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Zhang W.,Chongqing University | Chen J.,Chongqing University | Keyhani N.O.,University of Florida | Zhang Z.,Chongqing University | And 4 more authors.
BMC Genomics | Year: 2015

Background: The migratory locust, Locusta migratoria manilensis, is an immensely destructive agricultural pest that forms a devastating and voracious gregarious phase. The fungal insect pathogen, Metarhizium acridum, is a specialized locust pathogen that has been used as a potent mycoinsecticide for locust control. Little, however, is known about locust immune tissue, i.e. fat body and hemocyte, responses to challenge by this fungus. Methods: RNA-seq (RNA sequencing) technology were applied to comparatively examine the different roles of locust fat body and hemocytes, the two major contributors to the insect immune response, in defense against M. acridum. According to the sequence identity to homologies of other species explored immune response genes, immune related unigenes were screened in all transcriptome wide range from locust and the differential expressed genes were identified in these two tissues, respectively. Results: Analysis of differentially expressed locust genes revealed 4660 and 138 up-regulated, and 1647 and 23 down-regulated transcripts in the fat body and hemocytes, respectively after inoculation with M. acridum spores. GO (Gene Ontology) enrichment analysis showed membrane biogenesis related proteins and effector proteins significantly differentially expressed in hemocytes, while the expression of energy metabolism and development related transcripts were enriched in the fat body after fungal infection. A total of 470 immune related unigenes were identified, including members of the three major insect immune pathways, i.e. Toll, Imd (immune deficiency) and JAK/STAT (janus kinase/signal transduction and activator of transcription). Of these, 58 and three were differentially expressed in the insect fat body or hemocytes after infection, respectively. Of differential expressed transcripts post challenge, 43 were found in both the fat body and hemocytes, including the LmLys4 lysozyme, representing a microbial cell wall targeting enzyme. Conclusions: These data indicate that locust fat body and hemocytes adopt different strategies in response to M. acridum infection. Fat body gene expression after M. acridum challenge appears to function mainly through activation of innate immune related genes, energy metabolism and development related genes. Hemocyte responses attempt to limit fungal infection primarily through regulation of membrane related genes and activation of cellular immune responses and release of humoral immune factors. © 2015 Zhang et al.

Ming Y.,Chongqing University | Ming Y.,Chongqing Engineering Research Center for Fungal Insecticide | Ming Y.,Key Laboratory of Gene Function and Regulation Technologies under Chongqing Municipal Education Commission | Wei Q.,Chongqing University | And 8 more authors.
Applied Microbiology and Biotechnology | Year: 2014

The protein kinase sucrose non-fermenting-1(Snf1) regulates the derepression of glucose-repressible genes and plays a major role in carbon source utilization. In this study, MaSnf1, a sucrose non-fermenting protein kinase gene, has been identified from the entomopathogenic fungus Metarhizium acridum, which has a great potential as a biocontrol agent. The functions of MaSnf1 were characterized using gene disruption and complementation strategies. Disruption of MaSnf1 reduced the conidial yield and delayed the conidial germination on potato dextrose agar (PDA) medium. MaSnf1 is also important for response to ultraviolet radiation and heat shock stress and carbon source utilization in M. acridum. Bioassays by topical inoculation and intrahemocoel injection showed that the MaSnf1 deletion mutant exhibited greatly reduced pathogenicity. The reduced expression level of chitinase gene (Chi) and protease gene (Pr1A) in MaSnf1-disruption transformant (ΔMaSnf1) most likely affects the initial penetration into its host. Additionally, the reduced expression level of acidic trehalase gene (ATM1) probably causes a decline in growth rate in insect hemolymph. Inactivation of MaSnf1 led to a significant decrease in virulence, probably owing to reduction in conidial germination, and appressorium formation, impairment in penetration, and decrease in growth rate in insect hemolymph. © 2014, Springer-Verlag Berlin Heidelberg.

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