Ectopic expression of antifreeze protein gene from Ammopiptanthus nanus confers chilling tolerance in maize  

作　　者：Yuanyuan Zhang Yang Cao Hongying Zheng Wenqi Feng Jingtao Qu Fengling Fu Wanchen Li Haoqiang Yu 

机构地区：[1]Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region,Ministry of Agriculture,Maize Research Institute,Sichuan Agricultural University,Chengdu 611130,Sichuan,China [2]College of Life Science&Biotechnology,Mianyang Teachers’College,Mianyang 621000,Sichuan,China

出　　处：《The Crop Journal》2021年第4期924-933,共10页作物学报（英文版）

基　　金：supported by National Key Science and Technology Special Project(2016ZX08003-004);Sichuan Science and Technology Program(2018JY0470)。

摘　　要：Improved chilling tolerance is important for maize production. Previous efforts in transgenics and marker-assisted breeding have not achieved practical results. In this study, the antifreeze protein(AnAFP) from the super-xerophyte Ammopiptanthus nanus was aligned to KnS-type dehydrins.Phosphorylation in vitro and subcellular localization showed that AnAFP was phosphorylated by maize casein kinase II and translocated from nucleus to cytoplasm under chilling stress. AnAFP also increased lactate dehydrogenase activity. A parent line of commercial maize hybrids was transformed with the AnAFP gene. Based on thermal asymmetric interlaced PCR, one hemizygous and two homozygous integration sites were identified in one T_(1) line. Ectopic expression of AnAFP in transgenic lines was confirmed by quantitative real-time PCR, RNA-seq, and Western blotting. After chilling treatment, the transgenic lines showed significantly improved phenotype, higher kernel production, survival rate and biomass, and lower relative electrolyte leakage and superoxide dismutation than the untransformed line. Thus, ectopic expression of AnAFP gene improved chilling tolerance in the transgenic lines, which could be used to apply for further safety assessment for commercial breeding.

关 键 词：Antifreeze protein Chilling tolerance DEHYDRIN Ectopic expression Homozygosity identification 

分 类 号：S513[农业科学—作物学]