乙烯利诱导‘库尔勒香梨’萼片离区细胞程序性死亡特征及调控信号研究

doi:10.16420/j.issn.0513-353x.2024-0372

摘要/Abstract

摘要：

为明确乙烯利诱导‘库尔勒香梨’萼片脱落的细胞程序性死亡特征及调控信号，以‘库尔勒香梨’为研究材料，于大蕾期喷施300 mg · L^-1乙烯利和等量清水（对照），在离区形成前期、形成期、形成后期采集幼果，观察萼片脱落外部形态特征和离区细胞显微结构及超微结构，测定萼片脱落过程中脱萼果离区和宿萼果离区对应部位乙烯浓度、活性氧水平和Ca²⁺含量。结果表明：乙烯利诱导下‘库尔勒香梨’萼片脱落过程中脱落外部形态特征、萼片离区细胞显微结构和超微结构与对照表现一致；离区形成前期和形成期，脱萼果离区和宿萼果离区对应部位乙烯浓度显著高于对照，分别高137.38%、15.02%（脱萼果）和294.95%、110.16%（宿萼果）；在离区形成期，脱萼果离区羟自由基清除速率、超氧阴离子产生速率和Ca²⁺含量均显著高于对照，分别高33.88%、9.38%和15.02%。综上所述，乙烯利处理通过增加脱萼果离区乙烯浓度、活性氧水平以及Ca²⁺含量，从而加速‘库尔勒香梨’萼片细胞程序性死亡进程，促进萼片脱落。

关键词: 库尔勒香梨, 乙烯, 萼片脱落, 细胞程序性死亡, 细胞结构, 活性氧, 钙离子

Abstract:

To clarify the characteristics of programmed cell death and the regulatory signals in the calyx abscission induced by ethephon in Korla fragrant pear，in this study，Korla fragrant pears were used as the research material，and applied 300 mg · L^-1 ethephon and equal volume of distilled water（the control）at the large bud stage. Young fruits were collected before abscission zone formation，during abscission zone formation，and after abscission zone formation. The external morphological characteristics of calyx abscission and the microscopic and ultrastructural features of the abscission zone cells under different treatments were observed. The ethylene concentration，reactive oxygen species levels，and Ca²⁺content in the abscission zone of decalyx fruits and corresponding parts of the abscission zone of persistent calyx fruits during the abscission process were measured. The results showed that under the ethephon treatment，the external morphological characteristics，microscopic structure，and ultrastructure of the abscission zone cells in Korla fragrant pears were consistent with those of the control during the calyx abscission. Before and during abscission zone formation，the ethylene concentration in the abscission zone of decalyx fruits and the corresponding parts of persistent calyx fruits induced by ethephon significantly increased compared to the control，with increases of 137.38% and 15.02%（decalyx fruit） and 294.95% and 110.16%（persistent calyx fruit），respectively. During the abscission zone formation period，the hydroxyl radical clearance rate，superoxide anion production rate，and Ca²⁺ content in the abscission zone of decalyx fruits under ethephon induction were all significantly higher than the control，being 33.88%，9.38%，and 15.02% of the control，respectively. In summary，exogenous ethephon accelerated the programmed cell death in calyx cells of Korla fragrant pears by increasing the ethylene concentration，reactive oxygen species levels，and Ca²⁺ content in the abscission zone of decalyx fruit，thereby promoting calyx abscission.

Key words: korla fragrant pear, ethylene, calyx abscission, programmed cell death, cellular structure, active oxygen, calcium ion

温玥, 邵白俊杰, 孙天雨, 张淑琴, 王春峰, 田嘉. 乙烯利诱导‘库尔勒香梨’萼片离区细胞程序性死亡特征及调控信号研究[J]. 园艺学报, 2025, 52(4): 959-972.

WEN Yue, SHAOBAI Junjie, SUN Tianyu, ZHANG Shuqin, WANG Chunfeng, TIAN Jia. Study on the Characteristics and Regulatory Signals of Programmed Cell Death in the Abscission Zone of Korla Fragrant Pear Induced by Ethephon[J]. Acta Horticulturae Sinica, 2025, 52(4): 959-972.

