摘要
以采集自北京香山公园加拿大杨（Populus canadensis）的槐耳子实体为材料,利用组织分离获得纯培养并保存,编号为XS-01.结合形态与ITS鉴定确定其分类地位,进一步开展菌丝最适生长条件和液体发酵产漆酶研究.通过PCR扩增,获得长度为598 bp的部分ITS序列（KY933481）,采用MEGA 6.0构建发育树、测定遗传距离,XS-01与槐生多年卧孔菌（Perenniporia robiniophila）相似性最高,为100%,遗传距离为0.000.进一步结合子实体和菌丝形态,确定XS-01为槐生多年卧孔菌.结合生长速度和长势,确定了该菌株的最适生长碳源为淀粉和麦芽糖,最适生长氮源为酵母浸粉,最适生长C/N比为30/1-60/1,最适生长温度为32℃,最适生长pH 7,最适生长因子为VB1.Cu^（2＋）诱导的胞外漆酶发酵研究表明,1.0 mmol/L Cu^（2＋）对漆酶产量有显著的促进作用,产量在96 h达到最大值,为417.5 U/mL,比对照组提高93.4%,而2.0 mmol/L Cu^（2＋）对漆酶产量有显著抑制作用,96 h酶活为79.0 U/mL,是对照组的36.6%.本研究获得具有高产胞外漆酶能力的槐耳菌株,具有潜在的应用开发价值.
In the present study, fruiting bodies of a wild medicinal mushroom, ‘Huaier,＇ were collected from Populus canadensis in the Beijing Xiangshan Park. The pure culture strain was obtained from fruiting bodies using the tissue isolation method. It was stored and numbered as XS-01. It was systematically classified using morphological and ITS identification. Further studies were focused on mycelia optimum culture conditions and laccase production by liquid fermentation. A 598-bp partial ITS region sequence（Gen Bank accession number KY93348） was obtained using PCR method. Phylogenetic tree and genetic distance analysis were performed using the MEGA 6.0 software. The present strain possessed the highest homology（100%） with Perenniporia robiniophila, and the genetic distances were 0.000. Based on the ITS sequencing and morphological characteristics of fruiting bodies and mycelia, XS-01 was identified as P. robiniophila. Based on mycelial growth rate and quality, mycelia optimum culture conditions were revealed to be as follows： the optimum carbon sources were starch and maltose, the optimum nitrogen source was yeast extracts, the optimum C/N ratio range was 30/1-60/1, the best growth temperature was 32 ℃, the optimum pH was 7, and the optimum growth factor was VB1. Further study of Cu2＋ on extracellular laccase production revealed that 1.0 mmol/L Cu^（2＋） could significantly enhance the enzyme production, with the highest activity of 417.5 U/m L when cultured for 96 h and an increase ratio of 93.4% to the control. On the other hand, 2.0 mmol/L Cu^（2＋） can markedly decrease the enzyme production laccase activity at 96 h to 79.0 U/mL, which was 36.6% of that of the control. In conclusion, a pure strain of T. robiniophila with high extracellular laccase activity was obtained, suggesting its potential application for artificial cultivation and laccase production.
作者
徐鑫
陈青君
胡渤洋
王寿南
张国庆
王珊珊
李兵
韩鹏
张宇轩
XU Xin;CHEN Qingjun;HU Boyang;WANG Shounan;ZHANG Guoqing;WANG Shanshan;LI Bing;HAN Peng;ZHANG Yuxuan(College of Biological Science and Engineering, Beijing University of Agriculture, Key Laboratory of Urban Agriculture （North China） of Ministry of Agriculture, Beijing 102206, China;Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China)
出处
《应用与环境生物学报》
CAS
CSCD
北大核心
2018年第3期570-575,共6页
Chinese Journal of Applied and Environmental Biology
基金
北京市自然科学基金项目(5162006)
北京市科技新星计划项目(xx2015B025)
农业部华北都市农业重点实验室开放课题(kf 2017012)
北京市本科生科学研究计划项目资助~~
关键词
槐生多年卧孔菌
ITS鉴定
培养条件
漆酶
Perenniporia robiniophila
ITS identification
culture condition
laccase
