【摘要】：香菇是世界上產(chǎn)量第二大食用菌,僅次于雙孢蘑菇。香菇產(chǎn)業(yè)在我國食用菌產(chǎn)業(yè)的整體發(fā)展中占據(jù)著舉足輕重的地位。我國幅員遼闊,野生香菇種質(zhì)資源極為豐富,這為香菇育種以及挖掘抗性變異提供了可能。香菇大多數(shù)農(nóng)藝性狀都是由微效多基因控制的數(shù)量性狀,易受環(huán)境影響,僅對于表型進(jìn)行遺傳改良效果并不明顯。以全基因關(guān)聯(lián)分析為途徑的分子標(biāo)記輔助選擇可以高效挖掘與農(nóng)藝性狀以及抗逆性狀相關(guān)的遺傳變異位點(diǎn),有助于解析性狀的遺傳基礎(chǔ),也為育種提供優(yōu)良的種質(zhì)材料。本研究以59個(gè)野生香菇菌株群體和126(96個(gè)野生+30個(gè)栽培)個(gè)香菇菌株混合群體為實(shí)驗(yàn)材料,測定了野生群體的11個(gè)農(nóng)藝性狀,混合群體的3個(gè)抗逆性狀,利用基于全基因組重測序開發(fā)的高密度SNP(單核酸多態(tài)性)標(biāo)記,對供試群體進(jìn)行全基因組關(guān)聯(lián)分析。主要結(jié)果如下:(1)對野生香菇群體包括生育期、子實(shí)體形態(tài)和產(chǎn)量在內(nèi)的子實(shí)體性狀考察分析,結(jié)果表明:在兩年的出菇試驗(yàn)中,產(chǎn)量性狀均表現(xiàn)出較高的變異系數(shù),其中2014年的菇數(shù)、單菇鮮重、鮮菇總重分別為81.13%、89.42%和70.57%;生育期次之,菇形性狀最小,介于16.77%-71.11%之間。相關(guān)性分析表明菇形性狀間存在著極顯著正相關(guān),鮮菇總重與菇數(shù)間存在極顯著正相關(guān),生育期與鮮菇總重、菌蓋直徑以及菌蓋厚度間存在極顯著負(fù)相關(guān)。除菇數(shù)外,其余10個(gè)性狀在兩年間均表現(xiàn)出顯著正相關(guān)。產(chǎn)量性狀和菇形性狀的基因型與年份互作的結(jié)果對表型有著極顯著影響。(2)對混合群體包括重金屬砷、鎘以及高溫在內(nèi)的香菇菌絲體抗逆性狀考察分析,結(jié)果表明:重金屬砷、鎘以及高溫抗逆性狀變異系數(shù)分別為49.53%、22.58%和31.64%。相關(guān)性分析表明,重金屬砷、鎘以及高溫抗逆性狀間無顯著性相關(guān)關(guān)系。顯著性分析表明:混合群體鎘抗逆性狀中,野生菌株和栽培菌株之間存在極顯著抗性差異。(3)基于全基因組重測序開發(fā)的高密度SNP標(biāo)記,對59個(gè)野生菌株進(jìn)行全基因組關(guān)聯(lián)分析結(jié)果表明:在兩年的出菇試驗(yàn)中,共發(fā)掘到與農(nóng)藝性狀相關(guān)的SNP標(biāo)記904個(gè)。其中2014年,檢測到與菇數(shù)顯著相關(guān)位點(diǎn)196個(gè),單菇鮮重顯著相關(guān)位點(diǎn)133個(gè),菌蓋重量相顯著關(guān)位點(diǎn)31個(gè),菌柄直徑顯著相關(guān)位點(diǎn)1個(gè),其中有3個(gè)位點(diǎn)分別同時(shí)與菇數(shù)和單菇鮮重顯著相關(guān)。在2015年中,檢測到與單菇鮮重顯著相關(guān)位點(diǎn)280個(gè),菌柄重量顯著相關(guān)位點(diǎn)158個(gè),菌蓋重量顯著相關(guān)位點(diǎn)46個(gè),采菇期顯著相關(guān)位點(diǎn)27個(gè),原基期顯著相關(guān)位點(diǎn)24個(gè),菌柄長度顯著相關(guān)位點(diǎn)5個(gè),鮮菇總重顯著相關(guān)位點(diǎn)3個(gè),其中有21個(gè)位點(diǎn)分別同時(shí)與原基期和采菇期顯著相關(guān),24個(gè)位點(diǎn)分別同時(shí)與菌蓋重量和菌柄重量顯著相關(guān),1個(gè)位點(diǎn)同時(shí)與菌蓋重量、菌柄重量以及單菇鮮重三個(gè)農(nóng)藝性狀顯著相關(guān)。在126個(gè)香菇菌株菌絲體抗逆性狀全基因組關(guān)聯(lián)分析中,檢測到一個(gè)SNP位點(diǎn)與重金屬砷抗性顯著相關(guān),但在其連鎖不平衡衰減距離內(nèi)并未發(fā)現(xiàn)存在候選基因;檢測到206個(gè)SNP位點(diǎn)與重金屬鎘抗性顯著相關(guān);而與高溫抗性顯著相關(guān)SNP位點(diǎn)未被檢測到。(4)2014和2015兩年秋栽檢測到與農(nóng)藝性狀顯著相關(guān)的位點(diǎn)共涉及到候選基因291個(gè),其中2014年為122個(gè),2015年為169個(gè);菌絲體鎘抗逆性狀相關(guān)位點(diǎn)共涉及候選基因39個(gè),利用Blast2go對候選基因進(jìn)行功能注釋表明,這些基因主要有與離子、小分子、有機(jī)環(huán)狀化合物、蛋白等結(jié)合功能,具有轉(zhuǎn)運(yùn)酶、水解酶以及氧化還原酶活性等分子功能。生物學(xué)過程注釋中主要參與代謝調(diào)節(jié)、細(xì)胞周期調(diào)節(jié),有機(jī)物的分解合成,逆境響應(yīng)、外界刺激應(yīng)答,細(xì)胞組分合成等生物學(xué)過程。
[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.
【學(xué)位授予單位】：華中農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：S646.12

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