吲哚丁酸诱导桃砧木插穗不定根形成的分子机理

doi:10.3864/j.issn.0578-1752.2026.12.012

摘要/Abstract

摘要：

【目的】桃砧木扦插繁育中普遍存在生根难等问题。探索外源吲哚丁酸（3-indole butyric acid，IBA）诱导桃砧木不定根（AR）形成，从生理和分子整合视角阐明IBA促根的多维调控机制，为桃砧木高效无性繁育提供理论支撑与技术参考。【方法】以桃抗重茬无性系砧木‘GF677’硬枝插穗为试材，以清水为对照（CK），分别用100、200、300、400、500、600、700、800、900和1 000 mg·L^-1 IBA浸泡插穗基部后扦插，解析其生根过程中生理生化变化及相关基因表达特性。【结果】300 mg·L^-1 IBA为最适生根浓度，生根率达91%。生根期21 d‘GF677’CT（300 mg·L^-1 IBA）吲哚乙酸（IAA）、细胞分裂素（CTK）含量以及IAA/CTK均达峰值，‘GF677’CK（清水浸泡插穗基部）IAA和CTK含量在整个试验期间均低于‘GF677’CT；‘GF677’CT全氮（TN）含量于28 d达峰值，全磷（TP）与全钾（TK）含量在21—28 d达峰值；过氧化物酶（POD）活性在7 d最高，吲哚乙酸氧化酶（IAAO）活性则在扦插期呈先降后升趋势；两处理可溶性糖含量呈曲线下降趋势，21 d后‘GF677’CT高于‘GF677’CK；扦插期‘GF677’CK丙二醛（MDA）含量高于‘GF677’CT组，并出现两次峰值（7和21 d），‘GF677’CT在35 d MDA含量达到最大（74.07 g·kg^-1）。转录组分析显示，IBA处理显著富集于植物激素信号转导、氮代谢及糖代谢等通路。在激素通路中，Aux/IAA、AUX1等生长素响应基因及B-ARR等细胞分裂素响应因子呈现差异化表达；脱落酸（ABA）信号通路关键基因（如PYR/PYL）显著下调；氮代谢通路中，氮转运蛋白基因（Nrt）显著上调，硝酸还原酶基因（NR）则下调；糖代谢通路中，己糖激酶基因（HK3）表达增强，而蔗糖磷酸合酶基因（SPS1）及淀粉合成相关基因（SS1、GBSS、SBE1）均显著下调，表明碳代谢重心由蔗糖与淀粉合成转向分解供能。【结论】300 mg·L^-1吲哚丁酸通过协同调控内源激素稳态、驱动矿质养分与碳源向插穗基部再分配，并整合激素信号、氮同化及糖酵解/分解代谢等多层级分子通路，系统促进‘GF677’不定根形成。

关键词: 桃砧木, 硬枝扦插, 吲哚丁酸, 不定根, 生理生化, 分子机理

Abstract:

