Cloning of the HvnALS Gene in Highland Barley (Hordeum vulgare var. nudum) and Its Expression Analysis Under Pyroxsulam Stress
QIN Yun-Zhuo1, WENG Hua2,*
1 College of Agriculture and Animal Husbandry, Qinghai University, Xining 810016, China; 2 Academy of Agricultural and Forestry Sciences, Qinghai University, Xining 810016, China
Abstract：Pyroxsulam as an acetolactate synthase (ALS) inhibitor for controlling weeds in highland barley (Hordeum vulgare var. nudum) fields, ALS plays an important role in herbicide resistance in highland barley through the ALS gene. To investigate the role of the HvnALS gene in conferring pyroxsulam resistance in highland barley, this study utilized a pyroxsulam resistant variety 'Qing 0306' (R) and a susceptible variety 'Qing 0160' (S). Primers were designed based on transcriptome sequences to clone the HvnALS gene, followed by corresponding bioinformatic analysis. The results of ALS activity assay showed that the relative ALS activity of the resistant variety 'Qing 0306' (R) was extremely significantly higher than that of the control from 1~4 d after pyroxasulfone treatment. qPCR was employed to analyze the expression levels of the HvnALS gene in different resistant naked barley varieties under pyroxsulam treatment. The results showed that, the full length of the HvnALS gene (GenBank No. KAE8811959.1) was 1 941 bp, encoding 646 amino acids. Compared to known ALS gene amino acid sequences, no amino acid mutation sites were found in either variety. Multiple protein sequence alignment and phylogenetic tree analysis indicated that the HvnALS protein possesses the PLN02470 domain and exhibits the closest genetic relationship to the HvALS protein of barley (Hordeum vulgare). qPCR analysis revealed that the expression of this gene rapidly increased after pyroxsulam stress. At 1 d after application, its expression in the resistant material 'Qing 0306' (R) was ectremely significantly higher than that in the susceptible variety (P<0.01). Analysis of HvnALS gene expression following pyroxsulam stress suggested that the high-level resistance in 'Qing 0306' (R) might be associated with the overexpression of the target enzyme, rather than mutations in the target enzyme gene locus. This overexpression was likely the primary mechanism leading to pyroxsulam cross-resistance in highland barley.This study provides a theoretical reference for the genetic improvement of pyroxsulam-tolerant highland barley varieties.
秦云卓, 翁华. 青稞HvnALS基因克隆及其在啶磺草胺胁迫下的表达分析[J]. 农业生物技术学报, 2025, 33(12): 2602-2611.
QIN Yun-Zhuo, WENG Hua. Cloning of the HvnALS Gene in Highland Barley (Hordeum vulgare var. nudum) and Its Expression Analysis Under Pyroxsulam Stress. 农业生物技术学报, 2025, 33(12): 2602-2611.
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