小果白刺HAK5基因亚家族鉴定及盐胁迫下的表达分析

徐国轩; 张会龙; 李荣; 段蓉风; 罗志斌; 杨秀艳

doi:10.12302/j.issn.1000-2006.202512030

小果白刺HAK5基因亚家族鉴定及盐胁迫下的表达分析

徐国轩, 张会龙, 李荣, 段蓉风, 罗志斌, 杨秀艳

南京林业大学学报（自然科学版） ›› 0

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南京林业大学学报（自然科学版） ›› 0 DOI: 10.12302/j.issn.1000-2006.202512030

小果白刺HAK5基因亚家族鉴定及盐胁迫下的表达分析

徐国轩^1,2, 张会龙^1,2, 李荣^1,2, 段蓉风^1,2, 罗志斌^1,2,*, 杨秀艳^1,2,*

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Identification and Expression Analysis of HAK5 Gene Subfamily in Nitraria sibirica Pall.

Xu Guoxuan^1,2, Zhang Huilong^1,2, Li Rong^1,2, Duan Rongfeng^1,2, Luo Zhibin^1,2,*, Yang Xiuyan^1,2,*

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摘要

【目的】鉴定小果白刺(Nitraria sibirica Pall.)高亲和K⁺转运蛋白NsHAK5亚家族成员,明确NsHAK5s基因响应盐胁迫的表达模式。【方法】使用TBtools等软件利用生物信息学方法对NsHAK5亚家族成员进行分析。对生长50d、状态良好、长势一致的无性系植株进行试验。试验苗在1/2 Hoagland营养液中进行水培,分别添加0mM、200 mM和400 mM NaCl,盐处理7d与14d后,分别收集植株的根、茎与叶组织样品。通过RT-qPCR技术分析NsHAK5亚家族成员的组织表达模式以及响应盐胁迫的表达特征。【结果】从小果白刺基因组中鉴定出6个HAK5基因,分布在4和9号染色体上。系统发育分析显示,NsHAK5亚家族分为2个进化分支。顺式作用元件分析发现NsHAK5s基因启动子区包含多个光响应、激素应答和非生物响应元件。NsHAK5s基因表达具有组织特异性,NsHAK5c、NsHAK5d、NsHAK5e与NsHAK5f在根特异性表达,NsHAK5a与NsHAK5b在叶中特异性表达。与对照相比,在盐胁迫下,NsHAK5c、NsHAK5d和NsHAK5e在根中的表达量显著升高;NsHAK5a与NsHAK5b在叶中表达量升高;而NsHAK5f在根、茎与叶中表达量均显著降低。【结论】鉴定出6个小果白刺NsHAK5基因,与对照相比,在盐胁迫下5个成员表达量上调;而另外1个成员的表达量下调。NsHAK5s基因的组织表达特异性说明NsHAK5s成员可能具有功能分化。本研究结果可为解析NsHAK5s基因调控小果白刺响应盐胁迫的分子机制提供基础。

Abstract

【Objective】To identify HAK5 subfamily members in Nitraria sibirica Pall. and analyze the expression patterns of NsHAK5s under NaCl treatments. 【Method】NsHAK5 subfamily members were analyzed using bioinformatics methods with software such as TBtools. Experiments were conducted on 50-day-old, healthy, and uniformly growing clonal plants. The test seedlings were hydroponically cultured in 1/2 Hoagland nutrient solution supplemented with 0 mM, 200 mM, and 400 mM NaCl. After 7 and 14 days of salt treatment, root, stem, and leaf tissue samples were collected from the plants. The tissue-specific expression patterns and salt stress-responsive expression characteristics of the NsHAK5 subfamily members were analyzed using RT-qPCR.【Results】Six HAK5s were identified in the genome of the N. sibirica Pall., which were distributed on the second and fourth chromosomes. Phylogenetic analysis revealed that the NsHAK5 subfamily members were segregated into two distinct clades. Analysis of cis-acting elements indicated that the promoter regions of NsHAK5s contained multiple elements which were associated with light, hormone and abiotic stress responses. The expression of NsHAK5s exhibited a tissue-specific pattern: NsHAK5c, NsHAK5d, NsHAK5e, and NsHAK5f were specifically expressed in the roots, while NsHAK5a and NsHAK5b were specifically expressed in the leaves. Under salt exposure condition, the expression levels of NsHAK5c, NsHAK5d, and NsHAK5e were significantly upregulated in the roots, the expression levels of NsHAK5a and NsHAK5b were increased in the leaves, but the expression level of NsHAK5f was significantly downregulated in all of the tissues in comparison with the expression levels of these genes in the plants treated without salinity.【Conclusion】Six NsHAK5s were identified in the genome of N. sibirica. Among them, five members exhibited upregulated expression under salinity, while one member showed the down-regulation. The tissue-specific expression patterns of NsHAK5s suggest a potential functional divergence among the members, indicating these members of N. sibirica may play distinct roles in responding to salinity. These findings provide insights for elucidating the molecular mechanisms of NsHAK5s underlying salt tolerance of N. sibirica.

