原生质体融合获得羊肚菌黑色栽培种与黄色非栽培种间杂交高产菌株  

Protoplast fusion to obtain high-yield strains from interspecific hybridization between black cultivated species and yellow uncultured species of Morchella

作　　者：廖武俊 钟啟萍 李荣光 霍达 霍光华[1] LIAO Wujun;ZHONG Qiping;LI Rongguang;HUO Da;HUO Guanghua(College of Bioscience and Bioengineeing,Jiangxi Agricultural University,Nanchang 330045,China;Nanchang Business College of Jiangxi Agricultural University,Jiujiang,Jiangxi 330300,China)

机构地区：[1]江西农业大学生物科学与工程学院,江西南昌330045 [2]江西农业大学南昌商学院,江西九江330300

出　　处：《江西农业大学学报》2025年第2期349-359,共11页Acta Agriculturae Universitatis Jiangxiensis

基　　金：江西省重点研发计划项目(20203BBF63036)。

摘　　要：【目的】目前限制江西地区羊肚菌产业发展的主要问题为产量低、高产难重复及优质菌种少,而解决这些问题的关键在于菌种。本研究旨在选育适合江西本土栽培的高产羊肚菌新菌株。【方法】根据遗传与环境互作理论,有针对性的选择江西本土采集到的野生黄色羊肚菌JFRL01-161和在江西鹰潭地区商业栽培的梯棱羊肚菌JFRL01-107、六妹羊肚菌JFRL01-217作为原始菌种,通过20 g/L溶壁酶+10 g/L蜗牛酶组成的复合酶溶解细胞壁制取原生质体,再通过高温和紫外双灭活获得2种灭活原生质体,将不同灭活的原生质体进行两两融合获得融合子。对得到的融合子进行拮抗试验,栽培品比以及区间简单重复序列标记(ISSR)鉴定确定种源关系。【结果】经原生质体融合共获得融合菌株16株,即A11、A37、A39、B7、B12、B17、B18、B25、B26、B29、C1、C2、C5、C6、C11和C12;其中菌株A11、A37、A39和B18出菇并形成产量,特别是融合菌株A11产量达到351.90g/m^(2),显著高于亲本JFRL01-107产量264.51g/m^(2),年度重复栽培验证试验确认A11产量可达527.8g/m^(2),亲本JFRL01-107产量为262.4 g/m^(2)。非加权算数平均聚类系统发生树(UPGMA)显示A11分别与亲本JFRL01-161(相似水平系数0.24)和JFRL01-107(相似系数水平0.696)处于不同分支,证明融合菌株A11是不同于两亲本的新菌株。【结论】融合菌株A11产量显著高于亲本,经过重复栽培试验确认其种质稳定,经ISSR确定是不同于亲本的菌株,因此融合菌株A11可作为适应江西本土栽培的高产羊肚菌新菌株。[Objective]At present,the main issues that restricting the development of Morchella industry in Jiangxi Province are low yield,difficulty in replicating high yields,and limited availability of high-quality strains,and the key to addressing these challenges lies in the improvement of strains.The purpose of this study is to breed a new strain of Morchella with high yield,which is suitable for local cultivation in Jiangxi.[Method]According to the theory of interaction between heredity and environment,targeted selection was made of the wild yellow Morchella strain JFRL01-161 collected locally in Jiangxi,along with the commercially cultivated Morchella importuna strain JFRL01-107 and Morchella sextelata strain JFRL01-217 from Yingtan region of Jiangxi Province,which were used as original strains,and protoplasts were prepared by dissolving the cell wall with a complex enzyme composed of 20 g/L lywallzyme and 10 g/L snail enzyme,and then two kinds of inactivated protoplasts were obtained by high temperature and ultraviolet double inactivation.The obtained fusants were tested by antagonism,cultivar ratio and Inter-Simple Sequence Repeat to determine the provenance relationship.[Result]Sixteen fusion strains were obtained by protoplast fusion,namely A11,A37,A39,B7,B12,B17,B18,B25,B26,B29,C1,C2,C5,C6,C11 and C12.Among them,strains A11,A37,A39 and B18 successfully fruited and produced yields,the yield of the fusion strain A11 reached 351.90 g/m^(2),which was significantly higher than that of the parent JFRL01-107(264.51 g/m^(2)).The annual repeated cultivation verification test confirmed that the yield of A11 could reach 527.8 g/m^(2),and that of the parent JFRL01-107 was 262.4 g/m^(2).The phylogenetic tree(UPGMA)of unweighted arithmetic mean clustering system showed that A11 was in different branches from its parents JFRL01-161(similarity coefficient 0.24)and JFRL01-107(similarity coefficient level 0.696),which proved that A11 was a new strain different from its parents.[Conclusion]The yield of fused strain A11 was signifi

关 键 词：羊肚菌 高产羊肚菌新菌株A11 紫外和热双灭活 原生质体融合 区间简单重复序列标记分析 

分 类 号：S332.5[农业科学—作物遗传育种]