单 位：
山东省应用真菌重点实验室/青岛农业大学生命科学学院
关键词：
长根菇;双单杂交;小奥德蘑酮;木霉抗病性;SNP位点分析
摘 要：
【目的】利用双单杂交育种技术选育长根菇新菌株,并建立双单杂交后代的快速准确鉴定方法,以获得活性成分含量高、抗病性强的长根菇优质菌株。【方法】利用双单杂交技术进行长根菇杂交育种,通过拮抗试验、细胞核染色观察和SNP位点分析对双单杂交后代进行鉴别,以液体深层发酵和比色法测定杂合菌株的小奥德蘑酮产量,并通过对峙培养鉴定杂合菌株的木霉抗性。【结果】通过对长根菇亲本菌株OuE进行单孢分离,获得14株单核菌丝后代,编号依次为OuE01～OuE14。将获得的单核菌丝后代与亲本双核体菌株OuC进行双单杂交,经拮抗试验和SNP位点分析共筛选获得14株双单杂交后代。经过与两个亲本菌株(OuC和OuE)比较发现,杂交后代OuCE02、OuCE08和OuCE12的菌丝平均生长速度分别为16.00、16.10和15.30 mm/d,均显著高于两个亲本菌株(P<0.05);杂交后代OuCE03、OuCE05、OuCE08和OuCE10的菌丝发酵液小奥德蘑酮含量较亲本菌株OuC分别提高6.07%、47.66%、4.67%和15.42%;杂交后代OuCE01和OuCE08显示出较亲本菌株更强的木霉抗性。其中,杂交后代OuCE08在菌丝生长速度、小奥德蘑酮产量和木霉抗性方面均优于两个亲本菌株。【结论】双单杂交是一种快速有效获得食用菌新菌株的遗传育种方法,通过该方法筛选出一株高产小奥德蘑酮、高抗木霉的长根菇新菌株OuCE08。长根菇双单杂交后代快速、准确的检测鉴定可先采用拮抗试验作为初筛手段,再利用SNP位点分析进行复筛。
译 名：
Breeding of high oudenone production and Trichoderma resistant Oudemansiella raphanipes strain by dikaryon-monokaryon mating
作 者：
YU Hao;GE Zhi-hao;XU Li-li;GUO Li-zhong;Shandong Province Key Laboratory of Applied Mycology/College of Life Science,Qingdao Agriculture University;
关键词：
Oudemansiella raphanipes;;dikaryon-monokaryon mating;;oudenone;;Trichoderma resistance;;SNP loci analysis
摘 要：
【Objective】To breed new Oudemansiella raphanipes(also known as Hymenopellis raphanipes or O. radicata)strain by dikaryon-monokaryon mating,construct fast and accurate hybrid offspring identification methods,and produce high oudenone yield and high Trichoderma resistant cultivar.【Method】Dikaryon-monokaryon mating was used to perform genetic breeding of O. raphanipes. Hybrid offsprings were identified by antagonism test,nucleus fluorescence microscope observation and single nucleotide polymorphism(SNP)analysis. Oudenone yield of heterozygous strains were detected by submerged fermentation and colorimetry,the resistance to Trichoderma was determined using plate confrontation test.【Result】In this study,14 monokaryotic strains were isolated from the basidiospores of dikaryotic strain OuE,and their numbers were OuE01-OuE14. Dikaryon-monokaryon matings were performed between monokaryotic strains and dikaryotic strain OuC. Fourteen hybrid dikaryon offsprings were identified by plate antagonism test and single nucleotide polymorphisms analysis. Among them,the average growth rates of hypha of OuCE02,OuCE08 and OuCE12 were 16.00,16.10 and 15.30 mm/d,respectively,which were significantly higher than those of the parental strains(P<0.05). The oudenone yield of hybrid offsprings OuCE03,OuCE05,OuCE08 and OuCE10 increased 6.07%,47.66%,4.67% and15.42%,respectively,compared with parental strain OuC. OuCE01 and OuCE08 showed higher Trichoderma resistance than both parental strains. The growth rate,oudenone yield and Trichoderma resistance of OuCE08 were all better than the two parental strains.【Conclusion】The rapid and accurate methods to identify dikaryon-monokaryon mating offsprings are constructed. A new O. raphanipes strain,OuCE08,is obtained,which grows fast,produces high yield of oudenone and has high Trichoderma resistance. This study proves that dikaryon-monokaryon mating offsprings can be firstly selected by antagonism test and then verified by SNP analysis. This study provides a high quality cultivar for O. raphanipes production.
