苹果幼苗劈接愈合进程可溶性糖动态积累研究

doi:10.16420/j.issn.0513-353x.2024-1062

摘要/Abstract

摘要：

以‘平邑甜茶’（Malus hupehensis）实生自根苗嫁接株系为试材，解析嫁接愈合进程中可溶性糖积累量变化与动态分布，分析糖影响嫁接愈合的关键时期与影响嫁接愈合的关键可溶性糖组分，探究嫁接时外源施加可溶性糖对嫁接愈合的调控作用。结果表明：环境湿度100%条件下嫁接成活率与接穗叶面积、嫁接口可溶性糖积累量显著正相关，而室内环境（环境湿度30%）条件下嫁接成活率只与可溶性糖积累量/蒸腾率比值显著正相关，接穗叶面积为2 ~ 4 cm²时，可溶性糖积累量/蒸腾率比值达到峰值，嫁接成活率最高（95%）；根据嫁接后愈伤组织发育状态与分化特征，将嫁接愈合进程划分为5个愈合时期：隔离层期、愈伤细胞发生期、愈伤组织融合期、维管束分化期和维管系统建连期；愈伤组织融合期起始阶段（嫁接后7 d）可溶性糖在嫁接口新生愈伤组织大量富集，总糖含量以及6种糖组分含量相对变化率均在愈合口处不同时期有显著波动。依据茎糖组分含量配置6种糖水溶液，分别在嫁接时对伤口与叶片施加外源处理，发现1%棉子糖水溶液处理伤口、1%蔗糖水溶液处理伤口与叶片均对促维管束建连有重要贡献（VIP值 > 1），以上3种处理方式均能显著加速愈伤组织的发育，从而促进嫁接愈合进程。

关键词: 平邑甜茶, 嫁接愈合, 可溶性糖, 外源喷施, 棉子糖, 蔗糖

Abstract:

The self-grafted seedlings of Malus hupehensis were used as test materials to analyze the changes and dynamic distribution of soluble sugar accumulation in the process of grafting healing. The key period of sugar affecting grafting healing and the key soluble sugar components affecting grafting healing were analyzed，and the regulation of exogenous application of soluble sugar on grafting healing was explored. The results showed that the survival rate of grafting was significantly positively correlated with the leaf area of scion and the soluble sugar accumulation of grafting under the condition of 100% environmental humidity，while the survival rate of grafting was only significantly positively correlated with the ratio of soluble sugar accumulation / transpiration rate under the condition of indoor environment （30% environmental humidity）. When the leaf area of scion was 2-4 cm²，the ratio of soluble sugar accumulation/transpiration rate reached the peak，and the survival rate of grafting was the highest（95%）. According to the development status and differentiation characteristics of callus after grafting，the healing process of grafting was divided into five healing periods：isolation layer period，callus cell occurrence period，callus fusion period，vascular bundle differentiation period and construction period of vascular system. At the initial stage of callus fusion stage（7 days after grafting），soluble sugars were enriched in the new callus of the grafting joint，and the relative change rate of total sugar content and six sugar components content fluctuated significantly at different stages of the healing joint. Based on the contents of sugar components in stems，six kinds of sugar aqueous solutions were prepared to apply exogenous treatment to wounds and leaves during grafting. It was found that 1% raffinose aqueous solution treatment of wounds，1% sucrose aqueous solution treatment of wounds and 1% sucrose aqueous solution treatment of leaves all contributed significantly to the establishment of vascular bundles（VIP value > 1）. All the three aforementioned treatment methods could significantly accelerate the development of callus，thereby facilitating the graft healing process.

Key words: Malus hupehensis, grafting healing, soluble sugar, exogenous spraying, raffinose, sucrose

毛冲, 袁梓文, 黄闻霆, 项凌, 张潇, 陈琦璐, 余芷寒, 王圣元, 李天忠. 苹果幼苗劈接愈合进程可溶性糖动态积累研究[J]. 园艺学报, 2026, 53(6): 1669-1683.

MAO Chong, YUAN Ziwen, HUANG Wenting, XIANG Ling, ZHANG Xiao, CHEN Qilu, YU Zhihan, WANG Shengyuan, LI Tianzhong. Study on the Dynamic Accumulation of Soluble Sugars in the Apple Cleft Grafting Healing Process[J]. Acta Horticulturae Sinica, 2026, 53(6): 1669-1683.

