草业学报 ›› 2023, Vol. 32 ›› Issue (8): 186-201.DOI: 10.11686/cyxb2022150
• 研究论文 • 上一篇
史先飞(), 高宇, 黄旭升, 周雅莉, 蔡桂萍, 李昕儒, 李润植(), 薛金爱()
收稿日期:
2022-04-06
修回日期:
2022-05-18
出版日期:
2023-08-20
发布日期:
2023-06-16
通讯作者:
李润植,薛金爱
作者简介:
E-mail: 306214803@qq.com基金资助:
Xian-fei SHI(), Yu GAO, Xu-sheng HUANG, Ya-li ZHOU, Gui-ping CAI, Xin-ru LI, Run-zhi LI(), Jin-ai XUE()
Received:
2022-04-06
Revised:
2022-05-18
Online:
2023-08-20
Published:
2023-06-16
Contact:
Run-zhi LI,Jin-ai XUE
摘要:
油莎豆亦称油莎草,地上部分是优质牧草,地下块茎可积累大量油脂,抗逆性强、适应性广,是一种新型饲草和油料作物。WRKY是一类数量庞大的转录因子超家族,广泛参与调控植物生长发育、物质代谢和胁迫应答等生理过程。然而,有关于油莎豆WRKY家族成员及其功能知之甚少。本研究基于转录组分析,鉴定出67条油莎豆WRKY转录因子。系统发育树结果显示,油莎豆CeWRKY与拟南芥AtWRKY 类似,分为I、Ⅱ、Ⅲ 3个亚家族。CeWRKYs均具有典型的WRKY结构域和锌指结构域,其中7条蛋白序列的WRKY结构域存在变异,可能与WRKY成员功能歧化相关。油莎豆CeWRKY蛋白的序列长度、相对分子量、等电点等存在较大差异,均为亲水性蛋白,不含跨膜结构,亚细胞定位预测均位于细胞核。CeWRKY基因在块茎发育时期的表达模式具有时空特异性。不同CeWRKY基因参与不同逆境胁迫响应,其中CeWRKY8基因分别在干旱、盐、碱胁迫处理的油莎豆幼苗中表达谱变化显著,可能是参与油莎豆逆境胁迫响应调控的一个主效转录因子。本研究为深入解析油莎豆CeWRKY转录因子的多样性生物学功能以及介导逆境胁迫应答的分子机制奠定了基础,亦为优质高抗油莎豆新品种培育提供科学参考。
史先飞, 高宇, 黄旭升, 周雅莉, 蔡桂萍, 李昕儒, 李润植, 薛金爱. 油莎豆CeWRKY转录因子响应非生物胁迫的功能表征[J]. 草业学报, 2023, 32(8): 186-201.
Xian-fei SHI, Yu GAO, Xu-sheng HUANG, Ya-li ZHOU, Gui-ping CAI, Xin-ru LI, Run-zhi LI, Jin-ai XUE. Functional characterization of Cyperus esculentus CeWRKY transcription factors in response to abiotic stress[J]. Acta Prataculturae Sinica, 2023, 32(8): 186-201.
亚家族 Sub-family | 基因 Gene | 正向引物 Forward primer sequence (5′→3′) | 反向引物 Reverse primer sequence (5′→3′) |
---|---|---|---|
I | CeWRKY8 | ACCAGAGCATCAAGCAAGAAC | CCAGTTGTATCCGTCGTTGTT |
CeWRKY9 | CTGCTGAGGATGGTTACAACTG | TCGGTGATGGCTTCTGATGG | |
CeWRKY17 | GGACAAGAGAACAGACCAATCG | TCACCTCACAACTCGGATGC | |
CeWRKY44 | TCTTCAGCATCTGTGGTTAGTG | TACTCTCGGTTCTCGGACTG | |
CeWRKY64 | CGGACGACGGTTACAACTG | GACTGCGGCTTAGGATGGT | |
Ⅱa | CeWRKY6 | CCAACTGCGAACAATGTGAAGG | GGGCGAAAGAGCATCTGAAGT |
CeWRKY11 | CCGTGGACCAGATGGAATGC | GGTGTGCTCGCCTTCGTATG | |
CeWRKY18 | ACCGATCCACCGAAGAATGTTC | ACTGAGCCGCCTTCTGATGA | |
CeWRKY49 | AAGATGGATACCAGTGGAGGAA | GGAAGTGCTGTTGGCAGTATA | |
Ⅱb | CeWRKY65 | GGATGGCTATGAGTGGAGGAA | CCGAACACTTGGTTGGTCAG |
Ⅱc | CeWRKY1 | TGTCAAGAACAGTCCGAACC | TGCCATGATAAGCCACATAACT |
CeWRKY58 | GACGACGGGTTCAAATGGA | GCATCTTGGCTGGTGTAATAC | |
CeWRKY61 | TCTCCTTGCCAGATTACAGTGA | AGCGGTAGTAGTTCCTTGGATT | |
Ⅱd | CeWRKY4 | CAAGATGGACGAGCAGATAGC | GCAACGGAAGCACTGTTACT |
CeWRKY46 | ATCATGCCTCTTCCTCCTCTTC | CGGTGCTGTTGTTCCTCCT | |
Ⅱe | CeWRKY14 | CTACACGGCTGAGCACAATC | TTATCATGCCTTCTTCCGACTC |
CeWRKY15 | GCCACCACAAGCTCCATTA | TGTAATCTTCTCGCTCTTCTCC | |
Ⅲ | CeWRKY24 | TTCTGAGCCATCATCGTCTGT | CCTCCAAGTATGTCCATCCTCT |
CeWRKY38 | ACCGCAACACGCAAGGAT | ATCTCAGGTTGGAACGGCTTA | |
CeWRKY40 | TTCTGAGCCATCATCGTCTGT | CCTCCAAGTATGTCCATCCTCT | |
CeWRKY47 | GATCCAACCGTGTTCATCATCA | GCACATCATCTTCACCGTCTT | |
18SrRNA | CTACGTCCCTGCCCTTTGTACA | ACACTTCACCGGACCATTCAA |
表1 引物序列
Table 1 Primer sequences
亚家族 Sub-family | 基因 Gene | 正向引物 Forward primer sequence (5′→3′) | 反向引物 Reverse primer sequence (5′→3′) |
---|---|---|---|
I | CeWRKY8 | ACCAGAGCATCAAGCAAGAAC | CCAGTTGTATCCGTCGTTGTT |
CeWRKY9 | CTGCTGAGGATGGTTACAACTG | TCGGTGATGGCTTCTGATGG | |
CeWRKY17 | GGACAAGAGAACAGACCAATCG | TCACCTCACAACTCGGATGC | |
CeWRKY44 | TCTTCAGCATCTGTGGTTAGTG | TACTCTCGGTTCTCGGACTG | |
CeWRKY64 | CGGACGACGGTTACAACTG | GACTGCGGCTTAGGATGGT | |
Ⅱa | CeWRKY6 | CCAACTGCGAACAATGTGAAGG | GGGCGAAAGAGCATCTGAAGT |
CeWRKY11 | CCGTGGACCAGATGGAATGC | GGTGTGCTCGCCTTCGTATG | |
CeWRKY18 | ACCGATCCACCGAAGAATGTTC | ACTGAGCCGCCTTCTGATGA | |
