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利用454焦磷酸测序技术分离和鉴定黄栌(漆树科)的微卫星标记

Isolation and characterization of microsatellite markers for Cotinus coggygria Scop. (Anacardiaceae) by 454 pyrosequencing.

作者信息

Wang Wei, Li Zhuo, Li Yong

机构信息

College of Forestry, Henan Agricultural University, Zhengzhou 450002, China.

College of Nature Conservation, Beijing Forestry University, Beijing 100083, China.

出版信息

Molecules. 2014 Mar 24;19(3):3813-9. doi: 10.3390/molecules19033813.

Abstract

Cotinus coggygria Scop. (Anacardiaceae) is a deciduous shrub or small tree that is native to a large area covering from southern Europe, east across central Asia, and the Himalayas in northern China. Shotgun 454 pyrosequencing was used to develop microsatellite markers from the genome of C. coggygria. In this study, 349 microsatellite loci were identified from 40,074 individual sequence reads produced by one-sixteenth run, and primer pairs were designed for these loci. To test the primer amplification efficiency, 50 microsatellite primer pairs were tested across 12 individuals from two C. coggygria populations (Wuzhi Mountain: 36°30'N, 113°39'E; Tianlong Mountain: 37°42'N, 112°26'E). Among the 50 tested primer pairs, eight were found to be polymorphic. The average allele number of the microsatellites was 3.5 per locus, with a range from two to five. The inbreeding coefficient ranged from -0.478 to 0.222. The observed and expected heterozygosities varied from 0.167 to 0.750 and from 0.163 to 0.743, respectively. This set of markers is potentially useful for assessing the genetic diversity, as well as for understanding the population structure and phylogeographical and landscape genetic patterns, of C. coggygria.

摘要

黄栌(漆树科)是一种落叶灌木或小乔木,原产于欧洲南部、横跨中亚东部以及中国北方喜马拉雅山脉的大片区域。利用鸟枪法454焦磷酸测序技术从黄栌基因组中开发微卫星标记。在本研究中,从十六分之一测序运行产生的40,074条个体序列读数中鉴定出349个微卫星位点,并为这些位点设计了引物对。为测试引物扩增效率,对来自两个黄栌种群(五指山:北纬36°30′,东经113°39′;天龙山:北纬37°42′,东经112°26′)的12个个体测试了50对微卫星引物。在50对测试引物中,发现8对具有多态性。微卫星的平均等位基因数为每个位点3.5个,范围从2到5个。近交系数范围为-0.478至0.222。观察到的杂合度和预期杂合度分别在0.167至0.750和0.163至0.743之间变化。这组标记对于评估黄栌的遗传多样性、理解其种群结构以及系统地理学和景观遗传模式可能是有用的。

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Microsatellites, from molecules to populations and back.微卫星:从分子到群体再到分子。
Trends Ecol Evol. 1996 Oct;11(10):424-9. doi: 10.1016/0169-5347(96)10049-5.
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