2种优质梨果实质地差异的显微观察及与细胞壁代谢酶的相关性

白牡丹; 刘晓宇; 杨盛; 郝国伟; 张晓伟; 王燕平; 付宝春

doi:10.19303/j.issn.1008-0384.2023.08.005

2种优质梨果实质地差异的显微观察及与细胞壁代谢酶的相关性

doi: 10.19303/j.issn.1008-0384.2023.08.005

山西农业大学果树研究所/果树种质创制与利用山西省重点实验室，山西　太原　030031

基金项目: 国家现代农业产业技术体系建设专项（CARS-28-29）；山西农业大学农业科技创新研究课题（YCX2020YQ09）；山西农业大学生物育种工程项目（YZGC036）；山西省科技厅基础研究计划项目（20210302124122）

详细信息

作者简介:
白牡丹（1988 —），女，硕士，助理研究员，主要从事梨新品种选育与果树栽培生理研究，E-mail：mudanbai@139.com

通讯作者:
郝国伟（1971 —），男，硕士，副研究员，主要从事梨新品种选育与果树栽培生理研究，E-mail：hgw1971@sohu.com

中图分类号: S661.1
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出版历程
- 收稿日期: 2023-03-09
- 修回日期: 2023-04-30
- 网络出版日期: 2023-08-16
- 刊出日期: 2023-08-28

Correlation with Cell Wall Metabolizing-related Enzymes and Microstructure Observation in Texture Differences of two Kinds of High-Quality Pear Fruit

Pomology Institute, Shanxi Agricultural University/Shanxi Key Laboratory of Germplasm Improvement and Utilization in Pomology, Taiyuan, Shanxi　030031, China

摘要

摘要: 目的通过对果实质地差异明显的2个梨品种进行质地差异相关研究，旨在发现导致梨品种果实质地差异的关键因素，为优良梨品种的选育提供理论依据。方法以玉露香梨和大果水晶梨为试材，观察发育过程中果肉细胞超显微结构，测定果实的硬度、脆度值、细胞壁代谢相关酶的活性和相关基因的相对表达量，并进行相关性分析。结果 ①果实生长发育过程中玉露香梨和大果水晶梨果实硬度逐渐下降，且花后60 d 2个梨品种脆度值逐渐下降，玉露香梨果实硬度和脆度值显著低于大果水晶梨。②在400×放大显微倍数下，幼果期玉露香梨果肉细胞比大果水晶梨的大；果实膨大期和成熟期，2个梨品种果肉细胞均明显增大，玉露香梨果肉细胞小于大果水晶梨；玉露香梨细胞断裂边缘整齐平滑，形成脆性断口，大果水晶梨断裂边缘粗糙而不整齐。③果实发育过程中玉露香梨果肉果胶甲酯酶（Pectin methylesterase，PME）活性极显著低于大果水晶梨，而多聚半乳糖醛酸酶（Polygalacturonase，PG）活性和β-半乳糖苷酶（β- Galactosidase，β-Gal）活性极显著高于大果水晶梨。④果实发育过程中玉露香梨果肉PME基因、PG基因和β-Gal基因相对表达量极显著低于大果水晶梨的表达量。相关性分析表明，玉露香梨和大果水晶梨的硬度与β-Gal 基因相对表达量呈显著正相关，脆度值与PG 基因相对表达量呈显著正相关。结论随着果实生长发育，玉露香梨和大果水晶梨果实质地的差异不仅表现在果肉细胞超显微结构上，还与细胞壁代谢相关酶密切相关，PG基因和β-Gal 基因可能在玉露香梨和大果水晶梨质地变化中发挥关键作用。
- 梨 /
- 质地差异 /
- 果肉细胞显微结构 /
- 细胞壁代谢相关酶
Abstract: Objective Factors contributing to the distinctively different texture of the fruits of two pear cultivars were investigated. Method Cellular microstructures of the pulp of Yuluxiangli and Daguoshuijingli pears were observed under an electron microscope SU810 . Textural firmness and brittleness, as well as contents of cell wall-modifying enzymes and relative expressions of the associated genes, of the fruits during develop stages were measured to correlate with the microstructure observations. Result ①The firmness of the pears of both varieties gradually declined during the fruit development. The force required to break into the fruit decreased 60d after flowering as the fruits were ripening. In comparison, Yuluxiangli pears were significantly softer and less brittle than Daguoshuijingli. ②At 400× magnification, the cells of Yuluxiangli pear pulp appeared larger than those of Daguoshuijingli at young stage. As the fruits expanded and matured, the cells of both varieties enlarged significantly but those of Yuluxiangli became smaller than those of Daguoshuijingli. And upon cutting, the edge of fractured Yuluxiangli pears were clean and smooth, while that of Daguoshuijingli rough and irregular. ③ In the matured fruits, the pectin methylesterase (PME) activity in Yuluxiangli pears were extremely significantly lower than that in Daguoshuijingli, whereas the polygalacturonase (PG) and β-galactosidase (β-Gal) activities extremely significantly higher. ④ The relative expressions of PME, PG, and β-Gal in Yuluxiangli were extremely significantly lower than those in Daguoshuijingli during the fruit development. And that of β-Gal was significantly positively related to the textural firmness, but that of PG to the brittleness, of the fruits for both pear varieties. Conclusion The unique texture of Yuluxiangli and Daguoshuijingli pears could not only be differentiated by their pulp cell ultrastructural differences but also closely related to the cell wall-modifying enzymes, such as, PG and β-Gal, in the fruits. Hence, the genes associated with those enzymes likely played a critical role in the texture of the pears.
- Pears /
- distinct texture /
- ultrastructure of pulp cell /
- cell wall-modifying enzymes

