【作者】 杨燕；

【导师】 田鸿； 彭卫红；

【作者基本信息】 四川农业大学 ， 农业推广硕士（专业学位）， 2018， 硕士

【摘要】 近年来,子囊菌门食用菌羊肚菌栽培取得较大进展,由于其子囊果不稳定,在人工栽培中,普遍存在菌种名称混乱、品种退化、同种异名的现象。本研究以野生和人工栽培的18株羊肚菌作为研究材料,从子囊果发育的形态初步分析,结合ITS和ITS+EF-1a+RPB1+RPB2四段基因联合分析,确定试验材料分类地位,以及采用ISSR技术分析羊肚菌的遗传多样性。并以此为基础,对18株羊肚菌在相同栽培条件下的出菇性状考察,筛选出产量稳定,性状优异的菌株;与此同时研究不同基料下羊肚菌农艺性状差异,为优化羊肚菌人工培养技术提供基础。论文的研究内容和结果如下:(1)供试羊肚菌菌株形态学分析:按照羊肚菌子实体大小、颜色、菌帽和菌柄形状等特征,初步判断18株供试菌株中同时含有黑色羊肚菌类群和黄色羊肚菌类群。(2)供试羊肚菌菌株分子学鉴定:基于r DNA ITS(Internal Transcribed Spacer)序列和多基因序列聚类分析表明,黑色羊肚菌支系包括:9株来自不同地区的人工栽培菌株(YD9a、YD13、YD7、YD2、YD1、cang1、YD16、YD17、YD9b)和野生菌株GS(采自陕西)为梯棱羊肚菌(Morchella importuna);2株人工栽培菌株(12wen、1212M)为六妹羊肚菌(Morchella sextelata);野生羊肚菌菌株1431B(采自成都大邑)、WL3(采自重庆武隆)和2株人工栽培菌株(cang2、YD6)为高羊肚菌(Morchella elata Fr.)。黄色羊肚菌支系包括:野生菌株LF(采自湖北恩施)和YD15(采自绵阳平武)为粗柄羊肚菌(Morchella crassipes)。(3)供试菌株遗传多样性:利用ISSR(inter-simple sequence repeat)分子标记技术对供试菌株进行遗传多样性分析,表明18株羊肚菌分为两大类,即YD9a、YD7、YD13、YD1、YD2、YD17、GS、YD16、cang1、YD9b、12wen、1212M、WL3为一类;LF、YD15、cang2、1431B、YD6为另一类。其中YD1和YD2同属人工栽培的梯棱羊肚菌两者亲缘关系最近,基本无遗传多样性;此外,GS和YD16同属梯棱羊肚菌,其中GS为野生菌株,同样两者亲缘关系较近,无明显遗传多样性;12wen和1212M同属人工栽培的六妹羊肚菌,两者相似系数高达95%;而梯棱羊肚菌中YD9a与高羊肚菌中YD17遗传关系最远,遗传距离最大。(4)18株羊肚菌栽培研究:LF、YD15为粗柄羊肚菌,属于黄色羊肚菌支系,且为野生菌株,未出菇。在4株高羊肚菌中,WL3为野生菌株,未能出菇,而1431B已经出菇,产量最低,YD6产量最高。在梯棱羊肚菌中,YD13的产量最高,其次YD1,YD7,在六妹羊肚菌中,12wen菌株的产量明显高于1212M,推测1212M菌株已表现退化现象。(5)不同基料对梯棱羊肚菌产子囊果的影响:将羊粪、鸡粪、平菇废料、腐殖土作为不同基料,研究其对羊肚菌农艺性状的影响。,一定施用量的羊粪、鸡粪、腐殖土能促进羊肚菌生长,提高羊肚菌产量,在施用量为1kg/m~2时,促进效果最好。而平菇废料会抑制羊肚菌生长。更多还原

【Abstract】 In recent years,great progress has been made in the cultivation of Morchella.Due to the instability of the ascocarp,there were some widespread confusions that the species naming was coufused and the strains were degraded In the process of cultivation.In this study,18 strains of wild and cultivated Morchella were used as research materials.The taxonomic statues of the tested strains was determined from the morphological analysis of the ascocarp development and the combined analysis of the ITS and ITS+EF-1a+RPB1+RPB2four-segment genes and the genetic diversity of Morchella was analyzed by ISSR technique.On the basis of this,under the same cultibation conditions,Eighteen Morchella strains were selected for the strain breeding process,and strains with stable yield and excellent traits were screened out.At the same time,and the differences in agronomic traits of Morchella under different substrates were studied to provide the basis for optimizing the artificial culture technology of Morchella.The main research contents and results of the paper were as follows:(1)Morphological study of the tested strains: according to the characteristics of the fruiting body size,color,cap and stipe shape,18 strains were preliminarily judged to contain both the black morel and the yellow morel.(2)Molecular identification of the test strains: clustering analysis based on the r DNA ITS(Internal Transcribed Spacer)sequence and mufti-gene sequence showed that,the black morel included:9 strains of artificial cultivated strains from different regions(YD9a,YD13,YD7,YD2,YD1,cang1,YD16,YD17,YD9b)and wild strain GS(from shanxi)were Morchella importunas;2 cultivated strains(12wen,1212M)were Morchella sextelatas;wild strain 1431B(from Chengdu Dayi),WL3(from Chongqing Wulong),and two cultivated strains(cang2,YD6)were Morchella elatas.The yellow morel include wild strain LF(from Hubei)and YD15(from Mianyang,Pingwu)that were the Morchella crassipes.(3)Genetic diversity of the tested strains: The genetic diversity of the tested strains was analyzed by ISSR(inter-simple sequence repeat)molecular marker technique,indicating that the 18 morels species were divided into two major categories,namely YD9 a,YD7,YD13.YD1,YD2,YD17,GS,YD16,cang1,YD9 b,12wen,1212 M,WL3 were one type;LF,YD15,cang2,1431 B,and YD6 were another type.Among them,YD1 and YD2 were the Morchella importuna,which were closely related to each other and had not almost genetic diversity.In addition,GS and YD16 had similar genetic relationships,and GS was belong to the wild strain.Significant genetic diversity;12wen and 1212 M were belong to the artificially cultivated Morchella sextelata,and the similarity coefficient was as high as 95%.YD9 a and YD17 had the longest genetic distance and the largest genetic distance.(4)18 Morels cultivation researches: LF and YD15 were Morchella crassipes,belonging to yellow morel,and they were failed to produce mushrooms.Among the Morchella elatas,WL3 was a wild strain that failed to produce mushrooms.and 1431 B had mushroomed,yield was lowest,and YD6 had the highest yield.In the Morchella importuna,the yield of YD13 was the highest,followed by YD1,YD7,and the yield of the 12 wen strain was significantly higher than that of 1212 M in the morchella sextelata,suggesting that the 1212 M strain has shown degeneration.(5)Effects of different base materials on YD7: Using sheep manure,chicken manure,Pleurotus ostreatus waste and humus as different base materials to study their effects on Morchella importuna.It was found that sheep manure,chicken manure,and humus at a certain concentration can promote the growth of morel and increase the production of morel.The three materials are all the best at promoting the concentration,while Pleurotus ostreatus waste inhibits the growth of morel.更多还原