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Genome-Wide Analysis of CDPK Family in Foxtail Millet and Determination of SiCDPK24 Functions in Drought Stress

机译:谷子CDPK家族的全基因组分析和干旱胁迫下SiCDPK24功能的测定

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摘要

Plant calcium-dependent protein kinases (CDPKs) were reported to play important roles in plant resistance to abiotic stress. Foxtail millet cultivation “H138” was used for RNA-seq analysis. The data from drought-induced de novo transcriptomic sequences of foxtail millet showed that CDPKs were up- or down-regulated by drought to different degrees. In this study, 29 foxtail millet CDPKs were classified into four subgroups. These genes were unevenly distributed on nine foxtail millet chromosomes, and chromosomes 2, 3, and 9 contained the most SiCDPK members. Analysis of putative cis-acting elements showed that most foxtail millet CDPK genes contained the ABRE, LTR, HSE, MYB, MYC, DRE, CGTCA-motif, and TGACG-motif cis-acting elements, which could be activated by abiotic stresses. Real-time PCR analysis indicated that 29 SiCDPK genes experienced different degrees of induction under drought and ABA stresses. SiCDPK24 had the highest expression levels at 6 and 12 h of drought treatment and was chosen for further analysis. SiCDPK24 localized to the cell membrane and the nucleus of Arabidopsis mesophyll protoplasts. Western blot analysis showed that SiCDPK24 protein had autophosphorylation activity. Overexpression of SiCDPK24 in Arabidopsis enhanced drought resistance and improved the survival rate under drought stress. It also activated the expressions of nine stress-related genes, namely RD29A, RD29B, RD22, KIN1, COR15, COR47, LEA14, CBF3/DREB1A, and DREB2A. These genes are involved in resistance to abiotic stresses in Arabidopsis. These results indicate that foxtail millet CDPK genes play important roles in resisting drought stress.
机译:据报道植物钙依赖性蛋白激酶(CDPKs)在植物对非生物胁迫的抗性中起重要作用。谷子栽培“ H138”用于RNA-seq分析。干旱引起的谷子从头转录序列的数据表明,干旱对CDPKs的上调或下调程度不同。在这项研究中,将29种谷子小米CDPK分为四个亚组。这些基因在9条狐尾粟染色体上分布不均,并且2、3和9号染色体包含最多的SiCDPK成员。推定的顺式作用元件的分析表明,大多数谷子小穗CDPK基因包含ABRE,LTR,HSE,MYB,MYC,DRE,CGGTA-motif和TGACG-motif顺式作用元件,可以通过非生物胁迫激活。实时PCR分析表明29个SiCDPK基因在干旱和ABA胁迫下经历了不同程度的诱导。 SiCDPK24在干旱处理6和12 h时具有最高表达水平,因此被选择用于进一步分析。 SiCDPK24定位于拟南芥叶肉原生质体的细胞膜和细胞核。 Western印迹分析表明,SiCDPK24蛋白具有自磷酸化活性。 SiCDPK24在拟南芥中的过表达增强了抗旱性并提高了干旱胁迫下的存活率。它还激活了九个应激相关基因的表达,即RD29A,RD29B,RD22,KIN1,COR15,COR47,LEA14,CBF3 / DREB1A和DREB2A。这些基因参与了拟南芥对非生物胁迫的抗性。这些结果表明,谷子小穗CDPK基因在抵抗干旱胁迫中起重要作用。

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