here - First Legume Society Conference (LSC1)

Book of Abstracts First Legume Society Conference 2013: A Legume Odyssey Novi Sad, Serbia, 9-11 May 2013
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An optimized RNA Extraction Protocol for leaves, pods and seeds from Phaseolus
vulgaris
JR Parreira 1 , P Fevereiro 2,3 , SS Araújo 1
1 Centro BIOTROP - Veterinária e Zootecnia, Instituto de Investigação Científica e Tropical, Lisboa, Portugal
2 Department of Plant Biology, Faculty of Science, University of Lisbon, Lisboa, Portugal
3 Plant Cell BiotechnologyLab, Instituto de Tecnologia Química e Biológica, Universida de Nova de Lisboa,
Portugal
Phaseolus vulgaris is a major staple food crop and the most important legume in human diet, being
a source of protein, fiber, complex carbohydrates, vitamins and minerals. Grain filling (GF) is the
seed maturation phase that plays a major role in the seed storage reserve composition.
Understanding GF and how it can be affected by water deficit (WD) is a topic of great
agronomical and economical relevance, especially for the improvement of selecting and breeding
of common bean lines with improved tolerance to this abiotic stress. We aim to use a
transcriptomic approach to obtain a global insight into what genes are being expressed in seeds
that have a major role in GF under WD and what metabolic pathways are activated or repressed
in this condition. Additionally, we want to explore the role that surrounding organs, such as
leaves and pods might have in this biological process.Gene expression analysis requires a large
quantity of high quality RNA but routine RNA extraction protocols reveal to fail when applied to
pod and seed tissues collected from common bean. Here we present reliable RNA extraction
protocols that allow recovering high quantity and quality RNA suitable for other downstream
applications like Next Generation Sequencing or Quantitative PCR.
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