为探究甜瓜CIPK(CBL-interacting protein kinase)基因家族成员的生物学功能,以拟南芥中26个CIPKs的氨基酸序列为参照,用BLASTP方法鉴定了18个甜瓜CIPK家族成员,并对其理化性质、染色体分布、系统进化、基因结构、蛋白保守基序、顺式元件,以及基因的表达模式进行了分析。结果显示,CIPK基因家族成员的基因长度为1499~8499 bp,它们不均匀地分布在甜瓜的9条染色体上;根据进化关系可将其分为5个亚家族,分别包含了26、10、25、26和8个成员,并且其中有4个CmCIPK基因对发生了片段复制。基因结构分析表明,10个基因为内含子缺失型,8个基因为内含子富集型。蛋白保守基序分析发现,CmCIPK家族保守性较好,所有成员均含有CIPK家族的典型特征:N端激酶域中的激活环和C端调节域中的NAF/FISL结构域。在基因上游的2000 bp序列中存在多个与植物激素和逆境相关的顺式元件,显示可能有多种转录调控。转录组分析发现,CmCIPKs的组织表达量整体表现为叶>根>雄花>雌花>果。选择CmCIPK1-like和CmCIPK12-like基因验证其组织表达模式,显示分别在叶和雄花中的表达量最高。对其进行不同逆境处理,结果表明,这2个CmCIPK基因受NaCl和脱落酸(ABA)诱导表达;在干旱处理中,这2个基因经历短时间的上调或下调后恢复至最初表达水平。以上结果说明,CmCIPK1-like和CmCIPK12-like基因可能在ABA和非生物胁迫中发挥着重要作用,可为以后CmCIPK的功能研究提供参考。
Salinity is one of the most important abiotic stresses that adversely affects crop growth and productivity. A subtractive suppression hybridization(SSH) library were constructed from the roots of salt-sensitive Yulu cultivar melon seedlings under salt stress; 557 high-quality expressed sequence tags(ESTs) were randomly sequenced, with an average size of 428 bp, which assembled into 68 contigs and 315 singletons. Compared with our previous SSH library generated from the salt-tolerant Bingxuecui cultivar, the proportion of transcripts involved in metabolism, protein fate, cellular communication/signal transduction mechanisms, and cell rescue/defense were 4, 1.46, 0.94, and 0.4% higher, respectively, in the salt-tolerant cultivar than the in salt-sensitive cultivar. Quantitative real-time PCR analysis of eleven transcripts revealed temporal variations in their expression in the two cultivars under salt stress. One NAC gene(JZ477011) was heterologously expressed in yeast for functional characterization, and enhanced the sensitivity of yeast cells to high-salinity to salt stress and inhibited their growth. Information regards to their functions would aid in the understanding of response mechanisms to saline stress and in the development of molecular markers for selecting salt-tolerant melon cultivars.