图/表 8

图1 ‘库尔勒香梨’脱萼果与宿萼果采样示意图

Fig. 1 Sampling diagram for decalyx and persistent calyx fruits of Korla fragrant pear

图2 清水（对照）和乙烯利处理下‘库尔勒香梨’萼片脱落过程中外部形态特征变化

Fig. 2 Changes in external morphological characteristics of Korla fragrant pear during calyx abscission in the water（control）and under ethephon treatment

图3 清水（对照）和乙烯利处理下‘库尔勒香梨’脱萼果和宿萼果不同时期的萼筒横切显微结构 a ~ d分别为脱萼果不同时期（离区形成前期、形成期、形成后期）和脱落后；e ~ g分别为宿萼果不同时期（离区形成前期、形成期、形成后期）。DS：远轴端；PS：近轴端；VB：维管束；V：导管；ST：筛管。下同

Fig. 3 Cross-sectional microscopic structures of calyx tubes in different periods of decalyx fruit and persistent calyx fruit of Korla fragrant pear in the water（control）and under ethephon treatment a-d were the microstructures of the decalyx fruit at different periods（pre-zone formation，zone formation，and post-zone formation）detachment，respectively；e-g were the microstructures of the persistent calyx fruit at different periods（pre-zone formation，zone formation，post-zone formation），respectively. DS：Distal side；PS：Proximal side；VB：Vascular bundle；V：Vessel；ST：Sieve tube. The same below

图4 清水（对照）和乙烯利处理下‘库尔勒香梨’脱萼果和宿萼果不同时期的萼筒纵切显微结构 AZ：离区；PL：保护层

Fig. 4 Longitudinal section microscopic structures of calyx tubes in different periods of decalyx fruit and persistent calyx fruits of Korla fragrant pear in the water（control）and under ethephon treatment AZ：Abscission zone；PL：Protective layer

图5 清水（对照）和乙烯利处理下‘库尔勒香梨’萼片脱落过程中细胞程序性死亡的细胞超微结构 N：细胞核；Nu：核仁；S：淀粉粒；M：线粒体；GA：高尔基体；Chl：叶绿体；Pm：质膜；Pb：质体小球；CW：细胞壁

Fig. 5 Cellular ultrastructure of programmed cell death during calyx detachment of Korla fragrant pear in the water（control）and under ethephon treatment N：Nucleus；Nu：Nucleolus；S：Starch granule；M：Mitochondrion；GA：Golgi apparatus；Chl：Chloroplast；Pm：Plasma membrane；Pb：Plastid goblet；CW：Cell wall

图6 清水（对照）和乙烯利处理下‘库尔勒香梨’萼片细胞程序性死亡过程中乙烯浓度变化 ** 表示同一时期不同处理间在0.01水平上差异显著；* 表示同一时期不同处理间在0.05水平上差异显著。下同

Fig. 6 Changes in ethylene concentration during PCD in calyx of Korla fragrant pear in the water（control）and under ethephon treatment ** indicated that significant different at 0.01 level in the same period at different treatment；* indicated that significant different at 0.05 level in the same period at different treatment. The same below

图7 清水（对照）和乙烯利处理下‘库尔勒香梨’萼片细胞程序性死亡过程中活性氧水平变化

Fig. 7 Changes in reactive oxygen species levels during PCD in calyx of Korla fragrant pear in the water（control）and under ethephon treatment

图8 清水（对照）和乙烯利处理下‘库尔勒香梨’萼片细胞程序性死亡过程中Ca2+含量变化

Fig. 8 Changes in calcium ion content of korla fragrant pear during PCD in calyx in the water（control）and under ethephon treatment

