睡莲WRKY家族的全基因组鉴定和分子进化分析

doi:10.16420/j.issn.0513-353x.2021-0092

摘要/Abstract

摘要：

在睡莲中共鉴定出69个NcWRKY转录因子,将其分成3个亚家族,即Ⅰ、Ⅱ和Ⅲ,其中Ⅱ亚家族可进一步分为Ⅱa、Ⅱb、Ⅱc、Ⅱd和Ⅱe,与拟南芥中WRKY亚家族分类相同。Ⅱc亚家族中含有17个成员,是睡莲WRKY家族中成员数最多的亚家族。所有成员的蛋白质二级结构中无规则卷曲所占比例高达为39% ~ 78%。motif 1和motif 3被注释为WRKY保守结构。NcWRKY成员在睡莲14条染色体中均有分布,其中Ⅱa亚家族中所有成员都定位在Nc4号染色体。启动子顺式调控元件分析发现,NcWRKY成员广泛参与激素调控反应,通过不同组织中表达量分析筛选出在成熟叶柄中高表达的NcWRKY04与NcWRKY18,在花瓣中高表达的NcWRKY05和在雄蕊中高表达的NcWRKY61。基因复制事件分析发现,NcWRKY家族主要以片段复制事件为主。基因表达网络关系分析预测NcWRKY61和NcWRKY63可能与其他成员有共表达关系。

关键词: 睡莲, WRKY, 全基因组鉴定, 分子进化

Abstract:

Sixty-nine NcWRKY transcription factors were identified in Nymphaea colorata and can be divided into three large subfamilies（I,Ⅱ and Ⅲ）. The Ⅱ subfamily can be further classified into Ⅱa,Ⅱb,Ⅱc,Ⅱd and Ⅱe. The Ⅱc as subfamily has the highest number of NcWRKY members with 17 members. Random coil accounted for 39%-78% in the protein secondary structure of the NcWRKY members. Motif 1 and motif 3 were annotated as WRKY conserved domain. NcWRKY members were distributed in waterlily 14 chromosomes. All members of the Ⅱa subfamily were located on chromosome Nc4. NcWRKY members can be regulated by hormones through the cis-element analysis of their promoters. NcWRKY04 and NcWRKY18 had very high expression level in mature leafstalk,NcWRKY05 had very high expression level in flower and NcWRKY61 had very high expression level in stamen through tissue specific expression analysis. NcWRKY61 and NcWRKY63 may have a co-expression relationship with other members through the gene expression network.

Key words: Nymphaea colorata, WRKY, genomic identification, molecular evolution

中图分类号:

S682.32

潘鑫峰, 叶方婷, 毛志君, 李兆伟, 范凯. 睡莲WRKY家族的全基因组鉴定和分子进化分析[J]. 园艺学报, 2022, 49(5): 1121-1135.

PAN Xinfeng, YE Fangting, MAO Zhijun, LI Zhaowei, FAN Kai. Genomic Identification and Molecular Evolution of the WRKY Family in Nymphaea colorata[J]. Acta Horticulturae Sinica, 2022, 49(5): 1121-1135.

