中国香菇重要性状的全基因组关联分析

发布时间：2018-07-26 10:04

【摘要】：香菇是世界上产量第二大食用菌,仅次于双孢蘑菇。香菇产业在我国食用菌产业的整体发展中占据着举足轻重的地位。我国幅员辽阔,野生香菇种质资源极为丰富,这为香菇育种以及挖掘抗性变异提供了可能。香菇大多数农艺性状都是由微效多基因控制的数量性状,易受环境影响,仅对于表型进行遗传改良效果并不明显。以全基因关联分析为途径的分子标记辅助选择可以高效挖掘与农艺性状以及抗逆性状相关的遗传变异位点,有助于解析性状的遗传基础,也为育种提供优良的种质材料。本研究以59个野生香菇菌株群体和126(96个野生+30个栽培)个香菇菌株混合群体为实验材料,测定了野生群体的11个农艺性状,混合群体的3个抗逆性状,利用基于全基因组重测序开发的高密度SNP(单核酸多态性)标记,对供试群体进行全基因组关联分析。主要结果如下:(1)对野生香菇群体包括生育期、子实体形态和产量在内的子实体性状考察分析,结果表明:在两年的出菇试验中,产量性状均表现出较高的变异系数,其中2014年的菇数、单菇鲜重、鲜菇总重分别为81.13%、89.42%和70.57%;生育期次之,菇形性状最小,介于16.77%-71.11%之间。相关性分析表明菇形性状间存在着极显著正相关,鲜菇总重与菇数间存在极显著正相关,生育期与鲜菇总重、菌盖直径以及菌盖厚度间存在极显著负相关。除菇数外,其余10个性状在两年间均表现出显著正相关。产量性状和菇形性状的基因型与年份互作的结果对表型有着极显著影响。(2)对混合群体包括重金属砷、镉以及高温在内的香菇菌丝体抗逆性状考察分析,结果表明:重金属砷、镉以及高温抗逆性状变异系数分别为49.53%、22.58%和31.64%。相关性分析表明,重金属砷、镉以及高温抗逆性状间无显著性相关关系。显著性分析表明:混合群体镉抗逆性状中,野生菌株和栽培菌株之间存在极显著抗性差异。(3)基于全基因组重测序开发的高密度SNP标记,对59个野生菌株进行全基因组关联分析结果表明:在两年的出菇试验中,共发掘到与农艺性状相关的SNP标记904个。其中2014年,检测到与菇数显著相关位点196个,单菇鲜重显著相关位点133个,菌盖重量相显著关位点31个,菌柄直径显著相关位点1个,其中有3个位点分别同时与菇数和单菇鲜重显著相关。在2015年中,检测到与单菇鲜重显著相关位点280个,菌柄重量显著相关位点158个,菌盖重量显著相关位点46个,采菇期显著相关位点27个,原基期显著相关位点24个,菌柄长度显著相关位点5个,鲜菇总重显著相关位点3个,其中有21个位点分别同时与原基期和采菇期显著相关,24个位点分别同时与菌盖重量和菌柄重量显著相关,1个位点同时与菌盖重量、菌柄重量以及单菇鲜重三个农艺性状显著相关。在126个香菇菌株菌丝体抗逆性状全基因组关联分析中,检测到一个SNP位点与重金属砷抗性显著相关,但在其连锁不平衡衰减距离内并未发现存在候选基因;检测到206个SNP位点与重金属镉抗性显著相关;而与高温抗性显著相关SNP位点未被检测到。(4)2014和2015两年秋栽检测到与农艺性状显著相关的位点共涉及到候选基因291个,其中2014年为122个,2015年为169个;菌丝体镉抗逆性状相关位点共涉及候选基因39个,利用Blast2go对候选基因进行功能注释表明,这些基因主要有与离子、小分子、有机环状化合物、蛋白等结合功能,具有转运酶、水解酶以及氧化还原酶活性等分子功能。生物学过程注释中主要参与代谢调节、细胞周期调节,有机物的分解合成,逆境响应、外界刺激应答,细胞组分合成等生物学过程。
[Abstract]:Letinous edodes is the second largest edible fungus in the world, second only to the Agaricus bisporus. The letinous edodes industry occupies an important position in the overall development of the edible fungus industry in China. China has a vast territory, and the wild letinous edodes germplasm resources are very rich. This provides the possibility for the letinous edodes breeding and the variation of the resistance to the mining of resistance. Quantitative traits controlled by microsatellite are easily affected by the environment, and the effect of genetic improvement on phenotypes is not obvious. The molecular marker assisted selection based on full gene correlation analysis can efficiently excavate genetic variation loci related to agronomic traits and stress resistance, which helps to analyze the genetic basis of the traits and also for breeding. Excellent germplasm materials were provided. In this study, 59 wild letinous edodes strains and 126 (96 wild +30) letinous edodes strains were used as experimental materials to determine 11 agronomic traits of wild populations, 3 adversity resistance of mixed populations, and high density SNP (single nucleotide polymorphisms) based on whole genome re sequencing. The main results are as follows: (1) the analysis of the fruiting characteristics of the wild letinous edodes population including the growth period, the fruiting body shape and the yield, the results show that in the two years of the mushroom test, the yield traits all showed high coefficient of variation, including the number of mushrooms in 2014, the fresh weight of single mushroom and the total fresh mushroom. The weight was 81.13%, 89.42% and 70.57% respectively. The growth period was the next and the mushroom shape was the smallest. The correlation analysis showed that there was a very significant positive correlation between the mushroom shape traits. The total weight of fresh mushrooms had a significant positive correlation with the number of mushrooms. There was a significant negative correlation