Factor-dependent dissociation of wheat germ ribosomes

Ribosome dissociation factor has been found in wheat germ acetone powder extracts. Further purification of the crude extract by pH and ammonium sulfate fractionations, DEAE-cellulose and CM-Sephadex column chromatography has resulted in the separation of two active fractions. The possibility that ribosome dissociation activity exhibited by either fraction is due to a protease or nuclease is considered unlikely, based on results of experiments involving ribosome dissociation kinetics, subunit structural integrity, and treatment with a serine protease inhibitor. Wheat germ ribosome dissociation factor is not species-specific. Dissociation factor from both fractions will promote the dissociation of Escherichia coli 70-S as well as Artemia salina 80-S ribosomes. Although both dissociation factor activities show the same dependence on K⁺ and Mg²⁺ for optimal activity, the two activities exhibit significant differences in their sensitivity to sulfhydryl reagents and heat, and in their dependence on incubation temperature for activity. Certain properties of both factors suggest that neither factor is initiation factor eIF-3; however, the possibility that one or both factors are subunits of initiation factor eIF-3 remains to be determined.     

Amino acid incorporation by ribosomes and polyribosomes from wheat chloroplasts

Sucrose-gradient and analytical ultracentrifugation showed that chloroplast polyribosomes from 4-day-old seedlings had mono-, di-, tri-, tetra- and traces of penta-ribosomes, in contrast with those from 7-day-old seedlings in which only the mono-, di- and traces of tri-ribosomes were present. Without Mg(2+) the polyribosomes dissociated into ribosomal subunits. The rate of l-[U-(14)C]phenylalanine incorporation was threefold greater for preparations from 4- than from 7-day-old seedlings. Incorporation by the latter was stimulated by polyuridylic acid. The rates of incorporation were similar whether the reaction mixture contained chloroplast or wheat-germ transfer RNA and amino acid synthetases purified on methylated albumin-on-kieselguhr and Sephadex G-75 columns respectively. The cofactor requirement was the same as for isolated intact chloroplasts. Osmotic rupture of chloroplasts with and without Triton X-100 revealed the presence of free and bound ribosomes. Free single ribosomes isolated by osmotic shrinkage or prepared by pancreatic ribonuclease digestion of chloroplast polyribosomes had negligible incorporation activity. This activity was increased by washing or by polyuridylic acid, but was still only a fraction of that given by polyribosomes. A comparison of incorporation activity of chloroplast polyribosomes with those from the surrounding cytoplasm showed the former to be 20 times more active.     

Binding of wheat germ ribosomes to fragmented viral mRNA.

The specificity of binding of wheat germ ribosomes to mRNA was greatly altered by cleavage of the message. Fragmentation of reovirus mRNA allowed wheat germ ribosomes to bind and protect a variety of internal sequences which were not accessible to ribosomes in the intact message. In experiments using the polycistronic mRNA from bacteriophage R17, wheat germ ribosomes bound preferentially at the beginning of the lysis peptide and synthetase cistrons, and at a third site which may be derived from the C-terminal region of the A protein cistron. This result is similar to that reported previously in a mammalian translational system (J.F. Atkins et al., Cell 18:246-256, 1979) except that, in the present study, limited cleavage of the phage RNA was necessary to activate these sites. More extensive fragmentation of R17 RNA permitted wheat germ ribosomes to bind and protect a great many additional sites. Thus, presence of an (exposed) 5'-terminus on an RNA molecule appears to be necessary and sufficient for attachment of eucaryotic ribosomes.     

