Elucidation of the mating system and development of breeding technology in Agaricus blazei

姬松茸交配系统的阐明及育种技术的发展

• 批准号: 16580002
• 负责人: FUKUDA Masaki
• 金额: $ 1.02万
• 依托单位: Shinshu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16580002/
• 关键词: Agaricus blazei; Himematsutake; mating system; breeding; AFLP analysis

The development of Agaricus blazei cultivars with superior properties such as high fruiting body productivity will be very important for promoting commercial production. There is, however, little information available about the development of A.blazei cultivars. Therefore, fundamental genetic research on A.blazei takes on additional importance. This study was performed to examine genetic differences between heterokaryotic strains and their single-spore isolates (SSIs) and to confirm hybrid formation by pairing of SSIs based on AFLP profiles. Furthermore, stimulation conditions of the spore germination and de-heterokaryotization by the protoplast regeneration method.The total number of AFLP (Amplified Fragment Length Polymorphism) bands from individual SSIs was clearly less than those from their parental strains (heterokaryotic strains), indicating that the discrimination of heterokaryotic strain and its SSIs was possible by the AFLP profile. Many of polymorphic AFLP bands from the parental strains segregated in SSIs at a ratio of 1 : 1, suggesting that the SSIs are homokaryotic. Although it was very difficult to confirm hybrid formation by only colony morphologies of the isolates from paired colonies between SSIs, AFLP bands are suitable genetic makers for the confirmation of successful crosses between SSIs in A.blazei. Effects of several factors on basidiospore germination of A.blazei were examined : bark compost-extracts medium as basal medium, addition of succinic acid to medium, co-culture with living mycelia of A.blazei heterokaryotic strain, and heat treatment of basidiospores before culturing would be effective to stimulate spore germination. In addition, protoplast regeneration method seemed to be useful for de-heterokaryotization in A.blazei

因此，选育出具有高子实体生产力等上级性状的巴西蘑菇品种，对促进巴西蘑菇的商业化生产具有重要意义。然而，关于巴西曲霉栽培品种的发展信息很少。因此，对巴西曲霉的基础遗传学研究具有额外的重要性。本研究旨在通过AFLP分析来研究杂合菌株及其单孢分离物（SSI）之间的遗传差异，并通过配对SSI来确认杂种的形成。此外，还研究了原生质体再生法诱导孢子萌发和去异核化的条件，结果表明，单个SSR标记的AFLP（Amplified Fragment Length Polymorphism）条带总数明显少于其亲本菌株（杂合菌株），说明利用AFLP图谱可以区分杂合菌株及其SSR标记。亲本菌株的许多多态性条带以1：1的比例分离在SSI中，表明SSI是同核的。虽然这是非常困难的，以确认杂交形成的唯一的菌落形态的分离物从配对的菌落之间的SSI，AFLP带是合适的遗传标记，用于确认成功的杂交之间的SSI在A. blazei。研究了几种因素对巴西曲霉担孢子萌发的影响：以树皮堆肥浸提液为基本培养基，在培养基中添加琥珀酸，与巴西曲霉菌丝体共培养，培养前对担孢子进行热处理，均能有效促进孢子萌发。此外，原生质体再生方法也可用于巴西曲霉的去异核化