【Objective】Problems such as difficult rooting are common in the cutting propagation of peach rootstocks. The objective of this study is to explore the formation of adventitious roots (AR) of peach rootstocks induced by exogenous 3-indole butyric acid (IBA), and to clarify the multi-dimensional regulation mechanism of IBA promoting root formation from the perspective of physiological and molecular integration, so as to provide theoretical support and technical reference for efficient asexual breeding of peach rootstocks.【Method】Hardwood cuttings from the peach rootstock ‘GF677’ (a clonal line resistant to replanting) were used as experimental material. Water (CK) served as the control. Cuttings were immersed in IBA solutions at the concentrations of 100, 200, 300, 400, 500, 600, 700, 800, 900, and 1 000 mg·L^-1 at the base before being propagated by cuttings. The physiological and biochemical changes during rooting, along with related gene expression characteristics, were analyzed.【Result】300 mg·L^-1 IBA was the optimal rooting concentration, achieving a rooting rate of 91%. The indole acetic acid (IAA), cytokinin (CTK) contents and IAA/CTK in ‘GF677’ CT (treated with 300 mg·L^-1 IBA) reached the peak at 21 d of rooting stage, and the IAA and CTK contents in ‘GF677’ CK (cuttings with the base soaked in water) were lower than those in ‘GF677’ CT during the whole test period. The total nitrogen (TN) content of ‘GF677’ CT reached the peak at 28 d, and total phosphorus (TP) and total potassium (TK) contents reached the peak at 21-28 d. The peroxidase (POD) activity was highest at 7 d, while the indoleacetic oxidase (IAAO) activity decreased first and then increased at the cutting stage. The soluble sugar content of the two treatments showed a downward trend, and the ‘GF677’ CT was higher than the ‘GF677’ CK after 21 d. During the cutting period, the MDA content in the ‘GF677’ CK group was consistently higher than that in the ‘GF677’ CT group, with two peaks observed (at 7 and 21 d). In the ‘GF677’ CT group, MDA content peaked at 35 d (74.07 g·kg^-1). Transcriptome analysis revealed that IBA treatment significantly enriched pathways including plant hormone signaling, nitrogen metabolism, and sugar metabolism. In the hormone pathways, auxin-responsive genes such as Aux/IAA and AUX1, along with cytokinin response factors like B-ARR, exhibited differential expression; key genes in the abscisic acid (ABA) signaling pathway (e.g., PYR/PYL) were significantly downregulated. In the nitrogen metabolism pathway, nitrogen transporter genes (Nrt) were significantly up-regulated, while nitrate reductase genes (NR) were down-regulated. In the sugar metabolism pathway, hexokinase genes (HK3) showed enhanced expression, while sucrose phosphate synthase genes (SPS1) and starch synthesis-related genes (SS1, GBSS, SBE1) were significantly down-regulated, indicating a shift in carbon metabolism from sucrose and starch synthesis toward breakdown for energy supply.【Conclusion】300 mg·L^-1 IBA promotes adventitious root formation in ‘GF677’ by synergistically regulating endogenous hormone homeostasis, driving the redistribution of mineral nutrients and carbon sources toward the basal region of cuttings, and integrating multi-level molecular pathways including hormone signaling, nitrogen assimilation, and glycolysis/catabolism.

Key words: peach rootstock, hardwood cutting, 3-indole butyric acid (IBA), adventitious root, physiology and biochemistry, molecular mechanism

张帆, 王晨冰, 任家玄, 李宇. 吲哚丁酸诱导桃砧木插穗不定根形成的分子机理[J]. 中国农业科学, 2026, 59(12): 2697-2711.

ZHANG Fan, WANG ChenBing, REN JiaXuan, LI Yu. Molecular Mechanism of IBA-Induced Adventitious Root Formation in Peach Rootstock Cuttings[J]. Scientia Agricultura Sinica, 2026, 59(12): 2697-2711.

