植物研究 ›› 2026, Vol. 46 ›› Issue (5): 851-865.doi: 10.7525/j.issn.1673-5102.2026.05.007
董楚星1,2, 张梦圆1, 史鑫波1, 张岗2, 宋忠兴1, 王楠1, 焦红红1,3(
)
收稿日期:2026-04-22
出版日期:2026-09-20
发布日期:2026-10-09
通讯作者:
焦红红
E-mail:jhh211411@163.com
作者简介:董楚星(2004—),男,本科生,主要从事药物制剂研究。
基金资助:
Chuxing DONG1,2, Mengyuan ZHANG1, Xinbo SHI1, Gang ZHANG2, Zhongxing SONG1, Nan WANG1, Honghong JIAO1,3(
)
Received:2026-04-22
Online:2026-09-20
Published:2026-10-09
Contact:
Honghong JIAO
E-mail:jhh211411@163.com
摘要:
WRKY转录因子是植物类黄酮生物合成及生长发育的关键调控因子,但其在药用植物酸枣(Ziziphus jujuba var. spinosa)中的功能缺乏相关研究。本研究基于酸枣全基因组数据,采用生物信息学方法系统鉴定WRKY基因家族成员,并对其系统进化关系、染色体定位、保守结构域及启动子顺式作用元件进行分析。进一步通过外源脱落酸(abscisic acid,ABA)处理酸枣种子,结合类黄酮含量测定与基因表达谱分析,筛选与类黄酮积累显著相关的候选基因,并利用实时荧光定量PCR分析其在酸枣不同组织部位中的表达模式。结果表明:酸枣基因组中共鉴定出47个ZjWRKY基因,依据系统进化分析可分为Group Ⅰ、Group Ⅱ和Group Ⅲ共3个亚家族;ZjWRKY基因不均匀分布于酸枣11条染色体上,编码蛋白均含有保守的WRKYGQK结构域,主要定位于细胞核;89%的ZjWRKY基因启动子区含ABA响应元件(ABRE)。外源ABA可以促进萌发的酸枣种子中类黄酮的积累,且含量与12个ZjWRKY基因的表达模式相关;其中,与类黄酮含量相关的ZjWRKY10、ZjWRKY30及ZjWRKY45主要在酸枣种子、果肉中表达丰度较高,推测其可能协同调控酸枣果实和种子发育及ABA诱导的类黄酮生物合成。本研究结果为深入阐明ABA信号-WRKY转录因子-类黄酮合成调控网络提供了重要线索。
中图分类号:
董楚星, 张梦圆, 史鑫波, 张岗, 宋忠兴, 王楠, 焦红红. 酸枣WRKY转录因子家族鉴定及脱落酸介导类黄酮合成的关键基因筛选[J]. 植物研究, 2026, 46(5): 851-865.
Chuxing DONG, Mengyuan ZHANG, Xinbo SHI, Gang ZHANG, Zhongxing SONG, Nan WANG, Honghong JIAO. Genome-wide Identification of WRKY Transcription Factors and Key Genes for Abscisic Acid-mediated Flavonoid Synthesis in Sour Jujube (Ziziphus jujuba var. spinosa)[J]. Bulletin of Botanical Research, 2026, 46(5): 851-865.
