ABSTRACT: The striped stem borer, Chilo suppressalis Walker (Lepidoptera: Crambidae), is one of the most serious rice pests. Besides attacking rice, it also feeds on an economically important vegetable crop, water-oat Zizania latifolia. The species feeding on water-oat has higher growth and survival rate than those on rice, suggesting their success in adaptation to the new host plant. However, little is known about the molecular mechanisms of host plant adaptation. Here we investigated the midgut transcriptome responses of C. suppressalis larvae reared on rice and water-oat. A total of 1,633 differentially expressed genes were identified, with a greater number up-regulated on the more delicious new host. The up-regulation of most digestive and detoxification-related genes may be the result of adaptation to the changes in nutritional requirements and toxic chemicals during host shift. In contrast, down-regulation of ribosomal genes may be related to their better development performance when feeding on the new host. In conclusion, our results suggest that transcriptional regulation of genes related to digestion, detoxification and ribosome may play an important role in adaptation of C. suppressalis to a new host plant.
Project description:Chilo suppressalis (Walker, 1863) is a serious stem borer of rice and water-oat plants, and has phenotypically diverged into rice and water-oat populations. Insect gut microbiota plays an important role in the host life and understanding the dynamics of this complicated ecosystem may improve its biological control. The effect of diet and gut compartments on the gut microflora of divergent populations of C. suppressalis is not fully clear. Herein, we characterized the gut microbiota of C. suppressalis populations fed on two hosts (i.e., water-oats fruit pulps and rice seedlings), by sequencing the V3-V4 hypervariable region of the 16S rRNA gene using the Illumina MiSeq platform. Gut bacterial communities showed variation in relative abundance among C. suppressalis populations fed on water-oats fruit pulps or rice seedlings. Proteobacteria and Firmicutes became the predominant phyla, and Enterobacteriaceae, Enterococcaceae and Halomonadaceae were the predominant family in all C. suppressalis populations. The highest bacteria diversity was found in the midgut of the rice population fed on water-oat fruit pulps. Bacterial communities in the midgut were more diverse than those in the hindgut. The bacterial genera distribution showed great differences due to diet types and gut compartments among populations. Our results demonstrated that the host plants tested had a considerable impact on gut bacterial composition of C. suppressalis populations. Additionly, the unique gut morphology and physiological conditions (viz., oxygen content, enzymes) also contributed to variation in microbiomes. In conclusion, our study provided an important insight into investigation of insect-bacteria symbioses, and biocontrol of this species and other related lepidopterans.
Project description:Behavioral isolation in animals can be mediated by inherent mating preferences and assortative traits, such as divergence in the diel timing of mating activity. Although divergence in the diel mating time could, in principle, promote the reproductive isolation of sympatric, conspecific populations, there is currently no unequivocal evidence of this. We conducted different mate-choice experiments to investigate the contribution of differences in diel mating activity to the reproductive isolation of the rice and water-oat populations of Chilo suppressalis. The results show that inter-population difference in diel mating activity contributes to assortative mating in these populations. In the rice population, most mating activity occurred during the first half of the scotophase, whereas in the water-oat population virtually all mating activity was confined to the second half of the scotophase. However, when the photoperiod of individuals from the water-oat population was altered to more closely align their mating activity with that of the rice population, mate choice was random. We conclude that inter-population differences in diel mating time contribute to assortative mating, and thereby the partial reproductive isolation, of these host-associated populations of C. suppressalis.
Project description:Reproductive isolation between different host populations is often based on intraspecific sex pheromone differences. The mechanisms underlying these differences have not been thoroughly elucidated to date. Previous studies suggested that Chilo suppressalis has differentiated into rice and water-oat host populations, and these two populations manifest clear differences in sex pheromone titer and mating rhythm. Hence, this moth is an ideal model to investigate the endogenous mechanisms of intraspecific reproductive isolation. Here, we identified a series of putative genes associated with sex pheromone biosynthesis based on the C. suppressalis pheromone gland transcriptome data. Transcripts of most genes were at higher level in the rice population. Then we obtained 11 pivotal differentially expressed genes (DEGs). The expression levels of these DEGs exhibited a distinct increase in the rice population. Moreover, we also observed the expression rhythm of these DEGs is discrepant between two host populations. Our study offers a new understanding to elucidate the mechanisms of intraspecific reproductive isolation.