CeWRKY49 | AAGATGGATACCAGTGGAGGAA | GGAAGTGCTGTTGGCAGTATA | |
Ⅱb | CeWRKY65 | GGATGGCTATGAGTGGAGGAA | CCGAACACTTGGTTGGTCAG |
Ⅱc | CeWRKY1 | TGTCAAGAACAGTCCGAACC | TGCCATGATAAGCCACATAACT |
CeWRKY58 | GACGACGGGTTCAAATGGA | GCATCTTGGCTGGTGTAATAC | |
CeWRKY61 | TCTCCTTGCCAGATTACAGTGA | AGCGGTAGTAGTTCCTTGGATT | |
Ⅱd | CeWRKY4 | CAAGATGGACGAGCAGATAGC | GCAACGGAAGCACTGTTACT |
CeWRKY46 | ATCATGCCTCTTCCTCCTCTTC | CGGTGCTGTTGTTCCTCCT | |
Ⅱe | CeWRKY14 | CTACACGGCTGAGCACAATC | TTATCATGCCTTCTTCCGACTC |
CeWRKY15 | GCCACCACAAGCTCCATTA | TGTAATCTTCTCGCTCTTCTCC | |
Ⅲ | CeWRKY24 | TTCTGAGCCATCATCGTCTGT | CCTCCAAGTATGTCCATCCTCT |
CeWRKY38 | ACCGCAACACGCAAGGAT | ATCTCAGGTTGGAACGGCTTA | |
CeWRKY40 | TTCTGAGCCATCATCGTCTGT | CCTCCAAGTATGTCCATCCTCT | |
CeWRKY47 | GATCCAACCGTGTTCATCATCA | GCACATCATCTTCACCGTCTT | |
18SrRNA | CTACGTCCCTGCCCTTTGTACA | ACACTTCACCGGACCATTCAA |
图2 油莎豆CeWRKY转录因子WRKY域多序列比对Group I中 -N和 -C分别表示I亚族成员蛋白序列的N末端和C末端。-N and -C indicate N-terminal and C-terminal of the protein sequences of Group I CeWRKYs.
Fig.2 Multiple sequence alignment of WRKY domain of CeWRKY transcription factors
亚家族 Sub- family | 蛋白 Protein | 蛋白长度Length (aa) | 分子量 Molecular weight (kDa) | 理论等电点Isoelectric point (PI) | 不稳定系数Instability index | 亲水系数GRAVY | α-螺旋 α-helix (%) | β-转角 β-turn (%) | 延伸链 Extended strand (%) | 无规则卷曲 Random coil (%) |
---|---|---|---|---|---|---|---|---|---|---|
I | CeWRKY50 | 314 | 34.81318 | 7.72 | 56.94 | -1.139 | 14.65 | 4.78 | 14.01 | 66.56 |
CeWRKY7 | 389 | 43.66802 | 7.20 | 56.58 | -1.085 | 11.83 | 3.86 | 11.57 | 72.75 | |
CeWRKY8 | 448 | 49.63432 | 8.44 | 58.67 | -0.994 | 8.71 | 3.57 | 11.83 | 75.89 | |
CeWRKY30 | 459 | 49.00339 | 6.58 | 56.78 | -0.799 | 14.81 | 3.05 | 10.89 | 71.24 | |
CeWRKY44 | 479 | 52.80267 | 6.38 | 55.42 | -0.751 | 15.45 | 5.64 | 10.02 | 68.89 | |
CeWRKY64 | 572 | 62.51011 | 6.17 | 50.85 | -0.847 | 7.34 | 3.50 | 12.06 | 77.10 | |
CeWRKY17 | 588 | 63.76072 | 6.37 | 52.81 | -0.829 | 9.18 | 2.89 | 12.24 | 75.68 | |
CeWRKY9 | 728 | 78.87979 | 5.76 | 55.50 | -0.857 | 13.87 | 3.57 | 9.34 | 73.21 | |
IIa | CeWRKY49 | 161 | 18.06416 | 7.60 | 41.69 | -0.646 | 22.36 | 6.21 | 18.01 | 53.42 |
CeWRKY6 | 185 | 20.97675 | 9.05 | 56.29 | -0.841 | 34.59 | 2.70 | 16.76 | 45.95 | |
CeWRKY18 | 221 | 24.95229 | 8.98 | 52.85 | -0.802 | 24.43 | 3.17 | 14.48 | 57.92 | |
CeWRKY11 | 296 | 32.85796 | 8.13 | 52.07 | -0.776 | 26.69 | 2.03 | 13.18 | 58.11 | |
IIb | CeWRKY65 | 120 | 13.78119 | 6.28 | 41.76 | -0.963 | 27.50 | 3.33 | 15.00 | 54.17 |
CeWRKY5 | 133 | 14.84553 | 7.57 | 34.42 | -1.085 | 16.54 | 6.02 | 15.04 | 62.41 | |
CeWRKY16 | 274 | 30.45201 | 5.78 | 47.36 | -0.989 | 33.21 | 3.28 | 7.66 | 55.84 | |
CeWRKY52 | 346 | 36.12409 | 9.34 | 38.19 | -0.341 | 18.79 | 4.34 | 8.96 | 67.92 | |
CeWRKY10 | 538 | 56.84355 | 6.20 | 45.49 | -0.495 | 26.77 | 2.97 | 11.52 | 58.74 | |
IIc | CeWRKY36 | 116 | 13.78374 | 9.73 | 40.34 | -1.078 | 27.59 | 8.62 | 21.55 | 42.24 |
CeWRKY61 | 168 | 18.88298 | 9.02 | 37.08 | -0.717 | 15.48 | 7.74 | 17.86 | 58.93 | |
CeWRKY33 | 174 | 18.93200 | 6.82 | 47.98 | -0.856 | 21.26 | 5.75 | 12.64 | 60.34 | |
CeWRKY1 | 186 | 21.05430 | 7.00 | 40.16 | -0.901 | 20.43 | 3.76 | 13.98 | 61.83 | |
CeWRKY32 | 193 | 21.96437 | 9.45 | 62.34 | -1.016 | 21.24 | 5.70 | 17.62 | 55.44 | |
CeWRKY58 | 210 | 23.12745 | 5.66 | 47.32 | -0.696 | 19.52 | 6.67 | 13.33 | 60.48 | |
CeWRKY27 | 213 | 22.99072 | 7.79 | 55.08 | -0.569 | 23.94 | 5.63 | 14.08 | 56.34 | |
CeWRKY57 | 240 | 26.13286 | 9.32 | 50.55 | -0.886 | 5.83 | 3.75 | 10.00 | 80.42 | |
