摘要
中国板栗遗传多样性和遗传结构的AFLP分析
及其品种数据库的建立
中国板栗为壳斗科（Fagaceae）栗属（Castanea Miller）植物，在我国南北皆有分布，是我国的特色果树。
应用扩增性片段长度多态性(Amplified Fragment Length Polymorphism, AFLP)技术分析了中国板栗45个主栽品种的遗传多样性。应用9对引物组合，共产生260条条带，其中158条为多态性条带，多态性条带比率为60.77％；各品种间遗传距离差异很大，从0.1484到0.9225；平均遗传多样性h=0.1895，Shannon 信息指数为0.2894，结果表明板栗品种遗传多样性丰富，遗传多样性水平较高。在板栗4个居群中，湖北居群的遗传多样性最高，h=0.1830；而北京居群的遗传多样性最低，h=0.1631。居群内的遗传变异远高于居群间的遗传变异，居群间分化程度低。并且4个居群的基因流值都大于2，说明基因交流防止了由遗传漂变引起的居群间的遗传分化。
应用空间自相关分析方法对4个板栗居群AFLP遗传变异的空间结构进行研究，分析其遗传变异的分布特征。结果显示表明取样地的板栗居群缺乏空间结构，AFLP位点变异为随机分布的空间模式。
本试验优化了适合AFLP技术的板栗品种叶片的DNA提取、纯化方法，同时确立了应用AFLP技术研究板栗的反应体系。为今后进一步研究板栗的其他遗传性状奠定了基础。本研究表明AFLP技术稳定可靠、重复性好。
广泛收集板栗品种资源信息，初步确定以307个品种的信息为基础，利用Access 2000软件，建立了由表、查询、窗体及报表组成的中国板栗栽培品种资源数据库，分别记录了品种的产地、生物学特性、植物学特性、果实特性等要素的字段66个，为板栗工作者了解板栗品种资源情况，查询及利用提供了有效、方便快捷的工具。
Chestnut (Castanea mollissima Blume) is one of the most important trees that are indigenous to a large area of China and have long been cultivated，and is characteristic fruit tree in China, which can be found widely in northern and southern China.
Genetic diversity and population structures were assessed in cultivars of Chinese chestnut by AFLP (Amplified Fragment Length Polymorphism) analysis. 9 primer combinations were used , and a total of 260 bands were generated, including158 of the polymorphic bands, so the polymorphic percentage was 60.77%. Genetic distance ranged from 0.1484 to 0.9225 among cultivars, and averaged genetic diversity was found, being h=0.1895, which could fully showed that there was rich genetic diversity and higher one. Genetic diversity among populations ranged from 0.1631 to 0.1803, respectively for Beijing population and for Hubei population. The data of the relative magnitude of genetic differentiation among populations showed that the genetic variation within population was higher than that of among population, so the degree of the genetic variation among population was lower. And gene flows of 4 populations were over 2, which suggested that there existed sufficient genetic exchanges to prevent the genetic differentiation made by genetic drift among population. Meanwhile the distribution pattern among 45 cultivars was examined by spatial autocorrelation analysis, showing that the populations took on randomly distributed spatial pattern, but partial locus displayed some kind of spatial structure.
The reaction system of AFLP technique that was fit for chestnut was optimized, including the proccedure of drawing DNA, DNA purification and the optimal reaction condition for AFLP. And this can be basis for further analyse the genetic characteristic of chestnut. The result showed that AFLP technique was a reliable and highly informative one.
The database of varietal resource of chestnut was established via applying Microsoft 2000 in order to share resource information and to make it convenient for varietal intercourse. It included six forms of sixty-six fields, ten queries, twenty-one forms and several report forms and more than 300 varieties’ information was widely collected. Meanwhile it could be viewed as a nicer ministrant flat for researchers to learn and query what they want to know about chestnut’s information.
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