摘 要

青檀( Pteroceltis tatarinowii Maxim.)为我国特有第三纪残遗温带落叶树种，隶属于榆科( Ulmaceae) 唯一的单种属——青檀属( Pteroceltis) ，为国家三级保护植物。叶绿体微卫星是近年来发展起来的一种新型高效的分子标记技术。本研究以全国11个青檀野生居群为实验材料，根据前人已开发设计的cpSSR引物，挑选出了12对用于青檀cpSSR引物开发。结果筛选出NTCP13、NTCP3、NTCP40、ccmp5这4对引物可用于青檀cpSSR分析的引物，证明了这4对引物在青檀上的通用性。

关键词：青檀；叶绿体SSR；引物开发

The development of Pteroceltis tatarinowii Maxim cpSSR primer

ABSTRACT

Pteroceltis tatarinowii Maxim, a tertiary relict plant of a temperate deciduous species endemic to China, belonging to the only single species of the Ulmaceae-Pteroceltis. It has now been listed as a rare and endangered plant ( National Grand III) in China.Within the past decade chloroplast simple sequence repeats have been developed into a new efficient molecular marker. In this study, 11 P. tatarinowii populations were collected as experimental materials. According to the developed cpSSR primer, 12 pairs of primers has been picked out to test the efficiency in P. tatarinowii. Finally, 4 pairs of primers including NTCP13,NTCP3,NTCP40 and ccmp5 were proved as polymorphic, demonstrating the versatility of this 4 pairs of primers in P. tatarinowii .

Key words：Pteroceltis tatarinowii Maxim；chloroplast simple sequence repeat, cpSSR； development of primer

目 录

1.文献综述………………………………………………………………………………………1

1.1 叶绿体SSR的定义及用途……………………………………………………………...1

1.2 叶绿体SSR的研究进展………………………………………………………………...1

1.2.1 群体遗传结构………………………………………………….............................1

1.2.2 基因流动与进化………………………………………………….........................2

1.2.3 种质传播方式………………………………………………….............................2

1.2.4 物种演化………………………………………………….....................................3

1.2.5 胞质遗传特性………………………………………………….............................4

1.2.6 种群分类………………………………………………….....................................4

1.3 叶绿体SSR 应用的前景………………………………………………………………..5

1.3.1 SSR技术在植物中的应用研究及结论..................................................................5

1.4 针对青檀展开的研究……………………………………………………………………7

2.研究目的及意义………………………………………………………………………………8

2.1 研究目的............................................................................................................................8

2.2 研究意义............................................................................................................................8

3.研究方案………………………………………………………………………………............8

3.1 材料采集地点…………………………………………………………………………....8

3.2 研究方法…………………………………………………………………………...........10

3.2.1 DNA的提取............................................................................................................10

3.2.2 青檀cpSSR引物的获得........................................................................................10

3.2.3 PCR扩增..................................................................................................................11

3.2.4 PCR产物..................................................................................................................12

4.结果与分析………………………………………………………………………….................12

致谢……………………………………………………………....................................................16

参考文献………………………………………………………....................................................17

1.文献综述

1.1 叶绿体SSR的定义及用途

叶绿体微卫星是近年来发展起来的一种新型高效的分子标记技术，cpSSR全部是单碱基(A或T)重复类型,重复次数从10到16次不等。由于具有微卫星或称简单序列重复(simple sequence repeat, SSR)标记的优点又兼顾到叶绿体基因组(chloroplast DNA, cpDNA)的特点，所以cpSSR可用来鉴定种甚至亲缘关系很近的种，描述区域和个体水平的遗传差异，目前广泛用于植物群体遗传分析及系统发育分析研究，在更高分类水平和自然植物群体间基因漂移研究上特别有用。

1.2 叶绿体SSR的研究进展

由于具有SSR的优点，又具有叶绿体独立进化等特点，使得cpSSR技术成为一种研究生态群体遗传结构、基因流动、进化、物种演化、胞质遗传特性、种群分类等新的有用工具，得到了广泛的应用。

1.2.1 群体遗传结构