杨属植物HKT基因家族成员鉴定与盐胁迫下的表达模式分析

doi:10.3969/j.issn.1004-1524.20250023

浙江农业学报 ›› 2025, Vol. 37 ›› Issue (10): 2104-2115.DOI: 10.3969/j.issn.1004-1524.20250023

杨属植物HKT基因家族成员鉴定与盐胁迫下的表达模式分析

廖小龙¹(), 王兴胜¹, 陈勇¹, 李斌¹, 洪思丹², 梅利那², 国颖²^,^*()

1.伊犁哈萨克自治州林木良种繁育试验中心(伊犁哈萨克自治州平原林场),新疆伊犁 835311
2.南京林业大学林草学院,水土保持学院,江苏省杨树种质创新与品种改良重点实验室,南方现代林业协同创新中心,江苏南京 210037

收稿日期:2025-01-09 出版日期:2025-10-25 发布日期:2025-11-13
作者简介:廖小龙(1972—),男,四川遂宁人,硕士研究生,高级工程师,研究方向为林木良种选育。E-mail:632771570@qq.com
通讯作者: *国颖,E-mail:yingguo@njfu.edu.cn
基金资助:
科技创新2030-重大项目课题(2023ZD0405601)

Identification of the HKT gene family members in Populus species and analysis of their expression patterns under salt stress

LIAO Xiaolong¹(), WANG Xingsheng¹, CHEN Yong¹, LI Bin¹, HONG Sidan², MEI Lina², GUO Ying²^,^*()

1. Ili Experimental Center of Tree Breeding for Improved Varieties(Plain Forestry Farm), Ili Kazak Autonomous Prefecture, Ili 835311, Xinjiang, China
2. Jiangsu Key Laboratory for Poplar Germplasm Enhancement and Variety Improvement, Co-Innovation Center for the Sustainable Forestry in Southern China, College of Forestry and Grassland, College of Soil and Water Conservation, Nanjing Forestry University, Nanjing 210037, China

Received:2025-01-09 Online:2025-10-25 Published:2025-11-13

摘要/Abstract

摘要： 为探究高亲和性K⁺转运蛋白(high-affinity K⁺ transporter, HKT)在杨属植物盐胁迫响应中的功能,并筛选耐盐育种关键基因,对包括银腺杨84K在内的11个杨属(Populus)物种开展全基因组水平的HKT基因家族成员鉴定,系统分析这些成员的保守结构域、蛋白质理化性质、保守基序(motif)、染色体定位和顺式作用元件等,并结合盐胁迫下银腺杨84K的实时荧光定量PCR(qRT-PCR)和转录组测序(RNA-seq)数据解析HKT基因的时空表达谱。结果显示,共鉴定出30个HKT家族基因,其中29个基因定位于各物种的18号染色体。系统进化分析表明,这30个HKT基因均属于HKT1类,并划分为HKT1;4和HKT1;5两个亚组。顺式作用元件分析表明,启动子区域含有TC-rich repeats、MBS、G-box等重要的胁迫响应元件。PaghAHKT1;4在银腺杨84K中呈现出随着盐胁迫时间延长表达显著上调的趋势。综上,在杨属中共鉴定出30个HKT基因,这些基因在对盐胁迫的响应中发挥重要作用,其中PaghAHKT1;4可能是银腺杨84K耐盐育种的潜在靶基因。

关键词: 杨属, 高亲和性K⁺转运蛋白家族, 盐胁迫, 表达模式

Abstract:

To investigate the function of high-affinity K transporters (HKT) in the salt stress response of Populus species and to identify key genes for salt-tolerant breeding, a genome-wide identification of HKT gene family members was conducted in 11 Populus species, including Populus alba × P. glandulosa clone 84K (poplar clone 84K). Bioinformatics analyses were performed to characterize conserved domains, physicochemical properties, conserved motifs, chromosomal localization, and cis-acting elements. Additionally, the spatiotemporal expression profiles of HKT genes were analyzed using qRT-PCR and RNA-seq data from salt-treated poplar clone 84K. A total of 30 HKT family genes were identified, 29 of which were located on chromosome 18 across the species. Phylogenetic analysis revealed that all 30 HKT genes belong to the HKT1 subfamily and were further divided into two subgroups: HKT1;4 and HKT1;5. Cis-acting element analysis indicated that the promoter regions contained important stress-responsive elements, such as TC-rich repeats, MBS, and G-box. Notably, PaghAHKT1;4 exhibited a significant upregulation in expression with prolonged salt stress in poplar clone 84K. In conclusion, 30 HKT genes were identified in Populus, which play important roles in salt stress response. Among them, PaghAHKT1;4 might serve as a potential target gene for salt-tolerant breeding in poplar clone 84K.