CeWRKY51 | 254 | 28.82733 | 5.46 | 62.77 | -0.796 | 34.25 | 9.06 | 9.84 | 46.85 | |
CeWRKY60 | 254 | 28.37627 | 9.22 | 53.56 | -1.060 | 17.32 | 2.76 | 9.45 | 70.47 | |
CeWRKY20 | 319 | 35.62963 | 6.52 | 46.49 | -0.861 | 16.30 | 2.51 | 7.84 | 73.35 | |
CeWRKY31 | 369 | 40.91780 | 6.62 | 48.26 | -0.690 | 30.08 | 2.17 | 11.38 | 56.37 | |
IId | CeWRKY39 | 87 | 10.13063 | 9.12 | 58.77 | -0.591 | 6.90 | 10.34 | 21.84 | 60.92 |
CeWRKY55 | 133 | 15.15530 | 9.78 | 50.66 | -0.848 | 11.28 | 6.02 | 24.06 | 58.65 | |
CeWRKY59 | 187 | 20.95189 | 9.74 | 42.79 | -0.780 | 19.25 | 7.49 | 25.67 | 47.59 | |
CeWRKY2 | 289 | 31.14022 | 9.79 | 56.54 | -0.571 | 20.42 | 5.19 | 11.42 | 62.98 | |
CeWRKY4 | 321 | 34.23713 | 9.80 | 50.86 | -0.463 | 17.13 | 5.61 | 12.46 | 64.80 | |
CeWRKY28 | 332 | 37.20718 | 9.78 | 57.37 | -0.701 | 27.11 | 3.31 | 11.45 | 58.13 | |
CeWRKY46 | 381 | 42.20658 | 9.86 | 51.88 | -0.841 | 23.36 | 5.77 | 12.86 | 58.01 | |
IIe | CeWRKY48 | 160 | 17.18509 | 10.40 | 53.12 | -0.559 | 11.88 | 7.50 | 11.25 | 69.38 |
CeWRKY14 | 178 | 19.64963 | 4.75 | 38.07 | -0.866 | 18.54 | 6.74 | 14.61 | 60.11 | |
CeWRKY19 | 200 | 22.11250 | 8.96 | 92.13 | -1.132 | 12.00 | 5.50 | 13.00 | 69.50 | |
CeWRKY15 | 206 | 22.65692 | 9.53 | 58.76 | -0.609 | 24.76 | 2.91 | 10.19 | 62.14 | |
CeWRKY35 | 274 | 30.68032 | 5.13 | 44.70 | -0.615 | 22.63 | 4.74 | 11.68 | 60.95 | |
CeWRKY56 | 274 | 30.63824 | 5.13 | 44.55 | -0.624 | 24.82 | 3.28 | 10.58 | 61.31 | |
CeWRKY53 | 290 | 31.62324 | 4.82 | 47.58 | -0.572 | 15.86 | 7.24 | 14.14 | 62.76 | |
CeWRKY22 | 343 | 36.66320 | 7.06 | 45.77 | -0.442 | 27.70 | 5.54 | 8.75 | 58.02 | |
Ⅲ | CeWRKY29 | 150 | 17.04952 | 9.40 | 37.64 | -1.441 | 7.33 | 6.67 | 24.00 | 62.00 |
CeWRKY62 | 276 | 30.96083 | 6.09 | 41.47 | -0.539 | 30.80 | 4.71 | 15.22 | 49.28 | |
CeWRKY63 | 277 | 31.17213 | 6.09 | 41.13 | -0.542 | 29.60 | 3.61 | 11.19 | 55.60 | |
CeWRKY12 | 281 | 30.41290 | 8.28 | 52.10 | -0.455 | 23.84 | 6.41 | 16.73 | 53.02 | |
CeWRKY54 | 283 | 32.22986 | 5.75 | 52.35 | -0.865 | 26.15 | 6.01 | 15.55 | 52.30 | |
CeWRKY47 | 292 | 32.06679 | 5.20 | 53.90 | -0.541 | 29.45 | 1.71 | 9.93 | 58.90 | |
CeWRKY25 | 294 | 32.72820 | 7.72 | 60.10 | -0.840 | 24.83 | 6.46 | 19.39 | 49.32 | |
CeWRKY13 | 298 | 33.64385 | 6.11 | 44.73 | -0.707 | 24.83 | 4.70 | 10.74 | 59.73 | |
CeWRKY38 | 298 | 33.64380 | 5.92 | 46.14 | -0.706 | 22.82 | 3.69 | 9.40 | 64.09 | |
CeWRKY67 | 334 | 37.97822 | 5.75 | 37.75 | -0.736 | 24.85 | 4.49 | 15.27 | 55.39 | |
CeWRKY66 | 342 | 37.53184 | 5.95 | 65.22 | -0.916 | 20.47 | 3.51 | 14.62 | 61.40 | |
CeWRKY23 | 395 | 44.24915 | 5.30 | 52.64 | -0.427 | 24.56 | 7.34 | 26.08 | 42.03 | |
CeWRKY45 | 446 | 49.75952 | 6.30 | 55.45 | -0.416 | 26.01 | 7.85 | 22.65 | 43.50 | |
CeWRKY37 | 606 | 67.74975 | 5.50 | 60.91 | -0.584 | 35.48 | 7.26 | 14.85 | 42.41 | |
CeWRKY34 | 654 | 72.67170 | 5.22 | 59.34 | -0.604 | 33.33 | 5.96 | 14.22 | 46.48 | |
CeWRKY26 | 1186 | 133.44630 | 5.45 | 47.37 | -0.199 | 46.88 | 4.30 | 11.30 | 37.52 | |
CeWRKY40 | 1186 | 133.67930 | 5.34 | 49.92 | -0.218 | 47.64 | 3.79 | 11.30 | 37.27 | |
CeWRKY41 | 1186 | 133.34290 | 5.35 | 46.02 | -0.239 | 48.65 | 4.22 | 11.13 | 36.00 | |
CeWRKY24 | 1178 | 132.48820 | 5.50 | 45.29 | -0.208 | 47.79 | 4.75 | 12.56 | 34.89 | |
CeWRKY42 | 1178 | 132.38480 | 5.40 | 43.93 | -0.248 | 47.45 | 4.16 | 12.05 | 36.33 | |
CeWRKY43 | 1178 | 132.72110 | 5.38 | 47.86 | -0.228 | 47.03 | 4.67 | 11.46 | 36.84 | |
CeWRKY3 | 1193 | 128.04290 | 6.17 | 62.26 | -0.696 | 33.86 | 4.44 | 12.15 | 49.54 | |
