Sublethal doses of glyphosate [N-(phosphonomethyl)glycine] applied in a 50-μl drop in the partially furled third true leaf of 30-day-old sorghum [Sorghum bicolor (L.) Moench ‘Tophand’] seedlings induced basal stem swelling and bud release. Combining glyphosate with cycloheximide {3-[2-(3,5-dimethyl-2-oxycyclohexyl)-2-hydroxyethyl]glutarimide}, a cytokinin [N6 (2-isopentenyl)adenine], or L-phenylalanine significantly reduced the incidence of basal stem swelling. No such reductions was observed when indole-3-acetic acid or L-tyrosine was combined with glyphosate. The fresh weights of the seedlings given the combination treatments reflected the effects of glyphosate, except one combination that included tyrosine significantly reduced fresh weight and three combinations that included phenylalanine significantly increased fresh weight.

The effect of various growth regulators on seedling field bindweed (Convolvulus arvensis L. # CONAR) growth and control with herbicides was evaluated. Fourteen growth regulators were applied at 0.38 to 380 g/ha 7 days before herbicide application in greenhouse, growth chamber and field studies. Field bindweed seedlings pretreated with growth regulators were more effectively controlled by herbicides compared to those not pretreated in greenhouse evaluations. Abscisic acid [[R-(Z,E)]-(+)-5-(1-hydroxy-2,6-6-trimethyl-4-oxo-2-cyclohexane-1-yl)-3-methyl-2,4-pentadicenoic acid] (ABA), ancymidol [a-cyclopropyl-a-(p-methoxyphenyl)-5-pyrimidine methanol], cycloheximide {3-[2-(3,5-dimethyl-2-oxocylohexyl)-2-hydroxyethyl]glutarimide}, ethylenediamine tetraacetic acid [N,N′-1,2-ethanediyl-bis-(N-(carboxymethyl) glycine] (EDTA), and 1-naphthaleneacetic acid (NAA) increased field bindweed stem length. Ancymidol and gibberellic acid [ent-3α,10,13-trihydroxy-20-norgibberell-1,16-diene-7,19-dioc acid 19,10-lactone] (GA) increased root weight, and GA increased root length. Cycloheximide, ethephon [(2-chloroethyl)phosphonic acid] and MH (1,2-dihydro-3,6-pyridazinedione) decreased field bindweed root weight. Ancymidol, GA and IAA (indole-3-acetic acid) increased the number of shoots per plant of seedling field bindweed. Dicamba (3,6-dichloro-2-methoxybenzoic acid) efficacy was enhanced following field bindweed preconditioning with ethephon and IAA. Ancymidol and cycloheximide applied as pretreatments increased glyphosate [N-(phosphonomethyl)glycine] activity on seedling field bindweed. Ancymidol and IAA pretreatment improved field bindweed control with triclopyr {[(3,5,6-trichloro-2-pyridinyl)oxy] acetic acid}. Herbicide-growth regulator combination treatments that increased field bindweed control in greenhouse and growth chamber studies did not increase top growth control in the field.

Increasing the efficiency of intracytoplasmic sperm injection (ICSI) in domestic animals has been attempted by many researchers, however embryonic development to the blastocyst stage remains low compared with that of in vitro fertilization (IVF) embryos. One of the main problems observed in cattle is inadequate oocyte activation after ICSI. The present study compared the effect of cycloheximide (CHX), 6-dimethylaminopurine (DMAP), and anisomycin (ANY) on the fertilization rate, development, ploidy and quality of bovine embryos generated by ICSI. Although no differences were observed between treatments in terms of cleavage, higher blastocyst rates were observed for ANY (37.3%) compared with CHX (21.8%, P < 0.05) and DMAP (28.6%, P > 0.05) treatments. No differences were observed in the quality of embryos as assessed by the total number of cells, their distribution to the different embryo compartments [inner cell mass (ICM) and trophectoderm (TE)], the proportion of ICM cells to the total cell numbers and terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL)-positive cells. Similarly, no differences were observed in the normal ploidy of embryos (56, 67, and 55%) for ANY, CHX and DMAP, respectively. However, higher fertilization rates were observed for ANY (75%) and CHX (87%) treatments compared with DMAP (35%). In conclusion, ANY showed a superior developmental rate compared with CHX treatment. Although no significant differences were observed compared with an improved protocol of DMAP (2Io-DMAP), the lower fertilization rate recorded with DMAP strongly suggests that ANY could be a better alternative for oocyte activation than traditional chemical compounds used currently in ICSI.

