芝麻菜种质多样性研究与抗逆基因挖掘

发布时间：2018-06-01 10:49

本文选题：芝麻菜 + 农艺性状　；参考：《湖北大学》2016年博士论文

【摘要】：芝麻菜是十字花科重要的植物遗传资源,耐寒性较强并且耐盐碱、旱涝、瘠薄,对环境要求低,是北方干旱地区一种重要油料作物。在本研究中我们考察了 11份芝麻菜材料的农艺性状及各农艺性状之间的相关关系及其对单株产量的通径和回归关系。偏相关,逐步回归和通径分析表明全株角果数(X7),每角果粒数(X8),千粒重(X9)和主花序角果数(X6)对单株产量(Y)贡献为正且极显著,二次分枝数(X4)和株高(X1)贡献为正且显著,而分枝高(X2)对单株产量贡献为负且显著(P0.05)。农艺性状对单株产量(Y)的回归方程为 Y=-2.134 + 0.010X1-0.011X2 + 0.049X4+ 0.028X6+ 0.019X7 + 0.056X8 +0.465X9。全株角果数对单株产量直接贡献最大,其次是每角果粒数,主花序角果数,千粒重,二次分枝数和株高。二次分枝数通过全株角果数对单株产量间接影响最大,其次是株高、主花序角果数和每角果粒数。分枝高和千粒重通过全株角果数对单株产量表现轻微的负面影响。首次发现芝麻菜黄籽材料,筛选到一些单株产量高、植株高、抗病性好等优良性状的芝麻菜材料。研究发现在20%PEG处理下,芝麻菜根长受到促进,但是芽长和苗鲜重受到抑制,其他耐旱指数影响不大。芝麻菜RRL,RSL,RFSW,RSV比甘蓝型油菜品种中双9号高得多,但RSG,RMGT,RGV和RGDR差异不大。主成分分析表明前三个成分解释了85.464%的总变异。PC1解释了43.888%的总变异,PC2解释了27.846%的总变异,PC3解释了 13.73%的总变异。UPGMA聚类分析表明,测试芝麻菜材料可以聚类为5个大群。第一个大群基本上对干旱敏感;第二个大群总体是轻微-中度抗旱。第三大群包括具有较高的PC1和较低的PC2,总体比较抗旱。第四大群PC1总体偏低,对干旱高度敏感。第五大群包总体表现高度抗旱。采用SSR分子标记,NTSY聚类分析把199个芝麻菜品系和一个埃塞俄比亚芥品系分为7大类群,有4份芝麻菜品系和埃塞俄比亚芥游离在外。三维主坐标分析表明芝麻菜可以分为2大类群,其中C9、C86、C132游离在外。二维主坐标分析大致将芝麻菜分为4个主要的类群,其中C9、C14、C132游离在外。芝麻菜萌发5天后幼苗用液体MS/20%PEG溶液处理,提取总RNA进行转录组测序并分析。KEGG富集分析表明,在抗旱芝麻菜而不是敏感芝麻菜中,涉及到α-亚麻酸代谢、酪氨酸代谢、苯丙氨酸、酪氨酸、色氨酸生物合成,半乳糖代谢,异喹啉类生物碱生物合成,莨菪烷、哌啶及吡啶类生物碱生物合成,矿物质吸收等代谢途径的差异基因,全部上调表达;而涉及到核糖体,核糖体生物发生,嘧啶代谢,RNA降解,乙醛酸及二羧酸代谢,氨酰基-tRNA生物合成,柠檬酸循环,双组分系统,甲烷代谢,光合器官碳固定,细胞外基质受体相互作用等代谢途径的差异基因,全部下调表达。BLAST分析表明,PEG胁迫下在抗旱芝麻菜中有47个Unigene上调最显著(log2 ratio≥8),包括3个海藻糖磷酸磷酸酶基因,2个丙二烯氧化物酶基因,2个乙烯响应转录因子基因,2个低温诱导蛋白基因。与此同时,特异下调最显著(log2Fold Ratio≤-8)基因275个,其中包括38个核糖体蛋白基因,4个抗病蛋白基因,2个羧肽酶基因,2个细胞色素b5基因,4个丝/苏氨酸-蛋白激酶基因,4个热休克蛋白基因,3个谷氨酸脱氢酶基因,2个赖氨酸--tRNA连接酶基因,2个核苷二磷酸激酶基因,2个过氧化物酶基因,2个焦磷酸-活化液泡膜质子泵基因。克隆了芝麻菜NAS和DREB2A基因并构建过表达载体。建立了芝麻菜高效再生体系和遗传转化体系。上述研究结果对于芝麻菜种质资源利用,抗旱机制解析与抗旱基因挖掘,具有重要意义。
[Abstract]:Sesame is an important plant genetic resource of Cruciferae, with strong cold tolerance and salt tolerance, drought and waterlogging, barren and low environmental requirements. It is an important oil crop in the arid areas of northern China. In this study, the relationship between Agronomic Characters and agronomic characters of 11 sesame vegetables and the path to yield per plant were investigated in this study. Partial correlation, stepwise regression and path analysis showed that the total number of kernels (X7), the number of seed per kernels (X8), 1000 grain weight (X9) and the number of main inflorescence (X6) were positive and significant to the yield of single plant (Y), and the contribution of two branches (X4) and plant height (X1) was positive and significant, and the contribution of branch high (X2) to the yield of single plant was negative and significant (P0.05). The regression equation for single plant yield (Y) was Y=-2.134 + 0.010X1-0.011X2 + 0.049X4+ 0.028X6+ 0.019X7 + 0.056X8 +0.465X9. total fruit number directly contributed to yield per plant, followed by the number of kernels per plant, the number of main inflorescences, 1000 grain weight, two branches and plant height. The two branch number through the whole plant kernels per plant yield. The second was plant height, the number of main inflorescences and the number of kernels per plant, the number of branch height and 1000 grain weight had a slight negative effect on the yield of single plant. It was first found that sesame seed yellow seed material was found to have high yield, high plant and good disease resistance. The research found in 20%PEG Under treatment, the root length of sesame was promoted, but bud length and seedling fresh weight were inhibited, and other drought resistance index had little influence. Sesame