Transcriptional and Chemical Changes in Soybean Leaves in Response to Long-Term Aphid Colonization

Soybean aphids (Aphis glycines Matsumura) are specialized insects that feed on soybean (Glycine max) phloem sap. Transcriptome analyses have shown that resistant soybean plants mount a fast response that limits aphid feeding and population growth. Conversely, defense responses in susceptible plants are slower and it is hypothesized that aphids block effective defenses in the compatible interaction. Unlike other pests, aphids can colonize plants for long periods of time; yet the effect on the plant transcriptome after long-term aphid feeding has not been analyzed for any plant–aphid interaction. We analyzed the susceptible and resistant (Rag1) transcriptome response to aphid feeding in soybean plants colonized by aphids (biotype 1) for 21 days. We found a reduced resistant response and a low level of aphid growth on Rag1 plants, while susceptible plants showed a strong response consistent with pattern-triggered immunity. GO-term analyses identified chitin regulation as one of the most overrepresented classes of genes, suggesting that chitin could be one of the hemipteran-associated molecular pattern that triggers this defense response. Transcriptome analyses also indicated the phenylpropanoid pathway, specifically isoflavonoid biosynthesis, was induced in susceptible plants in response to long-term aphid feeding. Metabolite analyses corroborated this finding. Aphid-treated susceptible plants accumulated daidzein, formononetin, and genistein, although glyceollins were present at low levels in these plants. Choice experiments indicated that daidzein may have a deterrent effect on aphid feeding. Mass spectrometry imaging showed these isoflavones accumulate likely in the mesophyll cells or epidermis and are absent from the vasculature, suggesting that isoflavones are part of a non-phloem defense response that can reduce aphid feeding. While it is likely that aphid can initially block defense responses in compatible interactions, it appears that susceptible soybean plants can eventually mount an effective defense in response to long-term soybean aphid colonization.

[1]  Shingo Kikuta.  The Cytotoxic Effect of Genistein, a Soybean Isoflavone, against Cultured Tribolium Cells , 2020, Insects.

[2]  Erin M. Conlon.  Microarray Analysis , 2020, Definitions.

[3]  Li-Hua Zhu,et al.  Plant Genes Benefitting Aphids—Potential for Exploitation in Resistance Breeding , 2019, Front. Plant Sci..

[4]  M. Nepal,et al.  Molecular Basis of Soybean Resistance to Soybean Aphids and Soybean Cyst Nematodes , 2019, Plants.

[5]  Q. Song,et al.  Fine mapping of the soybean aphid-resistance genes Rag6 and Rag3c from Glycine soja 85-32 , 2017, Theoretical and Applied Genetics.

[6]  Q. Song,et al.  Fine mapping of the soybean aphid-resistance genes Rag6 and Rag3c from Glycine soja 85-32 , 2017, Theoretical and Applied Genetics.

[7]  Nik Kovinich,et al.  Distinct Mechanisms of Biotic and Chemical Elicitors Enable Additive Elicitation of the Anticancer Phytoalexin Glyceollin I , 2017, Molecules.

[8]  Nicole K Clay,et al.  SG2-Type R2R3-MYB Transcription Factor MYB15 Controls Defense-Induced Lignification and Basal Immunity in Arabidopsis , 2017, Plant Cell.

[9]  B. Cassone,et al.  Transcriptomic dynamics in soybean near-isogenic lines differing in alleles for an aphid resistance gene, following infestation by soybean aphid biotype 2 , 2017, BMC Genomics.

[10]  G. Lawrence,et al.  Components of the SNARE-containing regulon are co-regulated in root cells undergoing defense , 2016, Plant signaling & behavior.

[11]  C. Pieterse,et al.  Recognizing Plant Defense Priming. , 2016, Trends in plant science.

[12]  Chian Kwon,et al.  Arabidopsis immune secretory pathways to powdery mildew fungi , 2016, Plant signaling & behavior.

[13]  G. Jander,et al.  Abscisic acid deficiency increases defence responses against Myzus persicae in Arabidopsis. , 2016, Molecular plant pathology.

[14]  A. Agrawal,et al.  Mechanisms and evolution of plant resistance to aphids , 2016, Nature Plants.

[15]  G. Tylka,et al.  Soybean Aphid Infestation Induces Changes in Fatty Acid Metabolism in Soybean , 2015, PloS one.

