Maize Endochitinase Expression in Response to Fall Armyworm Herbivory

A large percentage of crop loss is due to insect damage, especially caterpillar damage. Plant chitinases are considered excellent candidates to combat these insects since they can degrade chitin in peritrophic matrix (PM), an important protective structure in caterpillar midgut. Compared to chemical insecticides, chitinases could improve host plant resistance and be both economically and environmentally advantageous. The focus of this research was to find chitinase candidates that could improve plant resistance by effectively limiting caterpillar damage. Five classes of endochitinase (I-V) genes were characterized in the maize genome, and we isolated and cloned four chitinase genes (chitinase A, chitinase B, chitinase I, and PRm3) present in two maize (Zea mays L.) inbred lines Mp708 and Tx601, with different levels of resistance to caterpillar pests. We also investigated the expression of these maize chitinases in response to fall armyworm (Spodoptera frugiperda, FAW) attack. The results indicated that both chitinase transcript abundance and enzymatic activity increased in response to FAW feeding and mechanical wounding. Furthermore, chitinases retained activity inside the caterpillar midgut and enzymatic activity was detected in the food bolus and frass. When examined under scanning electron microscopy, PMs from Tx601-fed caterpillars showed structural damage when compared to diet controls. Analysis of chitinase transcript abundance after caterpillar feeding and proteomic analysis of maize leaf trichomes in the two inbreds implicated chitinase PRm3 found in Tx601 as a potential insecticidal protein.

[1]  F. Huang.  Resistance of the fall armyworm, Spodoptera frugiperda, to transgenic Bacillus thuringiensis Cry1F corn in the Americas: lessons and implications for Bt corn IRM in China , 2020, Insect science.

[2]  G. Felton,et al.  Plant defenses interact with insect enteric bacteria by initiating a leaky gut syndrome , 2019, Proceedings of the National Academy of Sciences.

[3]  M. Whalon,et al.  Field-Evolved Resistance of the Fall Armyworm (Lepidoptera: Noctuidae) to Synthetic Insecticides in Puerto Rico and Mexico , 2018, Journal of Economic Entomology.

[4]  Yahaya M. Normi,et al.  Chitinase: diversity, limitations, and trends in engineering for suitable applications , 2018, Bioscience reports.

[5]  Lex E. Flagel,et al.  Mutational disruption of the ABCC2 gene in fall armyworm, Spodoptera frugiperda, confers resistance to the Cry1Fa and Cry1A.105 insecticidal proteins , 2018, Scientific Reports.

[6]  D. Okuma,et al.  Effective dominance of resistance of Spodoptera frugiperda to Bt maize and cotton varieties: implications for resistance management , 2016, Scientific Reports.

[7]  S. Shakeel,et al.  Turnabout Is Fair Play: Herbivory-Induced Plant Chitinases Excreted in Fall Armyworm Frass Suppress Herbivore Defenses in Maize1[OPEN] , 2016, Plant Physiology.

[8]  P. M. Dourado,et al.  Cross-Resistance between Cry1 Proteins in Fall Armyworm (Spodoptera frugiperda) May Affect the Durability of Current Pyramided Bt Maize Hybrids in Brazil , 2015, PloS one.

[9]  R. Luttrell,et al.  Evidence of multiple/cross resistance to Bt and organophosphate insecticides in Puerto Rico population of the fall armyworm, Spodoptera frugiperda. , 2015, Pesticide biochemistry and physiology.

[10]  W. Williams,et al.  Characterization of the Maize Chitinase Genes and Their Effect on Aspergillus flavus and Aflatoxin Accumulation Resistance , 2015, PloS one.

[11]  Pramod Kumar,et al.  Structural and functional evolution of chitinase‐like proteins from plants , 2015, Proteomics.

[12]  N. Price,et al.  Crystallographic structure of ChitA, a glycoside hydrolase family 19, plant class IV chitinase from Zea mays , 2014, Protein science : a publication of the Protein Society.

[13]  G. Felton,et al.  Herbivore cues from the fall armyworm (Spodoptera frugiperda) larvae trigger direct defenses in maize. , 2014, Molecular plant-microbe interactions : MPMI.

