斑玉蕈（Hypsizigus marmoreus）又称真姬菇，隶属担子菌亚门、层菌纲、伞菌目、白蘑科、玉蕈属，其形态精致，口感鲜嫩，因具有海蟹味，故又有“蟹味菇”之称。工厂化生产的斑玉蕈，其栽培周期相对较长，一般为80-110天，因此生产成本较高。有效缩短斑玉蕈栽培周期成为提高其生产效率、降低其栽培成本的关键。 本研究以斑玉蕈SIEF3133作为实验菌株，分别设计了含有0.01g/L、0.05g/L、0.1g/L、0.3g/L、0.5g/L、1g/L不同浓度的肥皂草素培养基以及对照组的培养基，从生理以及栽培性状、酶学以及分子生物学层面设计相关实验方案，将肥皂草素对斑玉蕈生长发育过程中生理变化的影响及原因，以及应用于工厂化栽培的技术可行性作为本研究的主要内容。 本论文的主要研究结果如下： 1. 在以羧甲基纤维素钠为主要碳源的土豆浸汁琼脂培养基中添加肥皂草素，使其在培养基中的浓度分别为0g/L（对照组）、0.01g/L、0.05g/L、0.1g/L、0.5g/L、1g/L、5g/L以及10g/L,对斑玉蕈菌落直径以及菌丝生长速度分别进行测定、分析。研究结果表明：肥皂草素具有抑制斑玉蕈菌丝生长速度的作用，且这种抑制程度与培养基中肥皂草素的浓度呈正相关；同时，当培养基中肥皂草素的浓度大于0.1g/L时，斑玉蕈菌丝不能进行生长。 2. 在对斑玉蕈菌落形态的比较分析中，发现肥皂草素造成斑玉蕈菌落边缘不规整，且菌丝较对照组浓密、洁白；在对斑玉蕈菌丝体显微观测中，发现含有肥皂草素的实验组其菌丝密度和菌丝体粗壮程度显著高于对照组。 3. 在斑玉蕈液体合成培养基中添加肥皂草素，使其在培养基中的浓度分别为0.01g/L，0.05g/L，0.1g/L以及0g/L（对照组），通过对不同培养阶段菌丝生理性状及其形态转变差异上的比较分析，研究结果发现：浓度为0.01g/L，0.05g/L，0.1g/L肥皂草素组以及对照组的斑玉蕈菌丝扭结时间分别为第66天， 70天， 68天与56天；其原基发生时间分别为第70天，74天，74天与84天；其原基数量随着培养基中肥皂草素浓度的升高而递减；浓度为0.1g/L的肥皂草素组在第85天可以形成子实体，浓度为0.05g/L的肥皂草素组在第110天可以形成子实体，其余组在研究过程中均未发现子实体的产生。研究结果表明：培养基中肥皂草素的添加延长了斑玉蕈营养生长的时间，缩短了其生殖生长的时间，并缩短了其整个培养周期，抑制了原基数量，促进了原基的发生、发育。 4. 分析了肥皂草素对斑玉蕈生长发育过程中胞外β-葡萄糖苷酶活力变化的影响，并对β-葡萄糖苷酶基因bgl相对表达量做了进一步的研究。研究中发现：在营养生长阶段，浓度为0.01g/L的肥皂草素组其胞外β-葡萄糖苷酶活力的变化与对照组差异不显著，酶活力的峰值均出现在第6天，浓度为0.05g/L以及0.1g/L的肥皂草素组胞外β-葡萄糖苷酶活力的变化水平相对于对照组受到不同程度的抑制，且其峰值出现在第10天；同时肥皂草素组其β-葡萄糖苷酶基因bgl相对于对照组表达量上的变化与胞外β-葡萄糖苷酶相对酶活力的变化基本一致；在斑玉蕈转入生殖生长后，胞外β-葡萄糖苷酶活力相对营养生长阶段出现较显著的提高并呈现双峰变化，对照组的峰值分别出现在第5天和第20天，肥皂草素组的峰值分别出现在第10天和第20天，且β-葡萄糖苷酶相对酶活力及其基因bgl相对表达量的测定结果均表现出前期抑制，后期促进。研究结果表明：营养生长阶段，肥皂草素对斑玉蕈胞外β-葡萄糖苷酶的抑制作用主要表现为对酶活力变化水平的抑制及其峰值出现时间上延滞，且β-葡萄糖苷酶活力及其基因bgl相对表达量的变化与斑玉蕈菌丝生物量的变化规律存在一定的关系；生殖生长阶段，肥皂草素对β-葡萄糖苷酶活力变化及其基因bgl相对表达量变化上的影响与生殖生长过程中斑玉蕈形态转变上的差异关系密切。 5. 在肥皂草素应用于斑玉蕈工厂化栽培技术可行性的研究中，分别进行了小规模结实性实验以及斑玉蕈规模化工厂栽培实验，通过向培养不同时间的斑玉蕈菌丝表面喷洒一定体积不同浓度的肥皂草素溶液，对子实体产量、性状进行测定分析，研究结果表明：肥皂草素对斑玉蕈子实体产量及其性状的影响与添加肥皂草素的浓度以及斑玉蕈的培养时间有着紧密关系；在斑玉蕈培养第60天喷洒浓度为0.01g/L或0.05g/L的肥皂草素溶液可缩短其25天的栽培时间，极大地提高了斑玉蕈的生产效率。

Hypsizigus marmoreus is named of Shimeji mushroomand attached to Basidiomycota, Hymenomycetes, Agaricales. It has both excellent appearance and fresh taste as it is also named of “crab falvor mushroom”. However, this kind of mushroom has to be cultivated for the longer period to get its fruiting bodies than most of other kinds’. It’s the target of our research on shorting the culture period so as to bring up its return. Using H.marmoreus SIEF3133 as our study subject, we designed several saponin levels(0.01g/L,0.05g/L,0.1g/L,0.5g/L,1g/L) in different media to investigate the effect on the hypha and fruiting body of H.marmoreus,and we selected β-glucosidase and bgl gene which could modulate this mushroom’s growth and development as factors. Finally, we had evaluated these results to make sure whether it was applicable for industry. The main results are as follows: By the results