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Identification and expression analysis of starch branching enzymes involved in starch synthesis during the development of chestnut (Castanea mollissima Blume) cotyledons.

ABSTRACT: Chinese chestnut (Castanea mollissima Blume) is native to China and distributes widely in arid and semi-arid mountain area with barren soil. As a perennial crop, chestnut is an alternative food source and acts as an important commercial nut tree in China. Starch is the major metabolite in nuts, accounting for 46 ~ 64% of the chestnut dry weight. The accumulation of total starch and amylopectin showed a similar increasing trend during the development of nut. Amylopectin contributed up to 76% of the total starch content at 80 days after pollination (DAP). The increase of total starch mainly results from amylopectin synthesis. Among genes associated with starch biosynthesis, CmSBEs (starch branching enzyme) showed significant increase during nut development. Two starch branching enzyme isoforms, CmSBE I and CmSBE II, were identified from chestnut cotyledon using zymogram analysis. CmSBE I and CmSBE II showed similar patterns of expression during nut development. The accumulations of CmSBE transcripts and proteins in developing cotyledons were characterized. The expressions of two CmSBE genes increased from 64 DAP and reached the highest levels at 77 DAP, and SBE activity reached its peak at 74 DAP. These results suggested that the CmSBE enzymes mainly contributed to amylopectin synthesis and influenced the amylopectin content in the developing cotyledon, which would be beneficial to chestnut germplasm selection and breeding.

SUBMITTER: Chen L

PROVIDER: S-EPMC5441625 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Identification and expression analysis of starch branching enzymes involved in starch synthesis during the development of chestnut (Castanea mollissima Blume) cotyledons.

Chen Liangke L Lu Dan D Wang Teng T Li Zhi Z Zhao Yanyan Y Jiang Yichen Y Zhang Qing Q Cao Qingqin Q Fang Kefeng K Xing Yu Y Qin Ling L

PloS one 20170523 5

Chinese chestnut (Castanea mollissima Blume) is native to China and distributes widely in arid and semi-arid mountain area with barren soil. As a perennial crop, chestnut is an alternative food source and acts as an important commercial nut tree in China. Starch is the major metabolite in nuts, accounting for 46 ~ 64% of the chestnut dry weight. The accumulation of total starch and amylopectin showed a similar increasing trend during the development of nut. Amylopectin contributed up to 76% of t ...[more]

PMID: 28542293

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Project description:UnlabelledBACKGROUND1471-2229-9-51: American chestnut (Castanea dentata) was devastated by an exotic pathogen in the beginning of the twentieth century. This chestnut blight is caused by Cryphonectria parasitica, a fungus that infects stem tissues and kills the trees by girdling them. Because of the great economic and ecological value of this species, significant efforts have been made over the century to combat this disease, but it wasn't until recently that a focused genomics approach was initiated. Prior to the Genomic Tool Development for the Fagaceae project, genomic resources available in public databases for this species were limited to a few hundred ESTs. To identify genes involved in resistance to C. parasitica, we have sequenced the transcriptome from fungal infected and healthy stem tissues collected from blight-sensitive American chestnut and blight-resistant Chinese chestnut (Castanea mollissima) trees using ultra high throughput pyrosequencing.ResultsWe produced over a million 454 reads, totaling over 250 million bp, from which we generated 40,039 and 28,890 unigenes in total from C. mollissima and C. dentata respectively. The functions of the unigenes, from GO annotation, cover a diverse set of molecular functions and biological processes, among which we identified a large number of genes associated with resistance to stresses and response to biotic stimuli. In silico expression analyses showed that many of the stress response unigenes were expressed more in canker tissues versus healthy stem tissues in both American and Chinese chestnut. Comparative analysis also identified genes belonging to different pathways of plant defense against biotic stresses that are differentially expressed in either American or Chinese chestnut canker tissues.ConclusionOur study resulted in the identification of a large set of cDNA unigenes from American chestnut and Chinese chestnut. The ESTs and unigenes from this study constitute an important resource to the scientific community interested in the discovery of genes involved in various biological processes in Chestnut and other species. The identification of many defense-related genes differentially expressed in canker vs. healthy stem in chestnuts provides many new candidate genes for developing resistance to the chestnut blight and for studying pathways involved in responses of trees to necrotrophic pathogens. We also identified several candidate genes that may underline the difference in resistance to Cryphonectria parasitica between American chestnut and Chinese chestnut.

Project description: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.

Dataset Information

Identification and expression analysis of starch branching enzymes involved in starch synthesis during the development of chestnut (Castanea mollissima Blume) cotyledons.

Publications

Identification and expression analysis of starch branching enzymes involved in starch synthesis during the development of chestnut (Castanea mollissima Blume) cotyledons.

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