图/表 9

参考文献 49

[1]	Cai C P, Niu E, Du H, Zhao L, Feng Y, Guo W. 2014. Genome-wide analysis of the WRKY transcription factor gene family in Gossypium raimondii and the expression of orthologs in cultivated tetraploid cotton. The Crop Journal, 2 (2-3):87-101. doi: 10.1016/j.cj.2014.03.001 URL
[2]	Cannon S B, Mitra A, Baumgarten A, Young N D, May G. 2004. The roles of segmental and tandem gene duplication in the evolution of large gene families in Arabidopsis thaliana. BMC Plant Biology, 4 (1):10. doi: 10.1186/1471-2229-4-10 URL
[3]	Chen C J, Chen H, Zhang Y, Thomas H R, Frank M H, He Y H, Xia R. 2020. TBtools:an integrative toolkit developed for interactive analyses of big biological data. Molecular Plant, 13 (8):1194-1202. doi: 10.1016/j.molp.2020.06.009 URL
[4]	Cline M S, Smoot M, Cerami E, Kuchinsky A, Landys N, Workman C, Christmas R, Avila-Campilo I, Creech M, Gross B, Hanspers K, Isserlin R, Kelley R, Killcoyne S, Lotia S, Maere S, Morris J, Ono K, Pavlovic V, Pico A R, Vailaya A, Wang P L, Adler A, Conklin B R, Hood L, Kuiper M, Sander C, Schmulevich I, Schwikowski B, Warner G J, Ideker T, Bader G D. 2007. Integration of biological networks and gene expression data using Cytoscape. Nature Protocols, 2 (10):2366-2382. doi: 10.1038/nprot.2007.324 URL
[5]	Eulgem T, Rushton P J, Robatzek S, Somssich I E. 2000. The WRKY superfamily of plant transcription factors. Trends Plant Science, 5 (5):199-206. doi: 10.1016/S1360-1385(00)01600-9 URL
[6]	Eulgem T, Somssich I E. 2007. Networks of WRKY transcription factors in defense signaling. Current Opinon in Plant Biology, 10 (4):366-371.
[7]	Fan K, Mao Z J, Zheng J X, Chen Y R, Lin W X. 2020. Molecular evolution and expansion of the KUP family in the allopolyploid cotton species Gossypium hirsutum and Gossypium barbadense. Frontiers in Plant Science, 11:545042. doi: 10.3389/fpls.2020.545042 URL
[8]	Fan K, Wang M, Miao Y, Ni M, Bibi N, Yuan S N, Li F, Wang X D. 2014. Molecular evolution and expansion analysis of the NAC transcription factor in Zea mays. PLoS ONE, 9 (11):e111837. doi: 10.1371/journal.pone.0111837 URL
[9]	Fan Z Q, Tan X L, Shan W, Kuang J F, Lu W J, Chen J Y. 2018. Characterization of a transcriptional regulator,BrWRKY6,associated with gibberellin-suppressed leaf senescence of Chinese flowering cabbage (Brassica parachinensis L.). Journal of Agricultural Food Chemistry, 66 (8):1791-1799. doi: 10.1021/acs.jafc.7b06085 URL
[10]	Gabarayeva N I, Rowley J R. 1994. Exine development in Nymphaea colorata(Nymphaeaceae). Nordic Journal of Botany, 14(6):671-691. doi: 10.1111/j.1756-1051.1994.tb01085.x URL
[11]	Guo H Y, Zhang Y T, Wang Z, Lin L M, Cui M H, Long Y H, Xing Z B. 2019. Genome-wide identification of WRKY transcription factors in the asteranae. Plants, 8 (10):393. doi: 10.3390/plants8100393 URL
[12]	He H S, Dong Q, Shao Y H, Jiang H Y, Zhu S, Cheng B J, Xiang Y. 2012. Genome-wide survey and characterization of the WRKY gene family in Populus trichocarpa. Plant Cell Report, 31(7):1199-1217.
[13]	Hui S, Wang P, Lin J Y, Zhao C, Bi Y, Wang X. 2016. Genome-wide identification and characterization of WRKY gene family in peanut. Frontiers in Plant Science, 7:534.
[14]	Huang S X, Gao Y F, Liu J K, Peng X L, Niu X L, Fei Z J, Cao S Q, Liu Y S. 2012. Genome-wide analysis of WRKY transcription factors in Solanum lycopersicum. Molecular Genetics and Genomics, 287(6):495-513. doi: 10.1007/s00438-012-0696-6 URL
[15]	Ishiguro S, Nakamura K. 1994. 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 β-amylase from sweet potato(Ipomoea batatas). Molecular & General Genetics Mgg, 244 (6):563-571.
[16]	Jiang C H, Huang Z Y, Xie P, Gu C, Li K, Wang D C, Yu Y Y, Fan Z H, Wang C J, Wang Y P, Guo Y H, Guo J H. 2016. Transcription factors WRKY70 and WRKY 11 served as regulators in rhizobacterium Bacillus cereus AR156-induced systemic resistance to Pseudomonas syringae pv. tomato DC3000 in Arabidopsis. Journal of Experimental Botany, 67 (1):157-174. doi: 10.1093/jxb/erv445 URL
[17]	Katoh K. 2002. MAFFT:a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Research, 30 (14):3059-3066. doi: 10.1093/nar/gkf436 URL
[18]	Kim H J, Nam H G, Lim P O. 2016. Regulatory network of NAC transcription factors in leaf senescence. Current Opinion in Plant Biology, 33:48-56. doi: 10.1016/j.pbi.2016.06.002 URL
[19]	Kim T, Kang K, Kim S H, An G, Paek N C. 2019. OsWRKY 5 promotes rice leaf senescence via senescence-associated NAC and abscisic acidaiosynthesis pathway. International Journal of Molecular Sciences, 20 (18):4437 doi: 10.3390/ijms20184437 URL
[20]	Krzywinski M, Schein J, Birol I, Connors J, Gascoyne R, Horsman D, Jones S J, Marra M A. 2009. Circos:An information aesthetic for comparative genomics. Genome Research, 19:1639-1645. doi: 10.1101/gr.092759.109 pmid: 19541911
[21]	Lam-Tung N, Schmidt H A, Arndt V H, Quang M B. 2015. IQ-TREE:a fast and effective stochastic algorithm for estimating maximum-likelihood phylogenies. Molecular Biology & Evolution, 32 (1):268-274.
[22]	Li J, Brader G, Kariola T, Palva E T. 2006. WRKY 70 modulates the selection of signaling pathways in plant defense. Plant Journal, 46 (3):477-491. doi: 10.1111/j.1365-313X.2006.02712.x URL
[23]	Li Xiaoying, Xu Hongxia, Chen Junwei. 2019. Identification and expression analysis of WRKY transcription factors in Eriobotrya japonica. Acta Horticulturae Sinica, 46 (5):939-954. (in Chinese)
	李晓颖, 徐红霞, 陈俊伟. 2019. 枇杷WRKY转录因子鉴定与表达分析. 园艺学报, 46 (5):939-954.
[24]	Lin Shengnan, Liu Jiewei, Zhang Xiaoni, Bao Manzhu, Fu Xiaopeng. 2021. Genome-wide identification and expression analysis of WRKY gene family in Dianthus caryophyllus. Acta Horticulturae Sinica, 48 (9):1768-1784. (in Chinese)
	林胜男, 刘杰玮, 张晓妮, 包满珠, 傅小鹏. 2021. 香石竹WRKY家族全基因组鉴定及其表达分析. 园艺学报, 48 (9):1768-1784.
[25]	Liu S, Kracher B, Ziegler J, Birkenbihl R P, Somssich I E. 2015. Negative regulation of ABA signaling by WRKY 33 is critical for Arabidopsis immunity towards Botrytis cinerea 2100. Elife, 4:e7295.