Project description:Chilo supprressalis is one of the most important rice pests that causes serious damage to production in the rice growth area of Asia. Vetiver grass (Vetiveria zizanioides) was previously found to effectively attract female adults of C. suppressalis laying eggs on vetiver leaves, while the larvae cannot complete their life cycles by feeding on vetiver, indicating a potential means of controlling this pest. In the present study, the transcriptomes of midguts of rice-fed and vetiver-fed C. suppressalis larvae were profiled, which aimed to clarify the molecular mechanism of vetiver as a dead-end trap plant preliminarily. We found that ingestion of vetiver provoked a robust transcriptional response in the larval midguts, and a total of 1,849 differentially expressed UniGenes were identified. We focused on 12 digestion-related genes, four immune-related genes and three detoxification-related genes. Most of these genes were significantly down regulated in the larval midguts at 6, 8, and 10 days after feeding on vetiver compared to on rice. Transcriptional dynamics suggested that these genes might be involved in toxicity responses following exposure to vetiver. Taken together, this study provides an initial molecular framework for developing biological control strategies for C. suppressalis in an effort to protect economically important rice crops.
Project description:BACKGROUND:Although the importance of host plant chemistry in plant-insect interactions is widely recognized, our understanding about the genetic basis underlying the relationship between changes in midgut proteins and adaptation of plant-feeding insects to novel host plants and habitats is very limited. To address this knowledge gap, the transcriptional profiles of midguts among three populations of the cicada Subpsaltria yangi Chen were compared. Among which, the Hancheng (HC) and Fengxiang (FX) populations occurring in the Loess Plateau feed on Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chow, while the population occurring in a much drier habitat in the Helan (HL) Mountains is locally specialized on a chemically divergent plant, Ephedra lepidosperma C. Y. Cheng. RESULTS:Based on comparative analysis, 1826 (HL vs HC) differentially expressed genes (DEGs) and 723 DEGs (HL vs FX) were identified between the populations utilizing different host plants, including 20, 36, 2, 5 and 2 genes related to digestion, detoxification, oxidation-reduction, stress response and water-deprivation response, respectively, and 35 genes presumably associated with osmoregulation. However, only 183 DEGs were identified between the HC and FX populations, including two genes related to detoxification, two genes related to stress response, and one gene presumably associated with osmoregulation. These results suggest that the weakest expression differences were between the populations utilizing the same host plant and occurring in the closest habitats, which may help explain the metabolic mechanism of adaptation in S. yangi populations to novel host plants and new niches. CONCLUSIONS:The observed differences in gene expression among S. yangi populations are consistent with the hypothesis that the host plant shift and habitat adaptation in the HL population was facilitated by differential regulation of genes related to digestion, detoxification, oxidation-reduction, stress response, water-deprivation response and osmoregulation. The results may inform future studies on the molecular mechanisms underlying the relationship between changes in midgut proteins and adaptation of herbivorous insects to novel host plants and new niches.
Project description:BACKGROUND:Chilo suppressalis is a widespread rice pest that poses a major threat to food security in China. This pest can develop resistance to Cry toxins from Bacillus thuringiensis (Bt), threatening the sustainable use of insect-resistant transgenic Bt rice. However, the molecular basis for the resistance mechanisms of C. suppressalis to Cry1C toxin remains unknown. This study aimed to identify genes associated with the mechanism of Cry1C resistance in C. suppressalis by comparing the midgut transcriptomic responses of resistant and susceptible C. suppressalis strains to Cry1C toxin and to provide information for insect resistance management. RESULTS:A C. suppressalis midgut transcriptome of 139,206 unigenes was de novo assembled from 373 million Illumina HiSeq and Roche 454 clean reads. Comparative analysis identified 5328 significantly differentially expressed unigenes (DEGs) between C. suppressalis Cry1C-resistant and -susceptible strains. DEGs encoding Bt Cry toxin receptors, aminopeptidase-P like protein, the ABC subfamily and alkaline phosphatase were downregulated, suggesting an association with C. suppressalis Cry1C resistance. Additionally, Cry1C resistance in C. suppressalis may be related to changes in the transcription levels of enzymes involved in hydrolysis, digestive, catalytic and detoxification processes. CONCLUSION:Our study identified genes potentially involved in Cry1C resistance in C. suppressalis by comparative transcriptome analysis. The assembled and annotated transcriptome data provide valuable genomic resources for further study of the molecular mechanisms of C. suppressalis resistance to Cry toxins.
Project description:The striped stem borer, Chilo suppressalis (Walker), is a notorious pest of rice that causes large losses in China. Breeding and screening of resistance rice cultivars are effective strategies for C. suppressalis management. In this study, insect-resistant traits of 47 rice cultivars were investigated by C. suppressalis artificial infestation (AI) both in field and greenhouse experiments, using the susceptible (S) cultivar 1665 as a control. Results suggest that two rice cultivars, namely 1688 and 1654, are resistant (R) and moderately resistant (MR) to C. suppressalis, respectively. Then, a comparative transcriptome (RNA-Seq) was de novo assembled and differentially expressed genes (DEGs) with altered expression levels were investigated among cultivars 1688, 1654, and 1665, with or without C. suppressalis infestation for 24 h. A total of 2569 and 1861 genes were up-regulated, and 3852 and 1861 genes were down-regulated in cultivars 1688 and 1654, respectively after artificial infestation with C. suppressalis compared to the non-infested control (CK). For the susceptible cultivar 1665, a total of 882 genes were up-regulated and 3863 genes were down-regulated after artificial infestation with C. suppressalis compared to the CK. Twenty four DEGs belong to proteinase inhibitor, lectin and chitinase gene families; plant hormone signal transduction and plant-pathogen interaction pathways were selected as candidate genes to test their possible role in C. suppressalis resistance. RT-qPCR results revealed that 13 genes were significantly up-regulated and 8 were significantly down-regulated in the resistant cultivar 1688 with C. suppressalis artificial infestation (1688AI) compared to the CK. Three genes, LTPL164, LTPL151, and LOC Os11g32100, showed more than a 10-fold higher expression in 1688AI than in 1688CK, suggesting their potential role in insect resistance. Overall, our results provide an important foundation for further understanding the insect resistance mechanisms of selected resistant varieties that will help us to breed C. suppressalis resistant rice varieties.
