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The LysM Receptor-Like Kinase LysM RLK1 Is Required to Activate Defense and Abiotic-Stress Responses Induced by Overexpression of Fungal chitinases in Arabidopsis Plants

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Molecular Plant 2012年第5期5卷 1113-1124页

作者： Yariv Brotman Udi Landau Smadar Pnini Jan Lisec Salma Balazadeh Bernd Mueller-Roeber Aviah Zilberstein Lothar Willmitzer Ilan Chet Ada Viterbo Genes and Small Molecules AG Willmitzer Max-Planck-lnstitut of Molecular Plant Physiology Am Muhlenberg 1 D-14476 Potsdam-Golm Germany Department of Plant Pathology and Microbiology The Smith Faculty of Agriculture Food and Environment The Hebrew University of Jerusalem Rehovot 76100 Israel Department of Plant Science Tel Aviv University Tel Aviv 69978 Israel University of Potsdam Institute of Biochemistry and Biology KarI-Liebknecht-Strae 24-25 Haus 20 14476 Potsdam-Golrn Germany

Application of crab shell chitin or pentamer chitin oligosaccharide to Arabidopsis seedlings increased toler- ance to salinity in wild-type but not in knockout mutants of the LysM Receptor-Like Kinasel （CERK1/LysM RLK1） gene, known to play a critical role in signaling defense responses induced by exogenous chitin. Arabidopsis plants overexpress- ing the endochitinase chit36 and hexoaminidase excyl genes from the fungus Trichoderma asperelleoides T203 showed increased tolerance to salinity, heavy-metal stresses, and Botrytis cinerea infection. Resistant lines, overexpressing fungal chitinases at different levels, were outcrossed to lysm rlkl mutants. Independent homozygous hybrids lost resistance to biotic and abiotic stresses, despite enhanced chitinase activity. Expression analysis of 270 stress-related genes, including those induced by reactive oxygen species （ROS） and chitin, revealed constant up-regulation （at least twofold） of 10 genes in the chitinase-overexpressing line and an additional 76 salt-induced genes whose expression was not elevated in the lysm rlkl knockout mutant or the hybrids harboring the mutation. These findings elucidate that chitin-induced signaling mediated by LysM RLK1 receptor is not limited to biotic stress response but also encompasses abiotic-stress signaling and can be conveyed by ectopic expression of chitinases in plants.

关键词： abiotic stress chitin-induced signaling chitinases LysM receptor kinase Trichoderma.

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Clustering and Expression Analysis of chitinases in Maize and Rice

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Journal of Life Sciences 2013年第3期7卷 244-251页

作者： Kui Xiang Wei-Tao Li Xue-Wei Chen Guang-Sheng Yuan Wei-Lan Chen Zhi-Ming Zhang Ya-Ou Shen Hai-Jian Lin Guang-Tang Pan Maize Research Institute Sichuan Agricultural University ChengDu 611130 China Rice Research Institute Sichuan Agricultural University ChengDu 611130 China

chitinases play an important role in regulating plant growth and development, especially defending themselves from fungal pathogens. It is important to do the biological analyses in crops. In this study, the result showed that the chitinases were distributed into the whole genome in rice, and nearly the whole genome in maize expect for Chromosome 9. The clustering results showed that one out of three chitinases from maize and rice belonged to new groups, which were separated from those in the conformed Classes I-VII. The identification of most amino acid sequences was very low among the chitinases from rice and/or maize. It was inferred that the chitinases with different functions were relatively stable during plant evolution. The relationship of chitinases expression between leaf blade and anther was positively significant in maize, but not significant in rice. Additionally, the ratio of chitinases with up- or down-regulated expression in sensitive maize under Fusarium moniliforme inoculation was different from that in sensitive rice under Magnaporthe grisea inoculation. It might be result from different tissues infected by different fungi. The number of chitinases from resistant maize was less than that from sensitive maize, which inferred that the resistant pathways on F. moniliforme should be not only chitin induced Pathogen-associated molecular PTI （patterns-triggered immunity） pathway, but also might include other PTI pathways that improve tolerance to F. moniliforme. The analysis of expression pattern of chitinases from maize and rice under fungi inoculation will be contributed into further research on the defense mechanism of fungi in crops.

关键词： chitinases fungi clustering expression maize rice.