导出引用

徐国轩, 张会龙, 李荣, 段蓉风, 罗志斌, 杨秀艳. 小果白刺HAK5基因亚家族鉴定及盐胁迫下的表达分析[J]. 南京林业大学学报（自然科学版）. 0 https://doi.org/10.12302/j.issn.1000-2006.202512030

Xu Guoxuan, Zhang Huilong, Li Rong, Duan Rongfeng, Luo Zhibin, Yang Xiuyan. Identification and Expression Analysis of HAK5 Gene Subfamily in Nitraria sibirica Pall.[J]. Journal of Nanjing Forestry University (Natural Sciences Edition）. 0 https://doi.org/10.12302/j.issn.1000-2006.202512030

中图分类号： S793.9

参考文献

[1] LI W, XU G, ALLI A, et al.Plant HAK/KUP/KT K+ transporters: Function and regulation[J]. Semin Cell Dev Biol, 2018, 74:133-41. DOI: 10.1016/j.semcdb.2017.07.009.
[2] WANG Y, Lü J, CHEN D, et al.Genome-wide identification, evolution, and expression analysis of the KT/HAK/KUP family in pear[J]. Genome, 2018, 61(10): 755-65. DOI: 10.1139/gen-2017-0254.
[3] EPSTEIN E.Dual pattern of ion absorption by plant cells and by plants[J]. Nature 1966, 212(5068): 1324-7. DOI: 10.1038/2121324a.
[4] SCHLEYER M, BAKKER E P.Nucleotide sequence and 3'-end deletion studies indicate that the K+-uptake protein kup from Escherichia coli is composed of a hydrophobic core linked to a large and partially essential hydrophilic C terminus[J]. J Bacteriol, 1993, 175(21): 6925-31. DOI: 10.1128/jb.175.21.6925-6931.1993.
[5] BAñUELOS M A, KLEIN R D, ALEXANDER-BOWMAN S J, et al. A potassium transporter of the yeast Schwanniomyces occidentalis homologous to the Kup system of Escherichia coli has a high concentrative capacity[J]. Embo J, 1995, 14(13): 3021-7. DOI: 10.1002/j.1460-2075.1995. tb07304. x.
[6] QUINTERO F J, BLATT M R.A new family of K+ transporters from Arabidopsis that are conserved across phyla[J]. Febs Letters, 1997, 415(2): 206-11. DOI: 10.1016/S0014-5793 (97)01125-3.
[7] FU H H, LUAN S.AtKUP1: A dual-affinity K+transporter from Arabidopsis[J]. Plant Cell, 1998, 10(1): 63-73. DOI: 10.1105/tpc.10.1.63.
[8] KIM E J, KWAK J M, UOZUMI N, et al.AtKUP1: An Arabidopsis gene encoding high-affinity potassium transport activity[J]. Plant Cell, 1998, 10(1): 51-62. DOI: 10.1105/tpc.10.1.51.
[9] LI Y, PENG L, XIE C, et al.Genome-wide identification, characterization, and expression analyses of the HAK/KUP/KT potassium transporter gene family reveals their involvement in K+ deficient and abiotic stress responses in pear rootstock seedlings[J]. Plant Growth Regul, 2018, 85(2): 187-98. DOI:10.1007/s10725-018-0382-8.
[10] RUBIO F, NIEVES-CORDONES M, ALEMáN F, et al. Relative contribution of AtHAK5 and AtAKT1 to K+ uptake in the high-affinity range of concentrations[J]. Physiol Plant, 2008, 134(4): 598-608. DOI: 10.1111/j.1399-3054.2008.01168.x.
[11] GUPTA M, QIU X, WANG L, et al.KT/HAK/KUP potassium transporters gene family and their whole-life cycle expression profile in rice (Oryza sativa)[J]. Mol Genet Genomics, 2008, 280(5): 437-52. DOI: 10.1007/s00438-008-0377-7.
[12] NIEVES-CORDONES M, RóDENAS R, CHAVANIEU A, et al. Uneven HAK/KUP/KT Protein Diversity Among Angiosperms: Species Distribution and Perspectives[J]. Front Plant Sci, 2016, 7: 127. DOI: 10.3389/fpls.2016.00127.
[13] 杨天元. 水稻高亲和钾离子转运蛋白基因OsHAK5的功能研究 [D], 南京: 南京农业大学, 2016. YANG T. Functional characterization of a high affinity potassium transporter geneOsHAK5in rice[D]. Nanjing: Nanjing Agricultural University, 2016.