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参考文献 31

[1]	何平, 李林光, 王海波, 常源升. 基于转录组分析不同着色桃果皮花青苷表达模式与转录因子. 植物生理学报, 2019, 55(3): 310-318.
	HE P, LI L G, WANG H B, CHANG Y S. Analysis of anthocyanin expression patterns and transcription factors in differently colored peach fruit peel based on transcriptome. Plant Physiology Journal, 2019, 55(3): 310-318. (in Chinese)
[2]	宋海岩, 赵科, 李靖, 郑健萍, 崔晓龙, 李景亚, 陈栋. 桃砧木GF677硬枝扦插繁殖技术的正交优化研究. 西昌学院学报(自然科学版), 2023, 37(4): 1-5.
	SONG H Y, ZHAO K, LI J, ZHENG J P, CUI X L, LI J Y, CHEN D. Research of orthogonal optimization of hardwood cutting propagation technology of peach rootstock GF677. Journal of Xichang University (Natural Science Edition), 2023, 37(4): 1-5. (in Chinese)
[3]	姜林, 张翠玲, 邵永春, 于福顺, 董军晓, 王博. 国内外桃树育苗技术研究进展. 北方果树, 2011(2): 1-5.
	JIANG L, ZHANG C L, SHAO Y C, YU F S, DONG J X, WANG B. Research progress on peach tree seedling technology in China and abroad. Northern Fruits, 2011(2): 1-5. (in Chinese)
[4]	贾志远, 葛晓敏, 唐罗忠. 木本植物扦插繁殖及其影响因素. 世界林业研究, 2015, 28(2): 36-41.
	JIA Z Y, GE X M, TANG L Z. Research progress in cutting propagation technology for woody plants and its affecting factors. World Forestry Research, 2015, 28(2): 36-41. (in Chinese)
[5]	REIGHARD G L, LORETI F. Rootstock development// The Peach: Botany, Production and Uses. Wallingford: CABI, 2008: 193-220.
[6]	张帆, 王鸿, 张雪冰, 陈建军. 吲哚丁酸对桃砧木GF677不定根形成过程中生理生化指标的影响. 云南农业大学学报(自然科学), 2023, 38(6): 1025-1031.
	ZHANG F, WANG H, ZHANG X B, CHEN J J. Effects of 3-indole butyric acid (IBA) on the physiological and biochemical indexes during adventitious root formation of peach rootstock GF677. Journal of Yunnan Agricultural University (Natural Science), 2023, 38(6): 1025-1031. (in Chinese)
[7]	张帆, 王鸿, 陈建军, 张雪冰. 桃无性系砧木不定根形成影响因子研究. 寒旱农业科学, 2023, 2(1): 59-65.
	ZHANG F, WANG H, CHEN J J, ZHANG X B. Study on influencing factors of adventitious root formation in peach clonal rootstocks. Journal of Cold-Arid Agricultural Science, 2023, 2(1): 59-65. (in Chinese)
[8]	张帆, 王鸿. 桃绿枝扦插研究进展. 中国果树, 2019(2): 20-25.
	ZHANG F, WANG H. Research progress on greenwood cutting of peach. China Fruits, 2019(2): 20-25. (in Chinese)
[9]	王小玲, 高柱, 余发新, 刘腾云, 王碧琴. 观赏羽扇豆离体培养生根关联酶活性及可溶性蛋白含量变化规律研究. 云南农业大学学报(自然科学版), 2010, 25(6): 835-839.
	WANG X L, GAO Z, YU F X, LIU T Y, WANG B Q. Study on changes of root-related enzyme activities and soluble protein content during in vitro rooting of ornamental lupin. Journal of Yunnan Agricultural University (Natural Science), 2010, 25(6): 835-839. (in Chinese)
[10]	鲁如坤. 土壤农业化学分析方法. 北京: 中国农业科学技术出版社, 2000.
	LU R K. Analytical Methods for Soil Agrochemistry. Beijing: China Agricultural Science and Technology Press, 2000. (in Chinese)
[11]	卢楠, 孟丙南, 孙宇涵, 李云, 王少明, 郭志民, 王青龙, 徐慧敏. 四倍体刺槐黄化嫩枝扦插生根过程生理生化分析. 东北林业大学学报, 2013, 41(11): 5-9.
	LU N, MENG B N, SUN Y H, LI Y, WANG S M, GUO Z M, WANG Q L, XU H M. Physiological and biochemical analysis during rooting of etiolated softwood cuttings in tetraploid Robinia pseudoacacia Journal of Northeast Forestry University, 2013, 41(11): 5-9. (in Chinese)
[12]	郭文啸, 孙强, 侯霄帆. 木槿嫩枝扦插生根技术研究. 上海农业科技, 2020(2): 72-74, 111.
	GUO W X, SUN Q, HOU X F. Study on rooting technology of Hibiscus syriacus softwood cuttings. Shanghai Agricultural Science and Technology, 2020(2): 72-74, 111. (in Chinese)
[13]	张焕欣, 董春娟, 李福凯, 王红飞, 尚庆茂. 植物不定根发生机理的研究进展. 西北植物学报, 2017, 37(7): 1457-1464.
	ZHANG H X, DONG C J, LI F K, WANG H F, SHANG Q M. Research advances in the mechanism of adventitious root formation in plants. Acta Botanica Boreali-Occidentalia Sinica, 2017, 37(7): 1457-1464. (in Chinese)
[14]	MURTHY H N, DANDIN V S, PAEK K Y. Tools for biotechnological production of useful phytochemicals from adventitious root cultures. Phytochemistry Reviews, 2016, 15(1): 129-145. doi: 10.1007/s11101-014-9391-z
[15]	ZHANG Y, GUO J, REN F, JIANG Q, ZHOU X, ZHAO J, LIU X. Integrated physiological, transcriptomic, and metabolomic analyses of the response of peach to nitrogen levels during different growth stages. International Journal of Molecular Sciences, 2022, 23(18): 10876. doi: 10.3390/ijms231810876