[26]	Liu Z Q, Shi L P, Yang S, Qiu S S, Ma X L, Cai J S, Guan D Y, Wang Z H, He S L. 2021. A conserved double-W box in the promoter of CaWRKY 40 mediates autoregulation during response to pathogen attack and heat stress in pepper. Molecular Plant Pathology, 22 (1):3-18. doi: 10.1111/mpp.13004 URL
[27]	Luo X Y, Li C, He X, Zhang X L, Zhu L F. 2020. ABA signaling is negatively regulated by GbWRKY1 through JAZ1 and ABI1 to affect salt and drought tolerance. Plant Cell Report, 39 (2):181-194.
[28]	Miao Y, Laun T, Zimmermann P, Zentgraf U. 2004. Targets of the WRKY 53 transcription factor and its role during leaf senescence in Arabidopsis. Plant Molecular Biology, 55 (6):853-867. pmid: 15604721
[29]	Nagaraju M, Kumar S A, Reddy P S, Kumar A, Rao D M, Kavi K P. 2019. Genome-scale identification,classification,and tissue specific expression analysis of late embryogenesis abundant(LEA)genes under abiotic stress conditions in Sorghum bicolor L. PLoS ONE, 14 (1):e209980.
[30]	Pasquini M, Bonfanti M, Martinazzo R. 2016. Genome-wide annotation of the soybean WRKY family and functional characterization of genes involved in response to Phakopsora pachyrhizi infection. Physical Chemistry Chemical Physics, 18 (9):6607-6617. doi: 10.1039/c5cp07272g pmid: 26868899
[31]	Pertea M, Kim D, Pertea G M, Leek J T, Salzberg S L. 2016. Transcript-level expression analysis of RNA-seq experiments with HISAT,StringTie and Ballgown. Nature Protocols, 11 (9):1650-1667. doi: 10.1038/nprot.2016.095 URL
[32]	Qin Z, Lv H J, Zhu X L, Meng C, Quan T Y, Wang M C, Xia G M. 2013. Ectopic expression of a wheat WRKY transcription factor gene TaWRKY71- 1 results in hyponastic leaves in Arabidopsis thaliana. PLoS ONE, 8 (5):e63033. doi: 10.1371/journal.pone.0063033 URL
[33]	Rushton P J, Bokowiec M T, Han S, Zhang H, Brannock J F, Chen X, Laudeman T W, Timko M P. 2008. Tobacco transcription factors:novel insights into transcriptional regulation in the Solanaceae. Plant Physiology, 147 (1):280-295. doi: 10.1104/pp.107.114041 pmid: 18337489
[34]	Rushton P J, Somssich I E, Ringler P, Shen Q J. 2010. WRKY transcription factors. Trends in Plant Science, 15 (5):247-258. doi: 10.1016/j.tplants.2010.02.006 pmid: 20304701
[35]	Song Y, Gao J. 2014. Genome-wide analysis of WRKY gene family in Arabidopsis lyrata and comparison with Arabidopsis thaliana and Populus trichocarpa. Chinese Science Bulletin, 59 (8):754-765. doi: 10.1007/s11434-013-0057-9 URL
[36]	Song Y, Jing S J, Yu D Q. 2009. Overexpression of the stress-induced OsWRKY 08 improves osmotic stress tolerance in Arabidopsis. Chinese Science Bulletin, 54 (24):4671-4678.
[37]	Ullah A, Sun H, Hakim, Yang X, Zhang X. 2018. A novel cotton WRKY gene,GhWRKY6-like,improves salt tolerance by activating the ABA signaling pathway and scavenging of reactive oxygen species. Physiologia Plantarum, 162 (4):439-454. doi: 10.1111/ppl.12651 URL
[38]	Wang C T, Ru J N, Liu Y W, Yang J F, Li M, Xu Z S, Fu J D. 2018. The maize WRKY transcription factor ZmWRKY 40 confers drought resistance in transgenic Arabidopsis. International Journal of Molecular Sciences, 19 (9):2580. doi: 10.3390/ijms19092580 URL
[39]	Wei K F, Chen J, Chen Y F, Wu L J, Xie D X. 2012. Molecular phylogenetic and expression analysis of the complete WRKY transcription factor family in maize(Zea mays L.). DNA Research, 19 (2):153-164. doi: 10.1093/dnares/dsr048 URL
[40]	Wei Y X, Shi H T, Xia Z Q, Tie W W, Ding Z H, Yan Y, Wang W Q, Hu W, Li K M. 2016. Genome-wide identification and expression analysis of the WRKY gene family in Cassava(Manihot esculenta Crantz). Frontiers in Plant Science, 7:25.
[41]	Xie T, Chen C J, Li C H, Liu J R, Liu C Y, He Y H. 2018. Genome-wide investigation of WRKY gene family in pineapple:evolution and expression profiles during development and stress. BMC Genomics, 19 (1):490. doi: 10.1186/s12864-018-4880-x URL
[42]	Xu Q, Feng W J, Peng H R, Ni Z F, Sun Q X. 2014. TaWRKY71,a WRKY transcription factor from wheat,enhances tolerance to abiotic stress in transgenic Arabidopsis thaliana. Cereal Research Communications, 42 (1):47-57. doi: 10.1556/CRC.2013.0051 URL
[43]	Ye Yujie. 2016. The Interaction between MaWRKY26 and MaVQ 5 invoving in the regulation of MeJA-Induced chilling tolerance of banana fruit [Ph. D. Dissertation]. Guangzhou: South China Agricultural University. (in Chinese)
	叶玉洁. 2016. MaWRKY26与MaVQ5蛋白互作调控MeJA诱导香蕉果实耐冷性的机制[博士论文]. 广州: 华南农业大学.
[44]	Yu Y, Wang L, Chen J, Liu Z, Park C M, Xiang F. 2018. WRKY 71 actsantagonistically against salt-delayed flowering in Arabidopsis thaliana. Plant Cell Physioloy, 59 (2):414-422.
[45]	Yu Yanchong. 2011. The molecular mechanism of WRKY 71 transcription factor in regulating flower and branch development of Arabidopsis thialiana[Ph. D. Dissertation]. Ji’nan: Shandong University. (in Chinese)
	于延冲. 2011. 拟南芥转录因子WRKY71对花和分枝发育的调控机制研究[博士论文]. 济南: 山东大学.
[46]	Zhang L S, Chen F, Zhang X T, Li Z, Zhao Y Y, Lohaus R, Chang X, Dong W, Ho S, Liu X, Song A, Chen J H, Guo W L, Wang Z J, Zhuang Y Y, Wang H F, Chen X Q, Hu J, Liu Y H, Qin Y, Wang K, Dong S S, Liu Y, Zhang S Z, Yu X X, Wu Q, Wang L S, Yan X Q, Jiao Y N, Kong H Z, Zhou X F, Yu C W, Chen Y C, Li F, Wang J H, Chen W, Chen X L, Jia Q Y, Zhang C, Jiang Y F, Zhang W B, Liu G H, Fu J Y, Chen F, Ma H, van de Peer Y, Tang H B. 2020. The water lily genome and the early evolution of flowering plants. Nature, 577 (7788):79-84. doi: 10.1038/s41586-019-1852-5 URL
[47]	Zhang Y, Wang L. 2005. The WRKY transcription factor superfamily:its origin in eukaryotes and expansion in plants. BMC Evolutionary Biology, 5:1. doi: 10.1186/1471-2148-5-1 URL
[48]	Zhen X, Zhang Z L, Zou X L, Huang J, Paul R, Daniel T, Shen Q X. 2005. Annotations and functional analyses of the rice WRKY gene superfamily reveal positive and negative regulators of abscisic acid signaling in aleurone cells. Plant Physiology, 137 (1):176-189. pmid: 15618416
[49]	Zeng Zexiang, Xiao Xianmei, Tan Xiaoli, Fan Zhongqi, Chen Jianye. 2021. Characteristics of the transcription factor BrWRKY57 and its regulation on BrPPH1 and BrNCED3. Acta Horticulturae Sinica, 48 (3):518-530. (in Chinese)
	曾泽湘, 肖显梅, 谭小丽, 范中奇, 陈建业. 2021. 菜薹BrWRKY57的特性及其对BrPPH1和BrNCED3的调控. 园艺学报, 48 (3):518-530.

