高卢蜜环菌几丁质酶基因的生物信息学分析与表达
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摘要
高卢蜜环菌是一种兼性寄生真菌,在天麻栽培过程中作为天麻的唯一营养源。几丁质酶作为一种糖苷水解酶,在高卢蜜环菌的生长发育及其与天麻的共生过程中发挥重要作用。高卢蜜环菌几丁质酶基因共有22条,对这些几丁质酶基因的氨基酸序列进行生物信息分析,发现有12条几丁质酶基因均含有糖苷水解酶18家族(GH18)结构域。对高卢蜜环菌、奥氏蜜环菌、天麻、酿酒酵母和哈茨木霉的几丁质酶氨基酸序列进行系统进化分析,进一步预测高卢蜜环菌中几丁质酶基因功能。利用独角金内酯类似物GR24诱导高卢蜜环菌分枝,并分析22条几丁质酶基因的表达模式,发现8条基因可能参与蜜环菌分枝、细胞壁降解和重塑等生理过程。
Armillaria gallica is a facultative parasitic fungus which is the only nutrient source of Gastrodia elata during its cultivation.Chitinase,as a glycosidic hydrolytic enzyme,plays an important role in the growth,development,stress tolerance and symbiotic signal transduction of A. gallica. There were 22 chitinase genes in A. gallica. Bioinformatics analysis of amino acid sequence of these chitinase genes revealed that 12 chitinase genes contained glycosidase 18 family( GH18) domain. Chitinase amino acid sequences of A. gallica,A. ostoyae,G. elata,Saccharomyces cerevisiae and Trichoderma harzianum were analyzed byclustering trees,so as to further predict the gene function of chitinase in A. gallica. Induction of A. gallica branching with strigolactone analogue GR24,high-throughput sequencing technology based on the induction of branch group( MHJ1),uninduced branch group( MHJ2) and blank control group( MHJ3) is used to detect the expression quantity,the transcription level data of 22 chitinase genes were obtained and the heat map was generated for expression pattern analysis. It was found that 8 genes may be involved in physiological processes such as A. gallica branching,cell wall degradation and remodeling. In this paper,the function of chitinase gene in A. gallica was just preliminarily analyzed and predicted.
Armillaria gallica is a facultative parasitic fungus which is the only nutrient source of Gastrodia elata during its cultivation.Chitinase,as a glycosidic hydrolytic enzyme,plays an important role in the growth,development,stress tolerance and symbiotic signal transduction of A. gallica. There were 22 chitinase genes in A. gallica. Bioinformatics analysis of amino acid sequence of these chitinase genes revealed that 12 chitinase genes contained glycosidase 18 family( GH18) domain. Chitinase amino acid sequences of A. gallica,A. ostoyae,G. elata,Saccharomyces cerevisiae and Trichoderma harzianum were analyzed byclustering trees,so as to further predict the gene function of chitinase in A. gallica. Induction of A. gallica branching with strigolactone analogue GR24,high-throughput sequencing technology based on the induction of branch group( MHJ1),uninduced branch group( MHJ2) and blank control group( MHJ3) is used to detect the expression quantity,the transcription level data of 22 chitinase genes were obtained and the heat map was generated for expression pattern analysis. It was found that 8 genes may be involved in physiological processes such as A. gallica branching,cell wall degradation and remodeling. In this paper,the function of chitinase gene in A. gallica was just preliminarily analyzed and predicted.
引文
[1]黄靖雯,赖长江生,袁媛,等.猪苓与蜜环菌化学成分研究的相关分析进展[J].中国中药杂志,2017,42(15):2905.
[2]Tsai C C,Wu K M,Chiang T Y,et al.Comparative transcriptome analysis of Gastrodia elata,(Orchidaceae)in response to fungus symbiosis to identify gastrodin biosynthesis-related genes[J].BMC Genomics,2016,17(1):212.
[3]Balestrini R,Bonfante P.Cell wall remodeling in mycorrhizal symbiosis:a way towards biotrophism[J].Front Plant Sci,2014,5:237.
