马褂木母树林与不同来源无性系的遗传多样性比较分析  

A Comparative Analysis of Genetic Diversity Between Seed-Tree Stand and Clones of Liriodendron spp.

作　　者：祝园青 白天道[1] 王晓波 胥猛[3] 蒋维昕[1] 黄寿先[1] ZHU Yuanqing;BAI Tiandao;WANG Xiaobo;XU Meng;JIANG Weixin;HUANG Shouxian(College of Forestry,Guangxi University,Nanning 530004,China;Xianshui Forest Farm,Quanzhou,Guangxi 541500,China;Co-Innovation Center for Sustainable Forestry in Southern China,Nanjing Forestry University,Nanjing 210037,China)

机构地区：[1]广西大学林学院/国家林业与草原局中南速生材繁育重点实验室,广西南宁530004 [2]全州县咸水林场,广西全州541500 [3]南京林业大学南方现代林业协同创新中心,江苏南京210037

出　　处：《江西农业大学学报》2021年第3期630-639,共10页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家林业和草原局中央预算内林业基本建设投资计划项目(林规发[2019]18号);广西财政林木良种专项基金(桂林计发[2018]23号);广西科技攻关项目(桂科攻10100012-2)。

摘　　要：【目的】马褂木(Liriodendron chinense(Hemsl.)Sarg.)为国家二级保护珍稀树种,因其叶形独特,干形通直,木材纹理清晰,为我国优良的用材及园林绿化树种。它与该属仅存的另一种北美鹅掌楸(L.tulipifera L.)均属古老的孑遗植物,具有重要的科学研究价值。开展马褂木天然实生苗群体和嫁接优树群体遗传结构研究对马褂木树种的保护以及开发利用具有十分重要的意义。课题利用EST-SSR分子标记技术分析中国马褂木嫁接无性系(LC)、北美鹅掌楸嫁接无性系(LT)和中国马褂木母树林(LCS)的遗传多样性,为马褂木优良无性系评价及良种繁育工作提供参考。【方法】以广西全州咸水林场的中国及北美嫁接无性系(共35株)和全州种源的马褂木采种母树林(53株)的新鲜叶片为实验材料,利用25对EST-SSR引物进行扩增,PCR产物经8%聚丙烯酰胺胶电泳及银染分析,计算其遗传多样性参数,遗传距离,开展遗传分化和遗传聚类分析,以比较不同材料的遗传多样性及亲缘关系。【结果】遗传多样性分析表明,LT的遗传多样性参数:等位基因数(na)为4.8400,有效等位基因数(ne)为3.3386,Shannon多样性指数(I)为1.2710,观测杂合度(Ho)为0.3993,期望杂合度(He)为0.6627,基因多样度(Nei)为0.6426,普遍高于LC(na=4.0400,ne=2.8289,I=1.1080,Ho=0.3596,He=0.6115,Nei=0.5910)及LCS(na=4.1600,ne=2.4502,I=0.9634,Ho=0.3135,He=0.5141,Nei=0.5088),Kruskal-Wallis检验显示组间在ne(P=0.029)、I(P=0.036)、He(P=0.012)、Nei(P=0.029)上达到显著水平;分子方差分析(AMOVA)结果显示,群体间、群体内的遗传变异分别占总变异量的14.3%和85.7%;基于个体共祖系数的遗传结构分析表明参试材料共祖群体数(K)为3,且基于Ward最小方差法对个体间遗传距离以分组数为3进行聚类分析,可将LCS、LC及LT较好地区分。【结论】参试材料总体上具有较高的遗传多样性,其中LT遗传多样性最高,其次为LC,而LCS最低;遗传[Objective]Liriodendron chinense(Hemsl.)Sarg.is a national second-class protected rare tree species,which is an excellent wood and landscaping tree in China due to its unique leaf shape,straight dry shape and clear wood texture.Both L.chinense(LC)and L.tulipifera(LT),the only two remaining plants in Liriodendron,are ancient relict plants of great scientific value.It is very important to study the genetic structure of natural seedling population and grafted tree populations of L.chinense.Given this,LC clones,LT clones and the seed-trees stand of L.chinense(LCS)from the provenance of Quanzhou were sampled and studied on their genetic diversity based on EST-SSR(expressed sequence tag-simple sequences repeats)markers technology,aiming to provide reference for superior clone selection and breeding of Liriodendron.[Method]In this study,fresh healthy leaves of a total of 35 grafted clones of L.chinense and L.tulipifera and 53 seed-trees of L.chinense stand were collected from Xianshui Forest Farm,Quanzhou,Guangxi Zhuang Autonomous Region.The total genomic DNAs of all the individuals were isolated and amplificated across 25 pairs of EST-SSR primers.The PCR products were electrophoresed by 8%polyacrylamide gel and stained with silver.The genetic diversity,genetic differentiation and genetic clustering were analyzed to reveal the genetic variation of and relationship among different tulip trees.[Result]The results showed that the genetic variation parameters of L.tulipifera(LT)clones were as follows:the number of alleles(na)was 4.8400,the number of effective alleles(ne)was 3.3386,Shannon diversity index(I)was 1.2710,observed heterozygosity(Ho)was 0.3993,expected heterozygosity(He)was 0.6627,gene diversity(Nei)was 0.6426,which were generally higher than those of L.chinense(LC)clones(na=4.0400,ne=2.8289,I=1.1080,Ho=0.3596,He=0.6115,Nei=0.5910)and LCS population(na=4.1600,ne=2.4502,I=0.9634,Ho=0.3135,He=0.5141,Nei=0.5088).Kruskal-wallis test showed significant levels in ne(P=0.029),I(P=0.036),He(P=0.012)and Nei(P=0.029)betwee

关 键 词：马褂木 北美鹅掌楸 嫁接无性系 SSR 遗传多样性 比较分析 

分 类 号：S792.21.04[农业科学—林木遗传育种]