彩色豆马勃是重要的林木外生菌根真菌,采用RT-PCR和cDNA末端快速扩增法(RACE)从彩色豆马勃中克隆得到硝酸还原酶基因,将其命名为ptnr(GenBank登录号为KC698974)。ptnr cDNA全长2 951 bp,ORF长2 754 bp,编码917个氨基酸,推测所得蛋白质的分子质量为102.06 ku,等电点为6.34。序列相似性检索和系统进化树分析结果显示:彩色豆马勃ptnr编码的氨基酸序列与担子菌皱木耳硝酸还原酶基因的氨基酸序列同源性最高为60%,与担子菌灰盖鬼伞的同源性为57%,与外生菌根真菌粘滑菇和双色蜡蘑的同源性分别为53%和56%。
Abstract
Pisolithus tinctorius is an important forest ectomycorrhizal fungus with wide application in root stimulations. Nitrate reductase gene, named as ptnr (GenBank accession number KC698974), was cloned from P. tinctorius, using reverse transcript-PCR(RT-PCR)and rapid amplification of cDNA ends(RACE). The annotated ptnr cDNA had a total length of 2 951 bp, with a 2 754 bp ORF, coding 917 amino acids. The encoded protein was estimated at the length of 102.06 ku and with an isoelectric point of 6.34. Both sequence similarity retrieval and phylogenetic tree analyses showed the amino acid sequence homology between P. tinctorius encoded nitrate reductase and Auricularia delicate encoded reductase was the highest, which could reach 60%, while the nitrate reductase sequence homology between P. tinctorius and Coprinopsis cinerea, or ectomycorrhizal fungi Hebeloma cylindrosporum, or Laccaria bicolor is 57%, 53% and 56%, respectively. Our study provides a basis for further research of genes involved in the symbiotic association between ectomycorrhizal fungi and forest.
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基金
收稿日期:2013-01-31 修回日期:2013-06-03
基金项目:国家林业公益性行业科研专项项目(201004061); “十二五”国家科技支撑计划(2012BAD19B0703); 江苏省普通高校研究生科研创新计划项目(CXZZ12_0547); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:周爱东,博士生。*通信作者:吴小芹,教授。E-mail:xqwu@njfu.com.cn。
引文格式:周爱东,吴小芹,王明生,等. 彩色豆马勃硝酸还原酶基因克隆与序列分析[J]. 南京林业大学学报:自然科学版,2013,37(6):22-26.
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