Gene Profiling of the Ascorbate Oxidase Family Genes under Osmotic and Cold Stress Reveals the Role of AnAO5 in Cold Adaptation in Ammopiptanthus nanus

Abstract

The uplift of the Qinghai Tibet Plateau has led to a drastic change in the climate in Central Asia, from warm and rainy, to dry and less rainfall. Ammopiptanthus nanus, a rare evergreen broad-leaved shrub distributed in the temperate desert region of Central Asia, has survived the drastic climate change in Central Asia caused by the uplift of the Qinghai-Tibet Plateau. Ascorbate oxidase (AO) regulates the redox status of the apoplast by catalyzing the oxidation of ascorbate acid to dehydroascorbic acid, and plays a key role in the adaptation of plants to environmental changes. Analyzing the evolution, environmental response, and biological functions of the AO family of A. nanus is helpful for understanding how plant genome evolution responds to climate change in Central Asia. A total of 16 AOs were identified in A. nanus, all of which contained the ascorbate oxidase domain, most of which contained transmembrane domain, and many were predicted to be localized in the apoplast. Segmental duplication and tandem duplication are the main factors driving the gene amplification of the AO gene family in A. nanus. Gene expression analysis based on transcriptome data and fluorescence quantitative PCR, as well as enzyme activity measurements, showed that the expression levels of AO genes and total enzyme activity decreased under short-term osmotic stress and low-temperature stress, but the expression of some AO genes (AnAO5, AnAO13, and AnAO16) and total enzyme activity increased under 7 days of cold stress. AnAO5 and AnAO11 are targeted by miR4415. Further functional studies on AnAO5 showed that AnAO5 protein was localized in the apoplast. The expression of AnAO5 in yeast cells and the transient expression in tobacco enhanced the tolerance of yeast and tobacco to low-temperature stress, and the overexpression of AnAO5 enhanced the tolerance of Arabidopsis seedlings to cold stress. Our research provides important data for understanding the role of AOs in plant adaptation to environmental change.

Chemical and Transcriptomic Analyses of Leaf Cuticular Wax Metabolism in Ammopiptanthus mongolicus under Osmotic Stress

“…Reverse transcription and qRT-PCR were performed according to the previously described method [32]. The eIF1 gene was used as the reference for qRT-PCR analysis of A. mongolicus genes [33].…”

The Characterization of R2R3-MYB Genes in Ammopiptanthus nanus Uncovers That the miR858-AnaMYB87 Module Mediates the Accumulation of Anthocyanin under Osmotic Stress

“…RNA extraction and qRT-PCR analysis of the R2R3-MYB genes, miRNAs, and miRNA precursors were conducted according to two previous reports [56,59]. The eIF1 gene and an snRNA, U6, were used as references for qRT-PCR analysis of genes and miRNAs, respectively [60].…”

Advances in Endangered Plant Research: Ammopiptanthus’s Responses to Biotic and Abiotic Stressors

“…The soil is often infertile, accompanied by salinity and alkalinity stress, and they are also susceptible to severe pest infestations [4]. Over the past decade, many researchers have devoted themselves to in-depth investigations of the physiological and molecular mechanisms by which Ammopiptanthus species respond to both biotic and abiotic stressors [6][7][8][9][10][11][12]. The primary objective of this article is to provide a comprehensive review of the defensive responses and stress resistance mechanisms in Ammopiptanthus, shedding light on the physiological and molecular patterns employed by Ammopiptanthus when facing environmental stressors, as well as their interplay in resistance mechanisms.…”