南京林业大学.doc

南京林业大学
本科毕业设计(论文)
题目:松属种间杂种子代的分子鉴定

学院: 森环院
专业: 林学
学号: 080101111
学生姓名: 马力
指导教师: 李火根
职称: 教授
二O一二年五月
松属种间杂种子代的分子鉴定
摘要:本文采用SSR分子标记对马尾松×湿地松、湿地松×马尾松、湿地松×加勒比松共12个杂交组合的62个子代DNA样本进行杂种鉴定,同时,筛选松属种间特异性引物。主要结果如下:
利用马尾松、湿地松、加勒比松DNA样品对248对SSR引物进行筛选,从中选取了10对通用性好的SSR引物用于杂交子代鉴定。选取种间差异明显、种内保守性强的1对引物(FJ164),分别以31个马尾松样本和27个湿地松样本对其进行检测,发现FJ164号引物在马尾松中均只扩增出320bp条带,在湿地松中只扩增出340bp条带,属种特异性引物,可用于马尾松与湿地松种间鉴别。
利用复筛选出的10对通用性好且具有种间特异性SSR引物对52个待鉴定杂交子代样本及其亲本进行检测。结果表明,所用待测子代在10个检测位点上均不完全是杂交基因型,因此可以初步认为所有的待测子代都不是种间杂交子代,其真实父本可能是在控制授粉过程中混入的外来污染花粉。
关键词:马尾松;湿地松;种间杂交;分子鉴定;SSR
Molecular identification of interspecies hybridization in Pinus
Abstract
In this paper, 62 filial generation progenies from bination including Pinus massoniana Lamb.×Pinus elliottii, Pinus elliottii ×Pinus massoniana. and Pinus massoniana Lamb.×Pinus caribaea Morelet were identified by SSR markers, and species-specific SSR markers which can be used in hybrid identification were also screened. The results are as follows.
10 out of 248 pairs SSR primers were screened to be high tranferability, and one pair of SSR primer (FJ164) is species-specific. This pair of primer amplified 320bp bands in Pinus massonianaLamb and 340bp bands in Pinus elliottii. So FJ164 primer was suitable for interspecies identification between Pinus massoniana and Pinus elliottii.
10 pairs of SSR primers were used to detect 52 progenies which was deemed to be inetrspecies hybridization and their parents. Unfortunately, no progeny was proved to be interspecies hybrid. The progeny’s male parent e from unexpected pollen.
Key words: Pinus massoniana; Pinus elliottii; interspecies hybridization; Molecular identification; SSR
目录
摘要 I
Abstract II
前言 1
1文献综述 2
、马尾松遗传改良进展 2
3
5
SSR标记的原理及特点 5
6
2材料与方法 6
6
7
DNA的提取与纯化 7
PCR扩增 9
***凝胶电泳(PAGE) 10
SSR引物筛选 11
杂交子代的鉴定方法 12
12
12
13
4讨