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

Sadržaj polifenola i antioksidativni kapacitet ekstrakata listova paradajza zaraženih plamenjačom

Late bligth is one of the main causes of the tomato yeild losses. In this paper, the biochemical response of tomato variety Plovdivski kasni in leaves infected with Phytopththora infestans was analyzed. It was found that there is a correlation between the degree of disease severity and the content of total polyphenols and flavonoids in the leaves. The different content of phenolic compounds also affected the antioxidant capacity of leaf extracts. By increasing the degree of infection, the content of the soluble polyphenolic compounds and the antioxidant capacity of the leaves decreased.Plamenjača je jedan od glavnih uzroka smanjenja prinosa paradajza što može dovesti do velikih ekonomskih gubitaka. U ovom radu je ispitan biohemijski odgovor listova paradajza sorte ‘Plovdivski kasni’ na infekciju prouzrokovačem plamenjače Phytopththora infestans. Utvrđeno je da postoji korelacija između stepena zaraženosti biljke i sadržaja ukupnih polifenola i flavonoida u listovima. Različit sadržaj fenolnih jedinjenja je uticao i na antioksidativni kapacitet ekstrakata listova. Povećanjem stepena infekcije sadržaj rastvorljivih polifenolnih jedinjenja i antioksidativni kapacitet listova se smanjivao