[14] MAIERHOFER T, SCHERZER S, CARPANETO A, et al.ArabidopsisHAK5 under low K+ availability operates as PMF powered high-affinity K+ transporter[J]. Nat Commun, 2024, 15(1): 8558.DOI: 10.1038/s41467-024-52963-6.
[15] VéRY A A, NIEVES-CORDONES M, DALY M, et al. Molecular biology of K+ transport across the plant cell membrane: What do we learn from comparison between plant species?[J]. J Plant Physio, 2014, 171(9): 748-69. DOI: 10.1016/j.jplph.2014.01.011.
[16] MA X H, RU D F, MORALES-BRIONES D F, et al. Genome sequence and salinity adaptation of the desert shrubNitraria sibirica(Nitrariaceae, Sapindales)[J]. DNA Res, 2023, 30(3). DOI: 10.1093/dnares/dsad011.
[17] VéRY A A, SENTENAC H. Molecular mechanisms and regulation of K+ transport in higher plants[J]. Annu Rev Plant Biol, 2003, 54: 575-603. DOI: 10.1146/annurev. arplant.54.031902.134831.
[18] ADAMS E, SHIN R.Transport, signaling, and homeostasis of potassium and sodium in plants[J]. J Integr Plant Biol, 2014, 56(3): 231-49. DOI: 10.1111/jipb.12159.
[19] FALHOF J, PEDERSEN J T, FUGLSANG A T, et al.Plasma Membrane H+-ATPase Regulation in the Center of Plant Physiology[J]. Mol Plant, 2016, 9(3): 323-37. DOI: 10.1016/j.molp.2015.11.002.
[20] LIANG X, LI J, YANG Y, et al.Designing salt stress-resilient crops: Current progress and future challenges[J]. J Integr Plant Biol, 2024, 66(3): 303-29. DOI: 10.1111/jipb.13599.
[21] YANG T, ZHANG S, HU Y, et al.The role of a potassium transporter OsHAK5 in potassium acquisition and transport from roots to shoots in rice at low potassium supply levels[J]. Plant Physiol, 2014, 166(2): 945-59. DOI: 10.1104/pp.114.246520.
[22] LI S J, WU G Q, LIN L Y.AKT1, HAK5, SKOR, HKT1;5, SOS1 and NHX1 synergistically control Na+ and K+ homeostasis in sugar beet (Beta vulgarisL.) seedlings under saline conditions[J]. J Plant Biochem. Biotechnol, 2022, 31(1): 71-84. DOI: 10.1007/s13562-021-00656-2.
[23] 王尚德, 康向阳, 李代丽. 唐古特白刺果用候选优株主分量分析及初选研究[J]. 北京林业大学学报, 2006,(05): 139-42. WANG S D, KANG X Y, LI D L. Evaluation and selection of plus trees ofNitraria tangutorumBobr. by PCA [J]. J Beijing For Univ, 2006, (05): 139-42. DOI: 10.13332/j.1000-1522.2006.05.025.
[24] 武香, 倪建伟, 张华新, 等. 盐胁迫对3种白刺渗透调节物质的影响 [J]. 东北林业大学学报, 2012, 40(01): 44-7+69. WU X, NI J W, ZHANG H X, et al. Effects of salt stress on osmotic adjustment substances in three species ofNitraria[J]. J Northeast For Univ, 2012, 40(01): 44-7+69. DOI: 10.13759/j.cnki.dlxb.2012.01.026.
[25] 闫永庆, 高彦博, 刘威, 等. 外源Ca²+对盐胁迫下唐古特白刺光合作用影响[J]. 东北农业大学学报, 2016, 47(04): 57-64.YANG Y Q, GAO Y B, LIU W, et al.Effect of exogenous Ca²+ on photosynthesis ofNitraria tangutorumduring salt stress [J]. J Northeast Agric Univ, 2016, 47(04): 57-64. DOI: 10.19720/j.cnki.issn.1005-9369.2016.04.008.
[26] 张雪, 贺康宁, 史常青, 等. 盐胁迫对柽柳和白刺幼苗生长与生理特性的影响[J]. 西北农林科技大学学报(自然科学版), 2017, 45(01): 105-11. ZHANG X, HE K N, SHI C Q, et al. Effects of salt stress on growth and physiological characteristics of tamarix chinensis andNitratia tangutorumSeedlings [J]. j.Northwest A&F Univ.(Nat. Sci. Ed.), 2017, 45(01): 105-11. DOI: 10.13207/j.cnki.jnwafu.2017.01.015.
