PagTMK10介导生长素信号途径影响84K杨高生长和径向生长

doi:10.11707/j.1001-7488.LYKX20240766

摘要/Abstract

摘要：

目的: 通过对84K杨PagTMK基因家族进行全基因组鉴定及生信分析，选择在幼茎、形成层和木质部均高表达的PagTMK10基因，探讨对杨树高生长和径向生长的影响，为进一步阐明生长素信号途径在茎生长中的机制提供帮助。方法: 利用生物信息学方法及相关软件，鉴定该基因家族成员；利用实时荧光定量PCR技术，分析PagTMK10基因在幼茎、幼叶、幼根、顶芽、成熟茎、老根、根尖、维管形成层、木质部和韧皮部中的表达；分别将PagTMK10启动子和编码区序列构建植物P_pagTMK10::GUS启动子载体和过表达35S::PagTMK10载体，利用农杆菌介导法创制84K杨转基因材料，鉴定PagTMK10基因对杨树高生长和径向生长的影响。结果: 在84K杨基因组中鉴定到10个TMK基因家族成员，其中PagTMK10与AtTMK1同源；PagTMK10基因在幼茎、形成层和木质部中相对表达量较高；P_pagTMK10::GUS转基因杨与P_pagDR5::GUS（生长素响应报告基因）转基因杨基因表达位置重合，主要表达部位为顶芽、形成层和初生木质部，推测PagTMK10基因通过生长素诱导影响高生长和径向生长；截顶土栽苗在萌芽数和生长速率上，PagTMK10-OE株系皆显著大于对照；对生长45天和6个月的过表达PagTMK10基因的转基因株系（#10、#49）进行分析，发现45天时第12茎节间形成层和初生木质部宽度、株高和地径分别比对照提高53.7%、40.3%、22.09%和20.12%，6个月时株高和地径分别比对照提高19.42%和16.53%。结论: PagTMK10基因与生长素表达部位关联，主要在顶芽、形成层和初生木质部中表达，影响高生长和径向生长。该研究可为进一步阐明PagTMK10基因参与84K杨茎生长的分子机制提供参考。

关键词: 84K杨, 生长素信号, PagTMK10, 高生长, 径向生长

Abstract:

Objective: In this study, through genome-wide identification and bioinformatics analysis of PagTMK gene family of Populus alba × P. glandulosa, PagTMK10 gene that is highly expressed in young stems, cambium and xylem was selected to explore its effect on height and radial growth. This study aims to help for further elucidating the mechanism of auxin signaling pathway in stem growth. Method: Bioinformatics and the related software were used to identify the PagTMK10 gene family members. The expression of PagTMK10 gene in young stems, young leaves, young roots, terminal buds, mature stems, old roots, root tips, vascular cambium, xylem and phloem was analyzed by real-time fluorescence quantitative PCR. The plant P_pagTMK10::GUS promoter vector and overexpressing 35S::PagTMK10 vector were constructed by using the PagTMK10 promoter and coding region sequences, respectively. The 84K poplar transgenic materials were created by Agrobacterium-mediated method and used to identify the effects of PagTMK10 gene on poplar height and radial growth. Result: A total of 10 TMK homologous genes were identified in 84K poplar genome, among which PagTMK10 gene was homologous to AtTMK1. The relative expression of PagTMK10 gene was higher in young stems, cambium and xylem. The expression location of P_pagTMK10::GUS transgenic poplar overlapped with that of P_pagDR5::GUS (auxin response reporter gene), and the main expression sites were in terminal buds, cambium and primary xylem. It is speculated that the PagTMK10 gene affects height and radial growth through auxin induction. The number of sprouts and growth rate of truncated PagTMK10-OE seedlings grown in soil were significantly greater than that of the control seedlings. It was found that the 12th internode cambium and primary xylem, plant height and basal diameter of 45 days old transgenic lines (#10, #49) that overexpressed PagTMK10 gene increased by 53.7%, 40.3%, 22.09% and 20.12%, respectively, compared to the control. The height and basal diameter of the 6 months old transgenic lines increased by 19.42 and 16.53%, respectively, compared to the control. Conclusion: PagTMK10 gene is associated with auxin expression site, and mainly expressed in terminal buds, cambium and primary xylem, affecting height and radial growth. This study can lay a foundation for further revealing the molecular mechanism of PagTMK10 gene involved in stem growth of 84K poplar.

Key words: Populus alba × P. glandulosa, auxin signaling, PagTMK10, height growth, radial growth

中图分类号:

S718.46

余浩森,曾智新,乔静,张琦琦,焦阳,杨雪鑫,张英睿,杨玉冰,赵禹森,舒文波. PagTMK10介导生长素信号途径影响84K杨高生长和径向生长[J]. 林业科学, 2025, 61(9): 113-122.

