Identification of AKR gene family members in Halogeton glomeratus and salt tolerance analysis of the root salt stress response gene HgAKR42639

doi:10.11686/cyxb2023335

Abstract

Abstract:

In order to explore the biological function of the AKR gene family and the salt tolerance of the root salt stress response gene HgAKR42639 in Halogeton glomeratus， the aldo-keto reductase gene （AKRs） was identified based on full-length transcriptome sequencing of H. glomeratus， and the protein sequence encoded by AKRs in H. glomeratus was analyzed by bioinformatics methods. The target gene expression， physiological indexes and sodium and potassium content in roots of H. glomeratus and the Arabidopsis thaliana HgAKR42639 gene were measured under 200 mmol·L^-1 NaCl stress for 0， 6， 12， 24 and 48 hours. We found that 23 HgAKRs were identified from the H. glomeratus transcriptome， encoding amino acid sequences ranging from 165 to 664 aa， and the prediction of subcellular localization was mainly in the cytoplasm. AKR protein conserved domains were highly similar with 3 motifs， and there were core elements， enhancing elements and stress response elements in the analysis of promoter cis-acting elements. The results of qRT-PCR showed that the relative expression of HgAKR42639 increased initially and then decreased in both H. glomeratus roots and A. thaliana， and the expression of HgAKR42639 peaked at 24 hours. With increase in exposure time to salt stress， the activities of catalase （CAT）， peroxidase （POD）， superoxide dismutase （SOD） and the proline content showed an increasing trend， while the contents of malondialdehyde （MDA） and soluble protein showed a downward trend， reaching their lowest levels at 24 hours. The content of Na⁺ and K⁺ decreased under the salt stress treatment for 24 hours， and K⁺/Na⁺ reached the maximum value. In summary， the AKR family genes of H. glomeratus were obtained in this study， and the results provide a theoretical basis for further research on the molecular mechanism of regulation of response to salt stress in AKR genes in H. glomeratus. This research provides a foundation for further verification of the HgAKR42639 gene role in salt tolerance.

Key words: Halogeton glomeratus, AKR gene family, bioinformatics analysis, salt stress

Zhen-huan ZHANG, Li-rong YAO, Jun-cheng WANG, Er-jing SI, Hong ZHANG, Ke YANG, Xiao-le MA, Ya-xiong MENG, Hua-jun WANG, Bao-chun LI. Identification of AKR gene family members in Halogeton glomeratus and salt tolerance analysis of the root salt stress response gene HgAKR42639[J]. Acta Prataculturae Sinica, 2024, 33(7): 68-83.

Figures/Tables 14

References 43

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引物名称Name of primers	引物序列Sequences of primers （5’→3’）	用途Purpose
HgAKR42639-F	AGGAAGCACATCGTTGAGGG	实时荧光定量PCR Real-time fluorescence quantitative PCR
HgAKR42639-R	TTCATTGCCCGGACAGTCTC	实时荧光定量PCR Real-time fluorescence quantitative PCR
HgActin-F	TGTTCTCAGTGGTGGTACAA	盐生草内参基因引物 Reference gene primers in H. glomeratus
HgActin-R	GTGCCACCACCTTAATCTTC	盐生草内参基因引物 Reference gene primers in H. glomeratus
AtActin-F	ACCCAAAGGCCAACAGAGAG	拟南芥内参基因引物 Reference gene primers in A. thaliana
AtActin-R	CACGTCCAGCAAGGTCAAGA	拟南芥内参基因引物 Reference gene primers in A. thaliana

引物名称Name of primers	引物序列Sequences of primers （5’→3’）	用途Purpose
HgAKR42639-F	AGGAAGCACATCGTTGAGGG	实时荧光定量PCR Real-time fluorescence quantitative PCR
HgAKR42639-R	TTCATTGCCCGGACAGTCTC	实时荧光定量PCR Real-time fluorescence quantitative PCR
HgActin-F	TGTTCTCAGTGGTGGTACAA	盐生草内参基因引物 Reference gene primers in H. glomeratus
HgActin-R	GTGCCACCACCTTAATCTTC	盐生草内参基因引物 Reference gene primers in H. glomeratus
AtActin-F	ACCCAAAGGCCAACAGAGAG	拟南芥内参基因引物 Reference gene primers in A. thaliana
AtActin-R	CACGTCCAGCAAGGTCAAGA	拟南芥内参基因引物 Reference gene primers in A. thaliana

序号Number	名称Name	基因登录号Gene accession ID	序号Number	名称Name	基因登录号Gene accession ID
1	HgAKR01	Hg02G012880.1	13	HgAKR13	Hg16G006890.1
2	HgAKR02	Hg02G012940.1	14	HgAKR14	Hg16G006900.1
3	HgAKR03	Hg03G002550.1	15	HgAKR15	Hg16G006910.1
4	HgAKR04	Hg07G000220.1	16	HgAKR16	Hg171G001710.2
5	HgAKR05	Hg09G006350.1	17	HgAKR17	Hg214G001090.1
6	HgAKR06	Hg09G010750.1	18	HgAKR18	Hg238G000730.2
7	HgAKR07	Hg09G010800.1	19	HgAKR19	Hg248G001020.1
8	HgAKR08	Hg102G000790.1	20	HgAKR20	Hg30G002300.1
9	HgAKR09	Hg15G004360.1	21	HgAKR21	Hg465G000090.1
10	HgAKR10	Hg16G006840.1	22	HgAKR22	Hg53G002170.1
11	HgAKR11	Hg16G006850.1	23	HgAKR42639	Hg02G042639.1
12	HgAKR12	Hg16G006870.1

