Identification and Expression Analysis of PIN Gene Family in Melon Under High Temperature Stress

doi:10.13560/j.cnki.biotech.bull.1985.2022-1196

Abstract

Abstract:

Auxin efflux carrier protein PIN（PIN-formed）is an important carrier element for regulating polar auxin transport in plants, thus it plays an important role in plant growth and development. However，it is still unclear about the members, properties, chromosomal distribution, phylogeny, promoters of PIN proteins in the genome of melon and expression characteristics of family members under high temperature stress. In the present study, 18 CmPINs members were screened and identified in the whole genome database of Cucumis melo using bioinformatics methods. The number of amino acids encoded by CmPIN family members was 51-642, the molecular weight was 5.19-70.30 kD and the instability index was 24.33-48.87. The results of subcellular localization prediction showed that CmPINs members were mainly distributed in the plasma membrane. There were the transmembrane structures for rest 16 PIN proteins except no transmembrane structure in CmPIN6 and CmPIN8. CmPINs family members were distributed on 9 chromosomes and contained 1-9 Motif. The genes of 18 CmPINs family members contained 1-10 exons and their secondary structure was mainly α-spiral and irregular curly structure. Based on the PIN protein evolutionary tree of CmPIN family members and Arabidopsis（Arabidopsis thaliana）, rice（Oryza sativa）, tomato（Solanum lycopersicum）, corn（Zea mays）, results showed that 18 CmPINs were clustered into 7 subfamilies, with the largest number in subgroup VI. The analysis of cis-acting elements of the promoter revealed that there were a large number of cis-acting basic elements in the melon CmPINs promoters, as well as cis-acting elements related to hormones, light signals and drought induction. Fluorescence quantitative analysis of melon radicles treated with different temperatures showed that the transcriptional expression of CmPIN13 and CmPIN18 genes with Yucasin+IAA treatment were significantly higher than those with Yucasin treatment under high temperature stress; moreover, the transcriptional expressions of CmPIN2, CmPIN13 and CmPIN18 genes with NPA+IAA treatment were significantly higher than those with NPA treatment, which suggested that CmPINs gene family were involved in the tolerance regulation during melon germination under high temperature stress.

Key words: melon, PIN gene family, gene characteristic, bioinformatics, high temperature stress, transcriptional expression

LI Jing-rui, WANG Yu-bo, XIE Zi-wei, LI Chang, WU Xiao-lei, GONG Bin-bin, GAO Hong-bo. Identification and Expression Analysis of PIN Gene Family in Melon Under High Temperature Stress[J]. Biotechnology Bulletin, 2023, 39(5): 192-204.

Figures/Tables 10

References 34

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基因 Gene	序列 Sequence（5'-3'）		退火温度 Annealing temperature Tm/℃	GC含量 GC content/%	产物长度 Product length/bp
Actin3	F	GGCAGTGGTGGTGAACATG	59.04	57.89	149
Actin3	R	TTCTGGTGATGGTGTGAGTC	57.16	50.00	149
CmPIN3	F	CTTCTCCTGTTTCTGATGTGT	51.40	42.90	123
CmPIN3	R	GTTCTGTCTTGGTGTCTCTTC	52.00	47.60	123
CmPIN13	F	CAGGTGATGAAAACTAAACGCCA	59.75	43.48	172
CmPIN13	R	CCGGTGCTCAAAATGTCAGG	59.48	55.00	172
CmPIN18	F	GAGTAGGAGGAACAGAGAAAG	50.10	47.60	155
CmPIN18	R	GACTAAGGACCAAGTGAGACC	52.70	52.40	155
CmPIN2	F	GGGTTTGGTGGTGGGTAAGA	59.52	55.00	172
CmPIN2	R	TTTTGAGTTTGACCGATGCAGG	59.71	45.45	172