The prokaryotic characteristics of Eimeria tenella ribosomes

• 1.1. Ribosomes were purified from the oocysts of Eimeria tenella and characterized. Those in the unsporulated oocysts were mostly in the form of polysomes which became gradually dissociated during the early phase of sporulation and were mostly in the monomeric form in sporulated oocysts.
• 2.2. The monomeric E. tenella ribosome had the size of about 80S and consisted of 60 and 40S subunits. It was readily and completely dissociated to subunits at high potassium concentrations, and its subunits could not hybridize with those of chicken liver ribosome.
• 3.3. The E. tenella ribosomal RNA was estimated to have sedimentation coefficients of 5, 16 and 23S.
• 4.4. The E. tenella ribosomal proteins had a gel electrophoretic pattern similar to that of E. coli.
• 5.5. The in vitro polypeptide synthesis mediated by E. tenella ribosomes was inhibitable by tetracycline and chloramphenicol which are also active against the parasite in vivo.
• 6.6. Chloramphenicol bound to E. tenella ribosomes with a dissociation constant of about 10⁻⁵ M.
• 7.7. All the evidence demonstrates prokaryotic characteristics of the E. tenella ribosome and suggests that the parasite may be a primitive eukaryote.

     

Temperature fluctuations during development reduce male fitness and may limit adaptive potential in tropical rainforest Drosophila

Understanding the potential for organisms to tolerate thermal stress through physiological or evolutionary responses is crucial given rapid climate change. Although climate models predict increases in both temperature mean and variance, such tolerances are typically assessed under constant conditions. We tested the effects of temperature variability during development on male fitness in the rainforest fly Drosophila birchii, by simulating thermal variation typical of the warm and cool margins of its elevational distribution, and estimated heritabilities and genetic correlations of fitness traits. Reproductive success was reduced for males reared in warm (mean 24 °C) fluctuating (±3 °C) vs. constant conditions but not in cool fluctuating conditions (mean 17 °C), although fluctuations reduced body size at both temperatures. Male reproductive success under warm fluctuating conditions was similar to that at constant 27 °C, indicating that briefly exceeding critical thermal limits has similar fitness costs to continuously stressful conditions. There was substantial heritable variation in all traits. However, reproductive success traits showed no genetic correlation between treatments reflecting temperature variation at elevational extremes, which may constrain evolutionary responses at these ecological margins. Our data suggest that even small increases in temperature variability will threaten tropical ectotherms living close to their upper thermal limits, both through direct effects on fitness and by limiting their adaptive potential.     

小麦温型现象研究

通过多年来用红外技术对不同品种小麦群体温度进行连续观测,结果发现,小麦具有温型现象.不同温度型的小麦在外部和内部性状上各有一些鲜明特点.冷型小麦的许多重要性状较暖型小麦为优.功能叶的功能期长10.8%,每株根数多20条,旗叶的叶绿素含量、蒸腾率和净光合率分别高出0.1389g·100g^-1·FW、0.0035mol·m^-2·s^-1和4.71μmol·m^-2·s^-1.由此可以认为,温度型和小麦的一系列重要性状密切相关.     

Synthetic hexaploid wheat enhances variation and adaptive evolution of bread wheat in breeding processes

Synthetic hexaploid wheat (SHW) that combines novel and elite genes from the tetraploid wheat Triticum turgidum L. and wild ancestor Aegilops tauschii Coss., has been used to genetically improve hexaploid common wheat. The abundant genetic diversity in SHW can effectively make breakthroughs in wheat genetic improvement through the inclusion of increased variation. In this paper, we reviewed the current advances in research and utilization of the primary SHW lines and SHW-derived wheat varieties that have enhanced evolution of modern wheat under conditions of natural and artificial selection in southwestern China. Using primary SHW lines, four high-yielding wheat varieties have been developed. In addition, using the SHW-derived varieties as breeding parents, 12 new wheat varieties were also developed. Results of genotype–phenotype and fingerprint analysis showed that the introgressed alleles from SHW lines have contributed a great number of elite characters to the new wheat varieties, and these elite characters include disease resistance, more spikes per plant, more grains per spike, larger grains, and higher grain-yield potential. We found that the primary SHW lines and SHW-derived varieties have identifiable effects to enhance genetic variation and adaptive evolution of modern hexaploid wheat, which significantly increased the grain yields of hexaploid wheat in recent years. These findings have significant implications in the breeding of high-yielding wheat varieties resistant to biotic and abiotic stresses using SHW as genetic resources.     

Interaction of elongation factor 2 from wheat germ with guanosine nucleotides and ribosomes.