名称 Name	位点 Locus	亚家族 Subfamily	蛋白质长度/aa Protein length	分子量 Molecular Weight	pI	α-螺旋 α-helix	延伸链 Extended strand	β-折叠 β-turn	无规则卷曲 Random coil
NcWRKY01	GWHPAAYW001268	Ⅱd	299	33 359.08	9.69	67	29	12	191
NcWRKY02	GWHPAAYW001468	Ⅲ	219	24 178.36	9.1	89	28	4	98
NcWRKY03	GWHPAAYW002553	Ⅱc	262	28 583.02	8.78	61	45	25	131
NcWRKY04	GWHPAAYW003267	Ⅲ	350	39 563.96	5.07	78	37	14	221
NcWRKY05	GWHPAAYW011370	Ⅱc	306	33 839.58	6.31	74	31	17	184
NcWRKY06	GWHPAAYW012017	Ⅰ	832	88 126.14	5.42	75	83	38	636
NcWRKY07	GWHPAAYW012284	Ⅱc	224	25 222.62	9.04	62	35	13	114
NcWRKY08	GWHPAAYW012407	Ⅲ	194	21 868.50	7.67	51	43	12	88
NcWRKY09	GWHPAAYW012566	Ⅰ	313	34 228.55	10.01	50	36	9	218
NcWRKY10	GWHPAAYW013009	Ⅰ	316	34 251.61	9.3	48	60	14	194
NcWRKY11	GWHPAAYW013286	Ⅱe	345	37 142.72	5.01	112	34	29	170
NcWRKY12	GWHPAAYW013314	Ⅱc	208	23 139.17	8.94	59	26	14	109
NcWRKY13	GWHPAAYW013362	Ⅰ	181	19 803.13	7.8	27	41	4	109
NcWRKY14	GWHPAAYW014542	Ⅰ	816	87 750.28	5.25	92	95	28	601
NcWRKY15	GWHPAAYW015340	Ⅱd	361	38 817.12	9.45	66	41	33	221
NcWRKY16	GWHPAAYW016607	Ⅱd	337	36 359.40	9.68	83	35	26	193
NcWRKY17	GWHPAAYW017157	Ⅱa	293	31 896.80	8.52	85	40	13	155
NcWRKY18	GWHPAAYW017158	Ⅱa	315	33 882.86	6.62	114	45	13	143
NcWRKY19	GWHPAAYW017159	Ⅱa	425	46 389.46	9.37	159	51	15	200
NcWRKY20	GWHPAAYW017160	Ⅱa	315	33 832.56	6.61	88	45	10	172
NcWRKY21	GWHPAAYW017161	Ⅱa	327	35 972.96	6.32	109	39	12	167
NcWRKY22	GWHPAAYW017408	Ⅰ	1 209	13 726.21	8.92	382	238	107	482
NcWRKY23	GWHPAAYW018263	Ⅱc	236	26 890.16	9.42	30	59	11	136
NcWRKY24	GWHPAAYW018963	Ⅱc	325	35 439.54	5.19	76	45	12	192
NcWRKY25	GWHPAAYW019317	Ⅰ	475	51 676.63	9.42	56	53	22	344
NcWRKY26	GWHPAAYW019476	Ⅱc	292	32 449.26	6.84	80	37	15	160
NcWRKY27	GWHPAAYW020087	Ⅱd	85	9 747.24	9.88	13	24	0	42
NcWRKY28	GWHPAAYW020328	Ⅲ	301	33 205.85	4.87	91	44	15	151
NcWRKY29	GWHPAAYW020329	Ⅲ	287	31 497.25	5.43	104	22	10	151
NcWRKY30	GWHPAAYW020535	Ⅱc	326	35 958.21	9.38	85	40	14	187
NcWRKY31	GWHPAAYW020992	Ⅱb	522	56 768.49	8.47	157	51	13	301
NcWRKY32	GWHPAAYW020993	Ⅱb	550	59 854.45	6.3	128	60	19	343
NcWRKY33	GWHPAAYW020994	Ⅱb	483	52 474.26	6.43	140	36	13	294
NcWRKY34	GWHPAAYW021005	Ⅱc	390	44 581.80	5.73	67	50	26	247
NcWRKY35	GWHPAAYW021006	Ⅱc	207	23 478.24	5.57	40	38	18	111
NcWRKY36	GWHPAAYW021007	Ⅱc	336	38 210.70	7.54	39	58	29	210
NcWRKY37	GWHPAAYW021021	Ⅱc	252	27 873.25	8.78	73	42	42	117
NcWRKY38	GWHPAAYW021561	Ⅱd	300	33 420.36	9.71	77	31	13	179
NcWRKY39	GWHPAAYW023351	Ⅰ	653	69 898.00	5.94	64	75	21	493
NcWRKY40	GWHPAAYW023893	Ⅱb	536	57 654.27	6.48	149	44	16	327
NcWRKY41	GWHPAAYW024187	Ⅱc	194	22 010.58	8.97	57	24	11	102
NcWRKY42	GWHPAAYW024283	Ⅰ	614	66 224.72	6.19	79	73	31	431
NcWRKY43	GWHPAAYW024356	Ⅰ	207	22 640.31	7.08	32	41	6	128
NcWRKY44	GWHPAAYW024753	Ⅰ	541	57 521.59	7.34	55	70	26	390
NcWRKY45	GWHPAAYW025120	Ⅰ	376	40 730.26	6.54	36	55	15	270
NcWRKY46	GWHPAAYW025126	Ⅰ	314	34 201.39	8.97	38	41	14	221
NcWRKY47	GWHPAAYW025139	Ⅰ	231	25 176.13	6.06	45	47	16	123
NcWRKY48	GWHPAAYW025454	Ⅱb	496	53 858.63	5.15	126	56	23	291
NcWRKY49	GWHPAAYW025832	Ⅰ	703	75 743.55	5.72	102	65	23	513
名称 Name	位点 Locus	亚家族 Subfamily	蛋白质长度/aa Protein length	分子量 Molecular Weight	pI	α-螺旋 α-helix	延伸链 Extended strand	β-折叠 β-turn	无规则卷曲 Random coil
NcWRKY50	GWHPAAYW026703	Ⅱe	94	10 236.55	9.99	22	19	9	44
NcWRKY51	GWHPAAYW027586	Ⅱd	289	31 308.83	9.85	58	26	12	193
NcWRKY52	GWHPAAYW027773	Ⅲ	366	40 600.28	5.83	92	37	10	227
NcWRKY53	GWHPAAYW004306	Ⅱb	473	51 009.43	5.11	153	61	20	239
NcWRKY54	GWHPAAYW005049	Ⅱb	540	57 498.21	6.36	134	57	18	331
NcWRKY55	GWHPAAYW005095	Ⅰ	605	65 201.75	7.75	57	59	12	477
NcWRKY56	GWHPAAYW005338	Ⅲ	263	29 278.76	5.46	86	26	14	137
NcWRKY57	GWHPAAYW005461	Ⅱe	255	28 218.34	5.41	41	35	12	167
NcWRKY58	GWHPAAYW005491	Ⅱc	315	34 590.63	6.27	80	27	8	200
NcWRKY59	GWHPAAYW006427	Ⅱc	317	35 052.79	6.42	57	30	11	219
NcWRKY60	GWHPAAYW006549	Ⅱe	431	47 359.75	5.77	78	32	19	302
NcWRKY61	GWHPAAYW007012	Ⅱb	557	59 663.54	6.11	156	39	11	351
NcWRKY62	GWHPAAYW008691	Ⅱd	1 144	126 555.82	8.82	383	213	90	458
NcWRKY63	GWHPAAYW009733	Ⅱd	73	8 382.87	9.82	9	15	4	45
NcWRKY64	GWHPAAYW009825	Ⅱe	122	13 701.70	10.19	24	16	12	70
NcWRKY65	GWHPAAYW009961	Ⅱc	199	21 660.89	6.94	46	27	6	120
NcWRKY66	GWHPAAYW009962	Ⅱe	290	31 222.80	5.11	72	19	11	188
NcWRKY67	GWHPAAYW010546	Ⅱc	230	25 695.79	6.59	47	40	9	134
NcWRKY68	GWHPAAYW010567	Ⅱb	1 076	114 597.89	5.94	404	153	63	456
NcWRKY69	GWHPAAYW010914	Ⅱe	445	47 724.61	5.12	87	35	21	302

