Identification and analysis of microRNAs responsible for brown planthopper resistance in BPH14/BPH15 pyramiding rice
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ABSTRACT: Background Rice farming faces a serious challenge from the brown planthopper (BPH), with the pyramiding of BPH14 and BPH15 genes delivering effective protection in elite rice strains. However, the molecular basis behind this resistance is still unclear. Results The study investigated miRNA levels in BPH14/BPH15 pyramiding line (B1415) and their recurrent parent (RP) under BPH infestation employing high-throughput sequencing and revealed 136 differentially expressed miRNAs (DEMs) among 550 known miRNAs. An integrated analysis highlighted that 587 miRNA-mRNA pairs linking 95 DEMs to 537 targeted genes were enriched in phenylpropanoid and lignin metabolism, circadian rhythms, and amino acid metabolism. The candidate DEMs, miR172d-3p, and miR396 family members were identified as negative regulators to decrease their target genes Os06g0708700 (encoding a nodulin-like protein) and Os11g0129700 (encoding an AP2 domain transcription factor), suggesting their key roles in rice against BPH. Conclusions Our investigation provides the first insights into miRNA-mediated defense mechanisms in the B1415. Identifying miRNAs and their target mRNAs in BPH resistance opens a new avenue for rice breeding programs, offering potential targets for improving pest resistance. Understanding these molecular interactions paves the way for developing more resistant rice cultivars, thereby contributing to sustainable rice production and food security.
Project description:Brown planthopper (BPH) is one of the most destructive pests in rice production. The pyramiding application of BPH-resistance genes BPH14 and BPH15 can effectively improve rice resistance to BPH, however, the molecular mechanisms underlying BPH14/BPH15 pyramiding lines are poorly understood. Here, a mRNA expression profiling analysis was performed on the near isogenic lines (NILs) containing the BPH14, BPH15 or BPH14/BPH15 and their recurrent parent (RP) Wushansimiao. A total of 14492 differentially expressed mRNAs (DEGs) were identified from 12 mRNA profiles of resistant NILs and RP at different feeding stages. In the transcriptome analysis, 531 DEGs appeared to be common among the resistant NILs compared to RP before and after BPH feeding, which were enriched in defense response, phosphorylation and salt stress response. In addition, 258 DEGs shared only in resistant NILs were obtained among the different feeding stages, which were enriched in oxidative stress response, karrikin response and chloroplast organization. 21 DEGs were further selected as candidates for BPH resistance. OsPOX8.1, a potential candidate DEG related to BPH resistance, increased reactive oxygen species levels in rice protoplast. Our results provide valuable information to further explore the defense mechanism of insect-resistant gene pyramiding lines and develop robust strategies for insect control.
Project description:MicroRNAs (miRNAs) regulate a spectrum of development and defense response processes in plants. The brown planthopper (BPH) is the most devastating insect pest of rice, BPH resistance gene BPH15 has a strong resistance to BPH. In this study, we analyzed six miRNA profiles of BPH15 introgression line (P15) and susceptible recipient line (PC) in 3 time points (0h, 6h and 48h) after BPH attacked, and identified 464 known miRNAs and 183 novel miRNAs. Before BPH feeding we identified 23 miRNAs expressed differently in P15 and PC. Then after BPH feeding, 104 miRNAs were found expressed differently in P15, and 80 miRNAs were found expressed differently in PC. Among them, miR167, miR444d, miR1846e, miR3979-3p, miR531 were found to be involved in BPH stress response. The response to BPH of P15 was much wider and more rapidly than PC. The levels of a subset of miRNAs were confirmed by qRT-PCR. The targets of miRNAs were predicted and validated by gene expression profiling. Additionally, the target genes of 2 differently expressed miRNAs (miR160f-5p and miR167a-5p) were confirmed by detecting YFP fluorescence and western blotting. Our data provide an important basis for evaluating the role of miRNA in the regulation of BPH interactions in rice.
Project description:Here, a combined small RNA and transcriptome sequencing analysis was performed on the BPH6-transgenic and Nipponbare plants sheath samples at the different feeding stage by BPH, and differentially expressed microRNAs (DEMs) and genes (DEGs) were identified and further analyzed for a possible involvement in BPH6 mediated resistance mechanism. A total of 217 DEMs and 7,874 DEGs were identified in the BPH6-transgenic and Nipponbare plants after BPH feeding. At the miRNA level, 29 miRNAs, including the members of miR160, miR166, miR169, miR1861, miR319 and miR390 family and other miRNAs, appeared opposite expression at early or late feeding stages between the two varieties, and 9 miRNAs specially expressed in the BPH6-transgenic plans, suggesting these miRNAs might play an important role in BPH6-mediated resistance to BPH. For the transcriptome, 949 DEGs appeared opposite expression at early or late feeding stages of two rice genotypes were identified and enriched in metabolic process, cellular development, cell wall organization, cellular component movement and hormone transport, and in involved in certain kinds of primary and secondary metabolite synthesis by GO and KEGG enrichment analysis. 24 genes were further selected to be considered as BPH resistance related gene candidates. Finally, integrated analysis of DEMs and DEGs showed that 34 miRNAs corresponding to 42 target genes were selected as BPH resistance related miRNA-mRNA interaction candidates.
