【作者】 张莹；

【导师】 辜运富；

【作者基本信息】 四川农业大学 ， 微生物学， 2025， 硕士

【摘要】 香菇(Lentinula edodes)是一种被人们广泛食用的真菌,属于真菌门担子菌纲伞菌目口蘑科香菇属,其子实体含有的蛋白质、氨基酸、维生素及矿质元素非常丰富,具有极大的营养和药用价值,在疾病预防与治疗中颇为重要,市场潜力巨大。为实现香菇栽培与环境保护及资源节约的协调发展,本研究通过筛选草腐配方主料及其颗粒度,并与不同比例的木屑混合进行出菇试验来优化栽培配方;同时对比草腐与木腐配方在子实体表型、单菇重量及含水量等方面的差异,并利用BSA分析和转录组测序技术,挖掘与培养料偏好相关的基因及差异表达基因的功能通路,为香菇栽培的可持续发展提供理论支持。试验结果如下:1.从189株菌株中随机筛选出10株用五种配方分别进行大试管栽培,结果显示不同菌株之间,在同一种配方中的菌丝生长速率有一定差异,但是这种差异也与配方主料的选择有关;不同配方之间,木屑配方施用下的菌丝平均生长速率最高,而玉米秸秆配方下的最低,其余配方的平均生长速率较为适中;各品种的菌丝色泽无较大差异均为洁白,不同菌株在同一种配方中的菌丝浓密程度有差异但不明显,不同配方之间的菌丝浓密程度有明显差别,以麦秸、木屑为主料的配方菌丝生长较浓密,以菌草、稻草和玉米秸秆为主料的配方的菌丝生长较稀疏。麦秸配方的菌丝生长速率适中且菌丝浓密,结合目前国内农副产品利用现状,顾选用麦秸作为与木屑栽培相对的草腐配方的主要材料。2.探究麦秸不同颗粒度对香菇生长速率影响,结果显示麦秸的颗粒度为5-8cm时菌丝生长速率最快,颗粒度为0.2 cm时菌丝生长速率最慢,而颗粒度为5-8cm时菌丝生长速率最快但其菌丝密度过于稀疏,会致使后续出菇试验成功率较低,综合来看1-3 cm为麦秸的最佳颗粒度。3.用不同颗粒度的麦秸和木屑以不同比例混合的配方栽培香菇,可以看出无论哪种颗粒度,菌丝生长速率均是随着麦秸含量的增加而递减,且三种颗粒度菌袋的满袋时间均是随着麦秸比例和颗粒度的增加而增加的,但在大试管栽培试验中麦秸颗粒度为1-3 cm时菌丝生长速率为三者中最高。4.用草腐配方和木腐配方分别对189株不同品种菌株进行栽培出菇,其中有25个菌株未能出菇,在两种培养料中均能出菇的有26株,仅能在木腐配方出菇的菌种有138株,仅能在草腐配方出菇的菌种有2株;出菇过程中,木腐配方的出菇速度明显快于草腐配方,两种配方的子实体表型也有明显差异;子实体含水量在两种配方均能出菇的这些品种中无显著差异,但草腐配方栽培出的香菇含水量略低于以木腐配方栽培出的香菇。5.通过BSA分析对目标性状相关基因区域进行定位后,发现基因在碳水化合物代谢、细胞壁组织和生物合成、信号转导等方面显著富集;随后利用CAZy酶注释,发现了多个与碳水化合物相关酶的基因有关的变异位点,包括GT1家族、GH37家族、GH17家族和GT4家族,这些变异位点会影响香菇在不同培养料中的生长和代谢能力。6.转录组测序结果显示,香菇在草腐和木腐培养料中生长时,存在显著的基因表达差异,其中330个基因上调,788个基因下调。这些差异表达基因主要涉及碳水化合物代谢、细胞壁合成和信号转导等生物学过程,表明这些基因可能与香菇对不同培养料的适应性有关。本研究通过优化草腐配方主料比例和颗粒度,为香菇栽培提供了一个更环保、更绿色的替代方案,同时在香菇菌株的培养料适应性机制方面有了更深入的研究;通过BSA分析和转录组测序,发现了与香菇生长和代谢相关的关键基因及通路,为后续培育适应性强、高产优质的香菇品种提供了理论依据和分子靶点,对推动香菇栽培的可持续发展及农业副产物的高效利用具有重要意义。更多还原