名称 Name	位点 Locus	亚家族 Subfamily	蛋白质长度/aa Protein length	分子量 Molecular Weight	pI	α-螺旋 α-helix	延伸链 Extended strand	β-折叠 β-turn	无规则卷曲 Random coil
NcWRKY01	GWHPAAYW001268	Ⅱd	299	33 359.08	9.69	67	29	12	191
NcWRKY02	GWHPAAYW001468	Ⅲ	219	24 178.36	9.1	89	28	4	98
NcWRKY03	GWHPAAYW002553	Ⅱc	262	28 583.02	8.78	61	45	25	131
NcWRKY04	GWHPAAYW003267	Ⅲ	350	39 563.96	5.07	78	37	14	221
NcWRKY05	GWHPAAYW011370	Ⅱc	306	33 839.58	6.31	74	31	17	184
NcWRKY06	GWHPAAYW012017	Ⅰ	832	88 126.14	5.42	75	83	38	636
NcWRKY07	GWHPAAYW012284	Ⅱc	224	25 222.62	9.04	62	35	13	114
NcWRKY08	GWHPAAYW012407	Ⅲ	194	21 868.50	7.67	51	43	12	88
NcWRKY09	GWHPAAYW012566	Ⅰ	313	34 228.55	10.01	50	36	9	218
NcWRKY10	GWHPAAYW013009	Ⅰ	316	34 251.61	9.3	48	60	14	194
NcWRKY11	GWHPAAYW013286	Ⅱe	345	37 142.72	5.01	112	34	29	170
NcWRKY12	GWHPAAYW013314	Ⅱc	208	23 139.17	8.94	59	26	14	109
NcWRKY13	GWHPAAYW013362	Ⅰ	181	19 803.13	7.8	27	41	4	109
NcWRKY14	GWHPAAYW014542	Ⅰ	816	87 750.28	5.25	92	95	28	601
NcWRKY15	GWHPAAYW015340	Ⅱd	361	38 817.12	9.45	66	41	33	221
NcWRKY16	GWHPAAYW016607	Ⅱd	337	36 359.40	9.68	83	35	26	193
NcWRKY17	GWHPAAYW017157	Ⅱa	293	31 896.80	8.52	85	40	13	155
NcWRKY18	GWHPAAYW017158	Ⅱa	315	33 882.86	6.62	114	45	13	143
NcWRKY19	GWHPAAYW017159	Ⅱa	425	46 389.46	9.37	159	51	15	200
NcWRKY20	GWHPAAYW017160	Ⅱa	315	33 832.56	6.61	88	45	10	172
NcWRKY21	GWHPAAYW017161	Ⅱa	327	35 972.96	6.32	109	39	12	167
NcWRKY22	GWHPAAYW017408	Ⅰ	1 209	13 726.21	8.92	382	238	107	482
NcWRKY23	GWHPAAYW018263	Ⅱc	236	26 890.16	9.42	30	59	11	136
NcWRKY24	GWHPAAYW018963	Ⅱc	325	35 439.54	5.19	76	45	12	192
NcWRKY25	GWHPAAYW019317	Ⅰ	475	51 676.63	9.42	56	53	22	344
NcWRKY26	GWHPAAYW019476	Ⅱc	292	32 449.26	6.84	80	37	15	160
NcWRKY27	GWHPAAYW020087	Ⅱd	85	9 747.24	9.88	13	24	0	42
NcWRKY28	GWHPAAYW020328	Ⅲ	301	33 205.85	4.87	91	44	15	151
NcWRKY29	GWHPAAYW020329	Ⅲ	287	31 497.25	5.43	104	22	10	151
NcWRKY30	GWHPAAYW020535	Ⅱc	326	35 958.21	9.38	85	40	14	187
NcWRKY31	GWHPAAYW020992	Ⅱb	522	56 768.49	8.47	157	51	13	301
NcWRKY32	GWHPAAYW020993	Ⅱb	550	59 854.45	6.3	128	60	19	343
NcWRKY33	GWHPAAYW020994	Ⅱb	483	52 474.26	6.43	140	36	13	294
NcWRKY34	GWHPAAYW021005	Ⅱc	390	44 581.80	5.73	67	50	26	247
NcWRKY35	GWHPAAYW021006	Ⅱc	207	23 478.24	5.57	40	38	18	111
NcWRKY36	GWHPAAYW021007	Ⅱc	336	38 210.70	7.54	39	58	29	210
NcWRKY37	GWHPAAYW021021	Ⅱc	252	27 873.25	8.78	73	42	42	117
NcWRKY38	GWHPAAYW021561	Ⅱd	300	33 420.36	9.71	77	31	13	179
NcWRKY39	GWHPAAYW023351	Ⅰ	653	69 898.00	5.94	64	75	21	493
NcWRKY40	GWHPAAYW023893	Ⅱb	536	57 654.27	6.48	149	44	16	327
NcWRKY41	GWHPAAYW024187	Ⅱc	194	22 010.58	8.97	57	24	11	102
NcWRKY42	GWHPAAYW024283	Ⅰ	614	66 224.72	6.19	79	73	31	431
NcWRKY43	GWHPAAYW024356	Ⅰ	207	22 640.31	7.08	32	41	6	128
NcWRKY44	GWHPAAYW024753	Ⅰ	541	57 521.59	7.34	55	70	26	390
NcWRKY45	GWHPAAYW025120	Ⅰ	376	40 730.26	6.54	36	55	15	270
NcWRKY46	GWHPAAYW025126	Ⅰ	314	34 201.39	8.97	38	41	14	221
NcWRKY47	GWHPAAYW025139	Ⅰ	231	25 176.13	6.06	45	47	16	123
NcWRKY48	GWHPAAYW025454	Ⅱb	496	53 858.63	5.15	126	56	23	291
NcWRKY49	GWHPAAYW025832	Ⅰ	703	75 743.55	5.72	102	65	23	513
名称 Name	位点 Locus	亚家族 Subfamily	蛋白质长度/aa Protein length	分子量 Molecular Weight	pI	α-螺旋 α-helix	延伸链 Extended strand	β-折叠 β-turn	无规则卷曲 Random coil
NcWRKY50	GWHPAAYW026703	Ⅱe	94	10 236.55	9.99	22	19	9	44
NcWRKY51	GWHPAAYW027586	Ⅱd	289	31 308.83	9.85	58	26	12	193
NcWRKY52	GWHPAAYW027773	Ⅲ	366	40 600.28	5.83	92	37	10	227
NcWRKY53	GWHPAAYW004306	Ⅱb	473	51 009.43	5.11	153	61	20	239
NcWRKY54	GWHPAAYW005049	Ⅱb	540	57 498.21	6.36	134	57	18	331
NcWRKY55	GWHPAAYW005095	Ⅰ	605	65 201.75	7.75	57	59	12	477
NcWRKY56	GWHPAAYW005338	Ⅲ	263	29 278.76	5.46	86	26	14	137
NcWRKY57	GWHPAAYW005461	Ⅱe	255	28 218.34	5.41	41	35	12	167
NcWRKY58	GWHPAAYW005491	Ⅱc	315	34 590.63	6.27	80	27	8	200
NcWRKY59	GWHPAAYW006427	Ⅱc	317	35 052.79	6.42	57	30	11	219
NcWRKY60	GWHPAAYW006549	Ⅱe	431	47 359.75	5.77	78	32	19	302
NcWRKY61	GWHPAAYW007012	Ⅱb	557	59 663.54	6.11	156	39	11	351
NcWRKY62	GWHPAAYW008691	Ⅱd	1 144	126 555.82	8.82	383	213	90	458
NcWRKY63	GWHPAAYW009733	Ⅱd	73	8 382.87	9.82	9	15	4	45
NcWRKY64	GWHPAAYW009825	Ⅱe	122	13 701.70	10.19	24	16	12	70
NcWRKY65	GWHPAAYW009961	Ⅱc	199	21 660.89	6.94	46	27	6	120
NcWRKY66	GWHPAAYW009962	Ⅱe	290	31 222.80	5.11	72	19	11	188
NcWRKY67	GWHPAAYW010546	Ⅱc	230	25 695.79	6.59	47	40	9	134
NcWRKY68	GWHPAAYW010567	Ⅱb	1 076	114 597.89	5.94	404	153	63	456
NcWRKY69	GWHPAAYW010914	Ⅱe	445	47 724.61	5.12	87	35	21	302