[16]	孙雪莲, 杨楚童, 胡亚楠, 邹显花. 植物扦插生根机理的研究进展. 农学学报, 2021, 11(10): 33-40. doi: 10.11923/j.issn.2095-4050.cjas2020-0016
	SUN X L, YANG C T, HU Y N, ZOU X H. Rooting mechanism of plant cuttings: A review. Journal of Agriculture, 2021, 11(10): 33-40. (in Chinese) doi: 10.11923/j.issn.2095-4050.cjas2020-0016
[17]	王因花, 燕丽萍, 孔雨光, 吴德军, 任飞, 梁静. 绒毛白蜡嫩枝扦插生根的解剖学特征与内源激素变化. 中南林业科技大学学报, 2023, 43(11): 28-35, 52.
	WANG Y H, YAN L P, KONG Y G, WU D J, REN F, LIANG J. Anatomical characteristics and endogenous hormone changes during softwood cutting rooting of Fraxinus velutina. Journal of Central South University of Forestry & Technology, 2023, 43(11): 28-35, 52. (in Chinese)
[18]	李斌. 长柄扁桃嫩枝扦插繁殖技术与生根机理研究[D]. 北京: 中国林业科学研究院, 2017.
	LI B. Research on softwood cutting propagation technology and rooting mechanism of Prunus pedunculata[D]. Beijing: Chinese Academy of Forestry, 2017. (in Chinese)
[19]	LIAO C. Advances on internal influence factors of adventitious rooting of forest trees. Agricultural Science & Technology, 2017, 18(7): 1168-1173.
[20]	CHAPMAN E J, ESTELLE M. Mechanism of auxin-regulated gene expression in plants. Annual Review of Genetics, 2009, 43: 265-285. doi: 10.1146/annurev-genet-102108-134148 pmid: 19686081
[21]	GUAN L, LI Y, HUANG K, CHENG Z M. Auxin regulation and MdPIN expression during adventitious root initiation in apple cuttings. Horticulture Research, 2020, 7: 143. doi: 10.1038/s41438-020-00364-3
[22]	FIDLER J, GRASKA J, GIETLER M, NYKIEL M, PRABUCKA B, RYBARCZYK-PŁOŃSKA A, MUSZYŃSKA E, MORKUNAS I, LABUDDA M. PYR/PYL/RCAR receptors play a vital role in the abscisic-acid-dependent responses of plants to external or internal stimuli. Cells, 2022, 11: 1352. doi: 10.3390/cells11081352
[23]	HWANG I, SHEEN J, MÜLLER B. Cytokinin signaling networks. Annual Review of Plant Biology, 2012, 63: 353-380. doi: 10.1146/annurev-arplant-042811-105503 pmid: 22554243
[24]	GUTIERREZ L, BUSSELL J D, PĂCURAR D I, SCHWAMBACH J, PĂCURAR M, BELLINI C. Phenotypic plasticity of adventitious rooting in Arabidopsis is controlled by complex regulation of auxin response factor transcripts and microRNA abundance. The Plant Cell, 2009, 21(10): 3119-3132. doi: 10.1105/tpc.108.064758
[25]	ZHANG Q, SHI M, TANG F, SU N, JIN F, PAN Y, CHU L, LU M, SHU W, LI J. Transcriptome analysis reveals the hormone signalling coexpression pathways involved in adventitious root formation in Populus. Forests, 2023, 14(7): 1436. doi: 10.3390/f14071436
[26]	张帆, 王鸿. 桃硬枝扦插生根机理研究进展. 植物生理学报, 2019, 55(11): 1595-1606.
	ZHANG F, WANG H. Research advances in the mechanism of hardwood cutting rooting in peach. Plant Physiology Journal, 2019, 55(11): 1595-1606. (in Chinese)
[27]	宋金耀, 刘永军, 宋刚, 王桂峰, 王贺山. 几个常见树种扦插生根过程中POD、IAAO活性及酚含量的变化. 江苏农业科学, 2007(6): 115-118.
	SONG J Y, LIU Y J, SONG G, WANG G F, WANG H S. Changes of POD, IAAO activities and phenolic content in cuttings of several common tree species during rooting. Jiangsu Agricultural Sciences, 2007(6): 115-118. (in Chinese)
[28]	翟亚芳, 刘贤德, 吕东, 赵明, 赵祜, 赵兴鹏, 王艺林, 赵玉红. 植物生长调节剂对鞑靼忍冬扦插生根及酶活性变化的影响. 中南林业科技大学学报, 2021, 41(7): 52-61.
	ZHAI Y F, LIU X D, LÜ D, ZHAO M, ZHAO H, ZHAO X P, WANG Y L, ZHAO Y H. Effects of plant growth regulators on rooting and enzyme activity changes in cuttings of Lonicera tatarica. Journal of Central South University of Forestry & Technology, 2021, 41(7): 52-61. (in Chinese)
[29]	LI C, HOU X, MOU K, LIU H, ZHAO Z, LIAO W. The involvement of abscisic acid in glucose promoted adventitious root development in cucumber. Scientia Horticulturae, 2022, 295: 110816. doi: 10.1016/j.scienta.2021.110816
[30]	TONG C, LI C, CAO X Y, SUN X D, BAO Q X, MU X R, LIU C Y, LOAKE G J, CHEN H H, MENG L S. Long-distance transport of sucrose in source leaves promotes sink root growth by the EIN3-SUC2 module. PLoS Genetics, 2022, 18(9): e1010424.
[31]	赵加欣, 张静亚, 尚庆茂, 谢露露, 董春娟. 西瓜接穗对嫁接苗根系糖代谢及生长发育的影响. 西北植物学报, 2020, 40(7): 1171-1179.
	ZHAO J X, ZHANG J Y, SHANG Q M, XIE L L, DONG C J. Effect of watermelon scion on sugar metabolism and growth in the roots of grafted seedlings. Acta Botanica Boreali-Occidentalia Sinica, 2020, 40(7): 1171-1179. (in Chinese)

