玉米CIPK基因家族的鉴定及ZmCIPK3的抗旱性功能研究

Identification of CIPK Gene Family Members and Investigation of the Drought Tolerance of ZmCIPK3 in Maize

钙调磷酸酶B类互作蛋白激酶(CIPK,CBL interacting protein kinases)是植物钙离子信号通路中响应非生物逆境胁迫的重要蛋白激酶之一。本研究以拟南芥和水稻中CIPK家族基因序列信息为基础,利用玉米参考基因组B73和生物信息学分析方法,全基因组范围内鉴定玉米CIPK基因家族成员,分析CIPK家族基因的进化关系、基因结构、基因表达模式和对干旱胁迫的响应。本研究共鉴定出44个玉米CIPK家族基因,并将其分为5个亚家族,每个亚家族有不同的外显子-内含子和UTR的结构特征;基于基因差异表达分析,筛选出5个与抗旱性相关的候选基因ZmCIPK3、ZmCIPK7、ZmCIPK44、ZmCIPK25和ZmCIPK28;进一步的遗传数据表明,干旱胁迫下ZmCIPK3拟南芥转基因株系的存活率明显高于野生型,提高了拟南芥的抗旱性;同时,干旱胁迫下ZmCIPK3拟南芥转基因株系中抗旱性相关生化指标过氧化物酶、超氧化物歧化酶活性显著高于野生型,而丙二醛和脯氨酸的含量显著低于野生型。本研究在玉米全基因组水平上鉴定了CIPK基因家族成员,分析了其在不同抗旱性材料、不同水分处理下的基因表达模式,明确了ZmCIPK3是一个抗旱性候选基因。

Calcineurin B-like interacting protein kinase CIPK(CBL interacting protein kinases)is one of important protein kinases in the plant calcium signal pathway in response to abiotic stresses.Based on the sequence information of the CIPK family genes in Arabidopsis and rice,the members of the maize CIPK gene family were identified from the maize reference genome B73.The evolution relationship,structure variation,transcriptional profile,and transcriptional re-programing under drought stress treatments of CIPK family genes were analyzed.44 maize CIPK family genes were identified and classified into five sub-families,each of which represented different characteristics on exon-intron and UTR structure.Based on the analysis of transcriptional profiles under drought stress condition,five candidate genes(ZmCIPK3,ZmCIPK7,ZmCIPK44,ZmCIPK25 and ZmCIPK28)involving into drought tolerance were identified.Furthermore,by over-expressing the ZmCIPK3 gene in Arabidopsis thaliana,under drought stress conditions the survival rate of ZmCIPK3-transgenic lines was significantly higher than that of the wild type,suggesting that ZmCIPK3 might increase the drought resistance.The activities of peroxidase and superoxide dismutase in transgenic lines were significantly higher than those of the wild type,while the contents of malondialdehyde and proline were significantly lower than those of wild type.This study identified the members of the maize CIPK gene family at the genome-wide level and their transcriptional profile upon drought stress treatments,and confirmed ZmCIPK3 as a candidate gene conferring drought tolerance.