Bioinformatics, expression analysis, and functional verification of allene oxide synthase gene HvnAOS1 and HvnAOS2 in qingke

Likun An; Ziao Wang; Yongmei Cui; Youhua Yao; Yixiong Bai; Yuehai Liu; Xin Li; Xiaohua Yao; Kunlun Wu

doi:10.1515/biol-2022-0855

Bioinformatics, expression analysis, and functional verification of allene oxide synthase gene HvnAOS1 and HvnAOS2 in qingke

Open Life Sci. 2024 Apr 25;19(1):20220855. doi: 10.1515/biol-2022-0855. eCollection 2024.

Authors

Likun An^{1

2

3

4}, Ziao Wang^{1

2

3

4}, Yongmei Cui^{1

2

3

4}, Youhua Yao^{1

2

3

4}, Yixiong Bai^{1

2

3

4}, Yuehai Liu^{1

2

3

4}, Xin Li^{1

2

3

4}, Xiaohua Yao^{1

2

3

4}, Kunlun Wu^{1

2

3

4}

Affiliations

¹ Academy of Agricultural and Forestry Sciences, Qinghai University, Xining, Qinghai 810016, China.
² Laboratory for Research and Utilization of Qinghai Tibet Plateau Germplasm Resources, Xining, Qinghai 810016, China.
³ Qinghai Key Laboratory of qingke Genetics and Breeding, Xining, Qinghai 810016, China.
⁴ Qinghai Subcenter of National qingke Improvement, Xining, Qinghai 810016, China.

Abstract

Allene oxide synthase (AOS) is a key enzyme involved in the jasmonic acid (JA) synthesis pathway in plants. To explore its function on the regulatory mechanism of JA synthesis, we screened and identified two AOS genes HvnAOS1 and HvnAOS2 in qingke. Both HvnAOS1 and HvnAOS2 contained conserved heme-binding motif, which is most closely related to AtsAOS2, indicating controlled dehydration of fatty acid hydroperoxides to allene oxides. Molecular docking simulations identified the key amino acid sites that were important for heme binding and interaction with 13(S)-HPOT, respectively. The expression pattern also indicated that HvnAOS1 and HvnAOS2 were highly induced by JA, abscisic acid, and salicylic acid. Subcellular localization of HvnAOS1 and HvnAOS2 using transient expression of Agrobacterium tumefaciens showed the green fluorescent protein signal in the cell cytoplasm of the N. benthamiana leaves. Overexpression of HvnAOS1 and HvnAOS2 in Arabidopsis aos mutant restored male fertility and plant resistance to Botrytis cinerea, indicating that HvnAOS1 and HvnAOS2 can restore the functions of AOS in Arabidopsis aos mutant.

Keywords: HvnAOS1 and HvnAOS2 genes; allene oxide synthase; bioinformatics analysis; molecular docking simulation; qingke.