[27] 朱礼明, 黎梦娟, 张景波, 等. 西伯利亚白刺基因组信息初探[J]. 林业科学研究, 2020, 33(01): 144-51. ZHU L M, LI M J, ZHANG J B, et al. A study onNitraria sibiricaPall. Genome [J]. For Res, 2020, 33(01): 144-51. DOI: 13275/j.cnki.lykxyj.2020.01.019.
[28] HU A S, YANG X Y, ZHU J F, et al.Selection and validation of appropriate reference genes for RT-qPCR analysis ofNitraria sibiricaunder various abiotic stresses[J]. BMC Plant Biol, 2022, 22(1): 592. DOI: 10.1186/s12870-022-03988-w.
[29] 杨秀艳, 李焕勇, 朱建峰, 等. NaCl胁迫下2种白刺光合特性适应性研究[J]. 核农学报, 2017, 31(10): 2047-54. YANG X Y, LI H Y, ZHU J F, et al. Study on Photosynthetic Characteristics ofNitrariaUnder NaCl Stress [J]. j.Nucl. Agric. Sci, 2017, 31(10): 2047-54. DOI: 1000-8551( 2017) 10-2047-08.
[30] CHEN C.W Y, LI J, et al. TBtools-II: A “one for all, all for one” bioinformatics platform for biological big-data mining[J]. Mol Plant, 2023, 16(11). DOI: 10.1016/j.molp.2023.09.010.
[31] 柴薇薇, 王文颖, 崔彦农, 等. 植物钾转运蛋白KUP/HAK/KT家族研究进展[J]. 植物生理学报, 2019, 55(12): 1747-61. CHAI W W, WANG W Y, CUI Y N, et al. Research progress of function on KUP/HAK/KT family in plants [J]. Plant Physiol, 2019, 55(12): 1747-61. DOI: 10.13592/j.cnki.ppj.2019.0133.
[32] KANNO S, MARTIN L, VALLIER N, et al.Xylem K+ loading modulates K+ and Cs+ absorption and distribution in Arabidopsis under K+-limited conditions[J]. Front Plant Sci, 2023, 14: :1040118. DOI: 10.3389/fpls.2023.1040118.
[33] OKADA T, YAMANE S, YAMAGUCHI M, et al.Characterization of rice KT/HAK/KUP potassium transporters and K+ uptake by HAK1 fromOryza sativa[J]. Plant Biotechnol (Tokyo), 2018, 35(2): 101-11. DOI: 10.5511/plantbiotechnology.18.0308a.
[34] RAGEL P, RóDENAS R, GARCíA-MARTíN E, et al. The CBL-Interacting Protein Kinase CIPK23 Regulates HAK5-Mediated High-Affinity K+ Uptake inArabidopsisRoots[J]. Plant Physiol, 2015, 169(4): 2863-73.DOI: 10.1104/pp.15.01401.
[35] JIANG C, BELFIELD E J, CAO Y, et al.AnArabidopsisSoil-Salinity-Tolerance Mutation Confers Ethylene-Mediated Enhancement of Sodium/Potassium Homeostasis[J]. The Plant Cell, 2013, 25(9): 3535-52. DOI: 10.1105/tpc.113.115659.
[36] TAKAHASHI R, NISHIO T, ICHIZEN N, et al.High-affinity K+ transporter PhaHAK5 is expressed only in salt-sensitive reed plants and shows Na+ permeability under NaCl stress[J]. Plant Cell Rep, 2007, 26(9): 1673-9.DOI: 10.1007/s00299-007-0364-1.
[37] MIAN A, OOMEN R J, ISAYENKOV S, et al.Over-expression of an Na+-and K+-permeable HKT transporter in barley improves salt tolerance[J]. Plant J, 2011, 68(3): 468-79.DOI: 10.1111/j.1365-313X.2011.04701.
[38] MA L, LI J R, LI J F, et al. Plant salt-tolerance mechanisms: Classic signaling pathways, emerging frontiers,future perspectives [J]. Mol Plant, 2025, 15:S1674-2052(25)00439-3. DOI: 10.1016/j.molp.2025.12.009.
[39] PANCHY N, LEHTI-SHIU M, SHIU S-H.Evolution of Gene Duplication in Plants[J]. Plant Physiology, 2016, 171(4): 2294-316. DOI: 10.1104/pp.16.00523.
[40] LYNCH M, CONERY J S.The evolutionary fate and consequences of duplicate genes[J]. Science, 2000, 290(5494): 1151-5. DOI: 10.1126/science.290.5494.1151.
[41] 张尚宏屈良鹄. 基因组的进化与内含子中的基因的进化[J]. 中山大学学报(自然科学版), 1999,(01): 51-5. ZHANG S H, QU L H. Genome Evolution and the Evolution of Genes in Introns [J]. J Sun Yat-sen Univ (Nat Sci), 1999, (01): 51-5. DOI: 10.1038/sj.cr.7290029.