Molecular Mechanism of Pyrroloquinoline Quinone-Mediated Rice Tolerance to Imidazolinone Herbicide Imazamox

Abstract

The Clearfield® technology is an useful tool for controlling weedy rice due to the effectiveness of imazamox and the cultivation of rice varieties resistant to imidazolines. However, residual imazamox in the soil probably causes phytotoxicity to subsequent non-resistant rice crops. We previously found that pyrroloquinoline quinone (PQQ), a bioactive elicitor, can enhance rice tolerance to imazamox. In this study, we explored the molecular mechanism of PQQ-mediated rice tolerance to imazamox by RNA-seq analysis, real-time quantitative PCR (RT-qPCR) assay, and enzyme activity assay. The results indicated that compared to imazamox at 66.7 mg a.i./L (IMA) alone, rice plants treated with imazamox at 66.7 mg a.i./L and PQQ at 0.66 mg a.i./L (IMA + PQQ) exhibited significantly reduced sensitivity to imazamox. Seven days post-treatment, IMA + PQQ-treated rice plants showed partial chlorosis and leaf curling, but IMA-treated rice plants had severe wilting and died. The fresh weight inhibition rate was 29.3% in the IMA + PQQ group, significantly lower than that of 56.6% in the IMA group alone. RNA-seq analysis showed differentially expressed genes were mainly involved in phenylpropanoid biosynthesis, diterpenoid biosynthesis, and MAPK signaling pathways in response to IMA + PQQ treatment. Both RNA-seq analysis and RT-qPCR assay showed that the expression of OsCATB gene in the catalase (CAT) gene family was upregulated at 12 h, the expression of OsGSTU1 gene was upregulated at 12, 24, and 48 h, while the expressions of OsABCB2, OsABCB11, and OsABCG11 genes were upregulated at 24 and 48 h. Enzyme activity assays revealed that the activity of superoxide dismutase in the IMA + PQQ group was increased by 47.45~120.31% during 12~72 h, compared to that in the IMA group. CAT activity in the IMA + PQQ group was increased by 123.72 and 59.04% at 12 and 48 h, respectively. Moreover, malondialdehyde levels indicative of oxidative damage were consistently lower in IMA + PQQ-treated plants, with a reduction of 46.29, 11.37, and 14.50% at 12, 36, and 72 h, respectively. Overall, these findings discover that PQQ has potential in reducing imazamox phytotoxicity in rice by enhancing antioxidant enzyme activities and regulating oxidative stress responses. They will provide valuable strategies for improving imazamox tolerance in crops.

Keywords

• imidazolinone herbicide
• phytotoxicity
• pyrroloquinoline quinone
• RNA-seq
• RT-qPCR
• enzymatic activity