Haosen Yu,Zhixin Zeng,Jing Qiao,Qiqi Zhang,Yang Jiao,Xuexin Yang,Yingrui Zhang,Yubing Yang,Yusen Zhao,Wenbo Shu. PagTMK10 Mediates Auxin Signaling Pathway Affecting Height and Radial Growth of Populus alba × P. glandulosa[J]. Scientia Silvae Sinicae, 2025, 61(9): 113-122.

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参考文献 0

	孔祥培, 张蒙悦, 丁兆军. 柳暗花明: 胞外生长素信号感受的新突破. 植物学报, 2023, 58 (6): 861- 865. doi: 10.11983/CBB23149
	Kong X P, Zhang M Y, Ding Z J. There is a way out-new breakthroughs in extracellular auxin sensing. Journal of Integrative Plant Biology, 2023, 58 (6): 861- 865. doi: 10.11983/CBB23149
	马　军, 徐通达. 植物非经典生长素信号转导通路解析. 生物技术通报, 2020, 36 (7): 15- 22.
	Ma J, Xu T D. Non-canonical auxin signaling pathway in plants. Biotechnology Bulletin, 2020, 36 (7): 15- 22.
	左雯腾, 孟佳慧, 卢孟柱, 等. 银腺杨生长素受体基因PagFBL3对茎生长发育的影响. 林业科学, 2023, 59 (11): 59- 67. doi: 10.11707/j.1001-7488.LYKX20230198
	Zuo W T, Meng J H, Lu M Z, et al. Effects of PagFBL3 gene on stem growth and development of Populus alba × P. glandulosa. Scientia Silvae Sinicae, 2023, 59 (11): 59- 67. doi: 10.11707/j.1001-7488.LYKX20230198
	舒文波, 赵树堂, 章晶晶, 等. 超量表达FBL1对84K杨根系和生长量影响研究. 林业科学研究, 2015, 28 (6): 871- 876. doi: 10.3969/j.issn.1001-1498.2015.06.017
	Shu W B, Zhao S T, Zhang J J, et al. Overexpressing FBL1 receptor led to root formation and growth of Populus alba × P. glandulosa cl. ‘84K’. Forest Research, 2015, 28 (6): 871- 876. doi: 10.3969/j.issn.1001-1498.2015.06.017
	徐慧芳, 陈　栩. 生长素研究现状及其在大豆育种中的应用. 中国科学: 生命科学, 2024, 54 (2): 247- 259. doi: 10.1360/SSV-2023-0069
	Xu H F, Chen X. Current opinions on auxin research and its application in soybean breeding. Scientia Sinica Vitae, 2024, 54 (2): 247- 259. doi: 10.1360/SSV-2023-0069
	张琦琦. 2024. ‘84K’杨内生菌脱菌体系及PagTMK家族部分形成层高表达基因功能研究. 武汉: 华中农业大学.
	Zhang Q Q. 2024. Endophytes debacterization system and functional study of some cambium high-expression genes in the PagTMK family of '84K'. Wuhan: Huazhong Agriculture University. ［in Chinese］
	Björklund S, Antti H, Uddestrand I, et al. Cross-talk between gibberellin and auxin in development of Populus wood: gibberellin stimulates polar auxin transport and has a common transcriptome with auxin. Plant Journal, 2007, 52 (3): 499- 511. doi: 10.1111/j.1365-313X.2007.03250.x
	Cao M, Chen R, Li P, et al. TMK1-mediated auxin signalling regulates differential growth of the apical hook. Nature, 2019, 568 (7751): 240- 243. doi: 10.1038/s41586-019-1069-7
	Chang C, Schaller G E, Patterson S E, et al. The TMK1 gene from Arabidopsis codes for a protein with structural and biochemical characteristics of a receptor protein kinase. Plant Cell, 1992, 4 (10): 1263- 1271.
	Chen Y R, Yordanov Y S, Ma C, et al. DR5 as a reporter system to study auxin response in Populus. Plant Cell Reports, 2013, 32 (3): 453- 463. doi: 10.1007/s00299-012-1378-x
	Cohen J D, Strader L C. An auxin research odyssey: 1989-2023. Plant Cell, 2024, 36 (5): 1410- 1428. doi: 10.1093/plcell/koae054
	Dai N, Wang W Y, Patterson S E, et al. The TMK subfamily of receptor-like kinases in Arabidopsis display an essential role in growth and a reduced sensitivity to auxin. PLoS One, 2013, 8 (4): e60990. doi: 10.1371/journal.pone.0060990
	Friml J, Gallei M, Gelová Z, et al. ABP1-TMK auxin perception for global phosphorylation and auxin canalization. Nature, 2022, 609 (7927): 575- 581. doi: 10.1038/s41586-022-05187-x
	Heidstra R. Asymmetric cell division in plant development. Progress in Molecular and Subcellular Biology, 2007, 45, 1- 37.
	Huang R F, Zheng R, He J, et al. Noncanonical auxin signaling regulates cell division pattern during lateral root development. Proceedings of the National Academy of Sciences of the United States of America, 2019, 116 (42): 21285- 21290.
	Kepinski S, Leyser O. The Arabidopsis F-box protein TIR1 is an auxin receptor. Nature, 2005, 435 (7041): 446- 451. doi: 10.1038/nature03542
	Li L X, Verstraeten I, Roosjen M, et al. Cell surface and intracellular auxin signalling for H⁺ fluxes in root growth. Nature, 2021, 599 (7884): 273- 277. doi: 10.1038/s41586-021-04037-6
	Lin W W, Zhou X, Tang W X, et al. TMK-based cell-surface auxin signalling activates cell-wall acidification. Nature, 2021, 599 (7884): 278- 282. doi: 10.1038/s41586-021-03976-4
	Marques-Bueno M M, Armengot L, Noack L C, et al. Auxin-regulated reversible inhibition of TMK1 signaling by MAKR2 modulates the dynamics of root gravitropism. Current Biology, 2021, 31 (1): 228- 237. doi: 10.1016/j.cub.2020.10.011
	Nilsson J, Karlberg A, Antti H, et al. Dissecting the molecular basis of the regulation of wood formation by auxin in hybrid aspen. Plant Cell, 2008, 20 (4): 843- 855. doi: 10.1105/tpc.107.055798
	Petrasek J, Friml J. Auxin transport routes in plant development. Development, 2009, 136 (16): 2675- 2688. doi: 10.1242/dev.030353
	Qiu D Y, Bai S L, Ma J C, et al. The genome of Populus alba x Populus tremula var. glandulosa clone 84K. DNA Research: an international journal for rapid publication of reports on genes and genomes, 2019, 26 (5): 423- 431. doi: 10.1093/dnares/dsz020
	Shu W B, Liu Y L, Guo Y H, et al. A Populus TIR1 gene family survey reveals differential expression patterns and responses to 1-naphthaleneacetic acid and stress treatments. Frontiers in Plant Science, 2015, 6, 719. doi: 10.3389/fpls.2015.00719
	Shu W B, Zhou H J, Jiang C, et al. The auxin receptor TIR1 homolog (PagFBL1) regulates adventitious rooting through interactions with Aux/IAA28 in Populus. Plant Biotechnology Journal, 2019, 17 (2): 338- 349. doi: 10.1111/pbi.12980
	Wang Q, Qin G C, Cao M, et al. 2020. A phosphorylation-based switch controls TAA1-mediated auxin biosynthesis in plants. Nature Communications, 11(1): 679.
	Xu C Z, Shen Y, He F, et al. Auxin-mediated Aux/IAA-ARF-HB signaling cascade regulates secondary xylem development in Populus. New Phytologist, 2019, 222 (2): 752- 767. doi: 10.1111/nph.15658
	Yang J, He H, He Y M, et al. 2021. TMK1-based auxin signaling regulates abscisic acid responses via phosphorylating ABI1/2 in Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 118(24): e2102544118.
	Yu Z P, Zhang F, Friml J, et al. Auxin signaling: research advances over the past 30 years. Journal of Integrative Plant Biology, 2022, 64 (2): 371- 392. doi: 10.1111/jipb.13225

