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TRANSCRIPTOME ANALYSIS OF GOSSYPIUM HIRSUTUM FLOWER BUDS INFESTED BY COTTON BOLL WEEVIL (ANTHONOMUS GRANDIS) LARVAE
Laser Capture Microdissection
Gossypium
RNA
Larvae
Transcriptome Sequencing
Biotic Stress
Author
Affilliation
Universidade Federal do Rio de Janeiro. Departamento de Genetica. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em DST e AIDS. Rio de Janeiro, RJ, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil / Escola de Medicina Faciplac. Departamento de Bioquímica e Biologia Molecular. Brasília, DF, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil
Universidade de São Paulo. Piracicaba, SP, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil / Universidade Católica de Brasília. Brasília, DF, Brasil
Universidade de São Paulo. Piracicaba, SP, Brasil
Universidade Federal do Rio de Janeiro. Departamento de Genetica. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Instituto Nacional de Infectologia Evandro Chagas. Laboratório de Pesquisa Clínica em DST e AIDS. Rio de Janeiro, RJ, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil / Escola de Medicina Faciplac. Departamento de Bioquímica e Biologia Molecular. Brasília, DF, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil
Universidade de São Paulo. Piracicaba, SP, Brasil
Embrapa Recursos Genéticos e Biotecnologia. Brasília, DF, Brasil / Universidade Católica de Brasília. Brasília, DF, Brasil
Universidade de São Paulo. Piracicaba, SP, Brasil
Universidade Federal do Rio de Janeiro. Departamento de Genetica. Rio de Janeiro, RJ, Brasil
Abstract
Background: Cotton is a major fibre crop grown worldwide that suffers extensive damage from chewing insects,
including the cotton boll weevil larvae (Anthonomus grandis). Transcriptome analysis was performed to understand
the molecular interactions between Gossypium hirsutum L. and cotton boll weevil larvae. The Illumina HiSeq 2000
platform was used to sequence the transcriptome of cotton flower buds infested with boll weevil larvae.
Results: The analysis generated a total of 327,489,418 sequence reads that were aligned to the G. hirsutum
reference transcriptome. The total number of expressed genes was over 21,697 per sample with an average length
of 1,063 bp. The DEGseq analysis identified 443 differentially expressed genes (DEG) in cotton flower buds infected
with boll weevil larvae. Among them, 402 (90.7%) were up-regulated, 41 (9.3%) were down-regulated and 432
(97.5%) were identified as orthologues of A. thaliana genes using Blastx. Mapman analysis of DEG indicated that
many genes were involved in the biotic stress response spanning a range of functions, from a gene encoding a
receptor-like kinase to genes involved in triggering defensive responses such as MAPK, transcription factors
(WRKY and ERF) and signalling by ethylene (ET) and jasmonic acid (JA) hormones. Furthermore, the spatial
expression pattern of 32 of the genes responsive to boll weevil larvae feeding was determined by “in situ” qPCR
analysis from RNA isolated from two flower structures, the stamen and the carpel, by laser microdissection (LMD).
Conclusion: A large number of cotton transcripts were significantly altered upon infestation by larvae. Among the
changes in gene expression, we highlighted the transcription of receptors/sensors that recognise chitin or insect
oral secretions; the altered regulation of transcripts encoding enzymes related to kinase cascades, transcription
factors, Ca2+ influxes, and reactive oxygen species; and the modulation of transcripts encoding enzymes from
phytohormone signalling pathways. These data will aid in the selection of target genes to genetically engineer
cotton to control the cotton boll weevil.
Keywords
TranscriptomeLaser Capture Microdissection
Gossypium
RNA
Larvae
Transcriptome Sequencing
Biotic Stress
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