Characterization of Annexin gene family and functional analysis of RsANN1a involved in heat tolerance in radish (Raphanus sativus L.)

Feng Shen; Jiali Ying; Liang Xu; Xiaochuan Sun; Jizhong Wang; Yan Wang; Yi Mei; Yuelin Zhu; Liwang Liu

doi:10.1007/s12298-021-01056-5

Characterization of Annexin gene family and functional analysis of RsANN1a involved in heat tolerance in radish (Raphanus sativus L.)

Physiol Mol Biol Plants. 2021 Sep;27(9):2027-2041. doi: 10.1007/s12298-021-01056-5. Epub 2021 Sep 14.

Authors

Feng Shen^#^{1

2}, Jiali Ying^#¹, Liang Xu^#¹, Xiaochuan Sun³, Jizhong Wang³, Yan Wang¹, Yi Mei², Yuelin Zhu¹, Liwang Liu¹

Affiliations

¹ National Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China) of MOAR, College of Horticulture, Nanjing Agricultural University, Nanjing, 210095 China.
² Jiangsu Coastal Area Institute of Agricultural Sciences, Yancheng, 224002 China.
³ Huaiyin Institute of Technology, Huaian, 223003 China.

^# Contributed equally.

Abstract

Plant annexins are a kind of conserved Ca²⁺-dependent phospholipid-binding proteins which are involved in plant growth, development and stress tolerance. Radish is an economically important annual or biennial root vegetable crop worldwide. However, the genome-wide characterization of annexin (RsANN) gene family remain largely unexplored in radish. In this study, a comprehensive identification of annexin gene family was performed at the whole genome level in radish. In total, ten RsANN genes were identified, and these putative RsANN proteins shared typical characteristics of the annexin family proteins. Phylogenetic analysis showed that the RsANNs together with annexin from Arabidopsis and rice were clustered into five groups with shared similar motif patterns. Chromosomal localization showed that these ten RsANN genes were distributed on six chromosomes (R3-R8) of radish. Several cis-elements involved in abiotic stress response were identified in the promoter regions of RsANN genes. Expression profile analysis indicated that the RsANN genes exhibited tissue-specific patterns at different growth stages and tissues. The Real-time quantitative PCR (RT-qPCR) revealed that the expression of most RsANN genes was induced under various abiotic stresses including heat, drought, salinity, oxidization and ABA stress. In addition, stress assays showed that overexpression of RsANN1a improved plant's growth and heat tolerance, while artificial microRNAs (amiRNA)-mediated knockdown of RsANN1a caused dramatically decreased survival ratio of Arabidopsis plants. These findings not only demonstrate that RsANN1a might play a critical role in the heat stress response of radish, but also facilitate clarifying the molecular mechanism of RsANN genes in regulating the biological process governing plant growth and development.

Supplementary information: The online version contains supplementary material available at 10.1007/s12298-021-01056-5.

Keywords: Gene function validation; Radish; RsANN1a; amiRNA; annexin gene family.