参考文献 29

[1]	Awwad F, Bertrand G, Grandbois M, Beaudoin N. 2019. Reactive oxygen species alleviate cell death induced by thaxtomin A in Arabidopsis thaliana cell cultures. Plants, 8 (9):332.
[2]	Chen Lei. 2023. Effect of early phenological irrigation on sepal abscission of balsam pear[M. D. Dissertation]. Alaer: Tarim University. (in Chinese)
	陈磊. 2023. 早期物候期灌水对香梨萼片脱落的影响[硕士论文]. 阿拉尔: 塔里木大学.
[3]	Chen Yan,Zhang Qian,Dong Yuzhong,Tao Shutian,Zhang Tianzheng,Bao Jianping. 2020. Effects of ethephon on the rate of calyx-abscission and fruit quality of‘Korla Fragrant Pear’and‘Xinli No.7’. Journal of Henan Agricultural University, 54 (6):949-955. (in Chinese)
	陈燕, 张倩, 董玉忠, 陶书田, 张天正, 包建平. 2020. 乙烯利对‘库尔勒香梨’和‘新梨7号’脱萼率及果实品质的影响. 河南农业大学学报, 54 (6):949-955.
[4]	Chen Yuanyuan, Li kai, Song Yuqin, Li na, Li Liulin. 2018. Effects of PBO on calyx abscission and quality of Yuluxiang pear fruit. South China Fruits, 47 (5):55-58. (in Chinese)
	陈园园, 李凯, 宋宇琴, 李娜, 李六林. 2018. PBO对玉露香梨果实脱萼及品质的影响. 中国南方果树, 47 (5):55-58.
[5]	Doniak M, Byczkowska A, Kaźmierczak A. 2016. Kinetin-induced programmed death of cortex cells is mediated by ethylene and calcium ions in roots of Vicia faba ssp. minor. Plant Growth Regulation,78:335-343.
[6]	Gang Minghui, Qiman · Yunus, Qin Weiming, Li Yanhong, Li Hui, Li Jiang. 2009. Effect of different plant growth regulators on set fruit rate, calyx dropping fruit rate,fruit shape and chlorophyll contents of Korla fragrant pear. Journal of Xinjiang Agricultural University, 32 (6):26-30. (in Chinese)
	刚明慧, 齐曼·尤努斯, 覃伟铭, 李艳红, 李惠, 李疆. 2009. 不同植物生长调节剂对库尔勒香梨坐果率、脱萼果率、果实形态及叶绿素含量的影响. 新疆农业大学学报, 32 (6):26-30.
[7]	Guan Junfeng, Li Guangmin. 2000. The physiological effects of calcium on ethylene biosynthesis and signal transduction in plants. Chinese Bulletin of Botany, 17 (5):413-418. (in Chinese)
	关军锋, 李广敏. 2000. 钙在植物乙烯生成及信号传递中的生理作用. 植物学通报, 17 (5):413-418.
[8]	Hao Zhichao, Wen Yue, Tian Jia, Shaobai Junjie, Yang Dinghua, Zhang Feng. 2022. Relationship between sepal abscission and phytohormones in different parts of Korla fragrant pear. Plant Physiology Journal, 58 (7):1369-1380. (in Chinese)
	郝志超, 温玥, 田嘉, 邵白俊杰, 杨丁花, 张峰. 2022. 库尔勒香梨萼片脱落与离区不同部位植物激素的关系. 植物生理学报, 58 (7):1369-1380.
[9]	Jia Bing, Yu Tao, Guo Guoling, Wang Youyu, Xu bo, Liu pu, Heng Wei, Zhu Liwu. 2021. Comparision of endogenous hormones content and expression difference of signal transduction genes between the calyx-persistence fruit and calyx-shedding fruit in‘Dangshansuli’pear. Journal of Nanjing Agricultural University, 44 (3):428-436. (in Chinese)
	贾兵, 余桃, 郭国凌, 王友煜, 许波, 刘普, 衡伟, 朱立武. 2021. ‘砀山酥梨’宿萼果与脱萼果内源激素含量比较及其信号传导基因的表达差异. 南京农业大学学报, 44 (3):428-436.
[10]	Levine A, Pennell R, Alvarez M, Palmer R, Lamb C. 1996. Calcium-mediated apoptosis in a plant hypersensitive disease resistance response. Current Biology, 6 (4):427-437.
[11]	Li Zhongguang, Du Chaokun, Gong Ming. 2005. Involvement of Ca²⁺ and calmodulin in the regulation of H₂O₂-induced heat tolerance in maize seedlings. Physiology and Molecular Biology of Plants, 31 (5):515-519. (in Chinese)
	李忠光, 杜朝昆, 龚明. 2005. Ca²⁺和钙调素对H₂O₂诱导的玉米幼苗耐热性的调控. 植物生理与分子生物学学报, 31 (5):515-519.
[12]	Lin Y, Lin H, Zhang S, Chen Y, Chen M, Lin Y. 2014. The role of active oxygen metabolism in hydrogen peroxide-induced pericarp browning of harvested longan fruit. Postharvest Biology and Technology,96:42-48.
[13]	Liu Yalong, Ou Chunging, Zhang Yanjie, Li Shuran, Ye Yu, Jiang Shuling, Wang Fei. 2024. Research progress on the mechanism of fruit abscission regulation in fruit trees. Plant Science Journal, 42 (1):125-133. (in Chinese)