[4]Free S J.Fungal cell wall organization and biosynthesis[J].Adv Genet,2013,81:33.
[5]程笑笑,冯自力,冯鸿杰,等.真菌源几丁质酶在植物抗真菌病害中的应用[J].植物保护,2017,43(3):29.
[6]潘华平.酵母细胞壁合成及其相关基因的研究[D].无锡:江南大学,2017.
[7]Adams D J.Fungal cell wall chitinases and glucanases[J].Microbiology,2004,150(7):2029.
[8]Yuan Y,Jin X,Liu J,et al.The Gastrodia elata genome provides insights into plant adaptation to heterotrophy[J].Nat Commun,2018,9(1):1615.
[9]Zwanenburg B,Tomá2 Pospí2il,Sanja C'avar Zeljkovic'.Strigolactones:new plant hormones in action[J].Planta,2016,243(6):1311.
[10]Ikegami T,Okada T,Hashimoto M,et al.Solution structure of the chitin-binding domain of Bacillus circulans WL-12 chitinase A1[J].J Biol Chem,2000,275(18):13654.
[11]Oyeleye A,Normi Y M.Chitinase:diversity,limitations,and trends in engineering for suitable applications[J].Biosci Rep,2018,38(4):1.
[12]Stringer M A,Dean R A,Sewall T C,et al.Rodletless,a new Aspergillus developental mutant induced by directed gene inactivation[J].Gene Dev,1991,5(7):1161.
[13]Joris B.Modular design of the Enterococcus hirae muramidase-2and Streptococcus faecalis autolysin[J].FEMS Microbiol Lett,1992,91(3):257.
[14]Simon A,Hawkins P T,Ellson C D,et al.The PX domain:a new phosphoinositide-binding module[J].J Cell Sci,2002,115(6):1099.
[15]Guo T,Wang H C,Xue W Q,et al.Phylogenetic analyses of Armillariareveal at least 15 phylogenetic lineages in China,seven of which are associated with cultivated Gastrodia elata[J].PLoSONE,2016,11(5):e0154794.
[16]Hoell I A,Klemsdal S S,Vaaje-Kolstad G,et al.Overexpression and characterization of a novel chitinase from Trichoderma atroviride strain P1[J].Biochim Biophys Acta,2005,1748(2):180.
[17]Lima L H C,Ulhoa C J,Fernandes A P,et al.Purification of a chitinase from Trichoderma sp.and its action on Scierotium rolfsii and Rhizoctonia solani cell walls[J].J Gen Appl Microbiol,1997,43(1):31.
[18]Sharma V,Bhandari P,Singh B,et al.Chitinase expression due to reduction in fusaric acid level in an antagonistic Trichoderma harzianum S17TH[J].Indian J Microbiol,2013,53(2):214.
[19]Carstens M,Vivier M A,Pretorius I S.The Saccharomyces cerevisiae chitinase,encoded by the CTS1-2 gene,confers antifungal activity against Botrytis cinerea to transgenic tobacco[J].Transgenic Res,2003,12(4):497.
[20]Pusztahelyi Tünde.Chitin and chitin-related compounds in plantfungal interactions[J].Mycology,2018,9(3):189.
[21]Gow N A R,Latge J P,Munro C A.The fungal cell wall:structure,biosynthesis,and function[J].Microbiol Spectr,2017,5(3):1.
[22]袁媛,于军,黄璐琦,等.基于同源基因功能分化的药用植物活性成分变异机制研究[J].中国中药杂志,2015,40(6):1023.
[23]Desaki Y,Miyata K,Suzuki M,et al.Plant immunity and symbiosis signaling mediated by Lys M receptors[J].Innate Immun,2017,24(2):92.
[2]Tsai C C,Wu K M,Chiang T Y,et al.Comparative transcriptome analysis of Gastrodia elata,(Orchidaceae)in response to fungus symbiosis to identify gastrodin biosynthesis-related genes[J].BMC Genomics,2016,17(1):212.
