摘要
该研究以‘桂粉葛1号’为材料,通过转录组测序的方法测得8.9Gb clean reads,组装成137 629个转录本,最终得到83 811个Unigene序列。进化树分析表明,葛根和苜蓿、花生聚为一支。用MISA软件在83 811个序列中检测到25 452个简单重复序列(SSR)位点,三核苷酸重复的SSR数量最多,其次是二核苷酸重复。三核苷酸重复中,(AAG)n是最普遍的重复单元(27.87%)。共设计了229对SSR引物,其中28对引物可以产生清晰的条带和丰富的多态性,被用于检测44份葛根资源的遗传多样性。在44个葛根资源的基因组DNA中共扩增出90个片段,其中89个条带有多态性,平均等位基因数为3.178 6。多态性信息量范围为0.083 0~0.774 2(平均数为0.4557)。聚类分析显示遗传相似性系数范围为0.266 7~1.000 0。这些结果提示所检测的葛根资源在DNA分子水平存在着丰富的遗传多样性。当阈值为0.58时,44个资源可以划分为2个类群,且44份资源的类群划分与地理来源之间没有直接关系,但这些标记将是葛根遗传多样性研究可用的基因组资源。
In this study,137 629 transcripts were assembled from 8.9 Gb of clean Illumina DNA sequencing read data,yielding 83 811 unigene sequences from Pueraria thomsonii ‘No.1’.A phylogenetic analysis indicated that Pueraria lobata and Medicago sativa clustered together with Arachis hypogaea (peanut).We detected 25 452 SSR loci in the 83 811 assembled unigenes using MISA software.Tri-nucleotide repeats were the most abundant followed by di-nucleotide repeats.Among the tri-nucleotide motifs,(AAG) n (27.87%),was the most common repeat unit.A total of 229 SSR primer pairs were designed,and 28 markers that gave clear,polymorphic amplification products were used to analyze the genetic diversity within a panel of 44 Pueraria accessions.Ninety SSR fragments,consisting of 89 alleles,were amplified from genomic DNA of the 44 accessions.The average allele number is 3.178 6.Polymorphic information content (PIC) values ranged between 0.083 0 and 0.774 2 (mean = 0.455 7).Cluster analysis showed that the genetic similarity coefficients among the accessions ranged from 0.266 7 to 1.000 0.These results suggest that the detected P.lobata resources have abundant genetic diversity at DNA molecular level.The in-group similarity coefficient (0.58) was observed in the 44 germplasm accessions,and all accessions could be clearly divided into two groups.The clustering results of tested P.lobata resource did not show clear correlation to their geographic origin.These markers are reliable genomic resource for genetic diversity analysis in Pueraria .
作者
肖亮
尚小红
曹升
谢向誉
曾文丹
严华兵
XIAO Liang;SHANG Xiaohong;CAO Sheng;XIE Xiangyu;ZENG Wendan;YAN Huabing(Cash Crops Research Institute,Guangxi Academy of Agricultural Sciences,Nanning 530007, China;Guangxi Crop Genetic Improvement and Biotechnology Key Lab,Nanning 530007, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2019年第1期59-67,共9页
Acta Botanica Boreali-Occidentalia Sinica
基金
广西科技计划工程(桂科AD17195072)
广西科技重要工程(桂科AA17202027-4)
广西自然科学基金青年基金(2018GXNSFBA294001)~~
关键词
葛
SSR标记
资源收集
遗传多样性
Pueraria
SSR markers
germplasm collection
genetic diversity