Functional characterization of Cyperus esculentus CeWRKY transcription factors in response to abiotic stress

doi:10.11686/cyxb2022150

Abstract

Abstract:

Cyperus esculentus is a new type of forage and oil crop， with its overground leaves providing high-quality herbage and its underground tubers accumulating high levels of oleic acid-enriched oils. C. esculentus also possesses strong tolerance against various stresses. WRKY transcription factor （TF） is one group among the large superfamily of transcription factors involved in growth and development， tissue metabolism， stress response and other physiological processes in the plant. However， there is limited knowledge about the WRKY family members and their functions in C. esculentus. Here， a total of 67 CeWRKY transcription factors were identified by mining transcriptome data. Phylogenetic analysis showed that CeWRKYs were classified into three groups which we designated I， Ⅱ and Ⅲ. CeWRKYs have both the typical WRKY domain and a zinc finger domain. Seven CeWRKYs exhibited mutation in the WRKY domains， which may be related to their functional divergence. The sequence length， molecular weight， and isoelectric point of CeWRKY proteins are predicted to vary greatly. CeWRKY proteins are predicted to be hydrophilic proteins without a transmembrane structure. These proteins were predicted to be localized in the nucleus. Temporal and spatial expression of CeWRKY genes were detected during C. esculentus tuber development.Different CeWRKY genes displayed differential expression patters in C. esculentus seedlings in response to different stresses. The expression profile of the CeWRKY8 gene changed significantly in C. esculentus seedlings exposed to drought， salt， or alkali stresses. This CeWRKY8 may be a major regulator of stress response in C. esculentus. The present study establishes the essential framework for complete understanding of the diverse biological function of CeWRKY transcription factors and molecular mechanisms mediating stress response in C. esculentus， providing a scientific pathway for developing new varieties of C. esculentus with high quality， enhanced biomass yield and strong stress resistance.

Key words: Cyperus esculentus, WRKY transcription factors, gene family identification, expression profiles, abiotic stress

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.

Figures/Tables 10

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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
Ⅱd	CeWRKY46	ATCATGCCTCTTCCTCCTCTTC	CGGTGCTGTTGTTCCTCCT
Ⅱe	CeWRKY14	CTACACGGCTGAGCACAATC	TTATCATGCCTTCTTCCGACTC
Ⅱe	CeWRKY15	GCCACCACAAGCTCCATTA	TGTAATCTTCTCGCTCTTCTCC
Ⅲ	CeWRKY24	TTCTGAGCCATCATCGTCTGT	CCTCCAAGTATGTCCATCCTCT
	CeWRKY38	ACCGCAACACGCAAGGAT	ATCTCAGGTTGGAACGGCTTA
	CeWRKY40	TTCTGAGCCATCATCGTCTGT	CCTCCAAGTATGTCCATCCTCT
	CeWRKY47	GATCCAACCGTGTTCATCATCA	GCACATCATCTTCACCGTCTT
	18SrRNA	CTACGTCCCTGCCCTTTGTACA	ACACTTCACCGGACCATTCAA

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
Ⅱd	CeWRKY46	ATCATGCCTCTTCCTCCTCTTC	CGGTGCTGTTGTTCCTCCT
Ⅱe	CeWRKY14	CTACACGGCTGAGCACAATC	TTATCATGCCTTCTTCCGACTC
Ⅱe	CeWRKY15	GCCACCACAAGCTCCATTA	TGTAATCTTCTCGCTCTTCTCC
Ⅲ	CeWRKY24	TTCTGAGCCATCATCGTCTGT	CCTCCAAGTATGTCCATCCTCT
	CeWRKY38	ACCGCAACACGCAAGGAT	ATCTCAGGTTGGAACGGCTTA
	CeWRKY40	TTCTGAGCCATCATCGTCTGT	CCTCCAAGTATGTCCATCCTCT
	CeWRKY47	GATCCAACCGTGTTCATCATCA	GCACATCATCTTCACCGTCTT
	18SrRNA	CTACGTCCCTGCCCTTTGTACA	ACACTTCACCGGACCATTCAA

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

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

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