ID	序列	长度/aa
ID	Sequence	Length
motif 1	ILDDGYRWRKYGQKVVKGNPYPRSYYRCT	29
motif 2	PGCPVRKQVZRASEDPSIVITTYEGKHNH	29
motif 3	RPSEDGYNWRKYGQKQVKGSEFPRSYYKCTHPNCQVKKKVERSHDGLVTEIIYKGTHNHPKPQPSRR	67
motif 4	NTNMQVSLNPVMLMPPNQIEVNRALPYSRGPDAVSLDLGVGISQAAENSSLHKQNQAEADSMLRQVHGGNAPSSGINCPVQSAGILPFYGNGIMGNRPDHYRVKEEDGQNFNF	113
motif 5	EREPRVAFQTMSEVD	15
motif 6	LKAEVGRVKEENERLKRMLSRICKSYKALQMQFASLIQS	39
motif 7	AAAVASAJASDPSFTAALAAAJSSIIG	27
motif 8	PLPVQATAMASTTSAAASMLLSGSSSST	28
motif 9	DVPTSSSTNRETSAPMAVQNL	21
motif 10	GRCHCSKRRKSRVKRVIRVPAISSKLAD	28
motif 11	MEEVEEANKAAVECCHRLIAVLAQPQEGLEWKGVMGLTGEALMKFRKVVSLLNHKAGHARFRRGSTQSPPRQKICMENPKLGLDPAAPKVYPEPPQCPPHPQAPNPHFQFLQQMKLQADLLYRRSNSSINLKFDNSSCTPTMSSTRSFMSSLSMDGSVANMDGKGFCLIGPASSQDRISHQQKRKCSGKGEDG	193
motif 12	MSSLNNFHRGTGRFMLGSEVDASHAQQFT	29
motif 13	ECPYVIYYITPQHDCDVGVISAAQIPFVSHGAPCAGRQGFY	41
motif 14	LAEPSPTTGTFSFRPCNAGFNSGLVSNAFSSSWNVSKKHYLGDGSGPGSFVFKPREEASSSVGALAVVNSNPEKNQNFGQCQGYSEPQASIKIEAMAVSAKDLSLSVATSEAPACGAASNSITPANPSSDELNVRRGPDAETQHVSLDQKGAPLTVVE	158
motif 15	SDSLGDEGDEGEPESKRRKME	21
motif 16	VCTDLPAYTPEIFPAAENDSCFQLQDILDFCDPLIDGELFEGLEMDGNLQIPTSDSNQPNKIGDSQAKKQRGECSSRKRK	80
motif 17	QGRAFNQLEABGTPELSSLSTSDDEGDED	29
motif 18	DQAQQPHAKKARVSVRARCD	20
motif 19	SSSASISVSPSYPTITLDLTQTSSPPZF	28
motif 20	GSPVALSLVMLKPPNHIENGRAPLFPTASNVVSLDLGVGIG	41

