果胶酶对灵武长枣果实发育中阿拉伯半乳糖蛋白分布的影响

doi:10.7525/j.issn.1673-5102.2024.01.009

摘要/Abstract

摘要：

以3种不同浓度的果胶酶处理灵武长枣（Ziziphus jujuba ‘Lingwu Changzao’）4个不同发育时期的果实，采用免疫细胞化学技术，在细胞水平上对果实阿拉伯半乳糖蛋白（AGPs）表位进行原位分析，探讨果胶酶对其不同发育时期的果实中AGPs分布的影响，为明确果胶酶对果实成熟软化的影响提供解剖学依据。结果表明：JIM13、JIM8和MAC204抗体所标记的抗原荧光强弱在各时期果实的不同组织存在一定的差异。当果肉组织用较低浓度的果胶酶0.028 U·mL^-1（E1）处理，果皮组织结构没有明显的变化，果皮及内部薄壁细胞细胞壁表面和细胞间隙中的AGPs抗原表位减少；0.056 U·mL^-1（E2）和0.084 U·mL^-1（E3）浓度果胶酶处理的果实，果实组织细胞壁解体程度增加，果皮及内部薄壁细胞细胞壁中AGPs抗原表位检测量逐渐降低，果胶酶浓度的增加导致AGPs在果实所有表位排列的更大影响和较低的荧光信号。经果胶酶最高浓度0.084 U·mL^-1（E3）处理后，Calcofluor White染色显示细胞壁区域荧光也不同程度减弱或降低，AGPs分布的紊乱和抗原表位的缺失与细胞中纤维素组装的变化有关。比较分析表明，不同发育时期的果实AGPs碳水化合物的分布不同，这与组织结构的变化有关；果胶酶作用下AGPs聚糖链的缺失导致细胞壁组分之间的相关性建立和细胞壁结构的重塑受阻，诱导了整个细胞壁结构的改变，影响了果实的成熟。

关键词: 果胶酶, 灵武长枣, 果实, 阿拉伯半乳糖蛋白, 免疫荧光定位

Abstract:

To explore the effects of pectinase on distribution of arabinogalactan proteins（AGPs） in fruits at different developmental stages，and to provide anatomical evidence for revealing the effects of pectinase on fruit ripening and softening， the fruits of Ziziphus jujuba ‘Lingwu Changzao’ at four different development stages were treated with three different concentrations pectinase， and AGPs epitopes analysis in situ at cellular level of fruits was performed using immunocytochemistry technique. The results showed that the fluorescence intensity of the antigens recognized by JIM13 and JIM8 and MAC204 antibodies was varied in different tissues during the development of fruit at each stage. When fruit tissues were treated with 0.028 U·mL^-1（E1） pectinase， there were no obvious changes in pericarp tissue structure， AGPs antigen epitopes in cell wall surface and intercellular space of pericarp and interior parenchyma cells decreased. When fruit tissues were treated with 0.056 U·mL^-1（E2） and 0.084 U·mL^-1（E3）pectinase， disintegration degree of cell wall in fruit tissue increased， the amount of AGPs epitopes detected in cell wall of pericarp and inner parenchyma cells decreased gradually， an increase in pectinase concentration resulted in a greater effect of AGPs on the arrangement of all epitopes in fruit and a lower fluorescence signal. After treatment with pectinase concentration up to 0.084 U·mL^-1（E3）， Calcofluor White staining revealed fluorescence attenuation or reduction to varying degrees in the cell wall region， disturbance of AGPs distribution and absence of antigen epitopes were associated with changes of cellulose assembly in cells. The results indicated that the distribution of AGPs carbohydrates differed in fruits at different stages， which was related to changes in tissue structure. The absence of AGPs glycan chains in the presence of pectinase led to the block of the establishment of correlations between cell wall components and cell wall structure remodeling， and the changes in the whole cell wall structures were induced and fruit ripening was affected.

Key words: pectinase, Ziziphus jujuba ‘Lingwu Changzao’, fruit, arabinogalactan proteins, immunofluorescence localization

中图分类号:

S665.1

王静, 章英才, 陶珊珊, 杨雪. 果胶酶对灵武长枣果实发育中阿拉伯半乳糖蛋白分布的影响[J]. 植物研究, 2024, 44(1): 62-74.

Jing WANG, Yingcai ZHANG, Shanshan TAO, Xue YANG. Effects of Pectinase on the Distribution of Arabinogalactan Proteins in Developing Fruit of Ziziphus jujuba ‘Lingwu Changzao’[J]. Bulletin of Botanical Research, 2024, 44(1): 62-74.

图/表 6

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