图1 CmCIPK基因在甜瓜染色体上的定位与共线性分析 A,CmCIPK家族成员在染色体上的分布。染色体大小由其相对长度表示;图中未显示不带有CmCIPK基因的染色体(第1、9和10号染色体)。B,CmCIPK基因的共线性分析。黑线表示CmCIPK家族中发生片段复制的基因对,灰线表示甜瓜基因组中所有发生片段复制的基因对。

Fig.1 Location of CmCIPK on the chromosomes of melon and collinearity analysis A, Specific distribution of members of the CmCIPKs on each chromosome. The size of the chromosome was represented by its relative length. The chromosomes without the CmCIPK(1st, 9th and 10th) were not shown in the figure. B, Collinearity analysis of CmCIPK. The black line represented the gene pair in the CmCIPK family where fragment replication occurred, and the gray line represented all the gene pairs in the melon genome where fragment replication occurred.

图2 甜瓜、拟南芥、水稻和黄瓜中CIPK蛋白的系统发育分析 使用MEGA 7.0软件将18个甜瓜CmCIPK蛋白(菱形)、26个拟南芥AtCIPK蛋白(圆形)、33个水稻OsCIPK蛋白(三角形),以及18个黄瓜CsCIPK蛋白(正方形)构建邻接树。5组分别标记为Ⅰ、Ⅱ、Ⅲ、Ⅳ、Ⅴ。

Fig.2 Phylogenetic analysis of CIPK proteins in melon, Arabidopsis, rice and cucumber 18 CmCIPK proteins (diamond), 26 AtCIPK proteins (circle), 33 OsCIPK proteins( triangle) and 18 CsCIPK proteins (square) were used to construct the neighbor tree by MEGA 7.0. The five groups were marked as Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ.

图3 甜瓜CmCIPK基因外显子-内含子结构分析 使用TBtool软件绘制基因结构,并根据系统发育树进行排列。黄色框代表外显子,绿色框代表UTR,线代表内含子。

Fig.3 Exon-intron structure of CmCIPKs Use TBtools software to draw gene structure, and arrange according to phylogenetic tree. The yellow boxes represented exons, the green boxes represented UTRs, and the lines represented introns.

图4 甜瓜CmCIPK蛋白基序分析 A,使用MEME程序鉴定保守基序,并根据系统发育树进行排列。每个基序均由一个彩色框表示,并在右侧给出其名称。B,编码已知功能域的基序序列注释。每个堆栈的总高度表示该位置的序列保守性,而每个堆栈中字母的高度代表基序中氨基酸的保守程度。

Fig.4 Motif analysis of CmCIPK proteins A, Use MEME to identify conserved motifs and arrange them according to the phylogenetic tree. Each motif was represented by a colored box, and its name was given on the right. B, Annotation of motif sequence encoding known functional domain. The total height of each stack indicated the sequence conservation at that position, and the height of the letters in each stack represented the degree of conservation of amino acids in the motif.

图5 甜瓜CmCIPK基因家族顺式作用元件分析 ABRE,脱落酸(ABA)响应元件;TGA-element,生长素响应元件;TC-rich repeats,防卫和逆境应答元件;LTR,冷胁迫应答元件;TCA-element,水杨酸应答元件;W-box,防卫应答元件;MBS,干旱胁迫应答元件;CGTCA-motif,茉莉酸甲酯(MeJA)响应元件。

Fig.5 Promoter cis-elements analysis of CmCIPK gene family ABRE, ABA-responsive element; TGA-element, auxin-responsive element; TC-rich repeats, defense and stress response element; LTR, low-temperature-responsive element; TCA-element, salicylic acid response element; W-box, defense and stress response element; MBS, drought stress response element; CGTCA-motif, Methyl jasmonate-responsive element.

图6 甜瓜CmCIPK基因在甜瓜不同组织中的转录丰度 使用Graphpad 8.2.1作热图,标度表示log2FPKM值的相对大小,绿色表示转录丰度较低,红色表示转录丰度较高。

Fig.6 Transcription abundance of CmCIPK in different tissues of melon Use Graphpad 8.2.1 to draw the heat map, the scale represented the relative size of the log2FPKM value, green represented low transcription abundance, and red represented high transcription abundance.

图7 CmCIPK1-like和CmCIPK12-like基因在甜瓜不同组织中的相对表达量 不同字母表示差异显著(P<0.05)。下同。

Fig.7 Expression level of CmCIPK1-like and CmCIPK12-like in different tissues of melon Different letters indicated significant differences (P<0.05). The same as below.

图8 100 μmol·L-1 ABA处理下CmCIPK1-like和CmCIPK12-like在甜瓜叶中的相对表达量

Fig.8 Relative expression level of CmCIPK1-like and CmCIPK12-like in leaves of melon under 100 μmol·L-1 ABA treatment

图9 NaCl处理5 d CmCIPK1-like和CmCIPK12-like在甜瓜根、茎、叶中的相对表达量

Fig.9 Relative expression level of CmCIPK1-like and CmCIPK12-like in roots, stems and leaves of melon under NaCl treatment for 5 days

图10 200 mmol·L-1 NaCl处理下CmCIPK1-like和CmCIPK12-like在甜瓜根、茎、叶中的相对表达量

Fig.10 Relative expression level of CmCIPK1-like and CmCIPK12-like in roots, stems and leaves of melon under 200 mmol·L-1 NaCl treatment

图11 15% PEG处理下CmCIPK1-like和CmCIPK12-like在甜瓜根、茎、叶中的相对表达量

Fig.11 Relative expression level of CmCIPK1-like and CmCIPK12-like in roots, stems and leaves of melon under 15% PEG treatment