Key words: Populus species, high-affinity K⁺ transporter(HKT) family, salt stress, expression pattern

中图分类号:

S792.11

廖小龙, 王兴胜, 陈勇, 李斌, 洪思丹, 梅利那, 国颖. 杨属植物HKT基因家族成员鉴定与盐胁迫下的表达模式分析[J]. 浙江农业学报, 2025, 37(10): 2104-2115.

LIAO Xiaolong, WANG Xingsheng, CHEN Yong, LI Bin, HONG Sidan, MEI Lina, GUO Ying. Identification of the HKT gene family members in Populus species and analysis of their expression patterns under salt stress[J]. Acta Agriculturae Zhejiangensis, 2025, 37(10): 2104-2115.

图/表 10

参考文献 39

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种/品种 Species/Cultivar	单倍型 Haplotype	组装方式 Assembly strategy
欧洲山杨P. tremula	/	基于参考基因组的基因组组装
毛果杨P. trichocarpa	/	Reference based assembly
胡杨P. euphratica	/	从头组装De novo assembly
小叶杨P. simonii	/
银白杨P. alba	/
新疆杨P. alba var. pyramidalis	/
美洲黑杨I69 P. deltoides I69	/
美洲黑杨2-2 P. deltoides 2-2	/
银腺杨(银腺杨84K)P. alba×P. tremula var. glandulosa clone 84K	单倍型A hapA
	单倍型G hapG
二倍体毛白杨P. tomentosa	单倍型A hapA
	单倍型D hapD
三倍体毛白杨P.×tomentosa Carr. clone 741	单倍型1 hap1
	单倍型2 hap2
	单倍型3 hap3

种/品种 Species/Cultivar	单倍型 Haplotype	组装方式 Assembly strategy
欧洲山杨P. tremula	/	基于参考基因组的基因组组装
毛果杨P. trichocarpa	/	Reference based assembly
胡杨P. euphratica	/	从头组装De novo assembly
小叶杨P. simonii	/
银白杨P. alba	/
新疆杨P. alba var. pyramidalis	/
美洲黑杨I69 P. deltoides I69	/
美洲黑杨2-2 P. deltoides 2-2	/
银腺杨(银腺杨84K)P. alba×P. tremula var. glandulosa clone 84K	单倍型A hapA
	单倍型G hapG
二倍体毛白杨P. tomentosa	单倍型A hapA
	单倍型D hapD
三倍体毛白杨P.×tomentosa Carr. clone 741	单倍型1 hap1
	单倍型2 hap2
	单倍型3 hap3

基因名 Gene name	正向引物序列 Forward primer sequence(5'-3')	反向引物序列 Reverse primer sequence(5'-3')
PaghHKT1;5	GGTGCTCTTCGTGGTTAT	CACAAAGATGGCTAAGGT
PaghAHKT1;4	GCCAGCATTCTTCTTATT	TGACCACAAACAGCACCA
PaghGHKT1;4	TCCCAACTCATTGTCTTC	CAAACAGCACCAAGATAG
UBQ	GTTGATTTTTGCTGGGAAGC	GATCTTGGCCTTCACGTTGT

基因名 Gene name	正向引物序列 Forward primer sequence(5'-3')	反向引物序列 Reverse primer sequence(5'-3')
PaghHKT1;5	GGTGCTCTTCGTGGTTAT	CACAAAGATGGCTAAGGT
PaghAHKT1;4	GCCAGCATTCTTCTTATT	TGACCACAAACAGCACCA
PaghGHKT1;4	TCCCAACTCATTGTCTTC	CAAACAGCACCAAGATAG
UBQ	GTTGATTTTTGCTGGGAAGC	GATCTTGGCCTTCACGTTGT