Project description:We determined whether plant volatiles help explain why Cotesia chilonis (a parasitoid of the target pest Chilo suppressalis) is less abundant in Bt than in non-Bt rice fields. Olfactometer studies revealed that C. chilonis females responded similarly to undamaged Bt and non-Bt rice plants. Parasitoids preferred rice plants damaged by 3(rd)-instar larvae of C. suppressalis, but did not differentiate between caterpillar-infested Bt and non-Bt plants. According to GC-MS analyses of rice plant volatiles, undamaged Bt and non-Bt rice plants emitted the same number of volatile compounds and there were no significant differences in the quantity of each volatile compound between the treatments. When plants were infested with and damaged by C. suppressalis larvae, both Bt and non-Bt rice plants emitted higher numbers and larger amounts of volatile compounds than undamaged plants, but there were no significant differences between Bt and non-Bt plants. These results demonstrate that the volatile-mediated interactions of rice plants with the parasitoid C. chilonis were not disrupted by the genetic engineering of the plants. We infer that parasitoid numbers are lower in Bt than in non-Bt fields because damage and volatile induction by C. suppressalis larvae are greatly reduced in Bt fields.
Project description:Background and Aim:Water is an increasingly scarce resource while some crops, such as paddy rice, require large amounts of water to maintain grain production. A better understanding of rice drought adaptation and tolerance mechanisms could help to reduce this problem. There is evidence of a possible role of root-associated fungi in drought adaptation. Here, we analyzed the endospheric fungal microbiota composition in rice and its relation to plant genotype and drought. Methods:Fifteen rice genotypes (Oryza sativa ssp. indica) were grown in the field, under well-watered conditions or exposed to a drought period during flowering. The effect of genotype and treatment on the root fungal microbiota composition was analyzed by 18S ribosomal DNA high throughput sequencing. Grain yield was determined after plant maturation. Results:There was a host genotype effect on the fungal community composition. Drought altered the composition of the root-associated fungal community and increased fungal biodiversity. The majority of OTUs identified belonged to the Pezizomycotina subphylum and 37 of these significantly correlated with a higher plant yield under drought, one of them being assigned to Arthrinium phaeospermum. Conclusion:This study shows that both plant genotype and drought affect the root-associated fungal community in rice and that some fungi correlate with improved drought tolerance. This work opens new opportunities for basic research on the understanding of how the host affects microbiota recruitment as well as the possible use of specific fungi to improve drought tolerance in rice.
Project description:The fungus Cochliobolus miyabeanus causes severe leaf spot disease on rice (Oryza sativa) and two North American specialty crops, American wildrice (Zizania palustris) and switchgrass (Panicum virgatum). Despite the importance of C. miyabeanus as a disease-causing agent in wildrice, little is known about either the mechanisms of pathogenicity or host defense responses. To start bridging these gaps, the genome of C. miyabeanus strain TG12bL2 was shotgun sequenced using Illumina technology. The genome assembly consists of 31.79 Mbp in 2,378 scaffolds with an N50 = 74,921. It contains 11,000 predicted genes of which 94.5% were annotated. Approximately 10% of total gene number is expected to be secreted. The C. miyabeanus genome is rich in carbohydrate active enzymes, and harbors 187 small secreted peptides (SSPs) and some fungal effector homologs. Detoxification systems were represented by a variety of enzymes that could offer protection against plant defense compounds. The non-ribosomal peptide synthetases and polyketide synthases (PKS) present were common to other Cochliobolus species. Additionally, the fungal transcriptome was analyzed at 48 hours after inoculation in planta. A total of 10,674 genes were found to be expressed, some of which are known to be involved in pathogenicity or response to host defenses including hydrophobins, cutinase, cell wall degrading enzymes, enzymes related to reactive oxygen species scavenging, PKS, detoxification systems, SSPs, and a known fungal effector. This work will facilitate future research on C. miyabeanus pathogen-associated molecular patterns and effectors, and in the identification of their corresponding wildrice defense mechanisms.