[16]  G. Stacey,et al.  Identification of Soybean Proteins and Genes Differentially Regulated in Near Isogenic Lines Differing in Resistance to Aphid Infestation. , 2015, Journal of proteome research.

[17]  I. Somssich,et al.  Negative regulation of ABA signaling by WRKY33 is critical for Arabidopsis immunity towards Botrytis cinerea 2100 , 2015, eLife.

[18]  Jiachen Zi,et al.  Investigation of the Chemical Interface in the Soybean-Aphid and Rice-Bacteria Interactions Using MALDI-Mass Spectrometry Imaging. , 2015, Analytical chemistry.

[19]  C. Ballaré,et al.  Soybean resistance to stink bugs (Nezara viridula and Piezodorus guildinii) increases with exposure to solar UV-B radiation and correlates with isoflavonoid content in pods under field conditions. , 2015, Plant, cell & environment.

[20]  H. Hirt,et al.  Signaling mechanisms in pattern-triggered immunity (PTI). , 2015, Molecular plant.

[21]  G. Sarath,et al.  Transcriptional responses of tolerant and susceptible soybeans to soybean aphid (Aphis glycines Matsumura) herbivory , 2015, Arthropod-Plant Interactions.

[22]  P. Thorpe,et al.  Plant immunity in plant–aphid interactions , 2014, Front. Plant Sci..

[23]  A. Michel,et al.  RNA-Seq reveals a xenobiotic stress response in the soybean aphid, Aphis glycines, when fed aphid-resistant soybean , 2014, BMC Genomics.

[24]  N. Yoshinaga,et al.  Insect-Induced Daidzein, Formononetin and Their Conjugates in Soybean Leaves , 2014, Metabolites.

[25]  J. Leach,et al.  The transcriptional network of WRKY53 in cereals links oxidative responses to biotic and abiotic stress inputs , 2014, Functional & Integrative Genomics.

[26]  R. Oelmüller,et al.  WRKY transcription factors , 2014, Plant signaling & behavior.

[27]  Karam B. Singh,et al.  Plant-aphid interactions with a focus on legumes. , 2013, Functional plant biology : FPB.

[28]  Suzanne M. Paley,et al.  The MetaCyc database of metabolic pathways and enzymes and the BioCyc collection of Pathway/Genome Databases , 2013, Nucleic Acids Res..

[29]  D. Nettleton,et al.  Transcriptome analyses and virus induced gene silencing identify genes in the Rpp4-mediated Asian soybean rust resistance pathway. , 2013, Functional plant biology : FPB.

[30]  K. Koehler,et al.  Performance and prospects of Rag genes for management of soybean aphid , 2013 .

[31]  L. Walling,et al.  Behavioral and Molecular‐Genetic Basis of Resistance against Phloem‐Feeding Insects , 2012 .

[32]  G. C. Macintosh,et al.  Phytohormone signaling pathway analysis method for comparing hormone responses in plant-pest interactions , 2012, BMC Research Notes.

[33]  S. Goławska,et al.  Antifeedant activity of luteolin and genistein against the pea aphid, Acyrthosiphon pisum , 2012, Journal of Pest Science.

[34]  K. Dancewicz,et al.  Intraspecific variation in alkaloid profile of four lupine species with implications for the pea aphid probing behaviour , 2012 .

[35]  E. Hodgson,et al.  Biology of the Soybean Aphid, Aphis glycines (Hemiptera: Aphididae) in the United States , 2011 .

[36]  Wenbin Li,et al.  QTL underlying the resistance to soybean aphid (Aphis glycines Matsumura) through isoflavone-mediated antibiosis in soybean cultivar ‘Zhongdou 27’ , 2011, Theoretical and Applied Genetics.

[37]  Murray Grant,et al.  Hormone crosstalk in plant disease and defense: more than just jasmonate-salicylate antagonism. , 2011, Annual review of phytopathology.

[38]  J. Bos,et al.  Effector proteins that modulate plant--insect interactions. , 2011, Current opinion in plant biology.

[39]  T. Yi,et al.  Secondary metabolites from Glycine soja and their growth inhibitory effect against Spodoptera litura. , 2011, Journal of agricultural and food chemistry.

[40]  I. Morkunas,et al.  Phytohormonal signaling in plant responses to aphid feeding , 2011, Acta Physiologiae Plantarum.

[41]  Xiangzong Meng,et al.  Phosphorylation of a WRKY Transcription Factor by Two Pathogen-Responsive MAPKs Drives Phytoalexin Biosynthesis in Arabidopsis[C][W] , 2011, Plant Cell.

