Identification of the HXK Gene Family in Cucumis melo and Their Expression Analysis under Abiotic Stresses

doi:10.13560/j.cnki.biotech.bull.1985.2024-0418

Abstract

Abstract:

【Objective】 To clarify the genetic characteristics and potential functions of the hexokinase family in melon（Cucumis melo）. 【Method】 The melon HXK gene family was identified at the whole genome level, and its protein's physicochemical properties, subcellular localization, systematic evolution, gene structure, chromosome localization, conserved motifs, protein structure, promoter cis acting elements, etc., were analyzed. Based on transcriptome data, the expression patterns of CmHXK gene in male flowers, female flowers, roots, leaves, fruits, and fruit development processes were analyzed. In addition, the expression patterns of CmHXK gene under abscisic acid, drought, salt, and low temperature stress were measured by RT-qPCR. 【Result】A total of six CmHXK gene family members were identified on chromosomes 2, 4, 5 and 8, containing 6-9 exons, 5-8 introns, with an average amino acid number of 462, an average molecular weight of 50.2 kD and an isoelectric point of 4.99-6.34. Systematic evolutionary analysis showed that the melon HXK members had the closest homology with cucumber. The main cis-acting elements in the melon CmHXK promoter were light responsive elements, hormone responsive elements, and stress responsive elements. The expression of CmHXKs was specific in different organs. Among them, CmHXK1, CmHXK2, and CmHXK5 expressed at high level in male flowers, female flowers, roots, leaves, and fruits during the development and maturation of fruits. However, the expressions of CmHXK3, CmHXK4, and CmHXK6 decreased with the development and maturation of fruits. The responses of CmHXKs to the four abiotic（abscisic acid, drought, salt, and cold）stresses differed. Among them, CmHXK1, CmHXK3, and CmHXK4 were strongly induced by ABA. CmHXK1, CmHXK3, and CmHXK6 were strongly induced by salt stress, and CmHXK1, CmHXK2, and CmHXK3 were strongly induced by drought stress. CmHXK1 and CmHXK6 were strongly induced by cold stress. 【Conclusion】 The CmHXK family are highly conserved and play important roles in the growth, development, and abiotic stress in melon. Among them, CmHXK1 is the most sensitive to four abiotic stresses.

Key words: melon, hexokinase, bioinformatics, abiotic stress, expression analysis

ZHANG Shuai-bo, YIN Jin-peng, WANG Ji-qing, XIAO Huai-juan, LI Meng. Identification of the HXK Gene Family in Cucumis melo and Their Expression Analysis under Abiotic Stresses[J]. Biotechnology Bulletin, 2024, 40(12): 102-112.

Figures/Tables 11

References 41

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基因Gene	正向引物序列Forward primer sequence（5'-3'）	反向引物序列Reverse primer sequence（5'-3'）
CmHXK1	GTTGGACAAGATGTGGTAGG	CCAGTTCCTAGAATCACAGC
CmHXK2 CmHXK3 CmHXK4 CmHXK5 CmHXK6	CAGGTGGTTGATGCTATGG GACAGATTCCACCTTTCTCC ATGGAGGAGAGTAGTTAGGG TTGCTGCTGTGATATTAGGC ATGGTTGATGATACTGTTGGG	CTTCAAAGTAAGAGACCTCTGG CAAGCAACAACATCTTTCCC CAGGGAGATTATCAACGTAGG CTTCTTCAGCCATCTTGACC GACTCTATATAGGCAGCATTGG
CmActin7	GTGATGGTGTGAGTCACACTGTTC	ACGACCAGCAAGGTCCAAAC

基因Gene	正向引物序列Forward primer sequence（5'-3'）	反向引物序列Reverse primer sequence（5'-3'）
CmHXK1	GTTGGACAAGATGTGGTAGG	CCAGTTCCTAGAATCACAGC
CmHXK2 CmHXK3 CmHXK4 CmHXK5 CmHXK6	CAGGTGGTTGATGCTATGG GACAGATTCCACCTTTCTCC ATGGAGGAGAGTAGTTAGGG TTGCTGCTGTGATATTAGGC ATGGTTGATGATACTGTTGGG	CTTCAAAGTAAGAGACCTCTGG CAAGCAACAACATCTTTCCC CAGGGAGATTATCAACGTAGG CTTCTTCAGCCATCTTGACC GACTCTATATAGGCAGCATTGG
CmActin7	GTGATGGTGTGAGTCACACTGTTC	ACGACCAGCAAGGTCCAAAC