This study evaluated the effects of reversible meiotic inhibition and different culture media (PZM3 or NCSU23) on production of porcine embryos by either in vitro fertilization (IVF) or parthenogenetic activation (PA). Oocytes from abattoir-derived ovaries were allocated into two groups for maturation: CHX (5 μg/ml cycloheximide for 10 h) or Control (no CHX). The percentage of metaphase II (MII) oocytes was determined at 36, 40 or 44 h of in vitro maturation. For IVF and PA, denuded oocytes were fertilized with purified sperm for 6 h or activated by electric stimuli. Zygotes were then subdivided into two culture groups: NCSU23 or PZM3. No effect of treatment with CHX and culture media was observed on cleavage (D3) and blastocyst (D7) rates in IVF and PA groups. There are no differences of quality or development rates between IVF-derived embryos cultured in NCSU23 or PZM3. However, we observed high quality PA embryos in PZM3 compared with NCSU23. Maturation arrest with CHX decreased the average blastocyst cell number in IVF while it was increased in PA embryos. As older oocytes are more effectively activated, CHX– blocked oocytes reached the mature stage faster than the control group. In conclusion, the CHX treatment for 10 h, followed by oocyte maturation for 40 h, is an efficient protocol to produce high quality parthenote embryos, especially when they are cultured in PZM3. However, this protocol is not satisfactory for IVF embryos production. In this case, a shorter maturation period could provide better embryo quality.

In the mouse, mature oocytes injected with prespermatozoal cell nuclei remain unactivated. Additional stimulation is needed to trigger oocyte activation leading to embryo development. We compared various electrical stimulations, treatment with cycloheximide alone or in combination with electrical stimulation, and injection of sperm-borne oocyte-activating factor (oscillogen) in terms of their oocyte activation and embryo development rates. Of all the treatments tested, a single electrical pulse (1.0 kV / cm, 128 μs) was the simplest, yet very effective, in allowing the development of the oocytes injected with spermatid nuclei.

Following fertilisation, the sperm triggers a series of intracellular changes which initiate oocyte activation and pronuclear formation. Oocyte activation can also be induced artificially by several chemicals, such as the calcium ionophore A23187. The sperm nucleus is transformed into the male pronucleus through the interaction of oocyte cytoplasmic factors. The profile of protein synthesis is different in bovine oocytes following fertilisation and parthenogenetic activation. The formation of male and female pronuclei was not blocked by the presence of the protein synthesis inhibitor cycloheximide. These results suggest that bovine oocyte activation by sperm and parthenogenetic activation induce different cytoplasmic responses for protein synthesis and that new protein synthesis is not required for male pronuclear formation in bovine zygotes.

In the course of the cell cycles of synchronous cultures of the chlorococcal alga Scenedesmus quadricauda, the following were monitored: total protein and RNA accumulation as a measure of growth processes, the timing of the commitment points at which the cells trigger the sequence of reproductive processes (DNA replication, nuclear and cellular division) and the course of the reproductive processes. The synchronous cultures were grown either under various lighting regimes, or in the temporary presence of specific inhibitors of either proteosynthesis (cycloheximide) or DNA replication (5-fluorodeoxyuridine). By adjusting the length of the light period, the cell cycle could be manipulated. Cell cycle patterns could be altered to give different numbers of sequences of reproductive processes. The extent of their mutual overlap could be influenced and the number of daughter cells produced could be altered. Schematic illustrations of various cell cycle patterns and comparisons with those of higher plants and other algal species are presented.

Sequence elements that can function as internal ribosome entry sites (IRES) have been identified in 5′ noncoding regions of certain uncapped viral and capped cellular mRNA molecules. However, it has remained largely unknown whether IRES elements are functional when located in their natural capped mRNAs. Therefore, the polysomal association and translation of several IRES-containing cellular mRNAs was tested under conditions that severely inhibited cap-dependent translation, that is, after infection with poliovirus. It was found that several known IRES-containing mRNAs, such as BiP and c-myc, were both associated with the translation apparatus and translated in infected cells when cap-dependent translation of most host-cell mRNAs was blocked, indicating that the IRES elements were functional in their natural mRNAs. Curiously, the mRNAs that encode eukaryotic initiation factor 4GI (eIF4GI) and 4GII (eIF4GII), two proteins with high identity and similar functions in the initiation of cap-dependent translation, were both associated with polysomes in infected cells. The 5′-end sequences of eIF4GI mRNA were isolated from a cDNA expression library and shown to function as an internal ribosome entry site when placed into a dicistronic mRNA. These findings suggest that eIF4G proteins can be synthesized at times when 5′ cap-dependent mRNA translation is blocked, supporting the notion that eIF4G proteins are needed in both 5′ cap-independent and 5′ cap-dependent translational initiation mechanisms.