[42]  F. Ausubel,et al.  Mining the plant–herbivore interface with a leafmining Drosophila of Arabidopsis , 2011, Molecular ecology.

[43]  J. Leach,et al.  Virus-induced gene silencing of WRKY53 and an inducible phenylalanine ammonia-lyase in wheat reduces aphid resistance. , 2010, Plant biotechnology journal.

[44]  A. Loraine,et al.  A regulon conserved in monocot and dicot plants defines a functional module in antifungal plant immunity , 2010, Proceedings of the National Academy of Sciences.

[45]  R. Dixon,et al.  Flavonoids and Isoflavonoids: From Plant Biology to Agriculture and Neuroscience , 2010, Plant Physiology.

[46]  A. Botha,et al.  Transcript Profiling of Wheat Genes Expressed During Feeding by Two Different Biotypes of Diuraphis noxia , 2010, Environmental entomology.

[47]  M. O'Neal,et al.  Constitutive and Induced Differential Accumulation of Amino Acid in Leaves of Susceptible and Resistant Soybean Plants in Response to the Soybean Aphid (Hemiptera: Aphididae) , 2010, Environmental entomology.

[48]  J. Bonnemain,et al.  Compatible plant-aphid interactions: how aphids manipulate plant responses. , 2010, Comptes rendus biologies.

[49]  F. Cellier,et al.  New insights into ferritin synthesis and function highlight a link between iron homeostasis and oxidative stress in plants. , 2010, Annals of botany.

[50]  K. Furtwängler,et al.  Flavone synthase II (CYP93B16) from soybean (Glycine max L.). , 2010, Phytochemistry.

[51]  D. Hyten,et al.  Fine mapping the soybean aphid resistance gene Rag1 in soybean , 2010, Theoretical and Applied Genetics.

[52]  Ping He,et al.  Differential innate immune signalling via Ca2+ sensor protein kinases , 2010, Nature.

[53]  S. Dhaubhadel,et al.  Differential expression of CHS7 and CHS8 genes in soybean , 2010, Planta.

[54]  T. Sakurai,et al.  Genome sequence of the palaeopolyploid soybean , 2010, Nature.

[55]  Steven B. Cannon,et al.  SoyBase, the USDA-ARS soybean genetics and genomics database , 2009, Nucleic Acids Res..

[56]  C. Pieterse,et al.  Networking by small-molecule hormones in plant immunity. , 2009, Nature chemical biology.

[57]  G. Delp,et al.  Microarray analysis of the interaction between the aphid Rhopalosiphum padi and host plants reveals both differences and similarities between susceptible and partially resistant barley lines , 2009, Molecular Genetics and Genomics.

[58]  Jonathan D. G. Jones,et al.  Role of plant hormones in plant defence responses , 2009, Plant Molecular Biology.

[59]  J. Bi,et al.  Constitutive and Induced Activities of Defense-Related Enzymes in Aphid-Resistant and Aphid-Susceptible Cultivars of Wheat , 2009, Journal of Chemical Ecology.

[60]  Kanako Sasaki,et al.  Molecular Cloning and Characterization of a cDNA for Pterocarpan 4-Dimethylallyltransferase Catalyzing the Key Prenylation Step in the Biosynthesis of Glyceollin, a Soybean Phytoalexin1[W] , 2008, Plant Physiology.

[61]  Dechun Wang,et al.  Inheritance of Soybean Aphid Resistance in PI 567541B and PI 567598B , 2008 .

[62]  G. Hartman,et al.  Evaluation of Artificial Diets for Rearing Aphis glycines (Hemiptera: Aphididae) , 2008, Journal of economic entomology.

[63]  G. Hartman,et al.  Soybean defense responses to the soybean aphid. , 2008, The New phytologist.

[64]  B. Asselbergh,et al.  Global switches and fine-tuning-ABA modulates plant pathogen defense. , 2008, Molecular plant-microbe interactions : MPMI.

[65]  G. Hartman,et al.  Discovery of Soybean Aphid Biotypes , 2008 .

[66]  G. Howe,et al.  Plant immunity to insect herbivores. , 2008, Annual review of plant biology.

[67]  L. Vodkin,et al.  Structural Features of the Endogenous CHS Silencing and Target Loci in the Soybean Genome , 2008 .

[68]  W. Boland,et al.  Recognition of Herbivory-Associated Molecular Patterns , 2008, Plant Physiology.

