岷江百合种质资源遗传多样性研究
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摘要
本文利用ISSR分子标记技术对分布于四川岷江流域的中国特有种—岷江百合(Lilium regale)的10个野生群体进行遗传多样性和遗传分化研究,主要结论如下:
(1)对165条引物进行筛选,筛出10条引物共检测到180个位点,其中179个位点是多态位点,多态位点百分率(PPL)为99.44%。
(2)在物种水平上,观察等位基因数(Na)为1.9944,有效等位基因数(Ne)为1.2916,Nei's基因多样度(h)为0.2029,Shannon多样性指数(I)为0.3398。
(3)在群体水平上,多态位点百分率(PPL)最高的是桃坪村群体(PPL=81.11%),最低的为光明乡群体(PPL=63.56%);有效等位基因数(Ne)最高的为飞虹阴坡群体(Ne=1.3211),最低的是光明乡群体(Ne=1.2011);Nei's基因多样度(h)最高的是飞虹阴坡群体(h=0.2018),最低的是光明乡群体(h=0.1397);Shannon多样性指数(I)最高的同样也是飞虹阴坡群体(I=0.3180),最低的是光明乡群体(I=0.2396)。
(4)经POPGEN32计算得到遗传分化系数(Gst=0.1894),AMOVA分子方差分析(PHIst=0.1934),基于Shannon多样性指数分析得到的遗传分化结果为((Hsp-Hpop)/Hsp=0.2038),都显示出大部分的遗传变异存在于群体之内。基因流(Nm)为2.1398,说明岷江百合野生群体间存在着一定的基因流。
(5)根据群体间的Nei's(1978)无偏遗传距离进行UPGMA聚类,10个群体可聚为三大类群:四川光明乡群体为一类群;羊毛坪群体、飞虹阳坡群体、飞虹阴坡群体为一类群;石大关群体、桃坪村群体、色尔古群体、黄岩洞群体、薛城镇群体和塔子山群体为一类群。
(6)岷江百合群体间的遗传距离与地理距离的相关性系数r=0.0936,p＞0.05,说明遗传距离与地理距离的相关性没有达到显著水平。而Mantel Test结果(r=0.5259,p＜0.05)显示,岷江百合群体间遗传距离与海拔差距之间的相关性显著。
(7)表型多样性分析:根据马氏距离用UPGMA法进行聚类分析,10个群体被聚为5大类群:光明乡群体单独聚为一类;飞虹阳坡群体也单独聚为一类;桃坪村群体与塔子山群体聚为一类;飞虹阴坡群体、石大关群体、色尔古群体和薛城镇群体聚为一类;羊毛坪群体和黄岩洞群体聚为一类。
(8)利用TFPGA的Mantel Test对马氏距离与Nei's无偏遗传距离进行相关性分析,得出二者之间的相关性系数r=0.6800、p＜0.05,说明马氏距离与Nei's无偏遗传距离的相关性达到显著水平;对马氏距离与地理距离进行相关性分析,得出两者间的相关性系数r=0.3496,p＜0.01,两者的相关性已经达到显著水平。
Genetic diversity and genetic differentiation among ten populations of Lilium regale,an endemic species in China,were analyzed using Inter Simple Sequence Repeat(ISSR)molecular technique.The main results were as follows:
(1)There were 179 polymorphic loci among 180 ISSR loci of 10 primes selected from 165 ISSR primers.The total percentage of polymorphic loci(PPL)was 99.44%.
(2)On the species level,the observed number of alleles(Na)and effective number of alleles(Ne)of Lilium regale was 1.9944 and 1.2916,respectively. Nei's gene diversity(h)and Shannon's information index(I)of Lilium regale at species level was 0.2029 and 0.3398.These data indicated that the genetic diversity at species level of Lilium regale was relatively high.
(3)On the population level,the population of Taoping Cun had the highest percentage of polymorphic loci(PPL=81.11%),while the population of Guangming Xiang was the lowest(PPL=63.56%).The population of Feihong Yinpo had the highest(Ne=1.3211),and the population of Guangming Xiang was the lowest with the effective number of alleles(Ne=1.2011).The population of Feihong Yinpo had the highest Nei's gene diversity(h= 0.2018),while the population of Guangming Xiang was the lowest(h= 0.1397).The Shannon diversity index of Feihong Yinpo population was the highest(I=0.3180),while the Shannon diversity index of Guangming Xiang population was the lowest(1=0.2396).