RRL, RSL, RFSW, RSV were much higher than those of Brassica napus, but RSG, RMGT, RGV and RGDR were not very different. Principal component analysis showed that the first three components explained 85.464% of the total variation.PC1 explained 43.888%. The total variation, PC2 explained 27.846% of the total variation, PC3 explained 13.73% of the total variation.UPGMA cluster analysis showed that the test of sesame vegetable material could be clustered into 5 large groups. The first large group was basically sensitive to drought; second large groups were mild to moderate drought resistance. Third groups including higher PC1 and lower PC2, overall comparison of drought resistance. Fourth large groups of PC1 were low and sensitive to drought. Fifth large groups were highly drought resistant. Using SSR molecular markers, 199 sesame varieties and one Ethiopia mustard line were divided into 7 groups, 4 sesame vegetable lines and Ethiopia mustard were out. Three dimensional principal coordinate analysis showed that sesame vegetables were available. It is divided into 2 groups, of which C9, C86 and C132 are free. The two dimensional principal coordinate analysis generally divides the sesame vegetables into 4 main groups, of which C9, C14 and C132 are free. The seedlings were treated with liquid MS/20%PEG solution after 5 days of germination, and the total RNA was extracted from the transcriptional group and analyzed by.KEGG enrichment analysis. In sensitive sesame, it involves the metabolism of alpha linolenic acid, tyrosine metabolism, phenylalanine, tyrosine, tryptophan biosynthesis, galactose metabolism, biosynthesis of isoquinoline alkaloids, scopolane, biosynthesis of biosynthesis of biosynthesis of piperidine and pyridine alkaloids, mineral absorption and other metabolic pathways, all of which are up-regulated and involved in ribosomes and nuclei. Glyoxylic biogenesis, pyrimidine metabolism, RNA degradation, glyoxylic acid and two carboxylic acid metabolism, aminoacyl -tRNA biosynthesis, citric acid cycle, bicomponent system, methane metabolism, photosynthetic organ carbon fixation, extracellular matrix receptor interaction and other metabolic pathways, all down regulated.BLAST analysis showed that under PEG stress in drought resistant sesame vegetables 47 Unigene up-regulated (log2 ratio > 8), including 3 trehalose phosphate phosphatase genes, 2 prodiene oxide enzyme genes, 2 ethylene responsive transcription factor genes, 2 low temperature induced protein genes, and 275 of the most significant down-regulation (log2Fold Ratio < -8) genes, including 38 ribosome protein genes, 4 resistance genes. The disease protein gene, 2 carboxypeptidase gene, 2 cytochrome b5 gene, 4 silk / threonine protein kinase gene, 4 heat shock protein gene, 3 glutamic dehydrogenase gene, 2 lysine --tRNA ligase gene, 2 nucleoside two phosphate kinase gene, 2 peroxidase gene, 2 pyrophosphate activated vacuolar proton pump gene. The NAS and DREB2A genes of sesame vegetable and the expression vector were constructed. The high efficiency regeneration system and genetic transformation system of sesame vegetables were established. The results are of great significance to the utilization of germplasm resources of sesame vegetables, the analysis of drought resistance mechanism and the mining of drought resistant genes.
【学位授予单位】：湖北大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：Q943.2;S565.9

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