序号Number	名称Name	基因登录号Gene accession ID	序号Number	名称Name	基因登录号Gene accession ID
1	HgAKR01	Hg02G012880.1	13	HgAKR13	Hg16G006890.1
2	HgAKR02	Hg02G012940.1	14	HgAKR14	Hg16G006900.1
3	HgAKR03	Hg03G002550.1	15	HgAKR15	Hg16G006910.1
4	HgAKR04	Hg07G000220.1	16	HgAKR16	Hg171G001710.2
5	HgAKR05	Hg09G006350.1	17	HgAKR17	Hg214G001090.1
6	HgAKR06	Hg09G010750.1	18	HgAKR18	Hg238G000730.2
7	HgAKR07	Hg09G010800.1	19	HgAKR19	Hg248G001020.1
8	HgAKR08	Hg102G000790.1	20	HgAKR20	Hg30G002300.1
9	HgAKR09	Hg15G004360.1	21	HgAKR21	Hg465G000090.1
10	HgAKR10	Hg16G006840.1	22	HgAKR22	Hg53G002170.1
11	HgAKR11	Hg16G006850.1	23	HgAKR42639	Hg02G042639.1
12	HgAKR12	Hg16G006870.1

序号Number	基因编号 Genetic code	氨基酸数 Amino acid quantity （aa）	等电点 Isoelectric point （PI）	分子量 Molecular weight （Da）	分子式 Molecular formula	负电荷的残基总数Total number of negatively charged residues	正电荷的残基总数Total number of positively charged residues	脂肪系数 Aliphatic index	不稳定系数 Instability index	亲水性平均值Grand average of hydropathicity
1	HgAKR01	317	6.00	35036.93	C₁₅₇₉H₂₄₅₅N₄₁₉O₄₆₅S₉	34	30	89.53	43.19	-0.240
2	HgAKR02	316	5.57	34807.62	C₁₅₆₂H₂₄₃₆N₄₁₄O₄₆₇S₁₀	36	29	89.49	43.46	-0.234
3	HgAKR03	310	5.76	34512.44	C₁₅₃₆H₂₄₃₅N₄₂₁O₄₆₀S₁₁	41	34	92.81	27.01	-0.215
4	HgAKR04	311	5.84	34942.01	C₁₅₇₃H₂₄₇₀N₄₁₄O₄₆₅S₁₀	42	35	92.15	39.65	-0.336
5	HgAKR05	334	6.20	37468.25	C₁₆₈₅H₂₆₅₉N₄₄₇O₄₈₈S₁₅	38	35	93.65	45.44	-0.244
6	HgAKR06	324	6.34	35556.17	C₁₆₀₆H₂₅₄₁N₄₁₃O₄₆₇S₁₄	37	35	91.17	38.70	-0.126
7	HgAKR07	351	6.12	38937.70	C₁₇₁₅H₂₇₃₉N₄₆₉O₅₂₁S₂₁	43	39	84.96	34.53	-0.300
8	HgAKR08	350	5.66	38224.84	C₁₇₀₀H₂₇₂₃N₄₅₅O₅₁₇S₁₃	45	39	92.80	26.30	-0.174
9	HgAKR09	664	6.20	73942.76	C₃₂₉₃H₅₂₁₀N₈₉₈O₉₇₇S₂₉	90	80	85.78	40.27	-0.320
10	HgAKR10	338	5.70	37428.78	C₁₆₆₄H₂₆₂₆N₄₅₂O₄₉₉S₁₅	46	39	84.56	25.72	-0.275
11	HgAKR11	309	6.21	34774.88	C₁₅₆₂H₂₄₅₃N₄₁₅O₄₅₉S₁₂	43	39	90.91	22.12	-0.238
12	HgAKR12	165	6.20	18704.43	C₈₃₂H₁₃₁₃N₂₂₇O₂₄₇S₈	21	19	87.45	25.62	-0.250
13	HgAKR13	338	5.68	37512.86	C₁₆₆₈H₂₆₃₀N₄₅₂O₅₀₁S₁₅	46	38	85.12	29.62	-0.272
14	HgAKR14	343	5.57	37951.12	C₁₆₇₁H₂₆₅₂N₄₆₂O₅₁₆S₁₅	48	39	83.06	37.63	-0.385
15	HgAKR15	317	8.38	35411.57	C₁₅₈₂H₂₄₉₃N₄₂₉O₄₆₇S₁₃	35	38	87.03	34.98	-0.298
16	HgAKR16	371	9.07	41209.57	C₁₈₅₇H₂₉₂₆N₄₈₈O₅₃₈S₁₆	28	36	91.48	34.66	-0.149
17	HgAKR17	322	5.48	35141.09	C₁₅₆₃H₂₄₈₇N₄₁₅O₄₈₁S₁₁	42	34	94.50	33.28	-0.170
18	HgAKR18	328	7.59	36492.74	C₁₆₃₄H₂₅₆₉N₄₃₉O₄₈₄S₁₂	37	38	87.68	34.03	-0.295
19	HgAKR19	298	9.03	33006.76	C₁₄₈₇H₂₃₃₉N₃₉₉O₄₃₆S₇	31	37	89.36	12.19	-0.308
20	HgAKR20	323	6.24	36545.87	C₁₆₂₁H₂₅₅₂N₄₅₄O₄₇₅S₁₇	44	39	84.18	41.19	-0.401
21	HgAKR21	220	5.71	24337.91	C₁₀₈₀H₁₇₁₈N₂₉₆O₃₂₃S₁₀	30	25	94.41	36.36	-0.156
22	HgAKR22	384	7.57	42245.83	C₁₈₇₉H₂₉₇₄N₅₁₈O₅₇₅S₇	46	47	87.89	24.76	-0.396
23	HgAKR42639	317	6.25	35151.21	C₁₅₇₅H₂₄₇₃N₄₁₉O₄₆₈S₁₂	36	34	90.41	32.06	-0.219