功能Function	元件Element	CmHXK1	CmHXK2	CmHXK3	CmHXK4	CmHXK5	CmHXK6
脱落酸ABA	ABRE	0	0	0	3	0	3
生长素Auxin	TGA-element	1	1	0	0	1	0
乙烯ETH	ERE	1	0	2	6	6	3
赤霉素GA	P-box	0	1	1	0	0	1
赤霉素GA	w box	1	0	0	2	1	0
茉莉酸甲酯MeJa	CGTCA-motif	1	2	0	0	1	2
茉莉酸甲酯MeJa	TGACG-motif	1	2	0	0	2	2
水杨酸SA	As-1	1	1	0	1	1	0
	TCA	1	0	1	0	0	1
	TCA-element	2	0	1	0	0	0
	DRE	0	0	0	3	1	0
非生物胁迫Abiotic stresses	LTR	0	0	0	0	1	0
	STRE	2	1	0	3	1	2
	WRE3	1	0	1	1	0	0
	WUN-motif	2	2	0	0	0	0
	TC-rich repeats	0	0	0	0	1	0
光响应Light	3-AFlbinding site	0	0	0	1	0	1
	AE-box	0	0	0	0	1	2
	ATCT-motif	1	0	0	0	0	0
	Box 4	3	3	2	3	2	2
	Box II	0	1	0	0	0	0
	chs-CMAla	0	0	0	1	0	0
	GA-motif	0	0	0	0	0	1
	GATA-motif	1	1	0	2	1	1
	G-Box	0	0	0	1	0	2
	G-box	0	1	0	2	1	2
	GT1-motif	2	0	0	2	1	0
	I-box	1	0	0	0	2	0
	Sp1	0	0	1	0	0	0
	TCCC-motif	0	3	0	0	0	0
	TCT-motif	1	1	3	0	1	0

功能Function	元件Element	CmHXK1	CmHXK2	CmHXK3	CmHXK4	CmHXK5	CmHXK6
脱落酸ABA	ABRE	0	0	0	3	0	3
生长素Auxin	TGA-element	1	1	0	0	1	0
乙烯ETH	ERE	1	0	2	6	6	3
赤霉素GA	P-box	0	1	1	0	0	1
赤霉素GA	w box	1	0	0	2	1	0
茉莉酸甲酯MeJa	CGTCA-motif	1	2	0	0	1	2
茉莉酸甲酯MeJa	TGACG-motif	1	2	0	0	2	2
水杨酸SA	As-1	1	1	0	1	1	0
	TCA	1	0	1	0	0	1
	TCA-element	2	0	1	0	0	0
	DRE	0	0	0	3	1	0
非生物胁迫Abiotic stresses	LTR	0	0	0	0	1	0
	STRE	2	1	0	3	1	2
	WRE3	1	0	1	1	0	0
	WUN-motif	2	2	0	0	0	0
	TC-rich repeats	0	0	0	0	1	0
光响应Light	3-AFlbinding site	0	0	0	1	0	1
	AE-box	0	0	0	0	1	2
	ATCT-motif	1	0	0	0	0	0
	Box 4	3	3	2	3	2	2
	Box II	0	1	0	0	0	0
	chs-CMAla	0	0	0	1	0	0
	GA-motif	0	0	0	0	0	1
	GATA-motif	1	1	0	2	1	1
	G-Box	0	0	0	1	0	2
	G-box	0	1	0	2	1	2
	GT1-motif	2	0	0	2	1	0
	I-box	1	0	0	0	2	0
	Sp1	0	0	1	0	0	0
	TCCC-motif	0	3	0	0	0	0
	TCT-motif	1	1	3	0	1	0

蛋白名称 Protein name	α-螺旋 α-helix/%	延伸链 Extended strand/%	β转角 β-turn/%	无规卷曲 Random/%
CmHXK1	47.39	14.06	5.22	33.33
CmHXK2	43.00	15.78	5.52	35.70
CmHXK3	44.51	13.57	4.79	37.13
CmHXK4	43.46	13.29	5.49	37.76
CmHXK5	47.59	12.85	5.62	33.94
CmHXK6	51.52	12.12	6.4	29.97