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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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Effects of Plant Extracts and Beauveria bassiana on the Activity of Defense-Related Enzymes in Solanum lycopersicum L.during Interaction with Fusarium oxysporum f.sp.lycopersici

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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 L.infected with Fusarium oxysporum after the combined application of Beauveria bassiana and plant extracts.Solanum lycopersicum plantlets were transplanted 15 days after the emergency.Five days after transplanting,Beauveria bassiana spores were applied at a concentration of 1×10^(7)spores mL^(−1)onto soil(along with A.indica(N)and P.auritum(H)leaf extracts)where S.lycopersicum plants were planted.Eight days after transplanting,spores of F.oxysporum strain were applied at a concentration of 1×10^(6)spores mL^(−1)to soil where S.lycopersicum plants were growing.The development of S.lycopersicum 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 chitinases.The treatments combining B.bassiana and A.indica and P.auritum extracts,had a significant difference in enzymatic activity levels,thus contributing to better defense mechanisms and a greater protection to S.lycopersicum plants in the presence of F.oxysporum.The application of B.bassiana and plant extracts to the ground mitigated damage caused by F.oxysporum on S.lycopersicum plants.

关键词： Glucanases chitinases peroxidases catalase defense mechanisms

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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 f.***.lycopersici(Fol 17108)in tomato plants in greenhouse conditions.Inoculation of Bb 1205 was the most promising among the agronomic variables and expression of the activity of the enzymesβ-1,3-glucanases and chitinases.Inoculation 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 injection.Infection with Fol 17108 occurred with 1×106 spores·mL−1,which were added directly to the soil.Spectrophotometry was used for measuring agronomic parameters,namely activity of chitinases andβ-1,3-glucanases in foliage and roots.When Bb 1205 was added to the soil,the chlorophyll index and aerial part length showed significant differences.In 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 dpi.As for leaves,a 3.8-fold increase was found when the stem was inoculated.In 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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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(Chi Ls) 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 specific in the immunologic activities. The chitinases and chitinaselike proteins are expressed in response to different proinflammatory cues in various tissues by activated macrophages, neutrophils and in different monocytederived cell lines. The mechanism and molecular interaction of chitinases in relation to immune regulation embrace bacterial infection, inflammation, 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 inflammatory diseases.

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

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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 world.chitinases play important roles in the degradation of chitin.chitinases 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 vertebrates.But there are few reports about Saccharomyces cerevisiae chitinase(ScCTS1).The aim of this study was to realize the high level expression of ScCTS1.The ScCTS1 was cloned into the expression vector pPIC9K.The recombinant plasmid was linearized and transformed into competent Pichia pastoris GS115.After screening by G418 plate,the fermentation conditions were optimized.Ultimately,under the optimal fermentation conditions,ScCTS1 enzymatic activity reached up to 94.6 U/mL.This paper presents the first report on the heterologous expression of a full-length ScCTS1 with considerably high activity.The 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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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 in some diseases affecting plants.

关键词： Lipases Proteases chitinases Host Evolution Biocontrol Agent

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Antagonism of local isolates of Trichoderma spp. on citrus root rot disease by Fusarium solani in the mekong delta of vietnam

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浙江大学学报(农业与生命科学版) 2004年第4期 42-42页

作者： Duong Minh,Jozef Coosemans,Le Lam Cuong,Ester Vandersmissen2, ,Pham Van Kim Belgium Plant Protection Department College of Agriculture Cantho Univ. Cantho city Vietnam Plant Protection Department College of Agriculture Cantho Univ. Cantho city Plant Protection Department Vietnam College of Agriculture Cantho Univ. Cantho city Vietnam Lab. Phytopathology & Plant Protection Dep. Applied Plant Sciences Faculty of Agriculture and Applied Biological Sciences Catholic Univ. Leuven

The local isolates of Trichoderma spp. and Fusarium solani were colected from citrus orchards in the Mekong delta of Vietnam and isolated on PDA, PDB and TSM medium for antagonism and Koch's postulate testing. The results showed that the high chitinolytic enzymes content of Trichoderma isolates can antagonise with Fusarium solani isolates by preventing the germination of Fusarium macroconidia in in-vitro condition. There are five promising isolates of Trichoderma spp. having high antagonism with Fusarium solani. These Trichoderma isolates also grew well in rice straws, maize stems, weeds and water hyacinth biowaste materials. These results supply the promising trend for biological control of root rot disease on citrus orchards of the Mekong delta.

关键词： Trichoderma Fusarium solani antagonism citrus chitinolytic enzymes chitinases

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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 f.***.cucumerinum,the casual organism of fusarium wilt.chitinases(EC 3.***.1.14)are chitin-degrading enzymes that hydrolyze the β-(1,4)-linkages in chitin.Plants 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 pathogens.In this study,we performed a comprehensive genome-wide identification and analysis of the chitinase gene family in C.sativus,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 domain.They were distributed unevenly across six chromosomes in C.sativus.Eighteen chitinase genes had active sites required for chitinolytic activities and functional redundancy was observed within chitinase classes.An organ-specific expression of these genes was observed with class I and II chitinases having higher expression levels in the root.Seven genes showed significant responses following F.oxysporum inoculation,with a class I chitinases being significant up-regulated by 200-fold compared to control plants.The results offer an insight into the role of chitinase in C.sativus-F.oxysporum interaction.

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

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