Mao Peijun, Run Yonghang, Wang Hanghui, Han Changdong, Zhang Lijun, Zhan Kehui, Xu Haixia, Cheng Xiyong
Co-construction State Key Laboratory of Wheat and Maize Crop Science, Collaborative Innovation Center of Henan Grain Crops, College of Agronomy, Henan Agricultural University, Zhengzhou, China.
Front Genet. 2022 Mar 16;13:846795. doi: 10.3389/fgene.2022.846795. eCollection 2022.
In plants, chloride channels (CLC) are involved in a series of specific functions, such as regulation of nutrient transport and stress tolerance. Members of the wheat L. CLC (TaCLC) gene family have been proposed to encode anion channels/transporters that may be related to nitrogen transportation. To better understand their roles, TaCLC family was screened and 23 gene sequences were identified using a Hidden Markov Model in conjunction with wheat genome database. Gene structure, chromosome location, conserved motif, and expression pattern of the resulting family members were then analyzed. Phylogenetic analysis showed that the TaCLC family can be divided into two subclasses (I and II) and seven clusters (-a, -c1, -c2, -e, -f1, -f2, and -g2). Using a wheat RNA-seq database, the expression pattern of TaCLC family members was determined to be an inducible expression type. In addition, seven genes from seven different clusters were selected for quantitative real-time PCR (qRT-PCR) analysis under low nitrogen stress or salt stress conditions, respectively. The results indicated that the gene expression levels of this family were up-regulated under low nitrogen stress and salt stress, except the genes of TaCLC-c2 cluster which were from subfamily -c. The yeast complementary experiments illustrated that , , and all had anion transport functions for NO or Cl, and compensated the hypersensitivity of yeast GEF1 mutant strain YJR040w () in restoring anion-sensitive phenotype. This study establishes a theoretical foundation for further functional characterization of genes and provides an initial reference for better understanding nitrate nitrogen transportation in wheat.
在植物中，氯离子通道（CLC）参与一系列特定功能，如调节养分运输和胁迫耐受性。已有人提出小麦（Triticum aestivum L.）CLC（TaCLC）基因家族的成员编码可能与氮运输有关的阴离子通道/转运体。为了更好地了解它们的作用，利用隐马尔可夫模型结合小麦基因组数据库对TaCLC家族进行了筛选，并鉴定出23个基因序列。然后分析了所得家族成员的基因结构、染色体定位、保守基序和表达模式。系统发育分析表明，TaCLC家族可分为两个亚类（I和II）和七个簇（-a、-c1、-c2、-e、-f1、-f2和-g2）。利用小麦RNA测序数据库，确定TaCLC家族成员的表达模式为诱导型表达。此外，分别在低氮胁迫或盐胁迫条件下，从七个不同簇中选择七个基因进行定量实时PCR（qRT-PCR）分析。结果表明，除了来自-c亚家族的TaCLC-c2簇的基因外，该家族的基因表达水平在低氮胁迫和盐胁迫下均上调。酵母互补实验表明，TaCLC-f1;1、TaCLC-f2;1和TaCLC-g2;1对NO₃⁻或Cl⁻均具有阴离子转运功能，并补偿了酵母GEF1突变体菌株YJR040w（Δgef1）在恢复阴离子敏感表型方面的超敏感性。本研究为进一步对TaCLC基因进行功能鉴定奠定了理论基础，并为更好地理解小麦中硝态氮的运输提供了初步参考。