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New insights on chitinases immunologic activities

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World Journal of Immunology 2016年第2期6卷 96-104页

作者： Michelino Di Rosa Violetta Maria Brundo Lucia Malaguarnera Department of Biomedical and Biotechnology Sciences University of Catania Department of Biological Geological and Environmental Sciences University of Catania

Mammalian chitinases and the related chilectins （ChiLs） belong to the GH18 family, which hydrolyse the glycosidic bond of chitin by a substrate-assisted mechanism. Chitin the fundamental component in the coating of numerous living species is the most abundant natural biopolymer. Mounting evidence suggest that the function of the majority of the mammalian chitinases is not exclusive to catalyze the hydrolysis of chitin producing pathogens, but include crucial role specifc in the immunologic activities. The chitinases and chitinase-like proteins are expressed in response to different proinflammatory cues in various tissues by activated macrophages, neutrophils and in different monocyte-derived cell lines. The mechanism and molecular interaction of chitinases in relation to immune regulation embrace bacterial infection, infammation, dismetabolic and degenerative disease. The aim of this review is to update the reader with regard to the role of chitinases proposed in the recent innate and adaptive immunity literature. The deep scrutiny of this family of enzymes could be a useful base for further studies addressed to the development of potential procedure directing these molecules as diagnostic and prognostic markers for numerous immune and infammatory diseases.

关键词： chitinases Chitinase like proteins Chronic infammation Immune regulation Autoimmunity

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Antifungal Potential of Beauveria bassiana on Solanum lycopersicum L. Infected with Fusarium oxysporum f. sp. lycopersici

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Phyton-International Journal of Experimental Botany 2023年第4期92卷 1235-1255页

作者： Henry López-López Nancy Ruiz-Lau Rocío Meza-Gordillo Víctor Manuel Ruiz-Valdiviezo Joseph Galthier Robledo-Luchetti Carlos Alberto Lecona-Guzmán Juan JoséVillalobos-Maldonado Luc Dendooven Joaquín Adolfo Montes-Molina Laboratorio de Biotecnología vegetal Tecnológico Nacional de Mexico/Campus Tuxtla GutiérrezTuxtla Gutiérrez29050Mexico CONACYT-Tecnológico Nacional de Mexico/Campus Tuxtla Gutiérrez Tuxtla Gutiérrez29050México ABACUS CinvestavCiudad de México07360México

The objective of this work was to evaluate the effect of Beauveria bassiana(Bb 1205)on controlling Fusarium oxysporum ***(Fol 17108)in tomato plants in greenhouse *** of Bb 1205 was the most promising among the agronomic variables and expression of the activity of the enzymesβ-1,3-glucanases and *** of Bb 1205 occurred at a concentration of 1×108 conidia·mL−1,which was administered onto the leaves,directly into the soil and via *** with Fol 17108 occurred with 1×106 spores·mL−1,which were added directly to the *** was used for measuring agronomic parameters,namely activity of chitinases andβ-1,3-glucanases in foliage and *** Bb 1205 was added to the soil,the chlorophyll index and aerial part length showed significant *** addition,it was determined that root length,fresh weight of foliage,flower,and fruit count increased 82 days after inoculation(dai).Chitinase activity induced by Bb 1205 in leaves and roots of tomato plants infected with Fol 17108 was observed when injected into the stem at 32 dai(41.8 and 11.6-fold,respectively).Inoculation on the foliage showed a 10-fold increase ofβ-1,3-glucanases in the roots after 82 *** for leaves,a 3.8-fold increase was found when the stem was *** the different in vivo applications,Bb 1205 activated its defenses by expressing the chitinase enzymes andβ-1,3-glucanase,thus reducing the damage caused by Fol 17108,demonstrating increase plant growth thereafter.

关键词： Mycoparasitism agronomic parameters chitinases β-1,3-glucanases

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Effects of Plant Extracts and Beauveria bassiana on the Activity of Defense-Related Enzymes in Solanum lycopersicum *** Interaction with Fusarium oxysporum ***

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Phyton-International Journal of Experimental Botany 2023年第9期92卷 2503-2518页

作者： JoséAdrian Perez-Robles Carlos Alberto Lecona-Guzmán Víctor Manuel Ruíz-Valdiviezo Joaquín Adolfo Montes-Molina Laboratorio de Biotecnología Vegetal Tecnológico Nacional de MexicoCampus Tuxtla GutiérrezTuxtla Gutiérrez29050Mexico

The objective to this work was to evaluate the enzymatic activity in the culture of Solanum lycopersicum *** with Fusarium oxysporum after the combined application of Beauveria bassiana and plant *** lycopersicum plantlets were transplanted 15 days after the *** days after transplanting,Beauveria bassiana spores were applied at a concentration of 1×10^(7)spores mL^(−1)onto soil(along with ***(N)and ***(H)leaf extracts)where *** plants were *** days after transplanting,spores of *** strain were applied at a concentration of 1×10^(6)spores mL^(−1)to soil where *** plants were *** development of *** plants was monitored for 114 days,whereby a randomized complete block treatment design was used,the roots of the plants were crushed with liquid nitrogen,after which UV/VIS spectrophotometry was used to determine protein concentration,as well as the activity ofβ-1,3-glucanases,peroxidases(POX,EC1.***.1.7),catalase and *** treatments combining *** and *** and *** extracts,had a significant difference in enzymatic activity levels,thus contributing to better defense mechanisms and a greater protection to *** plants in the presence of *** application of *** and plant extracts to the ground mitigated damage caused by *** on *** plants.