[14]  G. Howe,et al.  Caterpillar attack triggers accumulation of the toxic maize protein RIP2. , 2014, The New phytologist.

[15]  Pedro M. Coutinho,et al.  The carbohydrate-active enzymes database (CAZy) in 2013 , 2013, Nucleic Acids Res..

[16]  Dan M. Bolser,et al.  Gramene 2013: comparative plant genomics resources , 2013, Nucleic Acids Res..

[17]  Koichiro Tamura,et al.  MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. , 2013, Molecular biology and evolution.

[18]  B. Hall,et al.  Building phylogenetic trees from molecular data with MEGA. , 2013, Molecular biology and evolution.

[19]  J. Marden,et al.  Maize toxin degrades peritrophic matrix proteins and stimulates compensatory transcriptome responses in fall armyworm midgut. , 2013, Insect biochemistry and molecular biology.

[20]  J. Musarrat,et al.  Chitinases: An update , 2013, Journal of pharmacy & bioallied sciences.

[21]  Z. Fei,et al.  Genome-Wide Identification and Analysis of the TIFY Gene Family in Grape , 2012, PloS one.

[22]  G. Thompson,et al.  Status of resistance to Bt maize in Spodoptera frugiperda: lessons from Puerto Rico. , 2012, Journal of invertebrate pathology.

[23]  Seung Ho Chung,et al.  Role of trichomes in defense against herbivores: comparison of herbivore response to woolly and hairless trichome mutants in tomato (Solanum lycopersicum) , 2012, Planta.

[24]  Á. Gaitán,et al.  In vitro production of two chitinolytic proteins with an inhibiting effect on the insect coffee berry borer, Hypothenemus hampei (Ferrari) (Coleoptera: Curculionidae) and the fungus Hemileia vastatrix the most limiting pests of coffee crops , 2012, AMB Express.

[25]  A. Grover.  Plant Chitinases: Genetic Diversity and Physiological Roles , 2012 .

[26]  Robert D. Finn,et al.  InterPro in 2011: new developments in the family and domain prediction database , 2011, Nucleic acids research.

[27]  Lisa C. Harper,et al.  MaizeGDB: curation and outreach go hand-in-hand , 2011, Database J. Biol. Databases Curation.

[28]  R. Sekhon,et al.  Genome-wide atlas of transcription during maize development. , 2011, The Plant journal : for cell and molecular biology.

[29]  D. Luthe,et al.  Aboveground to belowground herbivore defense signaling in maize , 2011, Plant signaling & behavior.

[30]  V. Gupta,et al.  Role of Chitinase in Plant Defense , 2011 .

[31]  E. Bosak.  Using a developmental comparison to decipher priming of induced defenses in maize and its effects on a generalist herbivore , 2011 .

[32]  D. Luthe,et al.  Proteomic analysis of the maize rachis: Potential roles of constitutive and induced proteins in resistance to Aspergillus flavus infection and aflatoxin accumulation , 2011, Proteomics.

[33]  J. Osborne.  Improving your data transformations: Applying the Box-Cox transformation , 2010 .

[34]  G. Thompson,et al.  Discovery and Characterization of Field Resistance to Bt Maize: Spodoptera frugiperda (Lepidoptera: Noctuidae) in Puerto Rico , 2010, Journal of economic entomology.

[35]  Fiona M. McCarthy,et al.  Proteome profile of the developing maize (Zea mays L.) rachis , 2010, Proteomics.

[36]  D. Luthe,et al.  Anti-fungal activity of maize silk proteins and role of chitinases in Aspergillus flavus resistance , 2010 .

[37]  J. Kelly,et al.  The Genetics of Phenotypic Plasticity in Plant Defense: Trichome Production in Mimulus guttatus , 2010, The American Naturalist.

[38]  J. Tumlinson,et al.  Plants on Constant Alert: Elevated Levels of Jasmonic Acid and Jasmonate-Induced Transcripts in Caterpillar-Resistant Maize , 2010, Journal of Chemical Ecology.

[39]  Dawn H. Nagel,et al.  The B73 Maize Genome: Complexity, Diversity, and Dynamics , 2009, Science.