图/表 9

图1 不同叶面积接穗分组与嫁接示例

Fig. 1 Grouping and grafting examples with different leaf area scions

图2 糖溶液影响嫁接愈合验证试验示意图

Fig. 2 The effect of sugar solution on grafting healing verification experiment schematic diagram

图3 饱和空气湿度（100%）下不同叶面积接穗的嫁接成活率、茎可溶性糖积累量及二者的相关性

Fig. 3 Grafting survival rate，stem soluble sugar accumulation with different leaf areas and the correlation between the two under saturated air humidity（100%）

图4 室内（空气湿度30%）不同叶面积接穗的嫁接成活率、茎可溶性糖积累量及二者的相关性

Fig. 4 Grafting survival rate，stem soluble sugar accumulation with different leaf areas and the correlation between the two under air humidity（30%）

图5 嫁接愈合进程5个特征时期的愈合口表型与解剖学观察白框箭头指示特征结构；紫色部位指示愈伤细胞分布区域；红色部位指示愈伤组织发育的特征结构。S：接穗；R：砧木；LLC：隔离层细胞；NCC：新生愈伤细胞；CCM：愈伤细胞团；SVE：小管状分子；NPC：新生韧皮部；NEC：新生表皮细胞；CC：形成层细胞；TS：管状结构

Fig. 5 Phenotypic and anatomical observation of healing site in five characteristic periods of grafting healing process The white box arrow indicates the feature structure；The purple part indicates the dispersed area of callus cells；The red part indicated the characteristic structure of callus development. S：Scion；R：Rootstock；LLC：Isolated layer cells；NCC：New callus cells；CCM：Callus cell mass；SVE：Small vessel element；NPC：New phloem cells；NEC：New epidermal cells；CC：Cambial cells；TS：Tubular structure

图6 不同愈合时期嫁接口处可溶性糖的分布与变化率过碘酸希夫染液染色，同一样品中着色度深浅表示糖类物质相对积累量的多少；黑色虚框指示了愈伤组织所在部位的糖积累。不同小写字母表示差异显著（P < 0.05）。下同

Fig. 6 Distribution and change rate of soluble sugar at grafting interface in different healing periods The color depth in the same sample indicates the relative accumulation of carbohydrates；the black frame indicated the sugar accumulation in the site of callus. Different lowercase letters represent significant differcences（P < 0.05）. The same below

图7 嫁接口处5个愈合时期6种糖含量的变化率以自嫁接前取下的茎段样品作为空白对照。正值代表增长，负值代表降低

Fig. 7 Evaluation of the change rate of the six contents in the five healing periods at the grafting interface Used the stem segment sample taken before self-grafting as a blank control. Positive values indicate increases，while negative values indicate decreases

图8 嫁接口处木质部连通率测定示例（A）与6种糖不同处理方式的木质部连通率（B） S：接穗；R：砧木

Fig. 8 The xylem connectivity rate measurement example（A）at the grafting interface and the xylem connectivity rate（B）of six different sugar treatments S：Scion；R：Rootstock

图9 影响较大的糖处理方式在对嫁接口的影响（嫁接后7 d）白色虚框指示了愈伤组织的分布部位。S：接穗；R：砧木

Fig. 9 The influence of the more influential sugar treatment methods on the graft at 7 DAG The white frame indicates the distribution site of callus. S：Scion；R：Rootstock