表1
qRT-PCR引物
基因名称 Gene name | 正向引物序列(5′→3′) Forward primer sequence(5′→3′) | 反向引物序列(5′→3′) Reverse primer sequence(5′→3′) |
|---|---|---|
| ZjH3 | CTTGGTCGGTCTGTTCGAGG | ATGCCGGATCTAAGCCCTCT |
| ZjWRKY1 | ATCCGAACTGCCTCGTGAAA | CATCTGGCCATGCTCATCCA |
| ZjWRKY2 | GTGTTGAGGAGCGTACTGGT | CAGCCTGAGCTGCAAATGTT |
| ZjWRKY10 | TCCAATGAAGCTCCTGCGAGAC | AGGGCTGTGTGTGTGTGTGT |
| ZjWRKY11 | TAGTGCTGATCATGGGGCTG | GTTAGGCGCTGGACTTGCTT |
| ZjWRKY12 | TGCTGCTACTGCCACATCAA | AGCTTGAGTCTTTTCCGTCCC |
| ZjWRKY24 | ATGTCTCAGTCATCCGCTGG | TTTGATGGGTTTTTGGCCGT |
| ZjWRKY25 | CAACCATCCTCACCCCTCAC | GAAATGTCGAACTTCCGGCG |
| ZjWRKY30 | TCTCGCTGATCAACCACCAC | TCGGGAACACCTAATTGGGG |
| ZjWRKY31 | GAGTCAGGGCTTGATGGACC | GGTATGCCGTCCTCGGTATG |
| ZjWRKY32 | TTGGAACGCAGAGAGGTTGT | CCTGCAAGGTTTTTCATCCG |
| ZjWRKY36 | CCTGTCTGCCTTGAGGTGTA | AGGAGGCGGAGAGAGAGGTA |
| ZjWRKY37 | TGGGAATAACTTGTGCCGCT | ACAGCTTTGGGATGGACAACT |
| ZjWRKY44 | TCTCCGCCATCTCTCTCTGTCTGA | ACCACTGATTGATCGCTGCT |
| ZjWRKY45 | TCTGAGAATTCCGGCGAGAG | GCTCGGCAACCTTGATCGTA |
| ZjWRKY47 | ATGAATTAGCTCCGCAGGCA | AACTGCACTCGGTGTTTCCA |
表2
酸枣 ZjWRKY 基因家族蛋白理化性质分析
基因ID Gene ID | 基因名称 Gene name | 蛋白长度 Protein length/aa | 相对分子质量 Relative molecular mass/kDa | 等电点 pI | 不稳定指数 Instability index | 脂溶性系数 Aliphatic index | 总疏水性 GRAVY | 亚细胞定位 Subcellular localization |
|---|---|---|---|---|---|---|---|---|
| ZspiChr4G00070360.1 | ZjWRKY1 | 501 | 55 053.62 | 6.40 | 44.30 | 58.64 | -0.992 | 细胞核 |
| ZspiChr2G00176960.1 | ZjWRKY2 | 592 | 64 619.35 | 7.21 | 60.56 | 46.81 | -0.928 | 细胞核 |
| ZspiChr1G00015850.1 | ZjWRKY3 | 521 | 57 263.14 | 5.12 | 63.06 | 59.52 | -0.930 | 细胞核 |
| ZspiChr12G00259750.1 | ZjWRKY4 | 477 | 52 053.75 | 9.09 | 47.12 | 57.88 | -0.895 | 细胞核 |
| ZspiChr12G00265570.1 | ZjWRKY5 | 373 | 41 018.42 | 5.16 | 70.12 | 50.75 | -0.724 | 细胞核 |
| ZspiChr10G00218060.1 | ZjWRKY6 | 310 | 34 331.69 | 5.65 | 58.67 | 54.68 | -0.949 | 细胞核 |
| ZspiChr11G00191570.1 | ZjWRKY7 | 184 | 21 092.86 | 9.20 | 42.74 | 71.52 | -0.899 | 细胞核 |
| ZspiChr12G00259570.1 | ZjWRKY8 | 207 | 23 355.69 | 9.26 | 54.66 | 62.56 | -0.771 | 过氧化物酶体 |
| ZspiChr1G00006330.1 | ZjWRKY9 | 336 | 37 387.94 | 6.37 | 52.64 | 46.46 | -0.976 | 细胞核 |
| ZspiChr4G00073720.1 | ZjWRKY10 | 345 | 39 158.92 | 6.76 | 62.93 | 47.48 | -1.101 | 细胞核 |
| ZspiChr9G00139950.1 | ZjWRKY11 | 195 | 22 341.12 | 9.34 | 36.81 | 50.51 | -0.921 | 细胞核 |