[40]  G. Felton,et al.  Plants on early alert: glandular trichomes as sensors for insect herbivores. , 2009, The New phytologist.

[41]  T. Wada,et al.  Jasmonic acid control of GLABRA3 links inducible defense and trichome patterning in Arabidopsis , 2009, Development.

[42]  Brandi L. Cantarel,et al.  The Carbohydrate-Active EnZymes database (CAZy): an expert resource for Glycogenomics , 2008, Nucleic Acids Res..

[43]  D. Hegedus,et al.  New insights into peritrophic matrix synthesis, architecture, and function. , 2009, Annual review of entomology.

[44]  M. McMullen,et al.  Genetics and Biochemistry of Insect Resistance in Maize , 2009 .

[45]  E. Birney,et al.  Pfam: the protein families database , 2013, Nucleic Acids Res..

[46]  Bindu Nanduri,et al.  Comparative Proteomic Analysis of Listeria monocytogenes Strains F2365 and EGD , 2008, Applied and Environmental Microbiology.

[47]  G. Harman,et al.  Genome-wide identification, expression and chromosomal location of the genes encoding chitinolytic enzymes in Zea mays , 2008, Molecular Genetics and Genomics.

[48]  K. Klepzig,et al.  Global and comparative protein profiles of the pronotum of the southern pine beetle, Dendroctonus frontalis , 2008, Insect molecular biology.

[49]  K. Hoover,et al.  Plant-mediated alteration of the peritrophic matrix and baculovirus infection in lepidopteran larvae. , 2008, Journal of insect physiology.

[50]  D. Luthe,et al.  A Naturally Occurring Plant Cysteine Protease Possesses Remarkable Toxicity against Insect Pests and Synergizes Bacillus thuringiensis Toxin , 2008, PloS one.

[51]  R. L. Harrison,et al.  Genomic sequence analysis of a fast-killing isolate of Spodoptera frugiperda multiple nucleopolyhedrovirus. , 2008, The Journal of general virology.

[52]  A. Álvarez,et al.  Fall Armyworm Strains (Lepidoptera: Noctuidae) in Argentina, Their Associate Host Plants and Response to Different Mortality Factors in Laboratory , 2008 .

[53]  G. Felton,et al.  Arthropod-Inducible Proteins: Broad Spectrum Defenses against Multiple Herbivores1 , 2008, Plant Physiology.

[54]  N. Eckardt.  Chitin Signaling in Plants: Insights into the Perception of Fungal Pathogens and Rhizobacterial Symbionts , 2008, The Plant Cell Online.

[55]  J. Libantova,et al.  Heavy-metal stress induced accumulation of chitinase isoforms in plants , 2008, Molecular Biology Reports.

[56]  M. Digilio,et al.  The Chitinase A from the baculovirus AcMNPV enhances resistance to both fungi and herbivorous pests in tobacco , 2008, Transgenic Research.

[57]  M. D. McMullen,et al.  Genetic Basis of Resistance to Fall Armyworm (Lepidoptera: Noctuidae) and Southwestern Corn Borer (Lepidoptera: Crambidae) Leaf-Feeding Damage in Maize , 2007, Journal of economic entomology.

[58]  G. Howe,et al.  Stability of Plant Defense Proteins in the Gut of Insect Herbivores1[C][W][OA] , 2007, Plant Physiology.

[59]  C. Pieterse,et al.  Significance of inducible defense-related proteins in infected plants. , 2006, Annual review of phytopathology.

[60]  Cathie Martin,et al.  Trichomes: different regulatory networks lead to convergent structures. , 2006, Trends in plant science.

[61]  N. Novak,et al.  Expression of Poplar Chitinase in Tomato Leads to Inhibition of Development in Colorado Potato Beetle , 2006, Biotechnology Letters.

[62]  D. Luthe,et al.  Degradation of the S. frugiperda peritrophic matrix by an inducible maize cysteine protease. , 2006, Journal of insect physiology.

[63]  G. Howe,et al.  Jasmonate-inducible plant enzymes degrade essential amino acids in the herbivore midgut. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[64]  Xiang Xiao,et al.  Putative Exposed Aromatic and Hydroxyl Residues on the Surface of the N-Terminal Domains of Chi1 from Aeromonas caviae CB101 Are Essential for Chitin Binding and Hydrolysis , 2005, Applied and Environmental Microbiology.