	刘亚龙, 欧春青, 张艳杰, 李舒然, 叶宇, 姜淑苓, 王斐. 2024. 果树果实脱落调控机理研究进展. 植物科学学报, 42 (1):125-133.
[14]	Ma Hongchao, Wang Yanling, Wen Xu, Qi Man, Li Jiang. 2011. Effects of different reagent treatments on the microstructure of calyx tube with or without calyx of Korla fragrant pear. Journal of Fruit Science, 28 (3):518-520. (in Chinese)
	马宏超, 王燕凌, 文旭, 齐曼, 李疆. 2011. 不同药剂处理对库尔勒香梨脱萼和宿萼果萼片显微结构的影响. 果树学报, 28 (3):518-520.
[15]	Ma Wenjie, Zhu Mengting, Liu Shangqian, Wan Xinling, Liu Yan. 2023. Plant programmed cell death research progress. Journal of Plant Physiology, 59 (11):1998-2010. (in Chinese)
	马文杰, 朱梦婷, 刘尚钱, 万映伶, 刘燕. 2023. 植物程序性细胞死亡研究进展. 植物生理学报, 59 (11):1998-2010.
[16]	Michèle C. 2000. Hypersensitive response-related death. Plant Molecular Biology,44:321-334.
[17]	Ren H, Zhao X, Li W, Hussain J, Qi G, Liu S. 2021. Calcium signaling in plant programmed cell death. Cells,10:1089.
[18]	Rodriguez J, Morcillo R, Vierheilig H, Ocampo J, Ludwig-Müller J, Garrido J. 2010. Mycorrhization of the notabilis and steins tomato mutants in relation to abscisic acid and ethylene content. Journal of Plant Physiology, 167 (8):606-613.
[19]	Shaobai Junjie, Tian Jia, Hao Zhichao, Wang Qian, Zhang Feng, Wen Yue. 2023. Effects of ethephon on fruit developmental process and quality of Korla fragrant pear. Acta Botanica Boreali-Occidentalia Sinica, 43 (2):265-275. (in Chinese)
	邵白俊杰, 田嘉, 郝志超, 王乾, 张峰, 温玥. 2023. 乙烯利对库尔勒香梨果实发育进程和品质的影响. 西北植物学报, 43 (2):265-275.
[20]	Steffens B, Sauter M. 2005. Epidermal cell death in rice is regulated by ethylene,gibberellin,and abscisic acid. Plant Physiology, 139 (2):713-721.
[21]	Tal B, Marina D, Aleš K, Magda T, Maruša P, Shimon M, Shaul B, Sonia P, Naomi O, Lilian S, Martin В, Amnon L. 2011. Programmed cell death occurs asymmetrically during abscission in tomato. The Plant Cell, 23 (11):4146-4163.
[22]	Tian Mimi. 2021. Analysis of the function and mechanism of catalase in the growth and development in Arabidopsis thaniala[M. D. Dissertation]. Hefei: Hefei University of Technology. (in Chinese)
	田蜜蜜. 2021. 过氧化氢酶在拟南芥生长发育中的功能及作用机制分析[硕士论文]. 合肥: 合肥工业大学.
[23]	Vittoria L, Laura G. 2018. Programmed cell death in plants:an overview. Methods in Molecular Biology(Clifton,N J),DOI: 10.1007/978-1-4939-7668-3_1.
[24]	Wolters H, Jürgens G. 2009. Survival of the flexible:hormonal growth control and adaptation in plant development. Nature Reviews Genetics, 10 (5):305-317.
[25]	Xiong Guosheng, Li Jiayang, Wang Yonghong. 2009. Progress in the regulation of phytohormones. Chinese Scientific Bulletin, 54 (18):2718-2733. (in Chinese)
	熊国胜, 李家洋, 王永红. 2009. 植物激素调控研究进展. 科学通报, 54 (18):2718-2733.
[26]	Young T, Gallie D. 1999. Analysis of programmed cell death in wheat endosperm reveals differences in endosperm development between cereals. Plant Molecular Biology, 39 (5):915-926.
[27]	Yue J, Jiao J, Wang W, Jie X, Wang H. 2023. Silencing of the calcium-dependent protein kinase TaCDPK27 improves wheat resistance to powdery mildew. BMC Plant Biology, 23 (1):134.
[28]	Zeng Shaomin, Huang Xinzhong, Chen Xiaoming, Hu Ningsan, Huang Baoping, Jiang Cuicui. 2024. Effects of calyx removal agents on fruit quality and fruit setting rate of Zheli No.6 pear. South China Fruit, 53 (1):197-201,206. (in Chinese)
	曾少敏, 黄新忠, 陈小明, 胡宁三, 黄保平, 姜翠翠. 2024. 脱萼剂对“浙梨6号”果实品质与着果率的影响. 中国南方果树, 53 (1):197-201,206.
[29]	Zhao Congcong, Guan Qinglin, Wang Tao, Zhou Yanling. 2021. Study on antioxidant and antibacterial activities of extracts from Osmanthus fragrans in vitro. Food Research and Development, 42 (10):31-36. (in Chinese)
	赵丛丛, 管庆林, 王涛, 周艳玲. 2021. 桂花子提取物体外抗氧化和抑菌活性的研究. 食品研究与开发, 42 (10):31-36.