家蚕不同品种对人工饲料摄食性遗传模式及食性相关基因定位研究

发布时间：2018-02-12 03:00

本文关键词： 家蚕摄食性遗传模式 SSR分子标记人工饲料食性相关基因　出处：《山东农业大学》2017年硕士论文　论文类型：学位论文

【摘要】：用人工饲料养蚕是今后我国蚕业发展的方向,但缺乏人工饲料适应性蚕品种严重制约了该技术的生产应用。探明家蚕对人工饲料摄食性的遗传方式及分子机理,对于选育人工饲料适应性蚕品种及改良饲料配方均具有重要的理论和实践意义。为此,本研究以对人工饲料摄食性不同的家蚕品种(品系)为材料,探讨了不同蚕品种对人工饲料摄食性的遗传模式,并利用SSR标记和图位克隆技术对家蚕食性相关的基因进行了定位,采用半定量PCR技术测定了有差异的分子标记间14个预测基因在广食性品种中广04(ZG04)与低食性品种鲁七(L7)间的表达差异。主要获得如下结果:1.利用对人工饲料摄食性存在极显著差异的3对家蚕品种(品系)为材料,分别组配成P1、P2、F1、F2及BC1共10个世代,以人工饲料育收蚁36?h疏毛率为摄食性指标,应用联合尺度检验等方法进行摄食性遗传模式研究。结果表明:通过多代歧化选择选育的菁松A、菁松B的高食性与低食性品系对人工饲料的摄食性符合加性遗传;广食性品种ZG04对低食性品种L7为显性-加性-母性遗传模式。2.用ZG04和L7作为亲本组建F1代及BC1F回交群体(ZG04×L7)×L7和BC2F回交群体[(ZG04×L7)×L7]×L7,利用SSR技术对人工饲料摄食性相关基因进行定位和连锁分析,结果显示在第7连锁群上发现2个与食性连锁的SSR多态性标记S0709和S0710,遗传距离为1.1 cM。利用SSR Hunter 1.3软件在S0709和S0710之间搜索潜在的SSR位点,并根据PCR扩增结果和摄食性表现型对比,结果找到5个新的多态性SSR标记,这5个标记在BC2F群体中的电泳图谱中,低食性个体与L7基因型与表现型一致。利用KAIKObase家蚕数据库中的有关信息分析S0709-S0710间的碱基序列及预测基因,结果在S0709与S0710间有14个预测基因。根据标记位点与基因位点,绘制出14个基因与7个分子标记的连锁图。遗传连锁图的遗传距离为226720 bp,Gene8与S3标记间距离最近,为241 bp。3.分别用ZG04与L7的蚁蚕、2龄起蚕、3龄起蚕、3龄蚕头部为材料,采用半定量PCR技术,测定在S0709-S0710之间预测出的13个基因的表达情况。结果表明,其中10个基因在ZG04与L7间的表达无显著差异,但Gene1和Gene3在ZG04的蚁蚕、2龄起蚕、3龄起蚕及3龄蚕头部中的表达水平均显著高于L7。Gene4在ZG04中不表达,但是在L7不同龄期均有表达。关于这些基因的具体功能及其与食性的关系有待进一步研究。
[Abstract]:Silkworm rearing with artificial feed is the direction of sericulture development in China in the future, but the lack of artificial feed adaptive silkworm varieties seriously restricts the production and application of this technology. The genetic mode and molecular mechanism of silkworm feeding on artificial feed are investigated. It has important theoretical and practical significance for breeding artificial feed adaptive silkworm varieties and improving feed formula. The genetic model of feeding on artificial diet of different silkworm varieties was discussed, and SSR markers and map cloning techniques were used to localize the genes related to silkworm feeding. Semiquantitative PCR technique was used to determine the expression differences of 14 predictive genes among differential molecular markers between Guang04ZG04 and L7. The main results were as follows: 1. Three pairs of silkworm varieties (lines) with very significant differences were used as materials. A total of 10 generations of P1P2P2F1F1F 2 and BC1 were prepared, and 36? H the hair thinning rate is a feeding index, The genetic model of ingestion was studied by means of joint scale test. The results showed that the high and low diet lines of Pinus cyanine B selected by multiple generation disproportionation were in accordance with additive inheritance. ZG04 was dominant additive-maternal inheritance model for low feeding variety L7. Using ZG04 and L7 as parents, F1 generation and BC1F backcross population (ZG04 脳 L7) 脳 L7 and BC2F backcross population [ZG04 脳 L7] 脳 L7 were established. Using SSR technique, the genes related to feed intake were studied. Location and linkage analysis, The results showed that two SSR polymorphic markers S0709 and S0710 were found in the 7th linkage group, with a genetic distance of 1.1 cm. SSR Hunter 1.3 software was used to search for potential SSR loci between S0709 and S0710. The results of PCR amplification and feeding phenotype were compared. Results five new polymorphic SSR markers were found in the electrophoretic map of BC2F population. The results showed that there were 14 predicted genes between S0709 and S0710 by using the relevant information in KAIKObase silkworm database, the results showed that there were 14 predicted genes between S0709 and S0710, according to the marker loci and gene loci, there were 14 predicted genes in S0709 and S0710, and there were 14 predicted genes in S0709 and S0710. A linkage map of 14 genes and 7 molecular markers was drawn. The genetic distance of genetic linkage map was 226720 BP Gene8 and S3 markers, the distance was 241bp.3.The ZG04 and L7 ants were used as materials from the 2nd instar to the 3rd instar of silkworm, and from the 3rd instar to the 3rd instar of silkworm. The expression of 13 genes between S0709-S0710 and S0709-S0710 was determined by semi-quantitative PCR. The results showed that there was no significant difference in the expression of 10 genes between ZG04 and L7. However, the expression levels of Gene1 and Gene3 were significantly higher in the second instar silkworm from the 2nd instar and the 3rd instar silkworm head than in L7.Gene4 in ZG04. However, the specific function of these genes and their relationship with diet need to be further studied.
【学位授予单位】：山东农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S883.9

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