[65]  W. Williams,et al.  Quantitative Trait Loci Conferring Resistance to Fall Armyworm and Southwestern Corn Borer Leaf Feeding Damage , 2005 .

[66]  W. Terra,et al.  Sequences of cDNAs and expression of genes encoding chitin synthase and chitinase in the midgut of Spodoptera frugiperda. , 2005, Insect biochemistry and molecular biology.

[67]  G. Felton,et al.  The host plant as a factor in the synthesis and secretion of salivary glucose oxidase in larval Helicoverpa zea. , 2005, Archives of insect biochemistry and physiology.

[68]  P. Jollès,et al.  What's new in chitinase research? , 1992, Experientia.

[69]  Steven M. Lalonde,et al.  Transforming Variables for Normality and Linearity – When , How , Why and Why Not ' s , 2005 .

[70]  R. Camarillo,et al.  Colloidal chitin stained with Remazol Brilliant Blue R®, a useful substrate to select chitinolytic microorganisms and to evaluate chitinases , 2004 .

[71]  J. Sanderson,et al.  Novel Chitinolytic Enzymes with Biological Activity Against Herbivorous Insects , 1998, Journal of Chemical Ecology.

[72]  M. Adang,et al.  Surface morphology of peritrophic membrane formation in the cabbage looper, Trichoplusia ni , 2004, Cell and Tissue Research.

[73]  P. Frendo,et al.  Heavy-metal-responsive genes in maize: identification and comparison of their expression upon various forms of abiotic stress , 2004, Planta.

[74]  J. Bernal,et al.  Survivorship and development of fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae), on conventional and transgenic maize cultivars expressing Bacillus thuringiensis Cry9C and Cry1A(b) endotoxins , 2003 .

[75]  D. Luthe,et al.  Insect feeding mobilizes a unique plant defense protease that disrupts the peritrophic matrix of caterpillars , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[76]  Hendrik Poorter,et al.  Avoiding bias in calculations of relative growth rate. , 2002, Annals of botany.

[77]  N. Bohorova,et al.  Novel synthetic Bacillus thuringiensiscry1B gene and the cry1B-cry1Ab translational fusion confer resistance to southwestern corn borer, sugarcane borer and fall armyworm in transgenic tropical maize , 2001, Theoretical and Applied Genetics.

[78]  S. Thompson,et al.  Identification and Characterization of a Chitinase Antigen from Pseudomonas aeruginosa Strain 385 , 2001, Applied and Environmental Microbiology.

[79]  R. Barbehenn,et al.  Chitinolytic enzymes from Streptomyces albidoflavus expressed in tomato plants: effects on Trichoplusia ni , 2001 .

[80]  D. Luthe,et al.  A Unique 33-kD Cysteine Proteinase Accumulates in Response to Larval Feeding in Maize Genotypes Resistant to Fall Armyworm and Other Lepidoptera , 2000, Plant Cell.

[81]  D. Luthe,et al.  Influence of Whorl Region from Resistant and Susceptible Corn Genotypes on Fall Armyworm (Lepidoptera: Noctuidae) Growth and Development , 2000, Journal of economic entomology.

[82]  J. Daniel Hare,et al.  No benefit of glandular trichome production in natural populations of Datura wrightii? , 2000, Oecologia.

[83]  F. M. Davis,et al.  Vegetative phase change in maize and its association with resistance to fall armyworm , 2000 .

[84]  E. Werker.  Trichome diversity and development , 2000 .

[85]  B. Henrissat,et al.  Classification of chitinases modules. , 1999, EXS.

[86]  A. Monzingo,et al.  The structure and action of chitinases. , 1999, EXS.

[87]  A. Herrera-Estrella,et al.  Chitinases in biological control. , 1999, EXS.

[88]  M. Mitsutomi,et al.  Biochemistry of chitinases. , 1999, EXS.

[89]  G. Gooday.  Aggressive and defensive roles for chitinases. , 1999, EXS.

