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Blueberry (Vaccinium corymbosum) is a major berry crop in the United States, and
one that has great nutritional and economical value. Next generation sequencing
methodologies, such as 454, have been demonstrated to be successful and
efficient in producing a snap-shot of transcriptional activities during an
organism’s developmental stage(s) or its response to biotic or abiotic stresses.
Such application of this new sequencing technique allows for high-throughput,
genome-wide experimental verification of known and novel transcripts. We have
applied a high-throughput pyrosequencing technology (454 EST sequencing) for
transcriptome profiling of blueberry during different stages of fruit
development to gain an understanding of the genes that are up or down regulated
during this process. We have also sequenced flower buds at four different stages
of cold acclimation to gain a better understanding of the genes and biochemical
pathways that are up- or down-regulated during cold acclimation, since extreme
low temperatures are known to reduce crop yield and cause major losses to US
farmers. We have also sequenced a leaf sample to compare its transcriptome
profile with that of bud and fruit samples. Over 500,000 sequences were
generated and assembled into approximately 15,000 contigs and were annotated and
functionally mapped to Gene Ontology (GO) terms. A database was developed to
house these sequences and their annotations. A web based interface was also
developed to allow collaborators to search\browse the data and aid in the
analysis and interpretation of the data. The availability of these sequences
will allow for future advances, such as the development of a blueberry
microarray to study gene expression, and will aid in the blueberry genome
sequencing effort that is underway. |