Stable reference genes are indispensable for ensuring the fidelity of determined gene expression levels. However, the expression levels of reference genes are unable to remain constant under all possible experimental conditions. Therefore, the stability determination of reference genes is necessary in an experimental system set. In the preset study, the stability of nine cucumber candidate reference genes (CsACT, CsUBQ, CsEF1α, CsCYP, CsαTU, CsCACS, CsTIP41, CsYSL8 and CsHEL) subjected to stresses from Phytophthora melonis(P. melonis) were determined using four different analysis methods, including Delta Ct, BestKeeper, NormFinder and GeNorm. The study results revealed that CsUBQ and CsCYP were the most stable genes suitable as internal control in cucumber plants under attack by P. melonis condition.
An isolate of zucchini yellow mosaic virus (ZYMV) was obtained from naturally infected squash fruits were grown in Abees region, Alexandria governorate. Disease symptoms were Showing mosaic, yellowing and blistering and absis symptoms. The identification was based on the symptoms developed on diagnostic hosts and serological reactions with antisera to cucumber mosaic cucumovirus (CMV), watermelon mosaic potyvirus 2 (WMV-2) and ZYMV. Squash fruit isolate of ZYMV was transmitted by Aphis gossypii, Aphis neri and Myzus persicae in non-persistent manner. The virus was purified by ultra-centrifugation and PEG. The purified virus had an ultraviolet absorption spectrum typical of a nucleoprotein with A260/280 and A280/260 being 1.1 and 0.91 respectively. The yield of purified virus was 1.62 mg/100g infected leaf tissues. Specific antiserum was prepared and found to have a titer of 1:409600 as determined by indirect ELISA.
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