参考文献 45

[29]	Miao Li, Li Yansu, Fan Xingqiang, He Chaoxing, Yu Xianchang. 2017. Research advances on mechanism of interface healing of plant grafting. Plant Physiology Journal, 53 (1):17-28. (in Chinese)
	苗丽, 李衍素, 范兴强, 贺超兴, 于贤昌. 2017. 植物嫁接体接口愈合机制的研究进展. 植物生理学报, 53 (1):17-28.
[30]	Moore R, Walker D B. 1983. Studies of vegetative compatibility-incompatibility in higher plants. Protoplasma, 115 (2):114-121. doi: 10.1007/BF01279803 URL
[31]	Nie X, Li P, Men S. 2017. Studies on long distance transport of plant sterols by grafting. Acta Scientiarum Naturalium Universitatis Nankaiensis, 50 (1):47-53.
[32]	Phillips N, Reynolds A, Di Profio F. 2015. Nonstructural carbohydrate concentrations in dormant grapevine scionwood and rootstock impact propagation success and vine growth. Horttechnology, 25 (4):536-550. doi: 10.21273/HORTTECH.25.4.536 URL
[33]	Pina A, Errea P, Martens H J. 2012. Graft union formation and cell-to-cell communication via plasmodesmata in compatible and incompatible stem unions of Prunus spp. Scientia Horticulturae,143:144-150.
[34]	Porro D, Stefanini M, Dorigatti C, Ramponi M, Voltolini J A. 2006. Ecophysiological response and nutritional status of grapevine in relation to two different methods of grafting//Proceedings of the Vth international symposium on mineral nutrition of fruit plants. Leuven:International Society for Horticultural Science:71.
[35]	Stoddard F L, McCully M E. 1979. Histology of the development of the graft union in pea roots. Canadian Journal of Botany, 57 (14):1486-1501. doi: 10.1139/b79-185 URL
[36]	Wang L, Ruan Y. 2013. Regulation of cell division and expansion by sugar and auxin signaling. Frontiers in Plant Science,4:163.
[37]	Wang Lei, Gao Fangsheng, Zhang Zhihuan, Li Weiqiang, Cao Bili, Xu Kun. 2024. Change of sugar in grafted tomato fruit and correlation analysis with nitrogen and phosphorus content. Acta Horticulturae Sinica, 51 (2):361-371. (in Chinese) doi: 10.16420/j.issn.0513-353x.2023-0850
	王磊, 高方胜, 张志焕, 李炜蔷, 曹逼力, 徐坤. 2024. 嫁接番茄果实糖分变化及其与氮磷含量相关性分析. 园艺学报, 51 (2):361-371.
[1]	Asante A K, Barnett J R. 2016. Effect of temperature on graft union formation in mango Mangifera indica L 401. Tropical Agriculture,75:401-404.
[2]	Bolouri-Moghaddam M R, Le Roy K, Xiang L, Rolland F, van den Ende W. 2010. Sugar signalling and antioxidant network connections in plant cells. The FEBS Journal, 277 (9):2022-2037. doi: 10.1111/ejb.2010.277.issue-9 URL
[3]	Button J. 1978. The Effects of some carbohydrates on the growth and organization of citrus ovular callus. Zeitschrift für Pflanzenphysiologie, 88 (1):61-68. doi: 10.1016/S0044-328X(78)80118-4 URL
[4]	Christiaens F, Canher B, Lanssens F, Bisht A, Stael S, de Veylder L, Heyman J. 2021. Pars pro toto:every single cell matters. Frontiers in Plant Science,12:656825.
[5]	Dabirian S, Miles C A. 2017. Increasing survival of splice-grafted watermelon seedlings using a sucrose application. HortScience, 52 (4):579-583. doi: 10.21273/HORTSCI11667-16 URL
[6]	Devi P, Miles C A. 2021. Increasing survival and efficacy of splice-grafted watermelon using sucrose and antitranspirant//2nd International Symposium on Vegetable Grafting: 73-78.
[7]	Díaz A O, García A M, Devincenti C V, Goldemberg A L. 2005. Ultrastructure and histochemical study of glycoconjugates in the gills of the white croaker(Micropogonias furnieri). Anatomia Histologia Embryologia, 34 (2):117-122. pmid: 15771674
[8]	Duan Y D, Zhang F, Meng X M, Shang Q M. 2024. Spatio-temporal dynamics of phytohormones in the tomato graft healing process. Horticultural Plant Journal, 10 (6):1362-1370. doi: 10.1016/j.hpj.2022.11.014 URL