CeWRKY21 | 1200 | 128.87780 | 6.12 | 62.27 | -0.694 | 34.08 | 4.58 | 12.33 | 49.00 |
表2 油莎豆WRKY蛋白序列特征分析
Table 2 Analysis of CeWRKY protein sequences characters in C. esculentus
亚家族 Sub- family | 蛋白 Protein | 蛋白长度Length (aa) | 分子量 Molecular weight (kDa) | 理论等电点Isoelectric point (PI) | 不稳定系数Instability index | 亲水系数GRAVY | α-螺旋 α-helix (%) | β-转角 β-turn (%) | 延伸链 Extended strand (%) | 无规则卷曲 Random coil (%) |
---|---|---|---|---|---|---|---|---|---|---|
I | CeWRKY50 | 314 | 34.81318 | 7.72 | 56.94 | -1.139 | 14.65 | 4.78 | 14.01 | 66.56 |
CeWRKY7 | 389 | 43.66802 | 7.20 | 56.58 | -1.085 | 11.83 | 3.86 | 11.57 | 72.75 | |
CeWRKY8 | 448 | 49.63432 | 8.44 | 58.67 | -0.994 | 8.71 | 3.57 | 11.83 | 75.89 | |
CeWRKY30 | 459 | 49.00339 | 6.58 | 56.78 | -0.799 | 14.81 | 3.05 | 10.89 | 71.24 | |
CeWRKY44 | 479 | 52.80267 | 6.38 | 55.42 | -0.751 | 15.45 | 5.64 | 10.02 | 68.89 | |
CeWRKY64 | 572 | 62.51011 | 6.17 | 50.85 | -0.847 | 7.34 | 3.50 | 12.06 | 77.10 | |
CeWRKY17 | 588 | 63.76072 | 6.37 | 52.81 | -0.829 | 9.18 | 2.89 | 12.24 | 75.68 | |
CeWRKY9 | 728 | 78.87979 | 5.76 | 55.50 | -0.857 | 13.87 | 3.57 | 9.34 | 73.21 | |
IIa | CeWRKY49 | 161 | 18.06416 | 7.60 | 41.69 | -0.646 | 22.36 | 6.21 | 18.01 | 53.42 |
CeWRKY6 | 185 | 20.97675 | 9.05 | 56.29 | -0.841 | 34.59 | 2.70 | 16.76 | 45.95 | |
CeWRKY18 | 221 | 24.95229 | 8.98 | 52.85 | -0.802 | 24.43 | 3.17 | 14.48 | 57.92 | |
CeWRKY11 | 296 | 32.85796 | 8.13 | 52.07 | -0.776 | 26.69 | 2.03 | 13.18 | 58.11 | |
IIb | CeWRKY65 | 120 | 13.78119 | 6.28 | 41.76 | -0.963 | 27.50 | 3.33 | 15.00 | 54.17 |
CeWRKY5 | 133 | 14.84553 | 7.57 | 34.42 | -1.085 | 16.54 | 6.02 | 15.04 | 62.41 | |
CeWRKY16 | 274 | 30.45201 | 5.78 | 47.36 | -0.989 | 33.21 | 3.28 | 7.66 | 55.84 | |
CeWRKY52 | 346 | 36.12409 | 9.34 | 38.19 | -0.341 | 18.79 | 4.34 | 8.96 | 67.92 | |
CeWRKY10 | 538 | 56.84355 | 6.20 | 45.49 | -0.495 | 26.77 | 2.97 | 11.52 | 58.74 | |
IIc | CeWRKY36 | 116 | 13.78374 | 9.73 | 40.34 | -1.078 | 27.59 | 8.62 | 21.55 | 42.24 |
CeWRKY61 | 168 | 18.88298 | 9.02 | 37.08 | -0.717 | 15.48 | 7.74 | 17.86 | 58.93 | |
CeWRKY33 | 174 | 18.93200 | 6.82 | 47.98 | -0.856 | 21.26 | 5.75 | 12.64 | 60.34 | |
CeWRKY1 | 186 | 21.05430 | 7.00 | 40.16 | -0.901 | 20.43 | 3.76 | 13.98 | 61.83 | |
CeWRKY32 | 193 | 21.96437 | 9.45 | 62.34 | -1.016 | 21.24 | 5.70 | 17.62 | 55.44 | |
CeWRKY58 | 210 | 23.12745 | 5.66 | 47.32 | -0.696 | 19.52 | 6.67 | 13.33 | 60.48 | |
CeWRKY27 | 213 | 22.99072 | 7.79 | 55.08 | -0.569 | 23.94 | 5.63 | 14.08 | 56.34 | |
CeWRKY57 | 240 | 26.13286 | 9.32 | 50.55 | -0.886 | 5.83 | 3.75 | 10.00 | 80.42 | |
CeWRKY51 | 254 | 28.82733 | 5.46 | 62.77 | -0.796 | 34.25 | 9.06 | 9.84 | 46.85 | |
CeWRKY60 | 254 | 28.37627 | 9.22 | 53.56 | -1.060 | 17.32 | 2.76 | 9.45 | 70.47 | |
CeWRKY20 | 319 | 35.62963 | 6.52 | 46.49 | -0.861 | 16.30 | 2.51 | 7.84 | 73.35 | |
CeWRKY31 | 369 | 40.91780 | 6.62 | 48.26 | -0.690 | 30.08 | 2.17 | 11.38 | 56.37 | |
IId | CeWRKY39 | 87 | 10.13063 | 9.12 | 58.77 | -0.591 | 6.90 | 10.34 | 21.84 | 60.92 |
CeWRKY55 | 133 | 15.15530 | 9.78 | 50.66 | -0.848 | 11.28 | 6.02 | 24.06 | 58.65 | |
CeWRKY59 | 187 | 20.95189 | 9.74 | 42.79 | -0.780 | 19.25 | 7.49 | 25.67 | 47.59 | |
CeWRKY2 | 289 | 31.14022 | 9.79 | 56.54 | -0.571 | 20.42 | 5.19 | 11.42 | 62.98 | |
CeWRKY4 | 321 | 34.23713 | 9.80 | 50.86 | -0.463 | 17.13 | 5.61 | 12.46 | 64.80 | |
CeWRKY28 | 332 | 37.20718 | 9.78 | 57.37 | -0.701 | 27.11 | 3.31 | 11.45 | 58.13 | |
