硬毛粗盖孔菌过氧化氢酶家族基因鉴定及表达分析

硬毛粗盖孔菌是一种结实能力强的耐热真菌,为了探究其过氧化氢酶(CAT)基因家族的基本特征和功能,分别对不同交配型单核体Ct001_29和Ct001_31基因组的CAT基因家族进行了鉴定,并分析了不同CAT家族基因在不同温度条件下培养的菌丝、原基和子实体中的表达特征。硬毛粗盖孔菌基因组上有3个CAT基因(Ctcat1-Ctcat3),编码510-743个氨基酸;Ctcat1、Ctcat2和Ctcat3的等位基因之间的结构及序列相对保守,但编码区也存在少量SNP变异(6-14个)。在25 ℃培养菌丝和35 ℃培养菌丝、原基及子实体中,35 ℃菌丝的CAT酶活最高,为278 U/mg蛋白,原基期CAT酶活最低,为4 U/mg蛋白。Ctcat2的表达丰度显著高于Ctcat1和Ctcat3,Ctcat1和Ctcat3具有相似的表达模式,在35 ℃菌丝、原基和子实体中均上调表达;而Ctcat2具有相反的表达模式,在35 ℃菌丝、原基和子实体中均下调表达。此外,Ctcat2在原基时期、Ctcat3在25 ℃菌丝和35 ℃菌丝中具有偏好表达,Ct29cat2和Ct29cat3的表达量均高于各自的等位基因。本研究所发现的CAT等位基因表达偏好为进一步揭示大型真菌CAT的基因功能奠定了基础。

Identification and expression analysis of the catalase family genes in Funalia trogii

Funalia trogii is a typical thermotolerant fungus with strong fruiting ability. To clarify the basic characteristics and functions of catalase (CAT) gene family, the identification of CAT in genomes of two monokaryons (Ct001_29 and Ct001_31) with different mating types was conducted, and the CAT expression profiles were examined in mycelia (cultured at different temperatures), primordia, and fruiting bodies. There were three CAT genes (Ctcat1-Ctcat3) in the genome of F. trogii, encoding 510-743 amino acids. The gene structure and sequences of the alleles of Ctcat1, Ctcat2, and Ctcat3 were relatively conserved, but a few SNPs (6-14) existed in their coding regions. CAT activity varied with culture temperature. The maximal CAT activity (278 U/mg protein) was detected in mycelia cultured at 35 °C, and the minimal CAT activity (4 U/mg protein) was detected in primordia. The expression abundance of Ctcat2 was significantly higher than that of Ctcat1 or Ctcat3. Ctcat1 and Ctcat3 had similar expression patterns, showing up-regulated expression in mycelia, primordia, and fruiting bodies. However, Ctcat2 had the opposite expression pattern, showing down-regulated expression in mycelia cultured at 35 °C, primordia, and fruiting bodies. Allelic expression bias was detected in Ctcat2 (in primordia) and Ctcat3 (in mycelia cultured at 25 °C and 35 °C). The expression profiles of Ct29cat2 and Ct29cat3 were both higher than those of their alleles. The allelic expression bias revealed in this study lays the foundation for studying the gene function of macrofungal CAT in the future.