ORCID IDs: 0000-0002-6747-4302 (J.F.); 0000-0003-4066-4875 (W.-M.W.).Circular RNAs (circRNAs) play roles in various biological processes, but their functions in the rice (Oryza sativa) response to Magnaporthe oryzae remain elusive. Here, we demonstrate that circRNAs are involved in the rice-M. oryzae interaction using comparative circRNA-sequencing and transgenic approaches. We identified 2932 high-confidence circRNAs from young leaves of the blast-resistant accession International Rice Blast Line Pyricularia-Kanto51-m-Tsuyuake (IR25) and the blast-susceptible accession Lijiangxin Tuan Heigu (LTH) under M. oryzae-infected or uninfected conditions; 636 were detected specifically upon M. oryzae infection. The circRNAs in IR25 were significantly more diverse than those in LTH, especially under M. oryzae infection. Particularly, the number of circRNAs generated per parent gene was much higher in IR25 than in LTH and increased in IR25 but decreased in LTH upon M. oryzae infection. The higher diversity of circRNAs in IR25 was further associated with more frequent 39 and 59 alternative back-splicing and usage of complex splice sites. Moreover, a subset of circRNAs was differentially responsive to M. oryzae in IR25 and LTH. We further confirmed that circR5g05160 promotes rice immunity against M. oryzae. Therefore, our data indicate that circRNA diversity is associated with different responses to M. oryzae infection in rice and provide a starting point to investigate a new layer of regulation in the rice-M. oryzae interaction.
Magnaporthe oryzae is the causative agent of rice blast, a devastating disease in rice worldwide. Based on the gene-for-gene paradigm, resistance (R) proteins can recognize their cognate avirulence (AVR) effectors to activate effector-triggered immunity. AVR genes have been demonstrated to evolve rapidly, leading to breakdown of the cognate resistance genes. Therefore, understanding the variation of AVR genes is essential to the deployment of resistant cultivars harboring the cognate R genes. In this study, we analyzed the nucleotide sequence polymorphisms of eight known AVR genes, namely, AVR-Pita1, AVR-Pii, AVR-Pia, AVR-Pik, AVR-Pizt, AVR-Pi9, AVR-Pib, and AVR-Pi54 in a total of 383 isolates from 13 prefectures in the Sichuan Basin. We detected the presence of AVR-Pik, AVR-Pi54, AVR-Pizt, AVR-Pi9, and AVR-Pib in the isolates of all the prefectures, but not AVR-Pita1, AVR-Pii, and AVR-Pia in at least seven prefectures, indicating loss of the three AVRs. We also detected insertions of Pot3, Mg-SINE, and indels in AVR-Pib, solo-LTR of Inago2 in AVR-Pizt, and gene duplications in AVR-Pik. Consistently, the isolates that did not harboring AVR-Pia were virulent to IRBLa-A, the monogenic line containing Pia, and the isolates with variants of AVR-Pib and AVR-Pizt were virulent to IRBLb-B and IRBLzt-t, the monogenic lines harboring Pib and Piz-t, respectively, indicating breakdown of resistance by the loss and variations of the avirulence genes. Therefore, the use of blast resistance genes should be alarmed by the loss and nature variations of avirulence genes in the blast fungal population in the Sichuan Basin.
The receptor-like kinase (RLK) FERONIA functions in immunity in Arabidopsis. Here, we systemically screened rice RLK genes encoding FERONIA-like receptor (FLRs) that may be involved in rice-Magnaporthe oryzae interaction. The expression of 16 FLR genes was examined in response to the infection of M. oryzae in different rice varieties. For each FLR gene, at least two independent mutants were generated by CRISPR/Cas9 gene-editing technology in rice variety Zhonghua 11 (ZH11). Blast disease assay identified that the mutants of FLR1 and FLR13 showed increased susceptibility, whereas the mutants of FLR2 and FLR11 displayed enhanced resistance. Consistently, the mutant of FLR1 enhanced, but the mutant of FLR2 delayed the M. oryzae infection progress, which might be associated with the altered expression of defense-related genes. Together, these data indicate that at least 4 FLR genes are involved in rice-M. oryzae interaction and thus are potentially valuable in blast disease resistance.
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