CeWRKY46 | 381 | 42.20658 | 9.86 | 51.88 | -0.841 | 23.36 | 5.77 | 12.86 | 58.01 | |
IIe | CeWRKY48 | 160 | 17.18509 | 10.40 | 53.12 | -0.559 | 11.88 | 7.50 | 11.25 | 69.38 |
CeWRKY14 | 178 | 19.64963 | 4.75 | 38.07 | -0.866 | 18.54 | 6.74 | 14.61 | 60.11 | |
CeWRKY19 | 200 | 22.11250 | 8.96 | 92.13 | -1.132 | 12.00 | 5.50 | 13.00 | 69.50 | |
CeWRKY15 | 206 | 22.65692 | 9.53 | 58.76 | -0.609 | 24.76 | 2.91 | 10.19 | 62.14 | |
CeWRKY35 | 274 | 30.68032 | 5.13 | 44.70 | -0.615 | 22.63 | 4.74 | 11.68 | 60.95 | |
CeWRKY56 | 274 | 30.63824 | 5.13 | 44.55 | -0.624 | 24.82 | 3.28 | 10.58 | 61.31 | |
CeWRKY53 | 290 | 31.62324 | 4.82 | 47.58 | -0.572 | 15.86 | 7.24 | 14.14 | 62.76 | |
CeWRKY22 | 343 | 36.66320 | 7.06 | 45.77 | -0.442 | 27.70 | 5.54 | 8.75 | 58.02 | |
Ⅲ | CeWRKY29 | 150 | 17.04952 | 9.40 | 37.64 | -1.441 | 7.33 | 6.67 | 24.00 | 62.00 |
CeWRKY62 | 276 | 30.96083 | 6.09 | 41.47 | -0.539 | 30.80 | 4.71 | 15.22 | 49.28 | |
CeWRKY63 | 277 | 31.17213 | 6.09 | 41.13 | -0.542 | 29.60 | 3.61 | 11.19 | 55.60 | |
CeWRKY12 | 281 | 30.41290 | 8.28 | 52.10 | -0.455 | 23.84 | 6.41 | 16.73 | 53.02 | |
CeWRKY54 | 283 | 32.22986 | 5.75 | 52.35 | -0.865 | 26.15 | 6.01 | 15.55 | 52.30 | |
CeWRKY47 | 292 | 32.06679 | 5.20 | 53.90 | -0.541 | 29.45 | 1.71 | 9.93 | 58.90 | |
CeWRKY25 | 294 | 32.72820 | 7.72 | 60.10 | -0.840 | 24.83 | 6.46 | 19.39 | 49.32 | |
CeWRKY13 | 298 | 33.64385 | 6.11 | 44.73 | -0.707 | 24.83 | 4.70 | 10.74 | 59.73 | |
CeWRKY38 | 298 | 33.64380 | 5.92 | 46.14 | -0.706 | 22.82 | 3.69 | 9.40 | 64.09 | |
CeWRKY67 | 334 | 37.97822 | 5.75 | 37.75 | -0.736 | 24.85 | 4.49 | 15.27 | 55.39 | |
CeWRKY66 | 342 | 37.53184 | 5.95 | 65.22 | -0.916 | 20.47 | 3.51 | 14.62 | 61.40 | |
CeWRKY23 | 395 | 44.24915 | 5.30 | 52.64 | -0.427 | 24.56 | 7.34 | 26.08 | 42.03 | |
CeWRKY45 | 446 | 49.75952 | 6.30 | 55.45 | -0.416 | 26.01 | 7.85 | 22.65 | 43.50 | |
CeWRKY37 | 606 | 67.74975 | 5.50 | 60.91 | -0.584 | 35.48 | 7.26 | 14.85 | 42.41 | |
CeWRKY34 | 654 | 72.67170 | 5.22 | 59.34 | -0.604 | 33.33 | 5.96 | 14.22 | 46.48 | |
CeWRKY26 | 1186 | 133.44630 | 5.45 | 47.37 | -0.199 | 46.88 | 4.30 | 11.30 | 37.52 | |
CeWRKY40 | 1186 | 133.67930 | 5.34 | 49.92 | -0.218 | 47.64 | 3.79 | 11.30 | 37.27 | |
CeWRKY41 | 1186 | 133.34290 | 5.35 | 46.02 | -0.239 | 48.65 | 4.22 | 11.13 | 36.00 | |
CeWRKY24 | 1178 | 132.48820 | 5.50 | 45.29 | -0.208 | 47.79 | 4.75 | 12.56 | 34.89 | |
CeWRKY42 | 1178 | 132.38480 | 5.40 | 43.93 | -0.248 | 47.45 | 4.16 | 12.05 | 36.33 | |
CeWRKY43 | 1178 | 132.72110 | 5.38 | 47.86 | -0.228 | 47.03 | 4.67 | 11.46 | 36.84 | |
CeWRKY3 | 1193 | 128.04290 | 6.17 | 62.26 | -0.696 | 33.86 | 4.44 | 12.15 | 49.54 | |
CeWRKY21 | 1200 | 128.87780 | 6.12 | 62.27 | -0.694 | 34.08 | 4.58 | 12.33 | 49.00 |
图4 油莎豆CeWRKY基因在块茎不同发育时期的表达模式热图绿-黑-红3色代表基因表达的强度The three colors of green-black-red represent the intensity of gene expression. 80 d: 播种后80 d(块茎形成初期) 80 days after sowing (initial stage of tuber formation); 125 d: 播种后125 d(块茎形成中期)125 days after sowing (metaphase of tuber formation); 170 d: 播种后170 d(块茎成熟期) 170 days after sowing (tuber maturity period).
Fig.4 Heat map of expression patterns of CeWRKY genes during tuber development in C. esculentus
图5 油莎豆CeWRKY基因在盐胁迫下的表达模式NC: 对照Normal conditions; SS: 盐胁迫Salt stress; 数值是平均值+标准误差Values are the mean+standard error; *, **: P<0.05, P<0.01. 下同The same below.
Fig.5 Expression profiles of CeWRKY genes in C. esculentus seedlings under salt stress
图6 油莎豆CeWRKY基因在碱胁迫下的表达模式AS: 碱胁迫Alkali stress. 下同 The same below.
