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[1]高升,张宇,伍圆圆,等.耐热毛栓孔菌S0301同核体菌株的快速鉴定及产漆酶能力比较[J].应用与环境生物学报,2019,25(01):151-155.[doi:10.19675/j.cnki.1006-687x.2018.01017]
 GAO Sheng,ZHANG Yu,WU Yuanyuan,et al.A rapid method for the identification of homokaryotic strains in Trametes trogii S0301 and their laccase-producing ability[J].Chinese Journal of Applied & Environmental Biology,2019,25(01):151-155.[doi:10.19675/j.cnki.1006-687x.2018.01017]
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耐热毛栓孔菌S0301同核体菌株的快速鉴定及产漆酶能力比较
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
25卷
期数:
2019年01期
页码:
151-155
栏目:
研究论文
出版日期:
2019-02-25

文章信息/Info

Title:
A rapid method for the identification of homokaryotic strains in Trametes trogii S0301 and their laccase-producing ability
作者:
高升 张宇 伍圆圆 杨恩 徐慧妮 伊日布斯 陈玉惠 严金平
1昆明理工大学生命科学与技术学院生物转化实验室 昆明 650500 2西南林业大学生命科学学院 昆明 650224
Author(s):
GAO Sheng1 ZHANG Yu1 WU Yuanyuan1 YANG En1 XU Huini1 IRBIS Chagan1 CHEN Yuhui2 & YAN Jinping1**
1 Laboratory of Bioconversion of Life Science and Technology College, Kunming University of Science and Technology, Kunming 650500, China 2 College of Life Science, Southwest Forest University, Kunming 650224, China
关键词:
毛栓孔菌漆酶原生质体制备交配型基因同核体菌株异核体菌株
Keywords:
Trametes trogii laccase protoplast regeneration mating-type gene homokaryotic strain heterokaryoteyotic strain
分类号:
Q936
DOI:
10.19675/j.cnki.1006-687x.2018.01017
摘要:
栓孔菌属(Trametes)真菌菌株漆酶同工酶种类多,产漆酶优势独特,是重要的工业漆酶来源之一. 为建立方便快捷的获得同核体菌株的方法,以高产漆酶耐热毛栓孔菌Trametes trogii S0301为研究对象,在利用原生质体再生技术得到再生单菌、通过PCR克隆获得该菌株交配型基因b1、b2位点序列的基础上,设计b1、b2交配型基因特异引物并对各再生菌落中不同交配型基因类型进行识别,结合经典的锁状联合显微镜观察,共从45个性状稳定的再生菌株中获得了19个b1b2型异核体菌株,26个b1型同核体菌株,同核体菌株检出率为57.8%,且PCR鉴定结果和锁状联合观察结果一致. 此外,还发现b1型同核体菌株菌丝细、菌落稀疏,有更快的蔓延速度,在不添加漆酶诱导剂Cu2+的固体培养基中表现出更强的产漆酶能力. 本研究表明PCR克隆交配型基因的方法适用于同核体菌株的快速鉴定,同时b1型同核体菌株具有产漆酶的优势. (图3 表3 参22)
Abstract:
This study used heat-resistant Trametes trogii S0301 with high yield of laccase as the research object. The protoplast regeneration technology was used to obtain single regenerated strains, and PCR cloning was used to obtain the mating-type genes (b1 and b2); next, the specific primers for b1 and b2 genes were designed to identify the mating genotype in different regenerated colonies. As a classic standard, the clamp connection in each regenerated strain was detected using microscopic observation. In all, 45 stable regeneration strains were selected for further study. Among them, 19 b1b2 heterokaryotic strains and 26 b1 homokaryotic strains were obtained using PCR detecting method, and the detection rate of homokaryotic strains was 57.8%, which was consistent with the results of the clamp connection observation. In this study, we also found that b1 homokaryotic strains showed different properties from b1b2 heterokaryotic strains, such as thinner hypha, more rapid spread, and sparser colonies on the solid medium. In particular, b1 homokaryotic strains showed higher laccase-producing ability on the solid medium without Cu2+ as the laccase inducer, compared with the b1b2 heterokaryotic strains. This study showed that the way of PCR cloning mating-type genes was suitable for the rapid identification of homokaryotic strains. Moreover, b1 homokaryotic strains had the advantage of producing laccase.

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更新日期/Last Update: 2019-02-25