Project description:Here, a combined small RNA and transcriptome sequencing analysis was performed on the BPH6-transgenic and Nipponbare plants sheath samples at the different feeding stage by BPH, and differentially expressed microRNAs (DEMs) and genes (DEGs) were identified and further analyzed for a possible involvement in BPH6 mediated resistance mechanism. A total of 217 DEMs and 7,874 DEGs were identified in the BPH6-transgenic and Nipponbare plants after BPH feeding. At the miRNA level, 29 miRNAs, including the members of miR160, miR166, miR169, miR1861, miR319 and miR390 family and other miRNAs, appeared opposite expression at early or late feeding stages between the two varieties, and 9 miRNAs specially expressed in the BPH6-transgenic plans, suggesting these miRNAs might play an important role in BPH6-mediated resistance to BPH. For the transcriptome, 949 DEGs appeared opposite expression at early or late feeding stages of two rice genotypes were identified and enriched in metabolic process, cellular development, cell wall organization, cellular component movement and hormone transport, and in involved in certain kinds of primary and secondary metabolite synthesis by GO and KEGG enrichment analysis. 24 genes were further selected to be considered as BPH resistance related gene candidates. Finally, integrated analysis of DEMs and DEGs showed that 34 miRNAs corresponding to 42 target genes were selected as BPH resistance related miRNA-mRNA interaction candidates, 18 pairs of which were identified by qRT-PCR, two pairs of which were further confirmed by protoplast fluorescence micrographs and western blotting.
Project description:The aim of this study was to analyze potential brown planthopper (BPH) resistant genes in Rathu Heenati (RHT) by Affymetrix whole rice genome array,BPH susceptible and resistant rice varieties of TN1(Taichung Native 1)as control. All the resistant related genes derived from RHT will be analyzed according to the SSR markers interval flanked on the chromosome 3, 4, 6 and 10. It will be benefit to the gene clone and marker assistant breeding for Bph3 gene in the near future. We used microarrays to detail the global differential gene expression before and after brown planthopper attack in two different varieties, and identified distinct classes of high enriched genes induced by BPH or constituent in RHT The 2nd to 3rd instar nymphs of BPH were transferred to tillering stage seedings (10 BPH nymphs per plant) in a box covered with nylon-mesh. Stems of the rice plant infected by BPH were collected at 0h (T0), 8h (T8), 24h (T24) after BPH attack, total RNA were extracted for the microarray hybirdlization.
Project description:The aim of this study was to analyze potential brown planthopper (BPH) resistant genes in Rathu Heenati (RHT) by Affymetrix whole rice genome array,BPH susceptible and resistant rice varieties of TN1(Taichung Native 1)as control. All the resistant related genes derived from RHT will be analyzed according to the SSR markers interval flanked on the chromosome 3, 4, 6 and 10. It will be benefit to the gene clone and marker assistant breeding for Bph3 gene in the near future. We used microarrays to detail the global differential gene expression before and after brown planthopper attack in two different varieties, and identified distinct classes of high enriched genes induced by BPH or constituent in RHT
Project description:Nilaparvata lugens, the brown planthopper (BPH) sucks the rice phloem sap containing high sucrose to obtain carbon source. The comparative gene expression analyses were perfomed during feeding against starvation in order to determine sugar transporter and other feeding related gene expression. Young BPH females that feed rice seedlings or feed-deprived (water-supplied) for 24 hours were prepared in triplicate. Gene expression was compared in these two groups: feeding and feed-deprived.
Project description:Infestation with white-backed planthopper (WBPH) to rice caused induced resistance to rice pathogens but brown planthopper (BPH) infestation induce weaker resistance to rice pathogens. We compared changes in gene expression in rice plants infested with WBPH and BPH to gain some insight into the WBPH-induced resistance to rice pathogens. An analysis, using microarrays, of gene expression in rice plants infested with these planthoppers revealed that WBPH infestation caused high induction of many defense-related genes including pathogenesis-related (PR) genes than BPH infestation. Furthermore, hydroperoxide lyase 2 (OsHPL2) which is an enzyme to produce C6 volatiles was induced by WBPH infestation, but not by BPH infestation. Keywords: response to herbivory; induced resistance
Project description:Nilaparvata lugens, the brown planthopper (BPH) sucks the rice phloem sap containing high sucrose to obtain carbon source. The comparative gene expression analyses were perfomed during feeding against starvation in order to determine sugar transporter and other feeding related gene expression.
Project description:Brown planthopper (BPH) is the most notorious insect pest to rice. Drought is the most commonly occurring global adversity. BPH infestation caused adaxially-rolled leaves and shrunk bulliform cells similar to drought. The bulliform-cell characteristic gene, ACL1, negatively regulated BPH resistance and drought tolerance, with decreased cuticular wax in ACL1-D, which resulted in quicker water losing. ACL1 was specifically expressed in epidermis. TurboID system and various biochemical assays revealed that ACL1 interacted with the epidermal-characteristic HD-Zip IV ROCs. ROC4 and ROC5 positively regulated BPH resistance and drought tolerance through modulating cuticular wax and bulliform cells respectively. Overexpression of ROC4 and ROC5 both rescued ACL1-D in various related phenotypes simultaneously. Moreover, ACL1 competed with ROC4 and ROC5 in homo-dimerization and hetero-dimerization. Altogether, we illustrated that ACL1-ROCs complex synergistically mediate drought tolerance and BPH resistance through regulating cuticular wax and bulliform cells in rice, a new mechanism which might facilitate BPH resistance breeding.