Abstract

Though numerous studies have focused on the cell wall disassembly of bananas during the ripening process, the modification of homogalacturonan (HG) during fruit development remains exclusive. To better understand the role of HGs in controlling banana fruit growth and ripening, RNA-Seq, qPCR, immunofluorescence labeling, and biochemical methods were employed to reveal their dynamic changes in banana peels during these processes. Most HG-modifying genes in banana peels showed a decline in expression during fruit development. Four polygalacturonase and three pectin acetylesterases showing higher expression levels at later developmental stages than earlier ones might be related to fruit expansion. Six out of the 10 top genes in the Core Enrichment Gene Set were HG degradation genes, and all were upregulated after softening, paralleled to the significant increase in HG degradation enzyme activities, decline in peel firmness, and the epitope levels of 2F4, CCRC-M38, JIM7, and LM18 antibodies. Most differentially expressed alpha-1,4-galacturonosyltransferases were upregulated by ethylene treatment, suggesting active HG biosynthesis during the fruit softening process. The epitope level of the CCRC-M38 antibody was positively correlated to the firmness of banana peel during fruit development and ripening. These results have provided new insights into the role of cell wall HGs in fruit development and ripening.

Highlights

  • 2.2.ResultsThestudy studywas wasperformed performedon onfruits fruitsof of00dd(just (justemerging emerging from fromthe the bunch), bunch), 35

  • Plant cell walls play a fundamental role in fruit development and ripening

  • The degree of pectin methylesterification (DM) of fruit peel at 0 d was the highest (59.07%) while the YM fruit showed the lowest one (47.60%), but no significant difference was observed with DM during fruit development and after ripening (Table 1)

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Summary

2.2.Results

Thestudy studywas wasperformed performedon onfruits fruitsof of00dd(just (justemerging emerging from fromthe the bunch), bunch), 35. The peel became yellow and the pulp yellowwhitish. Thefruit fruitsize size(indicated (indicatedby byfruit fruitfresh freshweight, weight,length, length,and andperimeter) perimeter)increased increased significantly with the development of fruits Significantly with the development of fruits (Figure 1B). Data represent an average of replicates three replicates treatment consisted of three replicates. 0 d: fruits just emerging from the range test at p < 0.05 after angular transformation of the data. 0 d: fruits just emerging from the bunch; bunch; 35 d: 35 day-old fruits; 60 d: 60 day-old fruits; GM: green matured fruits (at harvest, 85 day d:fruits); 35 day-old d: 60 day-old. GM:ethylene green matured fruits (at harvest, 85 day-old fruits); old YM: fruits; yellow60matured fruits fruits;. YM: yellow matured fruits (6 d after ethylene treatment)

Systemic Analysis of the Developmental Changes in Gene Expression Levels with RNA-Seq
The Developmental Changes in the Expression Levels of HG-Modifying Genes
Enzyme Activities and Fruit Properties
Histological Analysis
PMEs and the Epitope of 2F4
The Epitopes of JIM5 and JIM7 Antibodies
Changes in HG Metabolism during the Development of Banana Fruits
Changes in HG Metabolism in Banana Peel during the Fruit Ripening Process
Plant Materials
Firmness of Banana Peel
RNA-Seq Data Analysis
Activity Assay of HG Degradation Enzymes
Content of Pectin, Methanol, and DM
Immuno-Labeling of Antibodies Related to Pectin HG
Histological Study
Statistical Analysis and the Calculation of Correlation Coefficient
Conclusions

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