Abstract
Verticillium wilt is a major disease that severely affects eggplant production, and a new eggplant miRNA named miRm0002 identified through high-throughput sequencing was highly induced by Verticillium wilt infection. However, the miRm0002 function was still elusive. In this study, the sequence of the miRm0002 precursor was cloned and transgenic eggplants were constructed. In vivo inoculation test and in vitro fungistatic test showed that overexpressing miRm0002 lines were more resistant to Verticillium dahliae and inhibiting miRm0002 lines were more sensitive, compared to the wild-type (WT) control. Some physiological indicators were selected and the results showed that SOD, POD, and CAT activities were significantly increased in Verticillium wilt-infected overexpressing miRm0002 lines, indicating that the expression of miRm0002 activates the antioxidant system. QRT-PCR assay showed that the transcript expression of miRm0002 candidate target ARF8, a gene encoding auxin response factor was negatively related to miRm0002 in WT as well as transgenic eggplants. However, RLM-RACE mapping and degradome sequencing showed miRm0002 could not cleave the sequence of ARF8. Taken together, these data suggest that miRm0002 plays a positive role in the defense response of eggplant against Verticillium wilt.
Highlights
Verticillium wilt (V. wilt), a disease caused by soil-borne Verticillium dahliae, reduces up to 34.1–42.5% of eggplant production worldwide [1,2]
Org/mfold/applications/rna-folding-form.php accessed on 20 September 2021), and the results showed that the 19 nt of miRm0002 was located at the stem (Figure 1b). 194 bp pri-miRm0002 sequence was cloned for further research (Figure 1c)
Wilt, the sense and antisense sequences of miRm0002 precursor were cloned and introduced into pCAMBIA1304 expression vectors driven by the CaMV35S promoter (Figure 2a), which were transferred into eggplant cultivar Suqi (Figure 2b)
Summary
Verticillium wilt (V. wilt), a disease caused by soil-borne Verticillium dahliae, reduces up to 34.1–42.5% of eggplant production worldwide [1,2]. Over the last two decades, significant progress has been made by scientists and farmers in improving the resistance of eggplants to V. wilt. Introducing resistance genes such as StoVe1, StoL13a, StoNPR1, and StoCYP77A2 from the highly resistant wild eggplant Solanum torvum into potato or tobacco has significantly improved their resistance to V. wilt [3,4,5,6]. 11,696 upregulated and 5949 downregulated genes, which are candidates for further genetic improvement [7] All these studies only focused on screening or introducing the resistance genes, the molecular mechanism of how eggplant interacts with the disease agent fungus remains elusive. Previous studies have showed that some miRNAs are involved in regulating the defense responses of plants against pathogen attack
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