[3]Balestrini R,Bonfante P.Cell wall remodeling in mycorrhizal symbiosis:a way towards biotrophism[J].Front Plant Sci,2014,5:237.
[4]Free S J.Fungal cell wall organization and biosynthesis[J].Adv Genet,2013,81:33.
[5]程笑笑,冯自力,冯鸿杰,等.真菌源几丁质酶在植物抗真菌病害中的应用[J].植物保护,2017,43(3):29.
[6]潘华平.酵母细胞壁合成及其相关基因的研究[D].无锡:江南大学,2017.
[7]Adams D J.Fungal cell wall chitinases and glucanases[J].Microbiology,2004,150(7):2029.
[8]Yuan Y,Jin X,Liu J,et al.The Gastrodia elata genome provides insights into plant adaptation to heterotrophy[J].Nat Commun,2018,9(1):1615.
[9]Zwanenburg B,Tomá2 Pospí2il,Sanja C'avar Zeljkovic'.Strigolactones:new plant hormones in action[J].Planta,2016,243(6):1311.
[10]Ikegami T,Okada T,Hashimoto M,et al.Solution structure of the chitin-binding domain of Bacillus circulans WL-12 chitinase A1[J].J Biol Chem,2000,275(18):13654.
[11]Oyeleye A,Normi Y M.Chitinase:diversity,limitations,and trends in engineering for suitable applications[J].Biosci Rep,2018,38(4):1.
[12]Stringer M A,Dean R A,Sewall T C,et al.Rodletless,a new Aspergillus developental mutant induced by directed gene inactivation[J].Gene Dev,1991,5(7):1161.
[13]Joris B.Modular design of the Enterococcus hirae muramidase-2and Streptococcus faecalis autolysin[J].FEMS Microbiol Lett,1992,91(3):257.
[14]Simon A,Hawkins P T,Ellson C D,et al.The PX domain:a new phosphoinositide-binding module[J].J Cell Sci,2002,115(6):1099.
[15]Guo T,Wang H C,Xue W Q,et al.Phylogenetic analyses of Armillariareveal at least 15 phylogenetic lineages in China,seven of which are associated with cultivated Gastrodia elata[J].PLoSONE,2016,11(5):e0154794.
[16]Hoell I A,Klemsdal S S,Vaaje-Kolstad G,et al.Overexpression and characterization of a novel chitinase from Trichoderma atroviride strain P1[J].Biochim Biophys Acta,2005,1748(2):180.
[17]Lima L H C,Ulhoa C J,Fernandes A P,et al.Purification of a chitinase from Trichoderma sp.and its action on Scierotium rolfsii and Rhizoctonia solani cell walls[J].J Gen Appl Microbiol,1997,43(1):31.
[18]Sharma V,Bhandari P,Singh B,et al.Chitinase expression due to reduction in fusaric acid level in an antagonistic Trichoderma harzianum S17TH[J].Indian J Microbiol,2013,53(2):214.
[19]Carstens M,Vivier M A,Pretorius I S.The Saccharomyces cerevisiae chitinase,encoded by the CTS1-2 gene,confers antifungal activity against Botrytis cinerea to transgenic tobacco[J].Transgenic Res,2003,12(4):497.
[20]Pusztahelyi Tünde.Chitin and chitin-related compounds in plantfungal interactions[J].Mycology,2018,9(3):189.
[21]Gow N A R,Latge J P,Munro C A.The fungal cell wall:structure,biosynthesis,and function[J].Microbiol Spectr,2017,5(3):1.
[22]袁媛,于军,黄璐琦,等.基于同源基因功能分化的药用植物活性成分变异机制研究[J].中国中药杂志,2015,40(6):1023.
[23]Desaki Y,Miyata K,Suzuki M,et al.Plant immunity and symbiosis signaling mediated by Lys M receptors[J].Innate Immun,2017,24(2):92.