关键词： Glucanases chitinases peroxidases catalase defense mechanisms

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High level expression of Saccharomyces cerevisiae chitinase(ScCTS1)in Pichia pastoris for degrading chitin

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International Journal of Agricultural and Biological Engineering 2015年第5期8卷 142-150页

作者： Zhao Youxi Jiang Huihui Rao Zhiming Ji Yizhi Cheng Yanling Ma Yanhe Key Laboratory of Industrial Biotechnology Ministry of EducationSchool of BiotechnologyJiangnan UniversityWuxi 214122China Beijing Key Laboratory of Biomass Waste Resource Utilization Biochemical Engineering CollegeBeijing Union UniversityBeijing 100023China State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijing 100101China

Chitin is the second most abundant renewable biopolymer in the *** play important roles in the degradation of *** are produced by different organisms for different purposes,which are widely expressed in the three domains of life,ranging from archaea,bacteria,to fungi,yeasts,plants,insects,and even *** there are few reports about Saccharomyces cerevisiae chitinase(ScCTS1).The aim of this study was to realize the high level expression of *** ScCTS1 was cloned into the expression vector *** recombinant plasmid was linearized and transformed into competent Pichia pastoris *** screening by G418 plate,the fermentation conditions were ***,under the optimal fermentation conditions,ScCTS1 enzymatic activity reached up to 94.6 U/*** paper presents the first report on the heterologous expression of a full-length ScCTS1 with considerably high *** work will not only make a great stride towards its potential applications in biotechnology,but also facilitate elucidating the precise mechanism of yeast cell division.

关键词： chitinases heterologous expression Saccharomyces cerevisiae pichia pastoris biotechnology

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Green Synthesis and Physical and Chemical Characterization of Chitosans with a High Degree of Deacetylation, Produced by a Binary Enzyme System

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Journal of Life Sciences 2014年第3期8卷 276-282页

作者： Laidson Paes Gomes Cristina Tristao Andrade Joab Trajano Silva Eduardo Mere Del Aguila Vania Margaret Flosi Paschoalin Institute of Chemistry University Federal of Rio de Janeiro Rio de Janeiro 21941-909 R J Brazil Institute of Macromolecules Professora Eloisa Mano University Federal of Rio de Janeiro Rio de Janeiro 21945-598 R J Brazil

Chitosan is a biopolymer obtained from chitin, where the N-acetylglucosamine monomer is in its deacetylated form; this polymer is useful for a wide variety of industrial applications. The properties and uses of chitosan depend on its physical and chemical characteristics, which result from the treatments used for its production. In this study, we report the preparation and characterization ofchitosan oligosaccharides by a green synthesis from crystalline shrimp chitin, using a sequential enzyme treatment by chitinase and chitin deacetylase. chitinases were purified from grapes and used to rupture the crystalline shrimp chitin structure, modifying the crystallinity index from 57.6% to 15.9%. The resultant polymers were deacetylated using a recombinant chitin deacetylase from Saccharomyces cerevisiae, which was cloned and expressed in Pichia pastoris. The chitosans produced showed an estimated DA （degree of acetylation） of approximately 20%, and the molecular weights ranged from -7,600 to -3,700 after treatment in pH 3.0 and pH 6.0 for 10 min and 40 min, respectively. Physical and chemical characterization of the products indicated that enzyme fragmentation of chitin probably makes the acetamide groups more accessible to deacetylation, forming homogeneous polymers that are free of hazardous sub-products, have defined low molecular weights, and are highly deacetylated.

关键词： Chitosan binary enzyme system chitinases recombinant chitin deacetylase low acetylation degree low molecular weight.

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Enzymes of Entomopathogenic Fungi, Advances and Insights

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Advances in Enzyme Research 2014年第2期2卷 65-76页

作者： Lluvia de Carolina Sánchez-Pérez Juan Esteban Barranco-Florido Silvia Rodríguez-Navarro José Francisco Cervantes-Mayagoitia Miguel ángel Ramos-López Departamento de Producción Agrícola y Animal Universidad Autónoma Metropolitana México City México Departamento de Sistemas Biológicos Universidad Autónoma Metropolitana México City México Facultad de Química Universidad Autónoma de Querétaro Querétaro México