| ZspiChr8G00160490.1 | ZjWRKY12 | 198 | 21 938.16 | 6.73 | 49.17 | 41.31 | -0.934 | 细胞核 |
| ZspiChr6G00036910.1 | ZjWRKY13 | 173 | 19 750.96 | 5.59 | 49.56 | 57.40 | -0.842 | 细胞外 |
| ZspiChr5G00099850.1 | ZjWRKY14 | 199 | 23 062.84 | 9.21 | 45.42 | 50.90 | -1.012 | 细胞核 |
| ZspiChr9G00117920.1 | ZjWRKY15 | 335 | 37 135.37 | 6.43 | 58.37 | 58.00 | -0.728 | 细胞核 |
| ZspiChr10G00210320.1 | ZjWRKY16 | 192 | 21 894.40 | 9.43 | 45.48 | 49.17 | -0.961 | 细胞核 |
| ZspiChr9G00131150.1 | ZjWRKY17 | 503 | 55 917.06 | 5.50 | 49.71 | 60.70 | -0.856 | 细胞核 |
| ZspiChr8G00160700.1 | ZjWRKY18 | 242 | 27 914.17 | 8.66 | 63.80 | 54.38 | -0.973 | 细胞核 |
| ZspiChr1G00002140.1 | ZjWRKY19 | 328 | 35 300.64 | 9.57 | 56.26 | 56.19 | -0.605 | 细胞核 |
| ZspiChr11G00199490.1 | ZjWRKY20 | 299 | 33 456.13 | 5.36 | 56.80 | 66.49 | -0.764 | 细胞核 |
| ZspiChr9G00117330.1 | ZjWRKY21 | 507 | 55 356.96 | 7.41 | 51.06 | 65.11 | -0.641 | 细胞核 |
| ZspiChr8G00154750.1 | ZjWRKY22 | 329 | 35 931.61 | 9.53 | 42.72 | 70.27 | -0.568 | 细胞核 |
| ZspiChr6G00041700.1 | ZjWRKY23 | 366 | 39 473.46 | 9.64 | 50.56 | 60.27 | -0.578 | 细胞核 |
| ZspiChr6G00034290.1 | ZjWRKY24 | 371 | 40 189.57 | 9.57 | 58.58 | 66.82 | -0.558 | 细胞核 |
| ZspiChr1G00028160.1 | ZjWRKY25 | 316 | 35 076.43 | 8.68 | 45.86 | 64.84 | -0.734 | 细胞核 |
| ZspiChr9G00122400.1 | ZjWRKY26 | 361 | 40 626.82 | 9.65 | 59.45 | 65.37 | -0.855 | 细胞核 |
| ZspiChr1G00005320.1 | ZjWRKY27 | 312 | 34 716.29 | 5.60 | 56.82 | 43.43 | -0.777 | 叶绿体 |
| ZspiChr11G00191590.1 | ZjWRKY28 | 392 | 44 753.87 | 4.89 | 67.54 | 63.60 | -0.701 | 细胞核 |
| ZspiChr12G00259500.1 | ZjWRKY29 | 355 | 38 920.61 | 5.92 | 66.41 | 44.82 | -0.774 | 细胞核 |
| ZspiChr9G00118050.1 | ZjWRKY30 | 289 | 32 228.5 | 5.27 | 53.06 | 50.59 | -1.017 | 细胞核 |
| ZspiChr8G00145050.1 | ZjWRKY31 | 327 | 37 411.67 | 5.13 | 57.02 | 62.32 | -0.758 | 细胞核 |
| ZspiChr11G00190750.1 | ZjWRKY32 | 370 | 41 879.58 | 5.27 | 62.82 | 58.22 | -0.810 | 细胞核 |
| ZspiChr3G00239020.1 | ZjWRKY33 | 362 | 40 124.62 | 5.68 | 56.69 | 57.71 | -0.777 | 细胞核 |
| ZspiChr2G00185960.1 | ZjWRKY34 | 738 | 80 905.86 | 5.90 | 52.26 | 60.14 | -0.688 | 细胞核 |
| ZspiChr10G00217520.1 | ZjWRKY35 | 389 | 43 130.26 | 7.70 | 50.74 | 50.59 | -1.050 | 细胞核 |
| ZspiChr11G00202900.1 | ZjWRKY36 | 1 258 | 138 887.95 | 8.23 | 47.52 | 84.80 | -0.335 | 细胞核 |
| ZspiChr1G00020420.1 | ZjWRKY37 | 590 | 64 658.71 | 6.12 | 51.73 | 64.86 | -0.703 | 细胞核 |