基因 Gene	序列 Sequence（5'-3'）		退火温度 Annealing temperature Tm/℃	GC含量 GC content/%	产物长度 Product length/bp
Actin3	F	GGCAGTGGTGGTGAACATG	59.04	57.89	149
Actin3	R	TTCTGGTGATGGTGTGAGTC	57.16	50.00	149
CmPIN3	F	CTTCTCCTGTTTCTGATGTGT	51.40	42.90	123
CmPIN3	R	GTTCTGTCTTGGTGTCTCTTC	52.00	47.60	123
CmPIN13	F	CAGGTGATGAAAACTAAACGCCA	59.75	43.48	172
CmPIN13	R	CCGGTGCTCAAAATGTCAGG	59.48	55.00	172
CmPIN18	F	GAGTAGGAGGAACAGAGAAAG	50.10	47.60	155
CmPIN18	R	GACTAAGGACCAAGTGAGACC	52.70	52.40	155
CmPIN2	F	GGGTTTGGTGGTGGGTAAGA	59.52	55.00	172
CmPIN2	R	TTTTGAGTTTGACCGATGCAGG	59.71	45.45	172

基因 Gene	基因ID Gene ID	氨基酸数量 Number of amino acids	分子量 Molecular weight/kD	理论等电点 Theoretical pI	不稳定指数 Instability index	脂溶指数 Aliphatic index	总平均疏水指数 Grand average of hydropathicity	跨膜结构域数目 Transmembrane domain number	亚细胞定位 Subcellular location
CmPIN1	MELO3C001206.2	135	15.15	6.69	24.33	111.93	0.795	4	液泡膜 Tonoplast membrane
CmPIN2	MELO3C017414.2	623	68.44	9.22	41.57	85.47	-0.012	10	细胞质膜Cytomembrane
CmPIN3	MELO3C017357.2	591	64.52	8.5	43.16	88.76	0.064	10	细胞质膜Cytomembrane
CmPIN4	MELO3C009892.2	417	46.35	9.76	48.87	118.63	0.454	11	细胞质膜Cytomembrane
CmPIN5	MELO3C008726.2	402	43.62	6.23	34.38	113.26	0.501	9	液泡膜Tonoplast membrane
CmPIN6	MELO3C008744.2	51	5.25	6.51	30.81	139.61	1.208	0	细胞质Cytoplasm
CmPIN7	MELO3C031448.2	166	17.73	9.26	40.24	107.65	0.489	2	叶绿体Chloroplast
CmPIN8	MELO3C031243.2	51	5.19	7.98	30.81	141.57	1.275	0	细胞外基质Extracellular matrix
CmPIN9	MELO3C031244.2	109	12.42	7.65	27.69	102.02	0.693	3	液泡膜Tonoplast membrane
CmPIN10	MELO3C013710.2	297	32.35	7.11	45.14	114.81	0.405	6	内质网Endoplasmic reticulum
CmPIN11	MELO3C013711.2	428	46.24	5.02	40.85	117.78	0.576	10	细胞质膜Cytomembrane
CmPIN12	MELO3C017060.2	421	45.46	7.66	36.48	128.5	0.704	10	液泡膜Tonoplast membrane
CmPIN13	MELO3C019102.2	611	65.37	8.75	37.4	96.73	0.265	10	细胞质膜Cytomembrane
CmPIN14	MELO3C025264.2	417	45.67	8.86	34.71	123.72	0.748	10	细胞质膜Cytomembrane
CmPIN15	MELO3C005311.2	456	50.01	5.25	33.04	123.53	0.594	10	细胞质膜Cytomembrane
CmPIN16	MELO3C005326.2	636	69.50	7.15	36.26	91.42	0.111	10	细胞质膜Cytomembrane
CmPIN17	MELO3C018353.2	628	68.55	8.69	36.15	92.77	0.142	10	细胞质膜Cytomembrane
CmPIN18	MELO3C002132.2	642	70.30	9.01	36.31	83.99	-0.029	10	细胞质膜Cytomembrane