ID	序列	长度/aa
ID	Sequence	Length
motif 1	ILDDGYRWRKYGQKVVKGNPYPRSYYRCT	29
motif 2	PGCPVRKQVZRASEDPSIVITTYEGKHNH	29
motif 3	RPSEDGYNWRKYGQKQVKGSEFPRSYYKCTHPNCQVKKKVERSHDGLVTEIIYKGTHNHPKPQPSRR	67
motif 4	NTNMQVSLNPVMLMPPNQIEVNRALPYSRGPDAVSLDLGVGISQAAENSSLHKQNQAEADSMLRQVHGGNAPSSGINCPVQSAGILPFYGNGIMGNRPDHYRVKEEDGQNFNF	113
motif 5	EREPRVAFQTMSEVD	15
motif 6	LKAEVGRVKEENERLKRMLSRICKSYKALQMQFASLIQS	39
motif 7	AAAVASAJASDPSFTAALAAAJSSIIG	27
motif 8	PLPVQATAMASTTSAAASMLLSGSSSST	28
motif 9	DVPTSSSTNRETSAPMAVQNL	21
motif 10	GRCHCSKRRKSRVKRVIRVPAISSKLAD	28
motif 11	MEEVEEANKAAVECCHRLIAVLAQPQEGLEWKGVMGLTGEALMKFRKVVSLLNHKAGHARFRRGSTQSPPRQKICMENPKLGLDPAAPKVYPEPPQCPPHPQAPNPHFQFLQQMKLQADLLYRRSNSSINLKFDNSSCTPTMSSTRSFMSSLSMDGSVANMDGKGFCLIGPASSQDRISHQQKRKCSGKGEDG	193
motif 12	MSSLNNFHRGTGRFMLGSEVDASHAQQFT	29
motif 13	ECPYVIYYITPQHDCDVGVISAAQIPFVSHGAPCAGRQGFY	41
motif 14	LAEPSPTTGTFSFRPCNAGFNSGLVSNAFSSSWNVSKKHYLGDGSGPGSFVFKPREEASSSVGALAVVNSNPEKNQNFGQCQGYSEPQASIKIEAMAVSAKDLSLSVATSEAPACGAASNSITPANPSSDELNVRRGPDAETQHVSLDQKGAPLTVVE	158
motif 15	SDSLGDEGDEGEPESKRRKME	21
motif 16	VCTDLPAYTPEIFPAAENDSCFQLQDILDFCDPLIDGELFEGLEMDGNLQIPTSDSNQPNKIGDSQAKKQRGECSSRKRK	80
motif 17	QGRAFNQLEABGTPELSSLSTSDDEGDED	29
motif 18	DQAQQPHAKKARVSVRARCD	20
motif 19	SSSASISVSPSYPTITLDLTQTSSPPZF	28
motif 20	GSPVALSLVMLKPPNHIENGRAPLFPTASNVVSLDLGVGIG	41