Entomopathogenic fungi (EF) are recognized biological control agents of insects. Basically, the entomopathogenic fungi pathogen activity depends on the ability of its enzymatic equipment, consisting of lipases, proteases and chitinases, which are in charge of breaking down the insect’s integument. Lipases are the first enzymes synthesized by the entomopathogenic fungi. Recently, a cytochrome P450 subfamily, referred as CYP52XI and MrCYP52 has been identified in Beauveria bassiana and Metarhizium robertsii, respectively. These break down long-chain alkenes and fatty acids to become initial nutrients. Subsequently, subtilisin type (Pr1) proteases sintetize;these enzymes are considered as virulence indicators and they are regulated by a signal transduction mechanism activated by the protein kinase A (PKA) mediated by AMPc. Through the employment of genetic engineering, it has been possible to increase virulence producing Pr1 recombinants with Androctonus australis neurotoxins or with chitinases, reducing the insect’s time of death. In the course of time, the Pr1 protease gene has presented evolutionary adaptations by gene duplication or horizontal transfer infecting different orders of insects. In the same way, the entomopathogenic fungi chitinases have presented a functional diversification. Currently, these have been phylogenetically classified into three subgroups, in accordance to the catalytic site domain and the chitin binding domain. The chitinolytic activity has increased through a directed evolution processes and genetic recombination with Bombyx mori chitinase. Recently, enzymes have been employed as control agents for insects and phytopathogenic fungi (disease originator) opening new potentialities in order to improve the entomopathogenic fungi use. Solid state fermentation is a bioprocess that would produce at great scale enzymes and some other metabolites in grade of increasing the entomopathogenic fungi virulence, in the control of insects and potentially

关键词： Lipases Proteases chitinases Host Evolution Biocontrol Agent

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Effect of Chitinase-Producing Strain V-8 on Controlling Cotton Fusarium Wilt

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Agricultural Science & Technology 2012年第11期13卷 2306-2310页

作者：孙正祥周燚长江大学农学院湖北荆州434025

[Objective] This study aimed to screen endophytic bacteria which is antag- onistic to cotton Fusarium wilt. [Method] Fresh cotton plants collected from cotton- growing areas in Jingzhou City, Hubei Province were used as experimental materials to isolate endophytic bacteria. Through chitinase test and co-culturing both micro-or- ganisms side by side on the same PDA culture plate, antagonistic strains to cotton Fusarium wilt were screened. [Result] A total of 83 bacterial isolates were obtained from cotton plants grown in the fields, six of which were chitinase-productive bacte- ria. Through chitinase test and co-culturing both micro-organisms side by side on the same PDA culture plate, strain V-8 which had the strongest antagonistic effect on Fusarium oxysporum f. sp. vasinfectum was screened. Strain V-8 had a wider anti- fungal spectrum with certain inhibitory effect on all the six important pathogenic fungi including Fusarium oxysporum f. sp niveum; it colonized stably in the rhizospheric soil of cotton, with a colonization density of up to 6.2x10s cfu/g fifty days after inoc- ulation; the relative effect on controlling cotton Fusarium wilt in pot test was 73.2%. The Findings of this study suggested that strain V-8 had great potential for biological control of cotton Fusarium wilt and could be taken as a substantial material for the cloning of chitinase genes. [Conclusion] The results from this study provides bases for the control of cotton fusarium wilt, as well as the exploitation of endophytic bac- teria resources in cotton and the development of novel biological pesticides.

关键词： chitinases Endophytic bacteria Cotton Fusarium-wilt Colonization

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Genome-wide identification and expression pattern of chitinase gene family in Cucumis sativus reveals its role in fusarium wilt resistance

Genome-wide identification and expression pattern of chitina...

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中国园艺学会第八届黄瓜学术研讨会

作者： Ezra Bartholomew Yanling Cai Mingming Dong Shudan Xue Yaqi Zhang Xuling Zhail 刘兴旺 Huazhong Ren 中国农业大学园艺学院

Cucumber(Cucumis sativus L.) is an important vegetable crop cultivated worldwide that is severely impacted by Fusarium oxysporum ***,the casual organism of fusarium ***(EC 3.***.1.14)are chitin-degrading enzymes that hydrolyze the β-(1,4)-linkages in *** are devoid of chitin and it has been proposed that some plant chitinases may play a role in defense response by degrading the chitin in the cell walls of invading fungal *** this study,we performed a comprehensive genome-wide identification and analysis of the chitinase gene family in ***,emphasizing on the phylogenetic relationships and expression profile in response to fusarium wilt.A total of 28 putative chitinase and chitinase-like genes were identified and grouped into five classes(I-V) based on their conserved *** were distributed unevenly across six chromosomes in *** chitinase genes had active sites required for chitinolytic activities and functional redundancy was observed within chitinase *** organ-specific expression of these genes was observed with class I and II chitinases having higher expression levels in the *** genes showed significant responses following *** inoculation,with a class I chitinases being significant up-regulated by 200-fold compared to control *** results offer an insight into the role of chitinase in *** interaction.

关键词： Cucumber chitinases genome-wide identification expression fusarium wilt

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