[69]  L. Walling.  Avoiding Effective Defenses: Strategies Employed by Phloem-Feeding Insects1 , 2008, Plant Physiology.

[70]  Jun Abe,et al.  Functional analysis of soybean genes involved in flavonoid biosynthesis by virus-induced gene silencing. , 2007, Plant biotechnology journal.

[71]  F. Goggin,et al.  Plant-aphid interactions: molecular and ecological perspectives. , 2007, Current opinion in plant biology.

[72]  E. Hodgson,et al.  Economic Threshold for Soybean Aphid (Hemiptera: Aphididae) , 2007, Journal of economic entomology.

[73]  L. Hesler,et al.  Resistance to Aphis glycines (Hemiptera: Aphididae) in Various Soybean Lines Under Controlled Laboratory Conditions , 2007, Journal of economic entomology.

[74]  H. Hirt,et al.  Activation of members of a MAPK module in β-glucan elicitor-mediated non-host resistance of soybean , 2007, Planta.

[75]  C. Smith,et al.  The molecular bases of plant resistance and defense responses to aphid feeding: current status , 2007 .

[76]  G. Hartman,et al.  Soybean aphid resistance genes in the soybean cultivars Dowling and Jackson map to linkage group M , 2006, Molecular Breeding.

[77]  S. Clough,et al.  Transcriptome changes in the phenylpropanoid pathway of Glycine max in response to Pseudomonas syringae infection , 2006, BMC Plant Biology.

[78]  W. F. Tjallingii,et al.  Location of resistance factors in the leaves of potato and wild tuber‐bearing Solanum species to the aphid Myzus persicae , 2006 .

[79]  R. Tibshirani,et al.  On testing the significance of sets of genes , 2006, math/0610667.

[80]  K. Nishitani,et al.  An Isoflavone Conjugate-hydrolyzing β-Glucosidase from the Roots of Soybean (Glycine max) Seedlings , 2006, Journal of Biological Chemistry.

[81]  Mohamed A. Farag,et al.  Functional analysis of members of the isoflavone and isoflavanone O-methyltransferase enzyme families from the model legume Medicago truncatula , 2006, Plant Molecular Biology.

[82]  G. Hartman,et al.  Soybean Aphid Resistance in Soybean Jackson Is Controlled by a Single Dominant Gene , 2006 .

[83]  G. Hartman,et al.  A Single Dominant Gene for Resistance to the Soybean Aphid in the Soybean Cultivar Dowling , 2006 .

[84]  W. F. Tjallingii,et al.  Salivary secretions by aphids interacting with proteins of phloem wound responses. , 2006, Journal of experimental botany.

[85]  G. Thompson,et al.  Transcriptomics and functional genomics of plant defence induction by phloem-feeding insects. , 2006, Journal of experimental botany.

[86]  Joong-Hoon Ahn,et al.  Regiospecific methylation of naringenin to ponciretin by soybean O-methyltransferase expressed in Escherichia coli. , 2005, Journal of biotechnology.

[87]  Martin J. Mueller,et al.  Signal signature and transcriptome changes of Arabidopsis during pathogen and insect attack. , 2005, Molecular plant-microbe interactions : MPMI.

[88]  R. Dixon,et al.  Structural Elucidation of Chalcone Reductase and Implications for Deoxychalcone Biosynthesis* , 2005, Journal of Biological Chemistry.

[89]  C. B. Hoffmann-Campo,et al.  Are Chemical Compounds Important for Soybean Resistance to Anticarsia gemmatalis? , 2005, Journal of Chemical Ecology.

[90]  F. Cellier,et al.  Siderophore-mediated upregulation of Arabidopsis ferritin expression in response to Erwinia chrysanthemi infection. , 2005, The Plant journal : for cell and molecular biology.

[91]  Senthil Subramanian,et al.  Partial Reconstruction of Flavonoid and Isoflavonoid Biosynthesis in Yeast Using Soybean Type I and Type II Chalcone Isomerases1[w] , 2005, Plant Physiology.

[92]  T. Aoki,et al.  Molecular and Biochemical Characterization of 2-Hydroxyisoflavanone Dehydratase. Involvement of Carboxylesterase-Like Proteins in Leguminous Isoflavone Biosynthesis1[w] , 2005, Plant Physiology.

[93]  R. Gentleman,et al.  A Model-Based Background Adjustment for Oligonucleotide Expression Arrays , 2004 .

