Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Apr 14;11(8):1136.
doi: 10.3390/foods11081136.

Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the "Calcification" Process

Yu Chen  1 , Cancan Zhu  1 , Yuqiang Zhao  1 , Shijie Zhang  1 , Wu Wang  1
Affiliations

Transcriptomics Integrated with Changes in Cell Wall Material of Chestnut (Castanea mollissima Blume) during Storage Provides a New Insight into the "Calcification" Process

Yu Chen et al. Foods. .

Abstract

Chestnut "calcification" is the result of a series of physiological and biochemical changes during postharvest storage; however, the associated mechanisms are unclear. In this study, several potential calcification-related physicochemical parameters in chestnut, including moisture, cell wall materials, cellulose, lignin, and pectin, were measured. Transcriptome analysis was performed on chestnut seeds during different stages of storage. The results showed that the degree of calcification in the chestnut seeds was significantly negatively correlated with the moisture content (r = -0.961) at room temperature (20-25 °C) and a relative humidity of 50-60%. The accumulation of cell wall material in completely calcified seeds was 5.3 times higher than that of fresh seeds. The total content of cellulose and lignin increased during the storage process. Transcriptome analysis of 0% and 50% calcified chestnut was performed; a total of 1801 differentially expressed genes consisting of 805 up-regulated and 996 down-regulated genes were identified during the calcification process. Furthermore, response to water, water deprivation, and salt stress were most enriched by gene ontology (GO) and gene set enrichment analysis (GSEA). The Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways related to chestnut calcification included purine metabolism, RNA degradation, the mRNA surveillance pathway, starch and sucrose metabolism, arginine and proline metabolism, and fatty acid metabolism, and were detected. Most of the genes involved in cellulose synthase, lignin catabolism, and pectin catabolism were down-regulated, while only two important genes, scaffold11300 and scaffold0412, were up-regulated, which were annotated as cellulose and pectin synthase genes, respectively. These two genes may contribute to the increase of total cell wall material accumulation during chestnut calcification. The results provided new insights into chestnut calcification process and laid a foundation for further chestnut preservation.

Keywords: calcification; cellulose; chestnut; lignin; pectin; transcriptome.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Relationship between the degree of chestnut calcification and the moisture content. Each experiment included three replicates. All of the data were expressed as means ± standard deviations.
Figure 2
Figure 2
Changes in the cell wall content in different stages of chestnut calcification. (A) Degree of chestnut calcification, from left to right, 0%, 25%, 50%, 75%, and 100%. Scale bars, 1 cm. (B) Changes in cell wall material (CWM) in different stages of chestnut calcification. (C) Changes in lignin and cellulose contents in different stages of chestnut calcification. (D) Changes in the pectin content at different stages of chestnut calcification. All of the data were expressed as means ± standard deviations. Different letters (a–e) in the same composition indicate significant difference (p < 0.05, Duncan’s multiple range test).
Figure 3
Figure 3
Heat maps of the differentially expressed genes (DEGs) in the 0% and 50% stages of chestnut calcification. The fragments per kb per million (FPKM) read values of unigenes were used for hierarchical cluster analysis. Expression levels are shown in different colors; the redder or bluer the color, the higher or lower the expression, respectively. The two groups, T1a, T1b, T1c and T3a, T3b, T3c, represent 0 % and 50% stages of chestnut calcification, respectively.
Figure 4
Figure 4
Gene ontology (GO) classification of differentially expressed genes between the 0% and 50% stages of chestnut calcification.
Figure 5
Figure 5
Gene ontology (GO) enrichment analysis of differentially expressed genes (DEGs).
Figure 6
Figure 6
Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis of differentially expressed genes (DEGs).
Figure 7
Figure 7
Gene set enrichment analysis (GSEA) enrichment analysis was performed base on GO annotations.
Figure 8
Figure 8
Gene set enrichment analysis (GSEA) analysis of differentially expressed genes (DEGs) expressed between the 0% and 50% calcification subtypes using the gene ontology (GO) datasets. (A) organization or biogenesis; (B) regulation of biological quality; (C) response to osmotic stress; (D) response to salt stress.
Figure 9
Figure 9
Relationship between the gene expression and the moisture content of chestnut calcification. (A) The differential expression genes (DEGs) in water transport (GO: 0006833) pathway; (B) the differential expression genes (DEGs) in response to water deprivation (GO: 0009414) pathway. Blue line: moisture content, red bars: transcriptomics RNA-seq analysis (FPKM). The letters (a,b) indicate significant differences (p < 0.05).
Figure 10
Figure 10
QRT-PCR validations of RNA-seq data. Relative expression was determined using three biological replicates. Black line: qRT-PCR expression, black bars: transcriptomics RNA-seq analysis (FPKM). The letters (a,b) indicate significant differences (p < 0.05).
See this image and copyright information in PMC

References

    1. Liu G.H., Fang J.Y. Spatial patterns of chestnut (Castanea millissima) and its species geographical distribution in China. Acta Ecol. Sin. 2001;21:164–170.
    1. De Vasconcelos M.C., Bennett R.N., Rosa E.A., Ferreira-Cardoso J.V. Composition of European chestnut (Castanea sativa Mill.) and association with health effects: Fresh and processed products. J. Sci. Food Agric. 2010;90:1578–1589. doi: 10.1002/jsfa.4016. - DOI - PubMed
    1. Cui H., Li H., Wang Y., Li S., Xue C. Structural characterization and biological activity of galactoglucan from Castanea mollissima Blume. J. Carbohydr. Chem. 2019;38:398–411. doi: 10.1080/07328303.2019.1630838. - DOI
    1. Zhang Y., Yang Z., Liu G., Wu Y., Ouyang J. Inhibitory Effect of Chestnut (Castanea mollissima Blume) Inner Skin Extract on the Activity of α-Amylase, α-Glucosidase, Dipeptidyl Peptidase IV and In Vitro Digestibility of Starches. Food Chem. 2020;324:126847. doi: 10.1016/j.foodchem.2020.126847. - DOI - PubMed
    1. Wen X., Gu C., Zhu D., Liu P., Lai Y., Zeng Q. Water stress affects on cell membrane lipid oxidation and calcification of chestnut (Castanea mollissima Bl.) Postharvest Biol. Technol. 2017;126:34–39. doi: 10.1016/j.postharvbio.2016.12.005. - DOI

LinkOut - more resources