【作者】 刘萍；

【导师】 徐彦军；

【作者基本信息】 贵州大学 ， 农艺与种业， 2022， 硕士

【摘要】 杏鲍菇（Pleurotus eryngii）是一种珍稀食用菌,随着市场需求量不断增加,工厂化杏鲍菇扩能增产过程中产量及转化率不高、品质低下的问题也日益突出。施用营养素和液体肥是工厂化企业在提高杏鲍菇产量和质量中常用的栽培技术措施。本试验探索微量元素肥和维生素VB₁对杏鲍菇子实体形成与体内生理生化变化的关系。其中微量元素是一些酶的必要因子,能增强培养基基质中相关酶活性,激活菌丝体的生物学特性,使菌丝体中的代谢过程顺利进行,同时积累和储存更多的养分,促进大量子实体的形成和发生,促进了菌体内蛋白质、氨基酸、多糖的合成,进而提高产量和品质;维生素B₁在食用菌生长期间的新陈代谢中起着十分重要的作用,其与蛋白质、氨基酸、糖类及脂类的生物合成及呼吸链等许多重要代谢有密切关系。一旦缺少,酶就会失去活性,新陈代谢就会失调,导致菌体生长和发育异常。明确二者对杏鲍菇生长和品质形成的关键作用,为提高工厂化栽培过程中杏鲍菇产量和品质提供理论和实践支撑。1、采用2×2析因设计,对杏鲍菇母种培养基添加微量元素肥和VB₁的筛选试验,表明微量元素肥和VB₁对菌丝生长速度表现为协同增加的作用,处理4（添加0.5 g的微量元素肥和0.01 g的维生素B₁）菌丝长势最佳,满管时间相比处理1提前2 d,生长速度最快且显著快于其他处理,其菌丝密度与分支呈现增加的趋势。2、采用二次饱和D-最优设计,在液体培养基中添加微量元素肥与VB₁,基于3个不同培养时间的菌丝体生物量确定最佳培养时间（即培养144 h）,在此培养时间中处理T4、处理T5、处理T6菌丝体生物量显著高于CK,处理T5菌丝量最高为8.6992 g/L,同时测定此阶段4种相关酶活性:漆酶活性在处理T4中显著高于其他处理组,除去处理T2,其余处理均显著高于CK,处理T4酶活性最高为0.4380 U/m L;过氧化物酶活性在处理T4与处理T5中显著高于其他处理,处理T6显著高于CK与处理T3,在处理T5酶活性最高为20.3622 U/m L;丙酮酸脱氢酶活性在处理T5与处理T6中显著高于CK、处理1、处理T2和处理T3,处理T4显著高于CK、处理T1和处理T3,处理T5酶活性最高为5.39702U/m L;乳酸脱氢酶活性在处理T4中显著高于CK、处理T1、处理T2、处理T5和处理T6,CK乳酸脱氢酶活性最低;最后基于菌丝体生物量和酶活性筛选出杏鲍菇液体发酵培养添加微量元素肥和维生素B₁的最佳处理方案为:微量元素肥为83.945 mg/L、维生素B₁为8.3028 mg/L。3、在杏鲍菇菌棒阶段添加微量元素肥和VB₁筛选过程中,处理T1和处理T3菌丝长势最好,处理T2和处理T5菌丝长势差于对照组,处理T1满袋时间最早仅需24 d;菌棒菌丝生长阶段内,多酚氧化酶活性整体呈现先增加后下降的趋势,菌丝在长至10 d时,处理T1、处理T3、处理T4、处理T6和CK显著高于处理T2和处理T5,处理T1多酚氧化酶活性最高,菌丝长至15天,处理T1显著高于其他处理组,其活性达到103.44 U/g,菌丝在长至15 d到20 d内,处理T3酶活性显著高于其他处理,处理T2酶活性最低;菌丝在长至20 d到25 d内,各处理酶活性均呈下降趋势,CK、处理T1和处理T3多酚氧化酶活性显著高于处理T2、处理T4、处理T5和处理T6,处理T3酶活性最高,为91.64 U/g;原基分化阶段中性蛋白酶和淀粉酶得到了有效的激活,处理T1和处理T4的中性蛋白酶和淀粉酶活性显著高于CK,处理T2和处理T5活性则显著低于CK;菇蕾数量差异非常显著,处理T3菇蕾数量显著高于其他处理,处理T5菇蕾数量最低,平均仅为22.33个。杏鲍菇产量在处理T4显著高于CK、处理T2、处理T3和处理T6,产量最高可达1.1627 Kg,蛋白质和多糖中处理T4显著高于其他处理,处理T3和处理T4显著高于CK,CK显著高于处理T6、处理T5和处理T2;分析杏鲍菇对铁、锌、硼元素的富集得到铁元素在处理T6的菌盖富集含量最高,锌元素在处理T4的菌盖富集含量最高达到100.9060 mg/Kg,硼元素在处理T5的菌柄富集含量最高达到92.0313 mg/Kg;通过建立产量、蛋白质和多糖回归模型,发现微量元素肥是影响产量和品质的主要原因;最后基于杏鲍菇产量和品质筛选出添加微量元素肥和VB₁最佳处理方案为:微量元素肥为315.8mg/Kg、维生素B₁为14.3425 mg/Kg。更多还原