of colony diameter measured, we could conclude the saponin added into the medium could depress hypha growth much more as the saponin level increased in the medium. It also appeared that of the saponin with six levels in every medium, only 0.01g/L,0.05g/L,0.1g/L and the control group were fit for vegetative growth. By comparison of different hypha patterns on condition of different saponin levels in microscopic and macroscopic view, we found that the hyphae cultivated on saponin media were superior in quality than the control group. Their hyphae were whiter, denser and thicker, and their Rhizomorphs were stronger. We mixed the saponin with the liquid medium for SIEF3133 as different saponin levels to investigate the changes of H.marmoreus physiology. By the research of the vegetative phase, guttation phase, kinking mycelium phase, primodia phase and other physiological standards,we concluded that the saponin added into the medium could prolong the span of vegetative growth but short the span of reproductive growth and cut back the period of cultivation thoroughly. These appearances were constantly coherent with different batches. Due to the relationship of cellulase and wood-rotting fungi growth and development which was announced by various articles, we got the β-D-glucosidase activity and the relative expression of bgl gene during the cultivation to be our targets. Then we designed shake-flask media and compost for cultivating SIEF3133. By the research of this study, we drew several conclusions:(A)Saponin could depress β-glucosidase activity and reduce bgl expression at vegetative stage(B)Saponin could depress β-glucosidase activity and bgl expression at the beginning of reproductive stage but promote them at following stages(C)β-glucosidase activity and bgl gene relative expression was concerned with hyphae biomass and reproductive period. For evaluating whether the saponin was available for industrial mushroom producing, we chose 250ml bottle filled with 160g compost and 1.2L bottle filled with 450g compost as our experiment media, and we inoculated the hyphae onto the compost surface. When vegetative hyphae grew into post-mature stage, we targeted several different stages such as the 50th day, 60th days,70th days and others to perfuse saponin solution with different levels onto the surface of the compost. For about 3 weeks, we recorded some physiological datasuch as the gross of fruiting bodies, the stipe length and the biomass of fruiting bodies. We found the low saponin levels could stimulate the gross of mature fruiting bodies, stipe length and its biomass. Most of the high saponin levels led to the depression. The post-mature periods were more sensitive to the saponin levels.