基因 Gene	基因ID Gene ID	氨基酸数量 Number of amino acids	分子量 Molecular weight/kD	理论等电点 Theoretical pI	不稳定指数 Instability index	脂溶指数 Aliphatic index	总平均疏水指数 Grand average of hydropathicity	跨膜结构域数目 Transmembrane domain number	亚细胞定位 Subcellular location
CmPIN1	MELO3C001206.2	135	15.15	6.69	24.33	111.93	0.795	4	液泡膜 Tonoplast membrane
CmPIN2	MELO3C017414.2	623	68.44	9.22	41.57	85.47	-0.012	10	细胞质膜Cytomembrane
CmPIN3	MELO3C017357.2	591	64.52	8.5	43.16	88.76	0.064	10	细胞质膜Cytomembrane
CmPIN4	MELO3C009892.2	417	46.35	9.76	48.87	118.63	0.454	11	细胞质膜Cytomembrane
CmPIN5	MELO3C008726.2	402	43.62	6.23	34.38	113.26	0.501	9	液泡膜Tonoplast membrane
CmPIN6	MELO3C008744.2	51	5.25	6.51	30.81	139.61	1.208	0	细胞质Cytoplasm
CmPIN7	MELO3C031448.2	166	17.73	9.26	40.24	107.65	0.489	2	叶绿体Chloroplast
CmPIN8	MELO3C031243.2	51	5.19	7.98	30.81	141.57	1.275	0	细胞外基质Extracellular matrix
CmPIN9	MELO3C031244.2	109	12.42	7.65	27.69	102.02	0.693	3	液泡膜Tonoplast membrane
CmPIN10	MELO3C013710.2	297	32.35	7.11	45.14	114.81	0.405	6	内质网Endoplasmic reticulum
CmPIN11	MELO3C013711.2	428	46.24	5.02	40.85	117.78	0.576	10	细胞质膜Cytomembrane
CmPIN12	MELO3C017060.2	421	45.46	7.66	36.48	128.5	0.704	10	液泡膜Tonoplast membrane
CmPIN13	MELO3C019102.2	611	65.37	8.75	37.4	96.73	0.265	10	细胞质膜Cytomembrane
CmPIN14	MELO3C025264.2	417	45.67	8.86	34.71	123.72	0.748	10	细胞质膜Cytomembrane
CmPIN15	MELO3C005311.2	456	50.01	5.25	33.04	123.53	0.594	10	细胞质膜Cytomembrane
CmPIN16	MELO3C005326.2	636	69.50	7.15	36.26	91.42	0.111	10	细胞质膜Cytomembrane
CmPIN17	MELO3C018353.2	628	68.55	8.69	36.15	92.77	0.142	10	细胞质膜Cytomembrane
CmPIN18	MELO3C002132.2	642	70.30	9.01	36.31	83.99	-0.029	10	细胞质膜Cytomembrane

基因 Gene	α-螺旋 Alpha helix/%	延伸链 Extended strand/%	无规则卷曲 Random coil/%	β-折叠 Beta turn/%
CmPIN1	51.11	22.22	25.19	1.48
CmPIN2	31.14	16.05	48.48	4.33
CmPIN3	30.12	15.91	49.24	4.74
CmPIN4	52.52	15.35	25.66	6.47
CmPIN5	46.02	16.67	31.59	5.72
CmPIN6	50.98	25.49	13.73	9.80
CmPIN7	24.70	27.71	39.76	7.83
CmPIN8	49.02	21.57	19.61	9.80
CmPIN9	46.79	25.69	26.61	0.92
CmPIN10	39.39	15.49	42.76	2.36
CmPIN11	40.42	18.93	36.45	4.21
CmPIN12	38.00	20.19	37.29	4.51
CmPIN13	31.75	14.24	49.75	4.26
CmPIN14	40.29	18.94	36.69	4.08
CmPIN15	41.45	17.11	38.38	3.07
CmPIN16	32.08	15.41	47.80	4.72
CmPIN17	30.89	16.40	47.13	5.57
CmPIN18	29.75	15.42	49.53	5.30