[9]	Errea P, Felipe A, Herrero M. 1994. Graft establishment between compatible and incompatible Prunus spp. Journal of Experimental Botany, 45 (3):393-401. doi: 10.1093/jxb/45.3.393 URL
[38]	Yang Zhijian, Feng Jinling, Chen Hui. 2013. Study on the anatomical structures in development of the nurse seed grafted union of Camellia oleifera. Plant Science Journal, 31 (3):313-320. (in Chinese) doi: 10.3724/SP.J.1142.2013.30313 URL
	杨志坚, 冯金玲, 陈辉. 2013. 油茶芽苗砧嫁接口愈合过程解剖学研究. 植物科学学报, 31 (3):313-320.
[39]	Yildiz K, Yilmaz H, Balta F. 2003. The effect of soluble sugars,total flavan and juglone concentrations in walnut scions on graft survival. Journal of the American Pomological Society, 57 (4):173-176. doi: 10.71318/apom.2003.57.4.173 URL
[40]	Zhang A, Matsuoka K, Kareem A, Robert M, Roszak P, Blob B, Bisht A, De Veylder L, Voiniciuc C, Asahina M, Melnyk C W. 2022. Cell-wall damage activates DOF transcription factors to promote wound healing and tissue regeneration in Arabidopsis thaliana. Current Biology, 32 (9):1883. doi: 10.1016/j.cub.2022.02.069 URL
[41]	Zhang H, Yuan F, Guan D, Wang A, Wu J, Jin C J. 2017. A review on water transport in xylem of vascular plants and its affecting factors. Chinese Journal of Ecology,36:3281-3288.
[42]	Zhang J, Wang P, Tian H, Wang Y, Jiang H. 2019. Using a new hybrid rootstock significantly increases the grafted plant rate and watermelon yield. International Agrophysics, 33 (1):97-106. doi: 10.31545/intagr/104402
[43]	Zhou Naifu, Zhang Junpei, Liu Hao, Cha Weiwei, Pei Dong. 2018. New protocols for paraffin sections of heterogeneous tissues of woody plants. Chinese Bulletin of Botany, 53 (5):653-660. (in Chinese)
	周乃富, 张俊佩, 刘昊, 查巍巍, 裴东. 2018. 木本植物非均质化组织石蜡切片制作方法. 植物学报, 53 (5):653-660. doi: 10.11983/CBB18051
[44]	Zhu Jie, Li Lu, Sun Mingfei, Gao Meina, Liang Bowen, Xu Jizhong, Li Zhongyong. 2023. Effects of different growth type scions on root architecture of self-rooted young apple trees. Acta Horticulturae Sinica, 50 (10):2183-2191. (in Chinese) doi: 10.16420/j.issn.0513-353x.2022-0771
	朱杰, 李潞, 孙明飞, 高美娜, 梁博文, 徐继忠, 李中勇. 2023. 普通型与短枝型苹果接穗对嫁接幼树根系构型的影响. 园艺学报, 50 (10):2183-2191. doi: 10.16420/j.issn.0513-353x.2022-0771
[10]	Espen L, Cocucci M, Sacchi G. 2005. Differentiation and functional connection of vascular elements in compatible and incompatible pear/quince internode micrografts. Tree Physiology,25:1419-1425.
[11]	Fu M M, Xu Z, Ma H Y, Hao Y F, Tian J, Wang Y, Zhang X Z, Xu X F, Han Z H, Wu T. 2024. Characteristics of long-distance mobile mRNAs from shoot to root in grafted plant species. Horticultural Plant Journal, 10 (1):25-37. doi: 10.1016/j.hpj.2023.05.009 URL
[12]	Giannino F, Mele B H, de Micco V, Toraldo G, Mazzoleni S, Cartenì F. 2019. An individual based model of wound closure in plant stems. IEEE Access,7:65821-65827.
[13]	Goldschmidt E. 2014. Plant grafting:new mechanisms,evolutionary implications. Frontiers in Plant Science,5:727.
[14]	Gross K C, Pharr D M, Locy R D. 1981. Growth of callus initiated from cucumber hypocotyls on galactose and galactose-containing oligosaccharides. Plant Science Letters, 20 (4):333-341. doi: 10.1016/0304-4211(81)90248-0 URL
[15]	Guan C, Xu Y, Yue H, Qiao X, Chen G, Yuan M, Wang Y, Sun J. 2025. Difference in sucrose concentration between scion and rootstock influences the incompatibility of cucumber/pumpkin grafted plants. Horticultural Plant Journal, 11 (3):1166-1180. doi: 10.1016/j.hpj.2024.02.014 URL
[16]	Hare P D, Cress W A, van Staden J. 1998. Dissecting the roles of osmolyte accumulation during stress. Plant Cell and Environment, 21 (6):535-553. doi: 10.1046/j.1365-3040.1998.00309.x URL