between the total weight of fresh mushrooms and the total weight of fresh mushrooms, the diameter of the cap and the thickness of the cap. The other 10 traits showed significant positive correlation in two years. The genotype and annual interaction of yield and mushroom traits had a significant impact on the phenotype. (2) analysis of the adversity resistance of letinous edodes mycelia, including heavy metal arsenic, cadmium and high temperature, showed that heavy metal arsenic, cadmium and high The variation coefficient of temperature and stress resistance was 49.53%, 22.58% and 31.64%. correlation analysis showed that there was no significant correlation between heavy metal arsenic, cadmium and high temperature stress resistance. The significant analysis showed that there was significant difference in resistance between the wild strain and the cultivated strain in the cadmium resistance of mixed population. (3) based on the whole genome re sequencing The total genomic correlation analysis of 59 wild strains showed that 59 SNP markers related to agronomic traits were found in the two years. Among them, there were 904 markers related to agronomic traits. In 2014, there were 196 significant correlation sites with mushroom numbers, 133 significant phase sites of fresh weight of single mushroom, 31 significant sites of cap weight. There were 1 significant correlation sites of stalk diameter, of which 3 sites were significantly related to mushroom number and single mushroom fresh weight. In 2015, there were 280 significant related loci with fresh weight of single mushroom, 158 significant related loci of stalk weight, 46 significant related loci of cap weight, 27 significant related loci in mushroom period, and 24 significant correlation site of primordial period 24. There were 5 significant related loci of stalk length, 3 of the significant correlation loci of fresh mushroom total weight, of which 21 loci were significantly related to the original and mushroom periods, and the 24 loci were significantly related to the weight of the cap and the weight of the stalk, and the 1 sites were simultaneously with the weight of the cap, the weight of the stalk and the fresh weight of three agronomic traits. A significant correlation between a SNP site and heavy metal arsenic resistance was detected in a total genome association analysis of 126 letinous edodes mycelial hypha resistance, but the candidate genes were not found in the linkage disequilibrium distance, and the 206 SNP loci were significantly correlated with the cadmium resistance of heavy metals, while the SNP locus was significantly related to the high temperature resistance. Not detected. (4) there were 291 candidate genes related to agronomic traits in autumn and 2014 and 2015. In 2014, there were 291 candidate genes, 122 in 2014 and 169 in 2015; 39 of the mycelial cadmium resistance related sites involved the candidate genes. The functional annotation of the candidate genes showed that these genes were mainly related to the candidate genes. Ions, small molecules, organic ring compounds, protein and other binding functions, with the functions of transporter, hydrolase, and oxidoreductase activity. Biological processes are mainly involved in metabolic regulation, cell cycle regulation, organic matter decomposition and synthesis, adverse response, external stimulus response, cell composition synthesis and other biological processes.
【学位授予单位】：华中农业大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S646.12

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