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Peptidomics Mapping of Proteolysis Highlights Triple Activation of Sprouted Seeds by Germination, Homogenisation and Species Mixture.

ABSTRACT: We investigated how seed proteolysis was enhanced by germination, by subsequent homogenisation (disrupting sprout compartments), and by co-incubation of homogenates from different species. Mass spectrometry of released peptides tracked proteolytic signatures from chickpea, lentil, mung and broccoli proteins, in soaked seeds, in sprouted seeds, and after sprout homogenisation followed by incubation alone or in mixture with other sprouts. The proteolytic signatures differed markedly among the four species, and in the different treatment conditions. After homogenisation, legumain-like cleavage (after asparagine) increased in lentil, and proline-rich peptides increased in broccoli. For co-incubated homogenised sprouts, each species’ homogenate significantly contributed 6 to 57% of proteolytic patterns in peptides of other species, with chickpea and broccoli homogenates notably releasing metabolic protein peptides from mung and lentil. Thus, germination, homogenisation and homogenate species mixtures can each contribute to proteolysis of seed peptides, potentially increasing digestibility and reducing allergenicity

INSTRUMENT(S):

ORGANISM(S): Cicer Arietinum (chickpea) (garbanzo) Brassica Oleracea Var. Oleracea Pisum Sativum (garden Pea) Vigna Radiata

TISSUE(S): Seed

SUBMITTER: Indrani Bera

LAB HEAD: Denis Shields

PROVIDER: PXD072115 | Pride | 2026-05-05

REPOSITORIES: Pride

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			Action	DRS
	2023_03_03_IBERA_S1.10_S3-A4_8796.d.zip	Other
	2023_03_03_IBERA_S1.11_S3-A7_8799.d.zip	Other
	2023_03_03_IBERA_S1.12_S3-A10_8802.d.zip	Other
	2023_03_03_IBERA_S1.13_S3-B1_8805.d.zip	Other
	2023_03_03_IBERA_S1.14_S3-B4_8808.d.zip	Other

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Project description:Purpose: The goals of this study are to analyze the transcriptome of five time point in broccoli seed germination and sprout development and to find the putative glucosinolate metabolism genes in the stage. Methods: Total mRNA of germinated seeds, 3 day cotyledons, 7 day botyledons, 11 day cotyledons and 11 day euphyllas of wild-type broccoli were harvested. Each sample was harvested in three independent biological replicates with equal weight and subsequently pooled together for sequencing. The sequence reads that passed quality filters were de novo assembled using VELVET followed by OASES. Then the assembled unigenes were used for the abundance and functional analysis. Results: A total of ~85million 251bp reads were obtained. After de novo assembly and searching the assembled transcripts against the Arabidopsis thaliana and Nr databases, 19,441 top-hit transcripts were clustered as unigenes with an average length of 2,133bp. These unigenes were classified according to their putative functional categories. Cluster analysis of total unigenes with similar expression patterns and differentially expressed unigenes among different tissues,as well as transcription factor analysis were performed. We identified 25 putative glucosinolate metabolismgenes sharing 62.04-89.72% nucleotide sequence identity with the Arabidopsis orthologs. This established a broccoli glucosinolate metabolic pathway with high colinearity to Arabidopsis. Many of the biosynthetic and degradation genes showed higher expression after germination than in seeds; especially the expression of the myrosinaseTGG2 was 20-130 times higher.These results along with the previous reports that glucosinolate concentration decreased exponentially once after germination indicate the breakdown products of glucosinolates may play important roles in broccoli seed germination and sprout development. Conclusion: Our study provides the largest genetic resource of broccoli to date. These data will pave the way for further studies and genetic engineering of broccoli sprouts to develop functional vegetables containing high levels of the anticarcinogenic glucosinolates. They will also provide new insight into the genomic research of this species and its relatives. Wild-type broccoli mRNA profiles of seeds, 3 day cotyledons, 7 day botyledons, 11 day cotyledons and 11 day euphyllas were generated by deep sequencing, three biological replicates pooling together for each tissue, using Illumina Myseq platform.

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Peptidomics Mapping of Proteolysis Highlights Triple Activation of Sprouted Seeds by Germination, Homogenisation and Species Mixture.

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