[94]  Thomas L. Madden,et al.  BLAST: at the core of a powerful and diverse set of sequence analysis tools , 2004, Nucleic Acids Res..

[95]  John D. Storey,et al.  Strong control, conservative point estimation and simultaneous conservative consistency of false discovery rates: a unified approach , 2004 .

[96]  Erich Kombrink,et al.  SNARE-protein-mediated disease resistance at the plant cell wall , 2003, Nature.

[97]  C. B. Hoffmann-Campo,et al.  Flavonoid Increase in Soybean as a Response to Nezara viridula Injury and Its Effect on Insect-Feeding Preference , 2003, Journal of Chemical Ecology.

[98]  Rafael A Irizarry,et al.  Exploration, normalization, and summaries of high density oligonucleotide array probe level data. , 2003, Biostatistics.

[99]  W. F. Tjallingii,et al.  The role of sinigrin in host plant recognition by aphids during initial plant penetration , 2002 .

[100]  F. Ausubel,et al.  MAP kinase signalling cascade in Arabidopsis innate immunity , 2002, Nature.

[101]  D. Hogg,et al.  First Report of Transmission of Soybean mosaic virus and Alfalfa mosaic virus by Aphis glycines in the New World. , 2001, Plant disease.

[102]  M. Pfaffl,et al.  A new mathematical model for relative quantification in real-time RT-PCR. , 2001, Nucleic acids research.

[103]  O. Yu,et al.  Identification and expression of isoflavone synthase, the key enzyme for biosynthesis of isoflavones in legumes , 2000, Nature Biotechnology.

[104]  G. Kochs,et al.  Molecular characterization and functional expression of dihydroxypterocarpan 6a‐hydroxylase, an enzyme specific for pterocarpanoid phytoalexin biosynthesis in soybean (Glycine max L.) , 1998, FEBS letters.

[105]  T. L. Graham,et al.  Signaling in Soybean Phenylpropanoid Responses (Dissection of Primary, Secondary, and Conditioning Effects of Light, Wounding, and Elicitor Treatments) , 1996, Plant physiology.

[106]  G. Zúñiga,et al.  Effect of infestation by aphids on the water status of barley and insect development , 1995 .

[107]  D. Expert,et al.  Iron Deficiency Induced by Chrysobactin in Saintpaulia Leaves Inoculated with Erwinia chrysanthemi , 1993, Plant physiology.

[108]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[109]  S. Chattopadhyay,et al.  Flavonoids with mosquito larval toxicity , 1990 .

[110]  I. Kaloshian,et al.  Plant Immunity: Connecting the Dots Between Microbial and Hemipteran Immune Responses , 2016 .

[111]  G. C. Macintosh,et al.  Multiple phytohormone signals control the transcriptional response to soybean aphid infestation in susceptible and resistant soybean plants. , 2013, Molecular plant-microbe interactions : MPMI.

[112]  J. Bos,et al.  Toward understanding the role of aphid effectors in plant infestation. , 2013, Molecular plant-microbe interactions : MPMI.

[113]  G. Stacey,et al.  Ethylene-responsive element-binding factor 5, ERF5, is involved in chitin-induced innate immunity response. , 2012, Molecular plant-microbe interactions : MPMI.

[114]  Albino Maggio,et al.  HOS3, an ELO-like gene, inhibits effects of ABA and implicates a S-1-P/ceramide control system for abiotic stress responses in Arabidopsis thaliana. , 2009, Molecular plant.

[115]  O. Yu,et al.  Metabolic Engineering of Isoflavone Biosynthesis , 2005 .

[116]  Alex E. Lash,et al.  Gene Expression Omnibus: NCBI gene expression and hybridization array data repository , 2002, Nucleic Acids Res..

[117]  R. Hammerschmidt.  PHYTOALEXINS: What Have We Learned After 60 Years? , 1999, Annual review of phytopathology.

[118]  W. Goebel,et al.  Differential expression of , 1999 .

[119]  G. Zúñiga,et al.  Allelochemicals in wheat and barley: role in plant—insect interactions , 1992 .

[120]  H. Sharma,et al.  Chemical basis of resistance in soya bean to cabbage looper, Trichoplusia ni , 1991 .

[121]  Ewa Urbanczyk-Wochniak,et al.  BIOINFORMATICS ORIGINAL PAPER doi:10.1093/bioinformatics/btm040 Databases and ontologies MedicCyc: a biochemical pathway database for , 2022 .