1. The amino acid composition of wheat germ EF2 differs to some extent from that of elongation factors from mammals and bacteria. 2. The purified wheat germ EF2, similarly as the factors from other sources, is active in the: EF1-dependent polymerization of phenylalanine; ribosome-dependent GTP hydrolysis; binding of guanosine nucleotides; and ADP-ribosylation in the presence of diphtheria toxin. Fusidic acid at a concentration of 1 mM inhibits all these EF2-dependent reactions. 3. Diphtheria toxin in the presence of NAD+ inhibits polymerization of phenylalanine but does not effect GTP binding to EF2. 4. Binding of GDP to wheat germ EF2 is inhibited by ribosomes. During interaction with ribosomes, GTP in EF2-GTP complex is rapidly hydrolysed to GDP. Both GTP and 5'-guanylmethylenediphosphonate competitively inhibit formation of the ribosome-EF2-GDP complex due to the replacement of GDP from the complex. The latter is stabilized by fusidic acid.     

The effect of the genome of the variety on the adaptive characteristics of alloplasmic strains of winter wheat

The analysis of adaptability and genetical distances between 12 cytoplasms of Aegilops, Triticum and Haynaldia villosa for three winter wheat genomes showed an existence of genome-plasmon interactions. The plasmons of Ae. variabilis, Ae. cylindrica, Ae. squarrosa var. strangulata, T. dicoccoides appeared to be perspective in practical breeding for adaptability. Interactions are revealed as alteration of productivity and adaptability, and as genetic divergence.     

小麦温型现象研究

通过多年来用红外技术对不同品种小麦群体温度进行连续观测,结果发现,小麦具有温型现象.不同温度型的小麦在外部和内部性状上各有一些鲜明特点.冷型小麦的许多重要性状较暖型小麦为优.功能叶的功能期长10.8%,每株根数多20条,旗叶的叶绿素含量、蒸腾率和净光合率分别高出0.1389g·100g^-1·FW、0.0035mol·m^-2·s^-1和4.71μmol·m^-2·s^-1.由此可以认为,温度型和小麦的一系列重要性状密切相关.     

Temperature and growth-induced water potential

When the steins of dark-grown soybean [Glycine max (L.) Merr.] seedlings grew rapidly at favorable temperatures in saturating humidities, a water potential of about 0·2 MPa was induced by growth ($pS_o-$pS_w, where $pS_o is the water potential of the basal nonelongating tissue and $pS_w is the water potential of the elongating tissue). If this water potential was caused by high concentrations of solute in the apoplast, as has been proposed, lowering the temperature should have little effect on the potential. On the other hand, if the water potential was caused by apoplast tensions generated by growth, then the tensions should disappear as growth is inhibited by low temperatures. We observed that the growth-induced water potential became too small to detect when growth was inhibited by temperatures as low as 13—5 °C. The disappearance was observed as a rise in apoplast water potential using a thermocouple psychrometer for intact plants, a rise in cell turgor using a miniature pressure probe and a decrease in apoplast tensions using a pressure chamber. The disappearance was not caused by a loss of solute from the apoplast because the tensions fully accounted for the growth-induced water potential at all temperatures. The results are consistent with the lack of solute measured directly in the apoplast solutions at high temperatures (Nonami & Boyer 1987). Therefore, it was concluded that little solute was present in the apoplast at any temperature, and the growth-induced water potential was associated mostly with a tension that moved water from the xylem and into the surrounding cells to meet the demand of cell enlargement.     

Physicochemical characteristics of isolated 55-S mitochondrial ribosomes from rat-liver

A procedure for the preparative isolation of rat-liver mitochondrial ribosomes free of cytoplasmic contamination is described. The 55-S ribosome was characterized by electrophoretic analysis on acrylamide and agarose gels. It is concluded that its charge/mass ratio is very low and that its volume is larger than that of E. coli ribosomes and smaller than that of rat-liver cytoplasmic ribosomes. Therefore, the 55-S ribosome is not adequately described by the term “miniribosome”.