摘要
本文利用简并引物PCR和RACE技术,从黄瓜中克隆到CsMAPK cDNA的保守序列,运用NCBI中Blastx进行氨基酸同源比对结果表明,克隆的这个保守区序列与其他植物的MAPK基因有很高的同源性(>80%),其中与拟南芥ATMPK20基因同源性最高(90%)。同源性比对结果表明已成功克隆到了黄瓜CsMAPK保守序列。另外,利用软件做出的一个保守区同源进化树,同样说明克隆到了黄瓜CsMAPK保守序列。为进一步验证该序列的功能,基于病毒诱导基因沉默的原理,将构建烟草花叶病毒载体,建立黄瓜RNA干扰体系,对该基因进行功能验证。
由于长春花具有很强的观赏价值,开花时期长,品种多样,花色鲜艳,将广泛应用于2010年上海市世博会。本文利用分子标记分析了长春花种质资源遗传多样性,为长春花的遗传育种研究提供一定参考。本论文优化了长春花ISSR-PCR体系,并利用ISSR分子标记技术对40个长春花种质资源进行了遗传关系分析,多态性PCR扩增结果表明:在115个位点,其中多态位点78个,多态位点百分率为67.8%,多态性较高;POPGENE32软件计算结果表明,40个长春花品种平均有效等位基因数为1.6384, Nei遗传多样性指数平均为0.3735,平均Shannon信息指数为0.5546;应用NCSYS软件计算得到品种间的遗传相似系数介于0.150～0.900,平均为0.584。通过聚类分析将这40个长春花种质分成7组,该聚类结果与以前根据形态进行的分类结果有极大的相似性。该研究将为长春花品种鉴定和品种保护提供依据,为长春花种质资源亲缘关系的确定及杂交育种提供理论依据。
The conserved cDNA domain of CsMAPK was obtained in cucumber (Cucumis sativus L.) by using degenerate primer and RACE. Bioinformatic analysis showed show that this conserved domains sequence with the MAPK gene of the other plants has high homology (> 80%), which with the Arabidopsis genes ATMPK20 highest homology (90%). The results show that we have been successfully obtained conserved domains of CsMAPK gene in cucumber. In addition, the homology evolutionary tree showed the sequence is conserved domain of CsMAPK gene in cucumber.Further, to verify the functions of sequence, we will construct tobacco mosaic virus expresstion vector based on the principle of virus-induced gene silencing pre-built, and set up the transient expression system for verifying the function of gene sequence.
Because Catharanthus roseus has strong ornamental value, flowering period long, varieties diverse, will be used in 2010 Shanghai EXPO extensively. This research analyzes the genetic diversity of germplasm resources in Catharanthus roseus by molecular markers. It provides a reference for genetic breeding in Catharanthus roseus.
Genetic relationships between forty varieties of Catharanthus roseus which are suitably cultivated in Shanghai were analyzed by ISSR (inter-simple sequence repeat) markers. Genetic diversity and similarity were analyzed in molecular level and the purpose of this study is further perfecting the evaluation of species resource in Catharanthus roseus. The result showed that a total of 115 bands based on 20 primers were detected, out of which 78 were polymorphic (67.8%). As analyzed by POPGENE32, the average value of effective number of alleles, Nei’s gene diversity and Shannon’s information index were 1.6384, 0.3735, 0.5546, respectively. As analyzed by NCSYS, the Nei’s genetic similarity coefficient between cultivars ranged from 0.150 and 0.900, mean is 0.584. Clustering analysis showed that the 40 varieties of C. roseus could be distinctively classified into 7 groups. This is in accord with previous result made by morphological study. The study will provide a basis for Catharanthus roseus varieties identify and species protection, and supply a theoretical basis for determining the genetic relationship and crossbreeding of Catharanthus roseus germplasm resources.
引文
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