[17]	Hou Yixing. 2022. Preliminary study on graft compatibility of different stock-scion combinations of grapes[Ph. D. Dissertation]. Urumqi: Xinjiang Agricultural University. (in Chinese)
	侯毅兴. 2022. 葡萄不同砧穗组合嫁接亲和性初步研究[博士论文]. 乌鲁木齐: 新疆农业大学.
[18]	Ikeuchi M, Rymen B, Sugimoto K. 2020. How do plants transduce wound signals to induce tissue repair and organ regeneration? Current Opinion in Plant Biology,57:72-77.
[19]	Kawaguchi K, Notaguchi M, Okayasu K, Sawai Y, Kojima M, Takebayashi Y, Sakakibara H, Otagaki S, Matsumoto S, Shiratake K. 2024. Plant hormone profiling of scion and rootstock incision sites and intra- and inter-family graft junctions in Nicotiana benthamiana. Plant Signaling & Behavior, 19 (1):2331358.
[20]	Koch K. 2004. Sucrose metabolism:regulatory mechanisms and pivotal roles in sugar sensing and plant development. Current Opinion in Plant Biology, 7 (3):235-246. doi: 10.1016/j.pbi.2004.03.014 URL
[21]	Li T, Zhang Y M, Liu Y, Li X D, Hao G L, Han Q H, Dirk L, Downie A B, Ruan Y L, Wang J M, Wang G Y, Zhao T Y. 2020. Raffinose synthase enhances drought tolerance through raffinose synthesis or galactinol hydrolysis in maize and Arabidopsis plants. Journal of Biological Chemistry, 295 (23):8064-8077. doi: 10.1074/jbc.RA120.013948 URL
[22]	Liang H, Liu J F, Shi X F, Ge M H, Zhu J H, Wang D H, Zhou M B. 2024. An integrated analysis of anatomical and sugar contents identifies how night temperatures regulate the healing process of oriental melon grafted onto pumpkin. Plants-Basel, 13 (11):1506.
[23]	Lin Huangfang, Cheng Saichuan, Lin Biying, Chen Yanzhen, Ye Huilan, Shao Qingqing, Lin Honghui, Zhai Tingkai, Wang Kai, Shen Baoying. 2025. Research progress on the healing process of cucumber grafting and related influencing factors. Acta Horticulturae Sinica, 52 (10):2799-2815. doi: 10.16420/j.issn.0513-353x.2024-0688
	林黄昉, 程赛川, 林碧英, 陈琰臻, 叶惠澜, 邵卿卿, 林鸿辉, 翟挺楷, 王锴, 申宝营. 2025. 黄瓜嫁接愈合进程及相关影响因素研究进展. 园艺学报, 52 (10):2799-2815. doi: 10.16420/j.issn.0513-353x.2024-0688
[24]	Liu Minghu, Xin Zhiming, Xu Jun, Sun Fei, Dou Lijun, Li Yonghua. 2013. Influence of leaf size of plant on leaf transpiration and temperature in arid regions. Chinese Journal of Plant Ecology, 37 (5):436-442. (in Chinese) doi: 10.3724/SP.J.1258.2013.00045
	刘明虎, 辛智鸣, 徐军, 孙非, 窦立军, 李永华. 2013. 干旱区植物叶片大小对叶表面蒸腾及叶温的影响. 植物生态学报, 37 (5):436-442. doi: 10.3724/SP.J.1258.2013.00045
[25]	Loupit G, Fonayet J, Prigent S, Prodhomme D, Renault-Spilmont A, Hilbert G, Franc C, Revel G, Richard T, Ollat N, Cookson S. 2022. Identifying early metabolite markers of successful graft union formation in grapevine. Horticulture Research,9:uhab070.
[26]	Mackey M, Kurosky A, Robb E, Nazar R. 2018. A graft mimic strategy for Verticillium resistance in tomato. Molecular Biotechnology, 60 (9):665-669. doi: 10.1007/s12033-018-0101-8
[27]	Melnyk C W, Schuster C, Leyser O, Meyerowitz E M. 2015. A developmental framework for graft formation and vascular reconnection in Arabidopsis thaliana. Current Biology, 25 (10):1306-1318. doi: 10.1016/j.cub.2015.03.032 pmid: 25891401
[28]	Miao L, Li Q, Sun T, Chai S, Wang C L, Bai L Q, Sun M T, Li Y S, Qin X, Zhang Z H, Yu X C. 2021. Sugars promote graft union development in the heterograft of cucumber onto pumpkin. Horticulture Research, 8 (1):146. doi: 10.1038/s41438-021-00580-5 pmid: 34193850
[45]	Zhu Taifu, Wang Yun, Su Xiaolan, Li Xin, Wang Lianchun. 2022. Physiological characteristics of rootstock-scion and their relationship with graft affinity during the grafting union process of grapefruit. Jiangsu Agricultural Sciences, 22 (13):50. (in Chinese)
	朱太富, 王赟, 苏效兰, 李欣, 王连春. 2022. 葡萄柚嫁接愈合过程中砧穗生理物质特性及其与嫁接亲和性的关系. 江苏农业科学, 22 (13):50.