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图 1 不同梨品种果实发育过程中硬度、脆度值变化

** 表示差异极显著（P＜0.01），* 表示差异显著（P＜0.05）。图3、4同。

Figure 1. Changes in firmness and brittleness of pears during fruit development

** indicates very significant difference at 0.01 level, * indicates significant difference at 0.05 level. The same as Fig.3,4.

下载: 全尺寸图片幻灯片

图 2 不同梨品种果实发育过程中果肉细胞超显微结构

a~c：玉露香梨果实幼果期、膨大期、成熟期；A~C：大果水晶梨果实幼果期、膨大期、成熟期。

Figure 2. Ultrastructure of pear pulp cells during fruit development

a: Yuluxiangli pear at young stage; b: Yuluxiangli pear at fruit expanding stage; c: Matured Yuluxiangli pear; A: Daguoshuijingli pear at young stage; B: Daguoshuijingli pear at fruit expanding stage; C: Matured Daguoshuijingli pear.

下载: 全尺寸图片幻灯片

图 3 果实发育过程中细胞壁代谢相关酶的活性变化

Figure 3. Changes on activities of metabolism-related enzymes during fruit development

下载: 全尺寸图片幻灯片

图 4 果实发育过程中果肉细胞壁代谢相关基因的相对表达量

Figure 4. Relative expressions of PME, PG, and β-Gal during fruit development

下载: 全尺寸图片幻灯片

表 1 qPCR引物

Table 1. Primers for qPCR

基因 Gene	引物序列 Primer sequence	片段大小 Amplified fragment length/bp	GenBank 登录号 GenBank Accession No.
果胶甲酯酶PME	F：5′-CAAAGCAGGAAGGTACGAAGAG-3′	121	AB067681.1
果胶甲酯酶PME	R：5′-TTTCTGCGCGACGTTTTTC-3′	121	AB067681.1
β-半乳糖苷酶β-Gal	F：5′-GTGTATGGGTCGTTGGAGAATC-3′	102	AJ811694.1
β-半乳糖苷酶β-Gal	R：5′-CAGACCAACGGAAATGCTAAG-3′	102	AJ811694.1
多聚半乳糖醛酸酶PG	F：5′-GCAATAACTTGGTGGTGAAGAATC-3′	132	AJ504855.2
多聚半乳糖醛酸酶PG	R：5′-CGTCCGTATTAGGGCTATCCTC-3′	132	AJ504855.2
Actin	F：5＇-TGCTGAGCGGGAAATTGTG-3＇	145	AF386514.1
Actin	R：5′-TCCGATAGTGATTACCTGTCCATC-3′	145	AF386514.1

下载: 导出CSV

表 2 果实质地与细胞壁代谢相关酶活性及相关基因表达量的相关性分析

Table 2. Correlation between texture, cell wall metabolism-related enzyme activity, and related genes expressions of pears

品种 Variety	指标 Indicator	PME活性 Activity of PME	PG活性 Activity of PG	β-Gal活性 Activity of β-Gal	PME 相对表达量 PME relative expression	PG 相对表达量 PG relative expression	β-Gal 相对表达量 β-Gal relative expression
玉露香梨 Yuluxiangli	硬度 Firmness	0.974	0.997*	1.000**	0.963	0.945	1.000*
	脆度值 Fracturability	0.989	0.954	0.924	0.994	0.999*	0.936
大果水晶梨 Daguoshuijingli	硬度 Firmness	1.000*	0.992	0.985	1.000*	0.984	0.998*
	脆度值 Fracturability	0.977	0.994	0.998*	0.970	0.998*	0.984
** 表示极显著相关（ P ＜0.01），* 表示显著相关（ P ＜0.01）。 ** indicates extremely significant correlation（ P ＜0.01）; * indicates significant correlation （P ＜0.01）.

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