Fig.6 Expression profiles of CeWRKY genes in C. esculentus seedlings under alkali stress
图7 油莎豆CeWRKY基因在干旱胁迫下的表达模式DS: 干旱胁迫Drought stress. 下同 The same below.
Fig.7 Expression profiles of CeWRKY genes in C. esculentus seedlings under drought stress
图8 胁迫相应Marker基因(RD22 和SOS1)在非生物胁迫下油莎豆幼苗中的表达模式
Fig.8 Expression profiles of stress-responsive marker genes in C. esculentus seedlings under abiotic stresses
1 | Defelice M S. Yellow nutsedge Cyperus esculentus L.-snack food of the gods. Weed Technology, 2002, 16(4): 901-907. |
2 | Gao Y, Sun Y, Gao H, et al. Ectopic overexpression of a type-II DGAT (CeDGAT2-2) derived from oil-rich tuber of Cyperus esculentus enhances accumulation of oil and oleic acid in tobacco leaves. Biotechnology for Biofuels, 2021, 14(1): 76. |
3 | Rodrigues A C, Maranhao Estelita M E. Morphoanatomy of the stem in Cyperaceae. Acta Botanica Brasilica, 2009, 23(3): 889-901. |
4 | Onuoha N O, Ogbusua N O, Okorie A N, et al.Tigernut (Cyperus esculentus L.) milk’ as a potent nutri-drink’ for the prevention of acetaminophen-induced hepatotoxicity in a murine model. Journal of Intercultural Ethnopharmacology, 2017, 6(3): 290-295. |
5 | Kamala A, Middha S K, Karigar C S. Plants in traditional medicine with special reference to Cyperus rotundus L.: a review. Biotech, 2018, 8(7): 309. |
6 | Liu H Y, Wang L H, Liu Y Y. Development status of olea bean products. Farm Products Processing, 2021(14): 97-99. |
刘红艳, 王丽华, 刘意元. 我国油莎豆制品开发现状. 农产品加工, 2021(14): 97-99. | |
7 | Yan J, Du J, Li C, et al. Main components analysis of Cyperus esculentus in Jingbian,Shaanxi. China Oils and Fats, 2016, 41(3): 85-88. |
闫军, 杜静, 李聪, 等. 陕西靖边油莎豆主要成分分析. 中国油脂, 2016, 41(3): 85-88. | |
8 | Chinnusamy V, Schumaker K, Zhu J K. Molecular genetic perspectives on crosstalk and specificity in abiotic stress signalling in plants. Journal of Experimental Botany, 2004, 55(395): 225-236. |
9 | Gill S S, Tuteja N. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry, 2010, 48(12): 909-930. |
10 | Cao L X, Ma Y Z, Yang J Y. Plant transcription factors in response to abiotic stress. Journal of Northern Agriculture, 2005(5): 10-14. |
曹丽霞, 马有志, 杨俊英. 应答非生物胁迫的植物转录因子. 北方农业学报, 2005(5): 10-14. | |
11 | Cui R X, Zhang Y W, Chen X Q, et al. The latest research progress on the stress responses of bZIP involved in plants. Biotechnology Bulletin, 2019, 35(2):143-155. |
12 | Xi S, Yu W, Na S. Transcriptional regulation of bHLH during plant response to stress. Biochemical and Biophysical Research Communications, 2018, 503(2): 397-401. |
13 | Li X, Liang Y, Bei G, et al. Research advance of the plant stress-related DREB transcriptional factor genes. Molecular Plant Breeding, 2017, 15(7): 2612-2622. |
14 | Mawlong I, Ali K, Kurup D, et al. Isolation and characterization of an AP2/ERF-type drought stress inducible transcription factor encoding gene from rice. Journal of Plant Biochemistry & Biotechnology, 2014, 23(1): 42-51. |
15 | Chen N, Chi X Y, Pan L J, et al. Advances in MYB transcription factors during salt-stress regulation in plants. Plant Physiology Journal, 2015, 51(9): 1395-1399. |
16 | Lu M, Sun Q P, Zhang D F, et al. Identification of 7 stress-related NAC transcription factor members in maize (Zea mays L.) and characterization of the expression pattern of these genes. Biochemical & Biophysical Research Communications, 2015, 462(2): 144-150. |
17 | Kong W L, Yu K, Dan N Z, et al. Genome-wide identification and expression analysis of WRKY transcription factor under abiotic stress in Beta vulgaris. Scientia Agricultura Sinica, 2017, 50(17): 3259-3273. |