(4)As analyzed by POPGENE,the genetic differentiation coefficient(Gst) among populations was 0.1894.The Analysis of molecular variance (AMOVA)demonstrated that the genetic differentiation coefficient among populations 0.1934(PHIst=0.1934).And the genetic differentiation coefficient among populations((Hsp-Hpop)/Hsp)calculated by the Shannon diversity index was 0.2038.These data demonstrated that the genetic variation of within-population accounts for the most of genetic differentiation among Lilium regale populations.
(5)The dendrograms of genetic relationships among populations were constructed based on Nei's(1978)unbiased genetic distance by means of UPGMA showed that the 10 natural Lilium regale populations could be divided into 3 big groups,the population of Guangming Xiang was included in group 1,the populations of Yangmao Ping,Feihong Yangpo and Feihong Yinpo populations were in group 2,population Shida Guan,population Taoping Cun,population Seer Gu,population Huangyan Dong,population Xuecheng Zhen and population Tazi Shan were clustered into group 3.
(6)The results of correlation analysis(r=0.0936,p＞0.05),showed that there was no significant correlation existed between geographical distance and Nei's unbiased genetic distance of the 10 Lilium regale populations.However,The results of Mantel Test(r=0.5259,p＜0.05)showed that there was significant correlation between Nei's unbiased genetic distance of the Lilium regale populations and the distance of elevation.
(7)For phenotype diversity analysis,the 10 populations were divided into 5 groups according to Mahalanbis Distance by means of UPGMA.The population of Guangming Xiang was included in group 1,the population of Feihong Yangpo was included in group 2,the populations of Taoping Cun and Tazi Shan were included in group 3,the populations of Yangmao Ping, Huangyan Dong were clustered into group 4,population Feihong Yinpo, population Shida Guan,populations Seer Gu and population Xuecheng Zhen were clustered into group 5.
(8)The results of correlation analysis(r=0.6800,p＜0.05),showed that there was significant correlation existed between Mahalanbis Distance and Nei's unbiased genetic distance of the 10 Lilium regale populations by Mantel Test of TFPGA.At the same time,significant correlation was found between geographical distance and Mahalanbis distance of the 10 Lilium regale populations.
(1)对165条引物进行筛选,筛出10条引物共检测到180个位点,其中179个位点是多态位点,多态位点百分率(PPL)为99.44%。
(2)在物种水平上,观察等位基因数(Na)为1.9944,有效等位基因数(Ne)为1.2916,Nei's基因多样度(h)为0.2029,Shannon多样性指数(I)为0.3398。
(3)在群体水平上,多态位点百分率(PPL)最高的是桃坪村群体(PPL=81.11%),最低的为光明乡群体(PPL=63.56%);有效等位基因数(Ne)最高的为飞虹阴坡群体(Ne=1.3211),最低的是光明乡群体(Ne=1.2011);Nei's基因多样度(h)最高的是飞虹阴坡群体(h=0.2018),最低的是光明乡群体(h=0.1397);Shannon多样性指数(I)最高的同样也是飞虹阴坡群体(I=0.3180),最低的是光明乡群体(I=0.2396)。
(4)经POPGEN32计算得到遗传分化系数(Gst=0.1894),AMOVA分子方差分析(PHIst=0.1934),基于Shannon多样性指数分析得到的遗传分化结果为((Hsp-Hpop)/Hsp=0.2038),都显示出大部分的遗传变异存在于群体之内。基因流(Nm)为2.1398,说明岷江百合野生群体间存在着一定的基因流。
(5)根据群体间的Nei's(1978)无偏遗传距离进行UPGMA聚类,10个群体可聚为三大类群:四川光明乡群体为一类群;羊毛坪群体、飞虹阳坡群体、飞虹阴坡群体为一类群;石大关群体、桃坪村群体、色尔古群体、黄岩洞群体、薛城镇群体和塔子山群体为一类群。
(6)岷江百合群体间的遗传距离与地理距离的相关性系数r=0.0936,p＞0.05,说明遗传距离与地理距离的相关性没有达到显著水平。而Mantel Test结果(r=0.5259,p＜0.05)显示,岷江百合群体间遗传距离与海拔差距之间的相关性显著。
(7)表型多样性分析:根据马氏距离用UPGMA法进行聚类分析,10个群体被聚为5大类群:光明乡群体单独聚为一类;飞虹阳坡群体也单独聚为一类;桃坪村群体与塔子山群体聚为一类;飞虹阴坡群体、石大关群体、色尔古群体和薛城镇群体聚为一类;羊毛坪群体和黄岩洞群体聚为一类。
(8)利用TFPGA的Mantel Test对马氏距离与Nei's无偏遗传距离进行相关性分析,得出二者之间的相关性系数r=0.6800、p＜0.05,说明马氏距离与Nei's无偏遗传距离的相关性达到显著水平;对马氏距离与地理距离进行相关性分析,得出两者间的相关性系数r=0.3496,p＜0.01,两者的相关性已经达到显著水平。
Genetic diversity and genetic differentiation among ten populations of Lilium regale,an endemic species in China,were analyzed using Inter Simple Sequence Repeat(ISSR)molecular technique.The main results were as follows:
(1)There were 179 polymorphic loci among 180 ISSR loci of 10 primes selected from 165 ISSR primers.The total percentage of polymorphic loci(PPL)was 99.44%.
