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Objectives
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Fabrication of rice cDNA microarray chip.
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Study gene expression patterns at various
stages of rice grain development.
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Compare mRNA profiles of good and poor grain
filling varieties.
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Identify genes/gene clusters that are
associated with rapid grain filling.
Progress
cDNA clones: cDNA libraries were
constructed from rice panicles and flag leaves of MR211 and MR219 rice
varieties collected at different stages of panicle development. Four
subtraction libraries were constructed, from MR84, to represent the various
stages of grain filling. About 7000 plasmid clones were isolated from these
libraries. From these, 822 genes were sequenced and the sequences were
blasted against the NCBI and TIGR rice databases. Based on protein
functions, the ESTs were classified into those associated with metabolism,
storage, signaling, transport, stress response, gene expression, energy,
cell-structure maintenance, allergens and plant hormone.
Stages of rice
grain development
Microarray: PCR amplification and purification of the cDNA insert
was performed in 96-well plate followed by screening and gel analysis. A
total of 6240 clones including 6096 rice clones and 144 standards and
controls were included in the 384-well format for microarray printing using
an Affymetrix arrayer.
Samples: Total RNA was extracted from the panicles of MR84
(good grain fill) and MR219 (inferior grain fill) for comparative studies.
Presently mRNA isolation and labeling with fluorescent dyes for
hybridization is in progress. Samples are also being collected to study
expression at different developmental stages in MR84 and from MR185 for
comparison to MR84.
F
Posters
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