Project description:The aim of the experiment was to identify the transcriptional changes between wild Brassica oleraceae lines (Winspit) and 2 cultivated lines (purple sprouting broccoli and savoy cabbage) that show different biofumigation phenotypes. Fully expanded leaves were compared from 8 week old plants.
Project description:Plant microRNAs are short (~21 nt), non-coding molecules that regulate gene expression by targeting mRNA cleavage or protein translation inhibition. In this manner, they play many important roles in the cells of living organisms. One of the plant species in which the entire set of miRNAs has not been yet completely identified is Brassica oleracea var. capitata (cabbage). For this reason and for the economic and nutritional importance of this food crop, Illumina high-throughput small RNAs sequencing has been performed to discover novel and conserved miRNAs in the mature cabbage leaves. In this study, raw reads generated from three small RNA libraries were bioinformatically processed and further analyzed to select sequences homologous to known B. oleracea and other plant miRNAs. As a result of this step, 267 conserved miRNAs (belonging to 62 families) have been discovered. The remaining unannotated tags were used in the prediction and evaluation of novel miRNAs, which resulted in the 26 potential miRNAs proposal. The expression of 13 selected miRNAs was analyzed by northern blot hybridization. To determine the functions and processes that these molecules may potentially regulate in cabbage leaves, GO annotation, GO enrichment analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway mapping of the predicted targets were performed. Taken together, for the first time, the large set of miRNAs was identified in the mature cabbage leaves. Potential target designation for these miRNAs may suggest their important role in the plant growth, development and other primary biological processes. The presented study not only supplements the knowledge about B. oleracea miRNAs, but also may be used in other researches concerning the improvement of the cabbage cultivation.
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.
Project description:Morphotypes of Brassica oleracea are the result of a dynamic interaction between the genes that regulate the transition between vegetative and reproductive stages and those that regulate leaf morphology and plant architecture. In kales ornate leaf patterns, flowering delaying and nutrient quality are some of the characters were potentially selected by humans during domestication. Understanding candidate genes responsible for kale domestication is of importance to ultimately improve crop production. We aim to identify candidate genes that are responsible for kale leaf shape diversity and the evolution of domestic kale. Here we look at the global pattern of expressed genes during one single phase of development in kale, cabbage and TO1000 to gain an understanding of the genome-wide differences among some of the vegetative B. oleracea phenotypes. We identified gene expression patterns that are shared among the phenotypes and estimate the contribution of morphotype-specific gene expression patterns that set each of them apart. Differentially expressed developmental genes that regulate the vegetative to reproductive transition were abundant and present in all comparisons.
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.