Flow cytometry estimation of nuclear size and ploidy level of habituated calli of sugar beet

in Biologia Plantarum (1999), 42(3), 321-332

A fully habituated (auxin- and cytokinin-independent) self-regenerating (organogenic) sugar beet cell line (HO) and a fully habituated non-organogenic one (HNO) derived from the former one, were analyzed ... [more ▼]

A fully habituated (auxin- and cytokinin-independent) self-regenerating (organogenic) sugar beet cell line (HO) and a fully habituated non-organogenic one (HNO) derived from the former one, were analyzed as to their nuclear size and DNA content. Flow cytometry and image analysis were used and cells of certified diploid leaves of the same sugar beet strain served as controls. The HNO cells had been shown previously to have many characteristics of cancerous cells. The analyses made on leaves and HNO cells indicated the presence of only one population of cycling cells. In HO cells. two cycling populations were detected: the first one had the same DNA content as the leaves while the second one contained two fold more DNA than the first population. HNO cells showed the higher nuclear size and DNA content. HNO cells also showed evidence of aneuploidy. Thus, nuclear size, DNA content and ploidy level increase together with the neoplasic progression to culminate in HNO cells with the loss of organogenic totipotency. [less ▲]

Diurnal Rhythmicity in the Pattern of Mrnas in the Leaves of Sinapis Alba

in Plant Physiology (1990), 94(4), 1590-1597

Previous studies have shown that certain specific leaf mRNAs exhibit a diurnal rhythmicity in their quantity in higher plants. To determine whether this situation is restricted to a few mRNAs, or affects ... [more ▼]

Previous studies have shown that certain specific leaf mRNAs exhibit a diurnal rhythmicity in their quantity in higher plants. To determine whether this situation is restricted to a few mRNAs, or affects a large number, we have used in vitro translation and two-dimensional polyacrylamide gel electrophoresis to analyze the mRNA complement in leaves of Sinapis alba at different times during an 8-hour/16-hour day/night cycle. A method for the visual analysis of two-dimensional polyacrylamide gel electrophoresis was also developed. This method selected, at each sampling time, spots that were significant. It then selected, between two sampling times, intensity changes that were significant at the 0.02 confidence level. During a day/night cycle, complex rhythmic changes affected about 10% of the mRNAs. Nineteen different rhythm patterns were found. These 19 patterns fell into four main classes: mRNAs that increase during the light period and decrease during the dark, mRNAs that increase and then decrease during the light period, mRNAs that decrease during the light period and increase during the dark period, and mRNAs that increase and then decrease during the dark period. [less ▲]

The action of exogenous abscisic acid on malate synthase synthesis in germinating castor bean seeds

in Planta (1985), 166

Newly Synthesized mRNA Is Translated During the Initial Imbibition Phase of Germinating Maize Embryo

in Plant Physiology (1983), 73(2), 484-487

RNA synthesis is activated in the cells of the plant embryo very soon after the start of seed imbibition. We previously reported that mainly heterogeneous nuclear RNA is synthesized in the radicle of Zea ... [more ▼]

RNA synthesis is activated in the cells of the plant embryo very soon after the start of seed imbibition. We previously reported that mainly heterogeneous nuclear RNA is synthesized in the radicle of Zea mays embryo during the first hours of germination. The present study was undertaken in order to detect the time of appearance of the newly synthesized messenger RNA in the polysomes of germinating maize axes.Free polysomes were prepared from embryonic axes rehydrated for 2 hours in the presence of radioactively labeled uridine. These polysomes were shown to be labeled and to contain labeled particles sedimenting, after dissociation with EDTA, in the 10S to 40S region of a sucrose gradient. The labeled polysomal RNA migrates heterogeneously in a gel with a mean size corresponding to about 16S, and 60% of these molecules are polyadenylated.The data indicate that the newly synthesized RNA associated with the polysomes after 2 h of germination consists of messenger RNA molecules. Analysis of the polysomes prepared 0.5 and 1 h after the start of imbibition suggests that translation of the newly synthesized messenger RNA probably occurs within the 1st hour of imbibition of the isolated axis, thus well before the completion of the initial water uptake. [less ▲]