蛋白质ID Protein ID		基因ID Gene ID		氨基酸长度 Amino acid length/aa		分子量 Molecular weight/ku	等电点 Isoelectric point
PaghAHKT1;5		Pop_A18G018657		535		60.02	9.41
PaghAHKT1;4		Pop_A18G034182		470		52.95	9.01
PaghGHKT1;5		Pop_G18G019156		473		52.68	9.22
PaghGHKT1;4		Pop_G18G086088		206		23.22	9.49
PalbaHKT1;5		XP_034918867.1		535		60.02	9.41
PalbaHKT1;4		XP_034906209.1		521		58.94	9.02
PapyHKT1;4		GWHPACDA010899		621		69.63	9.03
PapyHKT1;5		GWHPACDA011033		451		50.37	9.04
PtomTh1HKT1;5		GWHPBJCQ043518		535		59.98	9.33
PtomTh3HKT1;5		GWHPBJCQ045429		535		59.79	9.26
PtomTh3HKT1;4		GWHPBJCQ045526		501		56.69	9.16
PdelHKT1;5b		EVM0018125.1		516		58.11	9.51
PdelHKT1;4b		EVM0019360.1		238		27.38	8.64
PdelHKT1;5a		KAH8481580.1		473		52.73	9.27
PdelHKT1;4a		KAH8481435.1		166		18.90	9.24
PeupHKT1;5		GWHPAAYU005564.1		535		59.96	9.29
PeupHKT1;4b		GWHPAAYU005664.1		521		58.86	9.13
PeupHKT1;4a		GWHPAAYU005662.1		422		47.86	8.85
PsiHKT1;5	Simonii00009649-RA		535		59.94		9.39
PsiHKT1;4c	Simonii00039973-RA		521		58.81		9.15
PsiHKT1;4b	Simonii00039952-RA		521		59.00		9.13
PsiHKT1;4a	Simonii00039964-RA		498		56.32		9.07
PtomhAHKT1;5	KAG6741040.1		535		60.04		9.41
PtomhAHKT1;4	KAG6741120.1		500		56.53		9.16
PtomhDHKT1;5	KAG6740023.1		473		52.60		9.16
PtreHKT1;5	Potra2n18c32115.1		535		59.79		9.45
PtreHKT1;4b	Potra2n8c18208.1		521		59.10		9.27
PtreHKT1;4a	Potra2n18c32004.1		397		44.62		9.43
PtriHKT1;5	Potri.018G132200.1.p		535		60.09		9.45
PtriHKT1;4	Potri.018G147501.2.p		168		19.40		9.62

杨属植物HKT基因家族成员鉴定与盐胁迫下的表达模式分析

Identification of the HKT gene family members in Populus species and analysis of their expression patterns under salt stress

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种/品种 Species/Cultivar	单倍型 Haplotype	同源组1 OG01	同源组2 OG02	同源组3 OG03	同源组4 OG04	同源组5 OG05
银腺杨84K P. alba×P. tremula var. glandulosa clone 84K	单倍型A hapA	1	1	0	0	0
	单倍型G hapG	1	1	0	0	0
银白杨P. alba	/	0	0	2	0	0
新疆杨P. alba var. pyramidalis	/	1	1	0	0	0
三倍体毛白杨P.×tomentosa Carr. clone 741	单倍型1 hap 1	0	0	0	1	0
三倍体毛白杨P.×tomentosa Carr. clone 741	单倍型3 hap 3	0	0	2	0	0
美洲黑杨2-2 P. deltoides 2-2	/	1	1	0	0	0
美洲黑杨Ⅰ69 P. deltoides I69	/	1	1	0	0	0
胡杨P. euphratica	/	2	1	0	0	0
小叶杨P. simonii	/	3	1	0	0	0
二倍体毛白杨P. tomentosa	单倍型A hap A	1	1	0	0	0
二倍体毛白杨P. tomentosa	单倍型D hap D	0	0	0	1	0
欧洲山杨P. tremula	/	2	1	0	0	0
毛果杨P. trichocarpa	/	1	1	0	0	0
水稻Oryza sativa	/	0	0	1	3	4
总计Total	/	14	10	5	5	4

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