Faculty of Agriculture, Tottori University, Tottori, Japan.
Appl Environ Microbiol. 2013 Aug;79(15):4520-7. doi: 10.1128/AEM.00561-13. Epub 2013 May 10.
The enormous number of spores produced by fruiting bodies during cultivation of mushrooms can lead to allergic reactions of workers, reduction of commercial value, spread of mushroom disease, pollution of facilities, and depletion of genetic diversity in natural populations. A cultivar harboring a sporulation-deficient (sporeless) mutation would be very useful for preventing these problems, but sporeless commercial cultivars are very limited in usefulness because sporeless traits are often linked with traits that are unfavorable for commercial cultivation. Thus, identifying a causal gene of a sporeless phenotype not linked to the adverse traits in breeding and cultivation is crucial for the establishment of sporeless breeding using a strategy employing targeting induced local lesions in genomes (TILLING) in cultivated mushrooms. We used a Pleurotus pulmonarius (Fr.) Quél. sporeless strain to identify and characterize the single recessive gene controlling the mutation. The 3,853-bp stpp1 gene encodes a protein of 854 amino acids and belongs to the MutS homolog (MSH) family associated with mismatch repair in DNA synthesis or recombination in meiosis. Gene expression analysis of the fruiting body showed that this gene is strongly expressed in the gills. Phenotypic analysis of disruptants formed by gene targeting suggested a reproducible sporeless phenotype. Mutants deficient in a functional copy of this gene have no unfavorable traits for sporeless cultivar breeding, so this gene will be an extremely useful target for efficient and versatile sporeless breeding in P. pulmonarius and various other cultivated mushrooms.
在蘑菇栽培过程中,子实体产生的大量孢子会导致工人过敏反应、商业价值降低、蘑菇病传播、设施污染以及自然种群遗传多样性减少。一个含有孢子缺陷(无孢子)突变的品种将非常有助于预防这些问题,但无孢子商业品种的用途非常有限,因为无孢子特性通常与不利于商业栽培的特性相关联。因此,在利用靶向诱导基因组局部突变(TILLING)策略进行无孢子育种时,在育种和栽培中识别与不良性状无关的无孢子表型的因果基因对于建立无孢子育种至关重要。我们使用了一个糙皮侧耳(Fr.)Quél.无孢子菌株来鉴定和表征控制该突变的单个隐性基因。3853bp 的 stpp1 基因编码一个由 854 个氨基酸组成的蛋白质,属于 MutS 同源物(MSH)家族,与 DNA 合成中的错配修复或减数分裂中的重组有关。子实体的基因表达分析表明,该基因在菌褶中强烈表达。通过基因靶向形成的突变体的表型分析表明,该基因具有可重复的无孢子表型。该基因功能缺失的突变体没有无孢子栽培品种育种的不利性状,因此该基因将成为糙皮侧耳和各种其他栽培蘑菇高效、通用的无孢子育种的极其有用的目标。