Abstract
Grapevine powdery mildew is one of the most damaging fungal diseases. Therefore, a precise understanding of the grapevine disease resistance system becomes a subject of significant importance. Plant microRNAs(miRNAs) have been implicated to play regulatory roles in plant biotic stress responses. In this study, high-throughput sequencing and miRDeep-P were employed to identify miRNAs in Chinese wild Vitis pseudoreticulata leaves following inoculation with Erysiphe necator. Altogether, 126 previously identified microRNAs and 124 novel candidates of miRNA genes were detected. Among them, 43 conserved miRNAs belong to 20 families and 23 non-conserved but previously-known miRNAs belong to 15 families. Following E. necator inoculation, 119 miRNAs were down-regulated and 131 were up-regulated. Furthermore, the expression changes occurring in 32 miRNAs were significant. The expression patterns of some miRNAs were validated by semi-quantitative RT-PCR and qRT-PCR. A total of 485 target genes were predicted and categorized by Gene Ontology (GO). In addition, 14 vvi-miRNAs were screened with 36 targets which may be involved in powdery mildew resistance in grape. Highly accumulated vvi-NewmiR2118 was detected from accession “Baihe-35-1,” whose targets were mostly NBS-LRR resistance genes. It was down-regulated rapidly and strongly in “Baihe-35-1” leaves after inoculated with E. necator, indicating its involvement in grape powdery mildew resistance. Finally, the study verified interaction between vvi-NewmiR2118 and RPP13 by histochemical staining and GUS fluorescence quantitative assay.
Highlights
Grapevine is one of the world’s oldest and most important fruit crops—culturally, nutritionally and economically
The growth of P. syringae was restricted on host plants, implying that miRNA-mediated suppression of auxin signaling enables enhanced disease resistance in plants (Navarro et al, 2006). microRNA398 was down-regulated in response to oxidative stress caused by abiotic or biotic stresses (Sunkar et al, 2006; Jagadeeswaran et al, 2009a)
Using susceptible cultivars of Piont Noir as control material, we showed that vvi-NewmiR2118 was down-regulated greatly in “Baihe-35-1” leaves after inoculated with E. necator, indicating its involvement in grape powdery mildew resistance
Summary
Grapevine is one of the world’s oldest and most important fruit crops—culturally, nutritionally and economically. The researchers of this study believed that alternative splicing might be a mechanism adopted by plants of attenuating miRNA-mediated gene regulation. Do natural miRNAs act in plant biotic stress regulatory networks, and some appropriately modified artificial miRNAs can confer resistance to pathogen susceptible plants, which is elegantly proved by Niu et al (2006). Transgenic plants expressing both artificial miRNAs from a modified dimeric miR159 precursor conferred resistance to both viruses. These results implicate the possibility to use amiRNA-mediated approaches to engineer resistance to multiple viruses in crop plants (Niu et al, 2006)
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