摘要
利用转录组数据开发SSR标记是一种经济高效的DNA分子标记开发策略。本研究利用高通量测序技术开发楸树(Catalpa bungei)EST-SSR标记,了解其在转录组序列中的分布类型,并利用41对多态性较好的引物对来自安徽(AH)、河南(HN)、湖北(HB)和山东(SD)的4个楸树居群的48个无性系的遗传多样性进行分析。结果表明:在大于1kb的14634条序列中,共鉴定出3999个SSR位点,580条序列包含2个及以上位点;单核苷酸重复(1957,48.94%)是最常见的SSR类型,其次是二核苷酸重复(1164,29.11%),三核苷酸重复(834,20.86%),四核苷酸重复(41,1.03%)和五核苷酸重复(3,0.08%);共扩增出243个等位基因,每个位点的等位基因数为3~13个,平均为4.85个,观测杂合度为0.14?1.00,平均为0.63,期望杂合度为0.42?0.84,平均为0.67,大部分位点偏离哈迪-温伯格平衡;AH和HN居群在有效等位基因数和期望杂合度的数值较高,表明其遗传多样性较丰富;分子方差分析(AMOVA)表明,遗传变异主要发生在居群内。本研究为楸树及同属其他树种种质资源鉴定、遗传多样性提供依据,且在指导楸树良种选育等方面具有重要的意义。
Development of SSR markers based on transcriptome data is a fast,cost-effective strategy for DNA molecular markers.In this study,we used the high-throughput sequencing technology to develop EST-SSR markers in Catalpa bungei.Distribution patterns of the markers were analyzed in transcriptome sequences.The genetic diversity of 48 clones of four populations in C.bungei collected from Anhui province(AH),He'nan province(HN),Hubei province(HB)and Shandong province(SD)was analyzed using 41 highly polymorphic makers.The results showed that:A total of 3999 SSR sites were identified from the 14634 sequences larger than 1 kb.580 sequences contained two or more SSRs.The most common SSR types were single-nucleotide repeats(1957,48.94%),followed by di-(1164,29.11%),tri-(834,20.86%),tetra-(41,1.03%),penta-(3,0.08%).A total of 243 alleles were amplified.The average number of alleles per locus was 4.85 with a range of 3?13.The range of observed heterozygosity was 0.14?1.00 with the mean of 0.63.The range of expected heterozygosity was 0.42?0.84 with the mean of 0.67.Most of the loci were deviated from Hardy-Weinberg balance.The number of effective alleles and expected heterozygosity in AH and HN population was highest indicating that their genetic diversity is relatively abundant.Analysis of molecular variance(AMOVA)showed that most of genetic variation resulted from within-population.This study provides a basis for the identification and genetic diversity of C.bungei and other tree species germplasm resources,and is of great significance in guiding the breeding of eucalyptus species.
作者
杨丹丹
马玲玲
李亚
王良桂
王鹏
Yang Dandan;Ma Lingling;Li Ya;Wang Lianggui;Wang Peng(College of Landscape Architecture,Nanjing Forestry University,Nanjing,210037;Institute of Botany,Jiangsu Province and Chinese Academy of Sciences,Nanjing,210014)
出处
《分子植物育种》
CAS
CSCD
北大核心
2020年第4期1216-1223,共8页
Molecular Plant Breeding
基金
国家自然科学基金项目(31200509)
国家“十三五”课题楸树良种选育与高效培育技术研究(2017YFD0600604)
江苏省重点研发计划(现代农业)项目(BE2016384,BE2017372)
江苏省林业三新工程(LYKJ[2017]05)共同资助。