引物名称 Primer name	引物序列 Primer sequence（5'–3'）
PagTMK10-F	CGCTGAATTGGCCGGGCACTGC
PagTMK10-R	GTCAGCAGATGTGAAAGACTCCGCA
PagUBQ-F	GTTGATTTTTGCTGGGAAGC
PagUBQ-R	GATCTTGGCCTTCACGTTGT

引物名称 Primer name	引物序列 Primer sequence（5'–3'）
TMK10-Pro-F	GCCCTCTTATTTCTTTATT
TMK10-Pro-R	TTTGTGGTGTTTTCTCAT
PMV2-JD-F（菌落鉴定Colony identification）	TATGACCATGATTACGCCAAGC
PMV2-F（同源臂Homologous arm）	TGCATCCAACGCGTTGGGAGCTC
PMV2-R（同源臂Homologous arm）	GCCTTCGCCATTCTAGACTCGAG
TMK10-CDS-F	ATGAGAAAACACCACAAAAAG
TMK10-CDS-R	CTACCGCCCATCAGCAGAAGT
2301-JD-R（菌落鉴定Colony identification）	TATGACCATGATTACGAATTCGG
2301-F（同源臂Homologous arm）	AGCTTTCGCGAGCTCGGTACC
2301-R（同源臂Homologous arm）	TCTAGAGGATCCCCGGGTACC

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[14]	段杰马履一贾黎明侯雅琴公宁宁 . 北京低山地区油松人工林立地指数表的编制及应用[J]. 林业科学, 2009, 12(3): 7-12.
[15]	郭明辉. 森林培育措施对红松人工林径向生长性质的影响[J]. 林业科学, 2003, 39(5): 100-104.