(2)On the species level,the observed number of alleles(Na)and effective number of alleles(Ne)of Lilium regale was 1.9944 and 1.2916,respectively. Nei's gene diversity(h)and Shannon's information index(I)of Lilium regale at species level was 0.2029 and 0.3398.These data indicated that the genetic diversity at species level of Lilium regale was relatively high.
(3)On the population level,the population of Taoping Cun had the highest percentage of polymorphic loci(PPL=81.11%),while the population of Guangming Xiang was the lowest(PPL=63.56%).The population of Feihong Yinpo had the highest(Ne=1.3211),and the population of Guangming Xiang was the lowest with the effective number of alleles(Ne=1.2011).The population of Feihong Yinpo had the highest Nei's gene diversity(h= 0.2018),while the population of Guangming Xiang was the lowest(h= 0.1397).The Shannon diversity index of Feihong Yinpo population was the highest(I=0.3180),while the Shannon diversity index of Guangming Xiang population was the lowest(1=0.2396).
(4)As analyzed by POPGENE,the genetic differentiation coefficient(Gst) among populations was 0.1894.The Analysis of molecular variance (AMOVA)demonstrated that the genetic differentiation coefficient among populations 0.1934(PHIst=0.1934).And the genetic differentiation coefficient among populations((Hsp-Hpop)/Hsp)calculated by the Shannon diversity index was 0.2038.These data demonstrated that the genetic variation of within-population accounts for the most of genetic differentiation among Lilium regale populations.
(5)The dendrograms of genetic relationships among populations were constructed based on Nei's(1978)unbiased genetic distance by means of UPGMA showed that the 10 natural Lilium regale populations could be divided into 3 big groups,the population of Guangming Xiang was included in group 1,the populations of Yangmao Ping,Feihong Yangpo and Feihong Yinpo populations were in group 2,population Shida Guan,population Taoping Cun,population Seer Gu,population Huangyan Dong,population Xuecheng Zhen and population Tazi Shan were clustered into group 3.
(6)The results of correlation analysis(r=0.0936,p＞0.05),showed that there was no significant correlation existed between geographical distance and Nei's unbiased genetic distance of the 10 Lilium regale populations.However,The results of Mantel Test(r=0.5259,p＜0.05)showed that there was significant correlation between Nei's unbiased genetic distance of the Lilium regale populations and the distance of elevation.
(7)For phenotype diversity analysis,the 10 populations were divided into 5 groups according to Mahalanbis Distance by means of UPGMA.The population of Guangming Xiang was included in group 1,the population of Feihong Yangpo was included in group 2,the populations of Taoping Cun and Tazi Shan were included in group 3,the populations of Yangmao Ping, Huangyan Dong were clustered into group 4,population Feihong Yinpo, population Shida Guan,populations Seer Gu and population Xuecheng Zhen were clustered into group 5.
(8)The results of correlation analysis(r=0.6800,p＜0.05),showed that there was significant correlation existed between Mahalanbis Distance and Nei's unbiased genetic distance of the 10 Lilium regale populations by Mantel Test of TFPGA.At the same time,significant correlation was found between geographical distance and Mahalanbis distance of the 10 Lilium regale populations.
引文
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