18 | Ulker B, Somssich I E, Lker B, et al. WRKY transcription factors: from DNA binding towards biological function. Current Opinion in Plant Biology, 2004, 7(5): 491-498. |
19 | Li C, Li D, Shao F, et al. Molecular cloning and expression analysis of WRKY transcription factor genes in Salvia miltiorrhiza. BMC Genomics, 2015, 16(1): 200. |
20 | Zou Z, Yang L F, Wang D H, et al. Gene structures, evolution and transcriptional profiling of the WRKY gene family in castor bean (Ricinus communis L.). PLoS One, 2016, 11(2): e0148243. |
21 | Xie T, Chen C J, Li C H, et al. Genome-wide investigation of WRKY gene family in pineapple: evolution and expression profiles during development and stress. BMC Genomics, 2018, 19(1): 490. |
22 | Rushton P J, Somssich I E, Ringler P, et al. WRKY transcription factors. Trends in Plant Science, 2010, 15(5): 247-258. |
23 | Ishiguro S, Nakamura K. Characterization of a cDNA encoding a novel DNA-binding protein, SPF1, that recognizes SP8 sequences in the 5' upstream regions of genes coding for sporamin and-amylase from sweet potato. Molecular and General Genetics, 1994, 244(6): 563-571. |
24 | Atif R M. Evolutionary insights in expansion of DOF and WRKY protein families in diploid cotton species Gossypium raimondii and Gossypium arboreum. Amsterdam Netherlands: Plant Genome Evolution Conference, 2015. |
25 | Song H, Wang P, Hou L, et al. Global analysis of WRKY genes and their response to dehydration and salt stress in soybean. Frontiers in Plant Science, 2016, 7: 9. |
26 | Kayum M A, Jung H J, Park J I, et al. Identification and expression analysis of WRKY family genes under biotic and abiotic stresses in Brassica rapa. Molecular Genetics and Genomics, 2014, 290(1): 79-95. |
27 | Raja K, Sriram V, Backiyarani S, et al. Evolutionary expansion of WRKY gene family in banana and its expression profile during the infection of root lesion nematode, Pratylenchus coffeae. PLoS One, 2016, 11(9): e0162013. |
28 | Ross C A, Yue L, Shen Q J. The WRKY gene family in rice (Oryza sativa). Journal of Integrative Plant Biology, 2010, 49(6): 827-842. |
29 | Li P L, Song A P, Gao C Y, et al. Chrysanthemum WRKY gene CmWRKY17 negatively regulates salt stress tolerance in transgenic chrysanthemum and Arabidopsis plants. Plant Cell Reports, 2015, 34(8): 96-97. |
30 | Liu H, Yang W, Liu D, et al. Ectopic expression of a grapevine transcription factor VvWRKY11 contributes to osmotic stress tolerance in Arabidopsis. Molecular Biology Reports, 2011, 38(1): 417-427. |
31 | Qin Y, Tian Y, Liu X. A wheat salinity-induced WRKY transcription factor TaWRKY93 confers multiple abiotic stress tolerance in Arabidopsis thaliana. Biochemical & Biophysical Research Communications, 2015, 464(2): 428-433. |
32 | Sun Y D, Yun D Q. Activated expression of AtWRKY53 negatively regulates drought tolerance by mediating stomatal movement. Plant Cell Reports, 2015, 34(8): 1295-1306. |
33 | Qiu Y, Yu D. Over-expression of the stress-induced OsWRKY45 enhances disease resistance and drought tolerance in Arabidopsis. Environmental and Experimental Botany, 2009, 65(1): 35-47. |
34 | Zhou L, Wang N N, Gong S Y, et al. Overexpression of a cotton (Gossypium hirsutum) WRKY gene, GhWRKY34, in Arabidopsis enhances salt-tolerance of the transgenic plants. Plant Physiology and Biochemistry, 2015, 96: 311-320. |
