莉,刘莉,刘翔,徐伟欣,张平,李志文.不同变种甜瓜果实成熟性状及其香气成分的多样性分析[J].华北农学报,2013,28(3):102-108Zhou L,Liu L,Liu X,Xu W X,Zhang P,Li Z W.Diversity analysis of aroma and other mature fruit characters among different varieties of Cucumis melo L[J].Acta Agriculturae Boreali-Sinica,2013,28(3):102-108(in Chinese)
[14]Korbin M,Kuras A,Straczynska K,Orzel A,Danek J.Biotechnologicaldirections in polish breeding of Rubus[C].In:ISHS Acta Horticulturae 585:9th International Rubus and Ribes Symposium.Pucón:International Society for Horticultural Science,2008(1):133-139
[15]Hong Y P,Kin M J,Hong K N.Genetic diversity in natural populations of two geographic isolates of Korean black raspberry[J].Journal of Horticultural Science and Biotechnology,2003,78(3):350-354
[16]李金璐,王硕,于婧,王玲,周世良.一种改良的植物DNA提取方法[J].植物学报,2013,48(1):72-78Li J L,Wang S,Yu J,Wang L,Zhou S L.A modified CTAB protocol for plant DNA extraction[J].Chinese Bulletin of Botany,2013,48(1):72-78(in Chinese)
[17]尹燕雷,苑兆和,冯立娟,招雪晴,王金政,王超.不同栽培条件下凯特杏果实发育过程中香气成分的GC/MS分析[J].林业科学,2010,46(7):92-98Yin Y L,Yuan Z H,Feng L J,Zhao X Q,Wang J Z,Wang C.GC/MS analysis of aromatic components in Katy apricot fruit in various developmental periods under different cultivation condition[J].Scientia Silvae Sinicae,2010,46(7):92-98(in Chinese)
[18]孔祥琪,施瑞城,张彦军,谭乐和,皋香.气相色谱质谱联用技术分析热处理前后番木瓜汁挥发性香气[J].食品与发酵工业,2016,42(1):189-194Kong X Q,Shi R C,Zhang Y,Tan L H,Gao X.HS-SPME/GCMS analysis of the aromatic components of papayajuice before and after the heat treatment[J].Food and Fermentation Industries Editorial Staff,2016,42(1):189-194(in Chinese)
[19]Nybom H.Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants[J].Molecular Ecology,2004,13:1143-1155
[20]和志娇,和加卫,程在全,杨正松,杨洪涛,和文佳.三叶悬钩子自然居群遗传多样的ISSR分析[J].园艺学报,2012,39(11):2142-2150He Z J,He J W,Cheng Z Q,Yang Z S,Yang H T,He W J.Genetic diversity in the natural populations of Rubus delavayi demonstrated by inter-simple sequence repeats[J].Acta Horticuhurae Sinic,2012,39(11):2142-2150(in Chinese)
[21]和志娇,和加卫,杨正松,程在全,杨燕林,李燕,黄杏娥.红泡刺藤居群的遗传多样性研究[J].西北植物学报,2011,31(12):2406-2411He Z J,He J W,Yang Z S,Cheng Z Q,Yang Y L,Li Y,Huang X E.ISSR anlysis of genetic diversity of Rubus niveus Thunb populations[J].Acta Botanica Boreali-Occidentalia Sinica,2011,31(12):2406-2411(in Chinese)
[22]Wright S.The genetical structure of populations[J].Annals of Eugenices,1951,15(4):323-354