2 research outputs found
Development and application of qRT-PCR for sugar beet gene expression analysis in response to in vitro induced water deficit
Abstract Sugar beet is a significant industrial crop, often grown in
the areas where summer drought can severely limit root yield and sugar
content. In order to improve development of sugar beet cultivars with
increased drought tolerance it is necessary to understand plant
response to water stress at the genomic level. Since recent research
efforts have focused on the molecular response of the plant in order to
identify water deficit inducible genes, the aim of this investigation
was to develop qRT-PCR methodology for the quantification of gene
expression in sugar beet under conditions of water deficiency in vitro.
Sugar beet genotypes, selected for different response to water deficit,
were grown and multiplied in vitro. Axilary shoots were placed on
micropropagation media with 0%, 3% and 5% PEG, for 28 days. To
determine reaction of sugar beet genotypes to in vitro induced water
deficit changes in number of axillary shoots, shoot fresh weight and
dry matter content were measured. Total RNA was extracted from leaves
and reverse transcribed into cDNA, which served as matrix in real-time
PCR reaction using TaqMan technology. The housekeeping gene for
glutamine synthetase was used as endogenous control, while the genes
for alpha amylase and osmotin-like protein were target genes. The
relative quantification values for each target gene were calculated by
the 2- 06 06Ct method. Selected candidate genes differed in
relative gene expression among genotypes and applied PEG treatments.
The obtained results indicated that qRT-PCR protocol was efficient and
accurate, showing the potential to be used in further expression
analysis of candidate genes involved in sugar beet reaction to water
stress