[90]  S. Lawrence,et al.  Chitinase accumulates systemically in wounded poplar trees , 1998 .

[91]  I. Chet,et al.  The molecular biology of chitin digestion. , 1998, Current opinion in biotechnology.

[92]  K. Kramer,et al.  Insect chitinases: molecular biology and potential use as biopesticides. , 1997, Insect biochemistry and molecular biology.

[93]  P. Wang,et al.  An intestinal mucin is the target substrate for a baculovirus enhancin. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[94]  G. Bellemare,et al.  Structural and Evolutionary Relationships Among Chitinases of Flowering Plants , 1997, Journal of Molecular Evolution.

[95]  F. White,et al.  Chitinases for Insect Control , 1997 .

[96]  M. Koziel,et al.  Advances in insect control: the role of transgenic plants. , 1997 .

[97]  M. Lehane.  Peritrophic matrix structure and function. , 1997, Annual review of entomology.

[98]  A. Bravo,et al.  Interactions of Bacillus thuringiensis crystal proteins with the midgut epithelial cells of Spodoptera frugiperda (Lepidoptera: Noctuidae). , 1996, Journal of invertebrate pathology.

[99]  C. Koncz,et al.  Synergistic activity of a Bacillus thuringiensis delta-endotoxin and a bacterial endochitinase against Spodoptera littoralis larvae , 1996, Applied and environmental microbiology.

[100]  B. Henrissat,et al.  Plant chitinases use two different hydrolytic mechanisms , 1996, FEBS letters.

[101]  K. Kramer,et al.  BACULOVIRUS-MEDIATED EXPRESSION OF A MANDUCA SEXTA CHITINASE GENE: PROPERTIES OF THE RECOMBINANT PROTEIN , 1995 .

[102]  J. Beintema.  Structural features of plant chitinases and chitin‐binding proteins , 1994, FEBS letters.

[103]  J. Widholm,et al.  Molecular Analysis of Two cDNA Clones Encoding Acidic Class I Chitinase in Maize , 1994, Plant physiology.

[104]  Michael Freeling,et al.  The Maize Leaf , 1994 .

[105]  J. Mikkelsen,et al.  Plant chitinases. , 2008, The Plant journal : for cell and molecular biology.

[106]  G. Harman,et al.  Detection and quantification of N-acetyl-beta-D-glucosaminidase, chitobiosidase, and endochitinase in solutions and on gels. , 1993, Analytical biochemistry.

[107]  J. Dow,et al.  pH GRADIENTS IN LEPIDOPTERAN MIDGUT. , 1992, The Journal of experimental biology.

[108]  D. Shah,et al.  Antifungal proteins from plants. Purification, molecular cloning, and antifungal properties of chitinases from maize seed. , 1992, The Journal of biological chemistry.

[109]  Lin Zhe-fu,et al.  Chitinases from Seeds of Zea mays and Coix lachryma-jobi L. Purification and Some Properties , 1992 .

[110]  G. Wagner.  Secreting glandular trichomes: more than just hairs. , 1991, Plant physiology.

[111]  E. Cabib,et al.  Malaria parasite chitinase and penetration of the mosquito peritrophic membrane. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[112]  R. Poethig,et al.  Phase Change and the Regulation of Shoot Morphogenesis in Plants , 1990, Science.

[113]  W. Hurkman,et al.  Solubilization of plant membrane proteins for analysis by two-dimensional gel electrophoresis. , 1986, Plant physiology.

[114]  D. Young,et al.  Purification and characterization of chitinase enzymes from healthy and Verticillium albo-atrum-infected tomato plants, and from V. albo-atrum , 1982 .

[115]  I. Polacheck,et al.  An endochitinase from wheat germ. Activity on nascent and preformed chitin. , 1979, The Journal of biological chemistry.

[116]  M. Adang,et al.  The peritrophic membrane: Ultrastructural analysis and function as a mechanical barrier to microbial infection in , 1978 .

[117]  M. M. Bradford.  A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. , 1976, Analytical biochemistry.

[118]  D. Levin.  The Role of Trichomes in Plant Defense , 1973, The Quarterly Review of Biology.

[119]  J. Hanway.  How a corn plant develops , 1966 .