| ZspiChr1G00010250.1 | ZjWRKY38 | 758 | 81 979.08 | 5.57 | 52.18 | 57.56 | -0.763 | 细胞核 |
| ZspiChr1G00011820.1 | ZjWRKY39 | 633 | 67 853.91 | 6.12 | 41.23 | 63.76 | -0.633 | 细胞核 |
| ZspiChr4G00067590.1 | ZjWRKY40 | 227 | 24 273.76 | 4.77 | 69.68 | 64.93 | -0.434 | 细胞核 |
| ZspiChr2G00185940.1 | ZjWRKY41 | 350 | 39 454.27 | 6.01 | 60.99 | 64.63 | -0.649 | 高尔基体 |
| ZspiChr11G00203510.1 | ZjWRKY42 | 319 | 36 035.82 | 5.25 | 59.92 | 64.80 | -0.762 | 细胞核 |
| ZspiChr3G00243560.1 | ZjWRKY43 | 391 | 43 607.65 | 5.90 | 50.63 | 52.61 | -0.859 | 细胞核 |
| ZspiChr4G00065800.1 | ZjWRKY44 | 571 | 61 532.69 | 5.98 | 45.34 | 59.39 | -0.768 | 细胞核 |
| ZspiChr3G00239030.1 | ZjWRKY45 | 321 | 36 025.77 | 5.27 | 47.54 | 59.22 | -0.763 | 细胞核 |
| ZspiChr11G00203440.1 | ZjWRKY46 | 324 | 36 876.17 | 5.79 | 47.93 | 71.88 | -0.687 | 细胞核 |
| ZspiChr6G00038170.1 | ZjWRKY47 | 316 | 34 698.02 | 8.64 | 42.63 | 72.97 | -0.542 | 细胞核 |
表3
酸枣 ZjWRKY 基因对 Ka/Ks 分析
基因对 Gene pairs | 同义突变率 Ka | 非同义突变率 Ks | 同义突变率/非同义突变率 Ka/Ks | 选择压力 Selection pressure |
|---|---|---|---|---|
| ZjWRKY9/ZjWRKY10 | 0.319 | 2.373 | 0.134 | 纯化选择 |
| ZjWRKY40/ZjWRKY27 | 0.417 | 1.516 | 0.275 | 纯化选择 |
| ZjWRKY44/ZjWRKY39 | 0.287 | 3.056 | 0.094 | 纯化选择 |
| ZjWRKY25/ZjWRKY47 | 0.362 | NaN | NaN | 无法计算 |
| ZjWRKY2/ZjWRKY36 | 0.319 | 1.991 | 0.160 | 纯化选择 |
| ZjWRKY43/ZjWRKY32 | 0.381 | NaN | NaN | 无法计算 |
| ZjWRKY43/ZjWRKY31 | 0.340 | 2.847 | 0.126 | 纯化选择 |
| ZjWRKY31/ZjWRKY32 | 0.415 | 2.461 | 0.169 | 纯化选择 |
| ZjWRKY14/ZjWRKY11 | 0.370 | 1.581 | 0.234 | 纯化选择 |
| ZjWRKY22/ZjWRKY24 | 0.216 | 1.987 | 0.108 | 纯化选择 |
表4
类黄酮生物合成关键酶基因表达水平
基因名称 Gene name | 基因ID Gene ID | ABA 24 vs CK | ABA 48 vs CK | ||
|---|---|---|---|---|---|
| log2(FoldChange) | Padj | log2(FoldChange) | Padj | ||
| CHI | ZspiChr1G00028950 | 1.231 96 | <0.001 | -0.488 99 | <0.001 |
| ZspiChr5G00109140 | 2.773 76 | <0.001 | 1.451 71 | <0.001 | |
| F3H | ZspiChr5G00095680 | 0.978 29 | <0.001 | 1.069 25 | <0.001 |
| CHS | ZspiChr9G00134850 | -2.898 93 | <0.001 | -2.688 15 | <0.001 |
| FLS | ZspiChr1G00006420 | -1.007 06 | <0.001 | -0.641 72 | <0.001 |
| CCR | ZspiChr1G00008930 | -0.744 16 | <0.001 | 2.402 25 | <0.001 |
| ZspiChr6G00030760 | 0.000 02 | <0.001 | 1.662 04 | <0.001 | |
| ANR | ZspiChr11G00190780 | 1.480 21 | <0.001 | 1.885 05 | <0.001 |
| ZspiChr1G00018920 | 1.501 35 | <0.001 | 1.024 85 | <0.001 | |
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