【Abstract】 Lentinula edodes is a widely consumed fungus,belonging to the family Tricholomataceae,order Agaricales,class Basidiomycetes,and phylum Fungi.It is highly appreciated for its unique aroma and tender texture.The fruiting body of L.edodes is rich in proteins,amino acids,vitamins,and mineral elements,and has great nutritional and medicinal value.It plays an important role in disease prevention and treatment and has a huge market potential.To achieve coordinated development between L.edodes cultivation and environmental protection as well as resource conservation,this study optimized the cultivation formula by screening the main ingredients and particle size of the grass rot formula and conducting fruiting experiments combined with sawdust.Meanwhile,the differences in fruiting body phenotype,single mushroom weight,and water content between grass rot and wood rot formulas were compared.Using BSA analysis and transcriptome sequencing technology,genes related to substrate preference and functional pathways of differentially expressed genes were mined,providing theoretical support for the sustainable development of L.edodes cultivation.The main results of the trial are as follows:1.10 strains were randomly selected from 189 strains and cultivated in large test tubes with five formulas(Juncao,straw,wheat straw,woods,corn straw),and the results showed that there were certain differences in the growth rate of mycelium in the same formula between different strains,but this difference was also related to the selection of the main ingredients of the formula,and the average rate of corn straw-based formula was the lowest,the average rate of woods-based formula was the highest,and the average growth rate of other formulas was moderate.There was no great difference in the color of the mycelium of each variety was white,the mycelial density in the same formula was different but not obvious between different strains,and there was a significant difference in the mycelial density between different formulas,the mycelium growth of the formula with wheat straw and woods as the main materials was denser,and the mycelium growth of the formula with Juncao,straw and corn straw as the main material was sparse.Combined with the current utilization status of domestic agricultural and sideline products,wheat straw was selected as the main material of straw formula relative to sawdust cultivation.2.To explore the effect of different particle sizes of wheat straw on the growth rate of L.edodes,the results showed that the mycelial growth rate was the fastest when the particle size was 5-8cm,the slowest mycelial growth rate when the particle size was 0.2cm,and the mycelial growth rate was the fastest but the lowest mycelial density when the particle size was 5-8 cm,which may greatly reduce the bioconversion rate.3.Cultivated with different particle sizes of wheat straw and woods mixed with different proportions,it can be seen that no matter what kind of particle size,the mycelial growth rate decreases with the increase of wheat straw content,and the full bag time of the three granularity bacterial bags increases with the increase of straw proportion and particle size.However,in the large test-tube cultivation experiment,when the particle size of wheat straw was 1-3 cm,the mycelial growth rate was the highest among the three.4.With straw-based formula and woods-based formula,189 strains of different varieties of L.edodes were cultivated,of which 25 strains failed to produce mushrooms,26 strains could produce mushrooms in both cultures,138 strains could only produce mushrooms in the woods-based formula,and 2 strains could only produce mushrooms in the straw-based formula;In the process of mushroom emergence,the mushroom production speed of the woods-based formula was significantly faster than that of the straw-based formula,and the fruiting body phenotypes of the two formulas were also significantly different.There was no significant difference in the water content of fruiting bodies among the two formulas,but the water content of L.edodes cultivated with straw-based formula was slightly lower than that of L.edodes cultivated with woods-based formula.5.After locating the candidate gene regions associated with the target trait through BSA analysis,significant enrichment was observed in genes related to carbohydrate metabolism,cell wall organization and biosynthesis,and signal transduction.Subsequently,CAZy enzyme annotation revealed multiple variant sites associated with genes encoding carbohydrate-active enzymes,including the GT1,GH37,GH17,and GT4families.These variants may influence the growth and metabolic capacity of Lentinula edodes in different cultivation substrates.6.Transcriptome sequencing results showed significant differences in gene expression when L.edodes grew on straw-based and woods-based substrates,with 330genes upregulated and 788 genes downregulated.The main biological processes affected by these differentially expressed genes included carbohydrate metabolism,cell wall synthesis,and signal transduction,indicating that these genes may be related to the adaptability of L.edodes to different substrates.This study provides a more environmentally friendly and resource-saving alternative for L.edodes cultivation by optimizing the formula and particle size of the substrate,while also revealing the adaptation mechanisms of strains to the culture medium.Through functional gene analysis and transcriptome sequencing,key genes and pathways related to the growth and metabolism of L.edodes were identified,providing a theoretical basis and molecular targets for the subsequent breeding of L.edodes varieties with strong adaptability,high yield,and superior quality.The research findings are of great significance for promoting the sustainable development of L.edodes cultivation and the efficient utilization of agricultural by-products.更多还原