35 | Eulgem T, Rushton P J, Robatzek S, et al. The WRKY superfamily of plant transcription factors. Trends in Plant Science, 2000, 5(5): 199-206. |
36 | Xie Z W, Wang L J, Chen J Y, et al. Studies on WRKY transcription factors and their biological functions in plants. Journal of Agricultural Science and Technology, 2016, 18(3): 46-54. |
37 | Yue H, Wang M, Liu S, et al. Transcriptome-wide identification and expression profiles of the WRKY transcription factor family in broomcorn millet (Panicum miliaceum L.). BMC Genomics, 2016, 17(1): 343. |
38 | Guo C, Guo R, Xu X, et al. Evolution and expression analysis of the grape (Vitis vinifera L.) WRKY gene family. Journal of Experimental Botany, 2014, 65(6): 1513-1528. |
39 | Wang Q, Wang M, Zhang X, et al. WRKY gene family evolution in Arabidopsis thaliana. Genetica, 2011, 139(8): 973-983. |
40 | Dou L L, Li G L, Pang Z Y, et al. Cloning and function analysis of GhWRKY11 in cotton (Gossypium hirsutum). Journal of Agricultural Biotechnology, 2016, 24(5): 625-636. |
窦玲玲, 李光雷, 庞朝友, 等. 棉花转录因子GhWRKY11的克隆及功能分析. 农业生物技术学报, 2016, 24(5): 625-636. | |
41 | Zhang H M, Liu X Q, Chen H T, et al. Subcellular localization and expression analysis of a soybean transcription factor GmWRKY58 in response to abiotic stresses. Acta Agriculturae Boreali-Sinica, 2018, 33(2): 49-57. |
张红梅, 刘晓庆, 陈华涛, 等. 大豆转录因子GmWRKY58亚细胞定位及在非生物胁迫下的表达分析. 华北农学报, 2018, 33(2): 49-57. | |
42 | Li J P. Cloning and analysis of resistant functions of transcription factor ZmWRKY25 from Zea mays. Changchun: Jilin University, 2012. |
李建萍. 玉米转录因子ZmWRKY25的克隆及其抗逆功能分析. 长春: 吉林大学, 2012. | |
43 | Fu Q T, Yu D Q. Expression profiles of AtWRKY25, AtWRKY26 and AtWRKY33 under abiotic stresses. Hereditas (Beijing), 2010, 32(8): 848-856. |
付乾堂, 余迪求. 拟南芥AtWRKY25、AtWRKY26和AtWRKY33在非生物胁迫条件下的表达分析. 遗传, 2010, 32(8): 848-856. | |
44 | Xiao D C, Zhang Z J, Xu Y W, et al. Cloning and functional analysis of Phyllostachys edulis MYB transcription factor PeMYB2. Hereditas (Beijing), 2013, 35(10): 1217-1225. |
肖冬长, 张智俊, 徐英武, 等. 毛竹MYB转录因子PeMYB2的克隆与功能分析. 遗传, 2013, 35(10): 1217-1225. |
[1] | 柴继宽, 琚泽亮, 赵桂琴. 低温和低pH胁迫下青贮乳酸菌OL77的内参基因筛选及CspP基因的表达模式分析[J]. 草业学报, 2023, 32(5): 147-158. |
[2] | 李艳鹏, 魏娜, 翟庆妍, 李杭, 张吉宇, 刘文献. 全基因组水平白花草木樨TCP基因家族的鉴定及在干旱胁迫下表达模式分析[J]. 草业学报, 2023, 32(4): 101-111. |
[3] | 李变变, 张凤华, 赵亚光. 刈割高度对油莎豆氮代谢及产量和品质的影响[J]. 草业学报, 2023, 32(2): 84-96. |
[4] | 张洁, 程凯, 王迎春. 长叶红砂钙依赖蛋白激酶RtCDPK16的非生物胁迫应答分析[J]. 草业学报, 2023, 32(2): 97-109. |
[5] | 田骄阳, 王秋霞, 郑淑文, 刘文献. 全基因组水平蒺藜苜蓿CPP基因家族的鉴定及表达模式分析[J]. 草业学报, 2022, 31(7): 111-121. |
[6] | 刘亚男, 于人杰, 高燕丽, 康俊梅, 杨青川, 武志海, 王珍. 蒺藜苜蓿膜联蛋白MtANN2基因的表达模式及盐胁迫下的功能分析[J]. 草业学报, 2022, 31(5): 124-134. |
[7] | 张国香, 郭卫冷, 毕铭钰, 张力爽, 王丹, 郭长虹. 紫花苜蓿CAX基因家族鉴定及其对非生物胁迫的响应分析[J]. 草业学报, 2022, 31(12): 106-117. |
[8] | 张家驹, 于洁, 李明娜, 康俊梅, 杨青川, 龙瑞才. 蒺藜苜蓿lncRNA167及其剪切产物miR167c的鉴定和功能分析[J]. 草业学报, 2022, 31(1): 164-180. |
[9] | 侯洁茹, 段晓玥, 李州, 彭燕. 白三叶TrSAMDC1克隆及表达分析[J]. 草业学报, 2020, 29(8): 170-178. |
[10] | 黄沁梅, 杨伊如, 陈丽飞, 尹航, 刘贺, 刘颖婕, 周蕴薇, 何淼. 转露地菊CgDREB22基因的烟草抗逆性分析[J]. 草业学报, 2020, 29(10): 109-118. |
[11] | 杨柳慧, 尹航, 黄沁梅, 张彦妮, 何淼, 周蕴薇. 细叶百合LpWRKY20基因对非生物胁迫的响应及抗旱性分析[J]. 草业学报, 2020, 29(1): 193-202. |
[12] | 高慧娟, 吕昕培, 王润娟, 任伟, 程济南, 汪永平, 邵坤仲, 张金林. 转录组测序在林草植物抗逆性研究中的应用[J]. 草业学报, 2019, 28(12): 184-196. |
[13] | 柯丹霞, 彭昆鹏, 夏远君, 朱玉莹, 张丹丹. 盐胁迫应答基因GmWRKY6的克隆及转基因百脉根的抗盐分析[J]. 草业学报, 2018, 27(8): 95-106. |
[14] | 谢志坚, 周春火, 贺亚琴, 宋涛, 于洋, 吴佳. 21世纪我国稻区种植紫云英的研究现状及展望[J]. 草业学报, 2018, 27(8): 185-196. |
[15] | 索雅飞,杜超,李宁宁,王燕,王迎春. 珍稀泌盐植物长叶红砂RtSOD基因的克隆及功能分析[J]. 草业学报, 2018, 27(4): 98-110. |
阅读次数 | ||||||
全文 |
|
|||||
摘要 |
|
|||||