【Abstract】 Pleurotus eryngii is a rare edible fungus that can be used for both medicine and food.With the increasing market demand of Pleurotus eryngii,the problems of low yield,low conversion rate and low quality in the process of expanding the capacity and increasing production of the factory made Pleurotus eryngii are also increasingly prominent.This experiment explored the relationship between trace element fertilizer and vitamin VB₁on the formation of fruiting bodies of Pleurotus eryngii and the physiological and biochemical changes in vivo.Among them,trace elements are necessary factors for some enzymes,which can enhance the activities of related enzymes in the medium matrix,activate the biological characteristics of the mycelium,make the metabolic process in the mycelium go smoothly,and accumulate and store more nutrients at the same time.The formation and occurrence of a large number of fruiting bodies promotes the synthesis of proteins,amino acids and polysaccharides in the bacteria,thereby improving the yield and quality;vitamin B₁plays a very important role in the metabolism of edible fungi during the growth period.It is closely related to many important metabolisms such as the biosynthesis of lipids and lipids and the respiratory chain.Once absent,the enzyme becomes inactive and metabolism becomes dysregulated,resulting in abnormal growth and development of the bacteria.To clarify the key roles of the two on the growth and quality of Pleurotus eryngii,and provide theoretical and practical support for improving the yield and quality of Pleurotus eryngii in the process of industrial cultivation.1.Used a 2×2 factorial design,the screening test of adding trace element fertilizer and VB₁to the parent culture medium of Pleurotus eryngii showed that the trace element fertilizer and VB₁showed a synergistic effect on the growth rate of mycelium.Treatment 4（added 0.5 g the trace element fertilizer and 0.01 g of vitamin B₁）had the best mycelial growth,the full tube time was 2 days earlier than that of the treatment 1,the growth rate was the fastest and significantly faster than other treatments,and the density and branching of mycelium showed an increasing trend.2.Adopt secondary saturated D-optimal design,add trace element fertilizer and VB₁to the liquid medium,and determine the optimal cultivation time（cultured for 144 h）based on the mycelial biomass of 3different cultivation times.During the incubation time,the mycelial biomass of treatment T4,treatment T5 and treatment T6 was significantly higher than that of CK,and the maximum mycelial amount of treatment T5was 8.6992 g/L.At the same time,the activities of 4 related enzymes were determined at this stage:the laccase activity was higher in treatment T4.Significantly higher than other treatment groups,except treatment T2,other treatments were significantly higher than CK,the highest enzyme activity of treatment T4 was 0.4380 U/ml;peroxidase activity was significantly higher in treatment T4 and treatment T5 than other treatments,treatment T6 significantly higher than CK and treatment T3,the highest enzyme activity in treatment T5 was 20.3622 U/ml;pyruvate dehydrogenase activity in treatment T5 and treatment T6 was significantly higher than CK,treatment T1,treatment T2 and treatment T3,treatment T4significantly higher than CK,treatment T1 and treatment T3,the highest enzyme activity of treatment 5 was 5.39702 U/ml;the lactate dehydrogenase activity in treatment T4 was significantly higher than that of CK,treatment T1,treatment T2,treatment T5 and treatment T6,CK lactate dehydrogenase activity was significantly higher than that in treatment T4.The hydrogenase activity was the lowest;Finally,based on the mycelial biomass and enzyme activity,the optimal treatment plan for adding trace element fertilizer and vitamin B₁to the liquid fermentation culture of Pleurotus eryngii was screened out:the trace element fertilizer was 83.945 mg/L,and the vitamin B₁was 8.3028 mg/L.3.In the process of adding trace element fertilizers and VB₁screening in the stage of Pleurotus eryngii,treatment T1 and treatment T3 had the best mycelial growth,treatment T2 and treatment T5 was worse than the control group,and treatment T1 had the earliest time to fill the bag,it only takes 24 days;during the growth stage of mycelium,the activity of polyphenol oxidase showed an overall trend of increasing first and then decreasing.When the mycelium grew to 10 days,treatment T1,treatment T3,treatment T4,treatment T6 and CK were significantly higher In treatment T2 and treatment T5,the activity of polyphenol oxidase in treatment T1 was the highest,and the hyphae grew to 15 days.Treatment T1 was significantly higher than other treatment groups,and its activity reached 103.44 U/g.The enzyme activity of treatment T3 was significantly higher than that of other treatments,and the enzyme activity of treatment T2 was the lowest;the enzymatic activity of each treatment showed a downward trend when the mycelium grew from 20 to 25 days,and the polyphenol oxidase activity of CK,treatment T1 and treatment T3 was significantly higher than that of treatment T3,treatment T2,treatment T4,treatment T5 and treatment T6,treatment T3 had the highest enzyme activity,which was 91.64 U/g;neutral protease and amylase were effectively activated in the primordium differentiation stage.The amylase activity was significantly higher than that of CK,while the activities of treatments T2 and T5 were significantly lower than those of CK.The difference in the number of mushroom buds was very significant.The number of mushroom buds in the treatment T3 was significantly higher than that in other treatments,and the number of mushroom buds in the treatment T5 was the lowest,with an average of only 22.33.The yield of Pleurotus eryngii in treatment T4 was significantly higher than that in CK,T2,T3 and T6,and the highest yield was 1.1627 Kg.In protein and polysaccharide,treatment T4 was significantly higher than other treatments,and treatment T3 and T4 were significantly higher than CK.CK was significantly higher than treatment T6,treatment T5 and treatment T2;analysis of the enrichment of Pleurotus eryngii to iron,zinc and boron elements obtained iron elements,the enrichment content of caps in treatment T6 was the highest,the enrichment content of zinc in the caps of treatment T4 was up to 100.9060 mg/Kg,and the enrichment content of boron in the stems of treatment T5 was up to 92.0313 mg/Kg,protein and polysaccharide regression model,it was found that trace element fertilizer was the main reason affecting yield and quality;finally,based on the yield and quality of Pleurotus eryngii,the optimal treatment plan of adding trace element fertilizer and VB₁was screened out:trace element fertilizer was315.8 mg/Kg,Vitamin B₁was 14.3425 mg/Kg.更多还原