分组 Grouping	样本 Sample	测序数据质控后总读长 Clean reads	质控后总碱基数 Clean bases	GC含量 GC content (%)	Q20 (%)	Q30 (%)
G-A0	G-0d-C1	48130898	7189962548	45.58	99.52	97.80
	G-0d-C2	45815524	6845394843	45.55	99.52	97.79
	G-0d-C3	43127264	6433008530	45.58	99.54	97.93
G-A1	G-7d-C1	44570274	6653185431	45.69	99.54	97.95
	G-7d-C2	45239118	6755163127	45.79	99.59	98.13
	G-7d-C3	44526412	6653526828	45.74	99.50	97.73
G-B1	G-7d-T1	44462268	6641804574	45.64	99.56	98.03
	G-7d-T2	47407368	7073820172	45.55	99.53	97.86
	G-7d-T3	43640848	6508022015	45.55	99.55	97.97
G-A2	G-14d-C1	43332016	6457823345	45.61	99.57	98.03
	G-14d-C2	49177942	7347009809	45.70	99.56	97.96
	G-14d-C3	44863076	6702573052	45.66	99.59	98.14
G-B2	G-14d-T1	45916926	6861470515	45.55	99.65	98.21
	G-14d-T2	48791798	7293256377	45.56	99.58	98.06
	G-14d-T3	47935658	7163207778	45.60	99.59	98.13
G-A3	G-21d-C1	48129584	7192980193	45.55	99.56	97.98
	G-21d-C2	44523382	6650245512	45.55	99.55	97.95
	G-21d-C3	43457842	6495042709	45.56	99.54	97.88
G-B3	G-21d-T1	44817512	6700792576	45.61	99.56	97.99
	G-21d-T2	46635968	6972283857	45.56	99.52	97.79
	G-21d-T3	49185990	7351281479	45.65	99.55	97.95
G-A4	G-28d-C1	47880586	7157271070	45.51	99.54	97.88
	G-28d-C2	39522204	5888832483	45.80	99.59	98.11
	G-28d-C3	46997642	7021070203	45.47	99.58	98.08
G-B4	G-28d-T1	40929878	6115913412	45.55	99.61	98.16
	G-28d-T2	47449228	7084928273	45.59	99.59	98.08
	G-28d-T3	42246048	6310988514	45.56	99.61	98.18
G-A5	G-35d-C1	52348502	7813615221	45.80	99.66	98.36
	G-35d-C2	49788944	7432809520	45.94	99.68	98.50
	G-35d-C3	47084246	7023737201	45.71	99.64	98.35
G-B5	G-35d-T1	43244576	6458406795	45.48	99.54	97.90
	G-35d-T2	43516188	6493553276	45.53	99.49	97.70
	G-35d-T3	46003980	6869384171	45.59	99.57	98.03