Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (7): 195-205.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1304

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Isolation and Functional Verification of Genes Responding to Iron and Cadmium Stresses in Lycium barbarum

YU Hui(), WANG Jing, LIANG Xin-xin, XIN Ya-ping, ZHOU Jun, ZHAO Hui-jun()   

  1. Key Laboratory of Ecological Protection of Agro-pastoral Ecotones in the Yellow River Basin, National Ethnic Affairs Commission of the People's Republic of China, College of Biological Science and Engineering, North Minzu University, Yinchuan 750021
  • Received:2022-10-24 Online:2023-07-26 Published:2023-08-17
  • Contact: ZHAO Hui-jun E-mail:1632570918@qq.com;zhaohuijun1022@163.com

Abstract:

In this paper, the effects of iron(Fe)and cadmium(Cd)stress on the expression patterns of genes of LbFRO(ferric reduction oxidase), LbZIP(zinc-regulated transporters and iron-regulated transporter-like protein)and LbNRAMP(natural resistance-associated macrophage protein)in Lycium barbarum L. were investigated for providing theoretical basis and technical support in preventing and controlling yellow leaf disease of wolfberry in Ningxia, cultivation of high-efficiency Fe-utilizing wolfberry varieties and low-cadmium accumulation varieties. The seedlings of L. barbarum L. No. 1 were used as experimental materials, and the genes related to iron(Fe)and cadmium(Cd)absorption and transport were isolated by transcriptome sequencing technology; the expression patterns of the target genes under Fe and Cd stress were verified by real-time fluorescence quantitative PCR; the full-length sequences of the target genes were cloned by conventional PCR method; and the yeast expression vector of those genes were constructed and transformed into Fe/Cd sensitive yeast strain ΔCCC1/YK44 for functional verification. The results showed that LbFRO, LbIRT1, LbIRT2 and LbNRAMP3 were significantly up-regulated under Fe and Cd stress, phylogenetic tree analysis showed that those genes of L. barbarum L. were closely related to rice and Arabidopsis thaliana, the functions of transgenic yeast further verified that LbFRO, LbIRT2 and LbNRAMP3 genes enhanced the sensitivity of yeast to iron(Fe)and cadmium(Cd), indicating that these genes may be involved in the absorption and transport of Fe and Cd in L. barbarum L.

Key words: Lycium barbarum L., iron stress, cadmium stress, transgenic yeast, real-time fluorescence quantitative PCR