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在森林腐殖质上形成的苏格兰松菌根际中,牛肝菌谷氨酰胺合成酶基因的组织、转录及酶活性

Suillus bovinus glutamine synthetase gene organization, transcription and enzyme activities in the Scots pine mycorrhizosphere developed on forest humus.

作者信息

Juuti Jarmo T, Jokela Sanna, Paulin Lars, Timonen Sari, Sen Robin

机构信息

Department of Bio- and Environmental Sciences, General Microbiology, PO Box 56 (Viikinkaari 9), FI-00014 University of Helsinki, Finland.

Institute of Biotechnology, DNA Sequencing Laboratory, PO Box 56 (Viikinkaari 4), FI-00014 University of Helsinki, Finland.

出版信息

New Phytol. 2004 Nov;164(2):389-399. doi: 10.1111/j.1469-8137.2004.01166.x.

Abstract

•  Glutamine synthetase (GS) expression and activity is of central importance for cellular ammonium assimilation and recycling. Thus, a full characterization of this enzyme at the molecular level is of critical importance for a better understanding of nitrogen (N) assimilation in the mycorrhizal symbiosis. •  Genomic and cDNA libraries of Suillus bovinus were constructed to isolate the GS gene, glnA, and corresponding cDNAs. The transcription initiation site was identified and transcription and enzyme activities were characterized in pure culture mycelium and mycorrhiza, and extramatrical mycelium samples harvested from Scots pine-Suillus bovinus microcosms grown on forest humus. •  Pure culture mycelium, mycorrhiza and extramatrical mycelium all exhibited equivalent levels of GS transcription, translation and enzyme activities. However, levels of transcription and enzyme activity did not correlate as a large majority of detectable transcripts showed specific 5'-end truncation. •  Our data suggest that GS is constitutively expressed and not directly affected by environmental conditions of the symbiotic N uptake. Any changes in the intracellular ammonium level are most likely handled by regulatory flexibility of GS at enzyme level.

摘要

• 谷氨酰胺合成酶(GS)的表达和活性对于细胞铵同化和循环至关重要。因此,在分子水平上全面表征这种酶对于更好地理解菌根共生中的氮(N)同化至关重要。

• 构建了牛肝菌的基因组文库和cDNA文库,以分离GS基因glnA和相应的cDNA。确定了转录起始位点,并在纯培养菌丝体、菌根以及从生长在森林腐殖质上的苏格兰松-牛肝菌微观体系中收获的根外菌丝体样品中对转录和酶活性进行了表征。

• 纯培养菌丝体、菌根和根外菌丝体均表现出同等水平的GS转录、翻译和酶活性。然而,转录水平和酶活性并不相关,因为大多数可检测到的转录本显示出特定的5'端截短。

• 我们的数据表明,GS是组成型表达的,不受共生氮吸收环境条件的直接影响。细胞内铵水平的任何变化很可能通过GS在酶水平上的调节灵活性来处理。

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