CAJ | 학술논문

尽管分子标记广泛用于玉米品种纯度鉴定,但由于互交种基因组具有相同的基因,现有的分子标记技术尚不能对互交种的混杂进行有效判定.本研究基于玉米互交种胚乳等位基因二倍剂量关系,利用Insertion/Deletion (InDel)标记在PCR指数扩增期对等位基因扩增相对量的观察值与理论值进行统计分析,通过概率对互交种进行判定.经过筛选,亲本B73、Mo17间存在共显性差异的8个InDel标记中,5个标记的PCR扩增质量满足互交种样本(正交种:B73×Mo17,反交种:Mo17×B73)的鉴定要求.这些标记以不同的循环数进行PCR扩增,发现37个循环是处于指数扩增期的最佳鉴定时期.同一标记在每个样本中以37个循环数进行3次扩增重复,统计分析发现:不同标记在杂交种叶片DNA中等位基因扩增相对量与理论比1的统计概率值从6.50E-08到1.000不等,证实标记在等位基因间的扩增效率并不完全一致.对互交种胚乳等位基因扩增相对量进行卡方测验,4个InDel标记能够对互交种样本进行准确区分,所得概率符合判定标准.标记110仅能准确判定正交种,对反交种样本的统计概率不满足反交种判定的概率标准.结果显示,该分子标记方法可成功用于玉米互交种的判定.
진관분자표기엄범용우옥미품충순도감정,단유우호교충기인조구유상동적기인,현유적분자표기기술상불능대호교충적혼잡진행유효판정.본연구기우옥미호교충배유등위기인이배제량관계,이용Insertion/Deletion (InDel)표기재PCR지수확증기대등위기인확증상대량적관찰치여이론치진행통계분석,통과개솔대호교충진행판정.경과사선,친본B73、Mo17간존재공현성차이적8개InDel표기중,5개표기적PCR확증질량만족호교충양본(정교충:B73×Mo17,반교충:Mo17×B73)적감정요구.저사표기이불동적순배수진행PCR확증,발현37개순배시처우지수확증기적최가감정시기.동일표기재매개양본중이37개순배수진행3차확증중복,통계분석발현:불동표기재잡교충협편DNA중등위기인확증상대량여이론비1적통계개솔치종6.50E-08도1.000불등,증실표기재등위기인간적확증효솔병불완전일치.대호교충배유등위기인확증상대량진행잡방측험,4개InDel표기능구대호교충양본진행준학구분,소득개솔부합판정표준.표기110부능준학판정정교충,대반교충양본적통계개솔불만족반교충판정적개솔표준.결과현시,해분자표기방법가성공용우옥미호교충적판정.
The molecular markers have been extensively employing in the purity identification of maize hybrid. However, the application in distinguishing the hybrids of direct and reciprocal crosses was limited because of the same genomic content contained in the two hybrids. In consideration of endosperm carrying two copies of the maternal alleles and one copy of the paternal allele in its genome, we intended to differentiate the maize hybrids of direct and reciprocal crosses using the statistical probability of chi-square test between the observed value of relative quantification of allelic amplified products (RQAAP) and the expected value indicated by Insertion/Deletion (InDel) in the exponential amplification phase of PCR. Five out of eight co-dominant polymorphic InDel markers between parents B73 and Mo17 were screened for identification of hybrids of B73×Mo17 and Mo17×B73. The amplification results of InDel markers with different PCR cycles showed that 37 cycles was the best time point for analysis during the exponential amplification stage with three technical replications. The statistical probability of RQAAP in the leaf genome of hybrid from 6.50E-08 to 1.000 exhibited that the amplification rates of different markers between alleles could be inconsistent. Statistical analyses of RQAAP in the hybrid endosperm genome of direct and reciprocal crosses revealed that four out of five markers could distinguish the direction of crosses accurately. The probability measured from the remaining one marker was capable for detecting the direct cross, but has its ineffectiveness for reciprocal cross. The results show that the proposed methodology is a promising strategy to identify maize hybrids of direct and reciprocal crosses using molecular markers.