香蕉Ca2+-ATPase基因家族的鉴定及功能分析

doi:10.3864/j.issn.0578-1752.2025.07.013

Abstract

Abstract:

【Objective】Ca²⁺-ATPase is a vital Ca²⁺ efflux channel in plants, which plays an important role in maintaining cell homeostasis and inducing plant response to stress. In this study, bioinformatics and molecular biology were used to analyze the members of Ca²⁺-ATPase family in banana A genome, detect the changes of gene expression and verify gene function, so as to provide reference for exploring the mechanism of banana salt tolerance.【Method】BLAST the members of banana A genome in banana genome database by gene family name combined with Arabidopsis and rice Ca²⁺-ATPase sequences, and use Pfam, ExPASy, Cell-PLoc, NCBI, MEGA-X, TBtools and other online websites or software to predict and analyze the protein physicochemical properties, subcellular localization, conservative domain and cis-acting elements of banana Ca²⁺-ATPase. The expression of the family genes was analyzed by qRT-PCR, and the subcellular localization and prokaryotic expression of the key genes were analyzed.【Result】In this study, 20 members of Ca²⁺-ATPase family were identified in banana A genome, including 13 MaACAs and 7 MaECAs; proteins analysis of physicochemical properties showed that Ca²⁺-ATPase contained 6-11 transmembrane structures, encoding amino acids between 857 and 1 103. Subcellular localization predicted that MaACAs might be located on plasma membrane, chloroplast, endoplasmic reticulum and tonoplast, while MaECAs was located in endoplasmic reticulum. The conserved motifs are highly consistent, including four conserved domains peculiar to the family except MaACA13, responses to light, hormone, defense and stress. qRT-PCR results showed that other members of Ca²⁺-ATPase gene family except MaECA3 were up-regulated in Brazilian banana cells treated with 100 mmol·L^-1 NaCl, in which MaACA5 and MaACA10 were up-regulated to a large extent, and the expression of these two genes was affected by Ca²⁺. The results of subcellular localization of tobacco leaves showed that MaACA5 and MaACA10 were located on the plasma membrane, and the recombinant plasmids pET28a-MaACA5, pET28a-MaACA10 and transformed into E. coli BL21 were superior to the control strains under the conditions of 800 mmol·L^-1NaCl, 800 mmol·L^-1 mannitol and 50 ℃. 【Conclusion】 20 Ca²⁺-ATPase family members were identified in banana A genome, the gene structure was highly conservative, including hormone, defense and stress-related responses, MaACA5 and MaACA10 related to banana salt tolerance were screened and prokaryotic expression to verify the gene function, which provides a reference for further study of banana salt tolerance.

Key words: brazil banana, salt stress, calcium, Ca²⁺-ATPase, expression analysis, gene cloning, subcellular localisation

TENG MengXin, XU Ya, HE Jing, WANG Qi, QIAO Fei, LI JingYang, LI XinGuo. Identification and Functional Analysis of Ca²⁺-ATPase Gene Family in Banana[J].Scientia Agricultura Sinica, 2025, 58(7): 1418-1433.

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References 34

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蛋白 Protein	氨基酸数量 Amino acids size (aa)	分子量 Mw (Da)	等电点 pI	不稳定系数 The instability index (II)	脂肪系数Aliphatic index	亲水性平均系数 Grand average of hydropathicity	跨膜结构Transmembrane domain	亚细胞定位 Subcellular localization
MaACA1	1095	119281.68	8.36	38.1	101.13	0.05	8	A
MaACA2	1090	119195.56	8.27	36.89	100.53	0.04	11	A
MaACA3	1034	112541.57	5.87	35.13	103.09	0.22	6	B、C、D
MaACA4	1082	118244.8	8.47	40.33	101.71	0.066	7	A
MaACA5	1017	111767.31	5.7	31.68	99.45	0.121	7	A
MaACA6	1020	111212.01	5.88	35.51	103	0.2	6	B、C、D
MaACA7	1020	111518.14	5.91	32.3	102.91	0.158	6	B、C、D
MaACA8	1034	114138.1	6.74	31.15	107.53	0.169	10	B、C、D
MaACA9	1019	111002.4	5.26	32.57	104.26	0.201	6	B、C、D
MaACA10	1024	112321.63	5.85	34.05	95.55	0.091	8	A
MaACA11	1035	114343.58	8.3	30.9	106.03	0.144	10	B、C、D
MaACA12	1103	120673.88	8.87	35.9	102.1	0.072	9	A
MaACA13	857	92860.11	6.01	37.29	109.78	0.266	6	A
MaECA1	1064	116317.6	5.28	35.82	98.15	0.089	7	C
MaECA2	1059	115754.37	5.36	37.3	99.53	0.095	9	C
MaECA3	1051	116158.69	5.4	33.61	100.35	0.131	8	C
MaECA4	1058	115980.52	5.19	36.63	99.5	0.078	7	C
MaECA5	1000	109753.56	5.87	36.37	99.62	0.222	8	C
MaECA6	1070	118771.74	5.85	35.3	97.03	0.107	8	C
MaECA7	1059	115636.05	5.26	36.29	98.32	0.098	9	C

蛋白Protein	α-螺旋Alpha-helix (%)	β-折叠Beta strand (%)	无规则卷曲Loop (%)
MaACA1	44.20	12.24	43.56
MaACA2	46.42	11.83	41.74
MaACA3	49.13	12.38	38.49
MaACA4	47.41	12.20	40.39
MaACA5	46.02	15.44	38.54
MaACA6	48.73	14.51	36.76
MaACA7	50.49	13.14	36.37
MaACA8	48.55	14.02	37.43
MaACA9	50.54	13.35	36.11
MaACA10	46.97	15.43	37.60
MaACA11	48.79	14.30	36.91
MaACA12	47.60	10.79	41.61
MaACA13	49.01	13.54	37.46
MaECA1	51.13	10.90	37.97
MaECA2	51.65	10.86	37.49
MaECA3	52.71	10.56	36.73
MaECA4	50.85	11.15	38.00
MaECA5	47.30	13.30	39.40
MaECA6	53.64	10.84	35.51
MaECA7	47.69	11.43	40.89

Identification and Functional Analysis of Ca²⁺-ATPase Gene Family in Banana

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Identification and Functional Analysis of Ca2+-ATPase Gene Family in Banana

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Identification and Functional Analysis of Ca²⁺-ATPase Gene Family in Banana