Cha Thye San, Anne-Marie Kaben, Chuah Tse Seng
School of Fundamental Science, Universiti Malaysia Terengganu, 21030, Kuala Terengganu, Terengganu, Malaysia,
Mol Biol Rep. 2014 Feb;41(2):823-31. doi: 10.1007/s11033-013-2922-7. Epub 2013 Dec 29.
Eleusine indica is one of the most common weed species found in agricultural land worldwide. Although herbicide-glyphosate provides good control of the weed, its frequent uses has led to abundant reported cases of resistance. Hence, the development of genetic markers for quick detection of glyphosate-resistance in E. indica population is imperative for the control and management of the weed. In this study, a total of 14 specific random amplified polymorphic DNA (RAPD) markers were identified and two of the markers, namely S4R727 and S26R6976 were further sequence characterized. Sequence alignment revealed that marker S4R727 showing a 12-bp nucleotides deletion in resistant biotypes, while marker S26R6976 contained a 167-bp nucleotides insertion in the resistant biotypes. Based on these sequence differences, three pairs of new sequence characterized amplified region (SCAR) primers were developed. The specificity of these primer pairs were further validated with genomic DNA extracted from ten individual plants of one glyphosate-susceptible and five glyphosate-resistant (R2, R4, R6, R8 and R11) populations. The resulting RAPD-SCAR markers provided the basis for assessing genetic diversity between glyphosate-susceptible and -resistant E. indica biotypes, as well for the identification of genetic locus link to glyphosate-resistance event in the species.
光头稗是全球农田中最常见的杂草物种之一。尽管除草剂草甘膦对该杂草有良好的防除效果，但其频繁使用已导致大量关于抗性的报道案例。因此，开发用于快速检测光头稗种群中草甘膦抗性的遗传标记对于该杂草的防治和管理至关重要。在本研究中，共鉴定出14个特异性随机扩增多态性DNA（RAPD）标记，其中两个标记，即S4R727和S26R6976，进一步进行了序列特征分析。序列比对显示，标记S4R727在抗性生物型中出现12个碱基对的核苷酸缺失，而标记S26R6976在抗性生物型中包含167个碱基对的核苷酸插入。基于这些序列差异，开发了三对新的序列特征扩增区域（SCAR）引物。这些引物对的特异性通过从一个草甘膦敏感种群的十株单株植物以及五个草甘膦抗性（R2、R4、R6、R8和R11）种群中提取的基因组DNA进一步验证。所得的RAPD-SCAR标记为评估草甘膦敏感型和抗性光头稗生物型之间的遗传多样性提供了基础，也为鉴定该物种中与草甘膦抗性事件相关的基因座提供了基础。