复制类型	复制基因1	复制基因2	亚家族	Ka	Ks	Ka/Ks
Duplicated type	Duplicated gene 1	Duplicated gene 2	Subfamily	Ka	Ks	Ka/Ks
片段复制	NcWRKY02	NcWRKY08	Ⅲ	0.6235	1.6690	0.3735
Segmental duplication	NcWRKY01	NcWRKY38	Ⅱd	0.0884	0.9775	0.0905
	NcWRKY03	NcWRKY37	Ⅱc	0.3228	2.0820	0.1549
	NcWRKY06	NcWRKY14	Ⅰ	0.2384	0.9139	0.2609
	NcWRKY09	NcWRKY25	Ⅰ	0.3501	1.0760	0.3252
	NcWRKY07	NcWRKY41	Ⅱc	0.2282	2.1670	0.1052
	NcWRKY05	NcWRKY59	Ⅱc	0.2031	1.0310	0.1969
	NcWRKY11	NcWRKY66	Ⅱe	0.2233	1.4330	0.1557
	NcWRKY12	NcWRKY65	Ⅱc	0.3091	1.8660	0.1655
	NcWRKY16	NcWRKY51	Ⅱd	0.1750	0.9797	0.1786
	NcWRKY30	NcWRKY58	Ⅱc	0.3375	0.9217	0.3662
	NcWRKY28	NcWRKY56	Ⅲ	0.3797	1.0670	0.3556
	NcWRKY31	NcWRKY68	Ⅱb	0.3438	1.4880	0.2309
	NcWRKY48	NcWRKY53	Ⅱb	0.2196	1.0670	0.2058
	NcWRKY50	NcWRKY64	Ⅱe	0.2693	1.3330	0.2020
串联复制 Tandem duplication	NcWRKY18	NcWRKY20	Ⅱa	0.1617	0.6405	0.2525