摘要
本研究旨在为蒙古莸在分子水平上提供理论依据,以便对濒危稀有种制定科学的保护策略。本研究下载GenBank数据库公布的蒙古莸叶绿体基因组信息,利用生物信息学对蒙古莸叶绿体基因组进行cpSSR分析。利用MISA软件全面搜索蒙古莸叶绿体基因组SSR分子标记,并对其分析;运用Primer Premier 3.0设计并筛选多态性较好的引物用于后续研究。在全长151 707 bp的蒙古莸叶绿体基因组中,共鉴定出31个cpSSR位点,位点间平均分布距离为4 893.77 bp,其中以单核苷酸重复为主,且主要分布于LSC区,占SSR位点总数的74.19%;其次是四核苷酸重复,分布于LSC及SSC两区,占SSR位点总数的19.35%,分布最少为三核苷酸,仅占总位点数的3.23%。将所有31个位点合成31对叶绿体微卫星引物,通过毛细管电泳检测,最终筛选出8对多态性引物。此外,挑选多态性好的马鞭草EST-SSR引物,扩增结果产生非特异性条带,且与马鞭草扩增片段范围相差较大。基于软件的参数设置,单核苷酸重复数量占比最多,三核苷酸重复占比最少;从31对引物筛选出8对多态性引物,扩增率达100%,多态率为26%;此外,马鞭草与蒙古莸的扩增条带范围相差较大,说明二者有较大差异的遗传背景。在此基础上,本研究为后续蒙古莸的种质鉴定、系统发育、群体遗传进化分析提供理论依据。
The purpose of this research is to provide theoretical basis for C. mongolica at the molecular level, so as to formulate scientific protection strategies for endangered and rare species. According to the chloroplast genome information of C. mongholica published in GenBank database and analyzing the chloroplast genome of C.mongholica by bioinformatics, in order to provide theoretical basis for germplasm identification, phylogenetic analysis and population genetic structure. The SSR molecular makers of C. mongholica in chloroplast genome of were searched and analyzed by MISA, and the primers with better polymorphism were designed by Primer Premier 3.0and screened for follow-up study. A total of 31 cpSSR loci were identified in the 151 707 bp chloroplast genome of C. mongholica. The average distance between the loci was 4 893.77 bp. Among them, the single nucleotide repeat were dominant and they were mainly distributed in the LSC region, occupying SSR sites 74.19% of the total loci.Tetranucleotide repeats were distributed in LSC and SSC, accounting for 19.35% of the total SSR loci, and were the least distributed in LSC and SSC, accounting for 3.23% of the total SSR loci. Thirty-one pairs of cpSSR primers were synthesized at all sites, and it was found by capillary electrophores is that the primers had bad polymorphisms,and eight pairs of polymorphic primers were screened out by capillary electrophoresis. In addition, selecting the EST-SSR primers of Verbena officinalis with good polymorphism, the amplification results produced non-specific bands, and the range of fragments amplified by V. officinalis was quite different. Based on the parameter settings of the software, the number of single nucleotide repetitions is the largest, and the Trinucleotide repetition is the least,eight pairs of primers with polymorphism were selected from 31 pairs of primers, and the amplification rate was 100% and the polymorphism rate was 26%. In addition, there is a big difference in the range of amplified bands between V. officinalis and C. mongolica, which indicates that they have quite different genetic backgrounds. The range of amplified bands between V. officinalis and C. mongholica is quite different, which proves that the two have quite different genetic backgrounds. Based on research, it provides a theoretical basis for the subsequent germplasm identification, phylogeny, and population genetic evolution analysis of C. mongolica.
作者
刘雅璐
叶冬梅
包文泉
林涛
范宇阳
张素毓
Liu Yalu;Ye Dongmei;Bao Wenquan;Lin Tao;Fan Yuyang;Zhang Suyu(College of Forestry,Inner Mongolia Agricultural University,Hohhot,010019)
出处
《分子植物育种》
CAS
北大核心
2024年第11期3646-3653,共8页
Molecular Plant Breeding
基金
内蒙古自治区科技计划项目(201802109)资助。
