Authors:Z. Zeng, T. Zhang, G. Li, C. Liu, and Z. Yang
A synthetic autopolyploid was developed from diploid Aegilops tauschii, D genome progenitor of common wheat (Triticum aestivum). The tetraploid Ae. tauschii displayed a markedly larger organ size than the diploid donor. Fluorescence in situ hybridization (FISH) and DNA marker analysis revealed that there is no clear variation at either the chromosomal or DNA level between the diploid and tetraploid plants. We analyzed the variation in cytosine methylation patterns between the diploid and tetraploid plants by methylation-sensitive amplified polymorphism (MSAP) and detected 228 and 232 methylated sites in diploid and tetraploid plants, respectively. Statistical comparison indicated that the tetraploid Ae. tauschii genotype displayed no significant difference in polymorphic methylation level compared to the diploid ones. Twenty-two different genomic fragments displaying different methylation behavior during the ploidy conversions were isolated and sequenced. It demonstrated that alterations in the level of methylation have the most profound effects on coding genes. We demonstrated that there are some genes expressions modified by DNA methylation may be correlated with phenotypic alteration after autotetraploidization.
Authors:L.J. Wu, Y. Shang, T. Liu, W.J. Chen, B.L. Liu, L.Q. Zhang, D.C. Liu, B. Zhang, and H.G. Zhang
In this study, the cDNA of homocysteine S-methyltransferase was isolated from Aegilops tauschii Coss., with the gene accordingly designated as AetHMT1. Similar to other methyltransferases, AetHMT1 contains a GGCCR consensus sequence for a possible zinc-binding motif near the C-terminal and a conserved cysteine residue upstream of the zinc-binding motif. Analysis of AetHMT1 uncovered no obvious chloroplast or mitochondrial targeting sequences. We functionally expressed AetHMT1 in Escherichia coli and confirmed its biological activity, as evidenced by a positive HMT enzyme activity of 164.516 ± 17.378 nmol min−1 mg−1 protein when catalyzing the transformation of L-homocysteine. Compared with the bacterium containing the empty vector, E. coli harboring the recombinant AetHMT1 plasmid showed much higher tolerance to selenate and selenite. AetHMT1 transcript amounts in different organs were increased by Na2SeO4 treatment, with roots accumulating higher amounts than stems, old leaves and new leaves. We have therefore successfully isolated HMT1 from Ae. tauschii and characterized the biochemical and physiological functions of the corresponding protein.
Authors:P.-J. Zhou, H.-T. Zhou, Y. Liu, S.-S. Qu, and Y.-G. Zhu
In this paper, the thermogenesis and differential scanning calorimetry (DSC) curves of energy release of the mitochondria
isolated from variant strains of cytoplasmic male sterile lines of rices have been determined by using an LKB2277 Bioactivity
Monitor and a DuPont 910 differential scanning calorimeter. The regularity and characteristics of the energy release of the
mitochondria at constant and changing temperature were investigated, the differences in shape of the curves and the thermodynamic
and kinetic characteristics of the thermogenesis of the mitochondria were compared, the thermodynamic and kinetic parameters
of energy release of the mitochondria in the thermogenesis increasing stage were calculated, and the experimental thermokinetic
equations describing the different thermogenesis processes were established.
Authors:L. N. Xu, F. Y. Gai, G. F. Mu, Y. Gao, H. T. Liu, and F. Luan
Formaldehyde in aquatic products was determined by micellar electrokinetic capillary chromatography (MEKC) after derivatization with 2,4-dinitrophenylhydrazine. Separation was carried out at 25 °C and 25 kV, using a fused silica capillary (75 µ internal diameter; 50.5 cm effective length) and an ultraviolet detector set at 360 nm. The optimal background electrolyte was 20 mM sodium tetraborate and 20 mM sodium dodecyl sulfate at pH 9.0 with 3 s hydrodynamic injection at 30 mbar. Electrophoretic analysis took approximately 6.5 min. The correlation coefficient of the calibration curve was 0.999 over the concentration range 2.0–100.0 mg L−1, and the LOD and LOQ values were 0.57 and 1.89 µg mL−1, respectively. The recoveries were from 83.7% to 97.2% with steam distillation as the sample pretreatment method.
Authors:J.Q. Xu, L. Wang, B.L. Liu, T.F. Xia, D.C. Liu, X. Chang, T.W. Zhang, H.G. Zhang, and Y.H. Shen
As one of the world’s earliest domesticated crops, barley is a model species for the study of evolution and domestication. Domestication is an evolutionary process whereby a population adapts, through selection; to new environments created by human cultivation. We describe the genome-scanning of molecular diversity to assess the evolution of barley in the Tibetan Plateau. We used 667 Diversity Arrays Technology (DArT) markers to genotype 185 barley landraces and wild barley accessions from the Tibetan Plateau. Genetic diversity in wild barley was greater than in landraces at both genome and chromosome levels, except for chromosome 3H. Landraces and wild barley accessions were clearly differentiated genetically, but a limited degree of introgression was still evident. Significant differences in diversity between barley subspecies at the chromosome level were observed for genes known to be related to physiological and phenotypical traits, disease resistance, abiotic stress tolerance, malting quality and agronomic traits. Selection on the genome of six-rowed naked barley has shown clear multiple targets related to both its specific end-use and the extreme environment in Tibet. Our data provide a platform to identify the genes and genetic mechanisms that underlie phenotypic changes, and provide lists of candidate domestication genes for modified breeding strategies.
Authors:J.W Guo, Q. Li, W.Q. Chen, X. Li, L.Q. Li, T.G. Liu, Z.L. Ren, and P.G. Luo
Leaf senescence is a notably important trait that limits the yield and biomass accumulation of agronomic crops. Therefore, determining the chromosomal position of the expression sequence tags (ESTs) that are associated with leaf senescence is notably interesting in the manipulation of leaf senescence for crop improvement. A total of 32 ESTs that were previously identified during the delaying leaf senescence stage in the stay-green wheat cultivar CN17 were mapped to 42 chromosomes, a chloroplast, a mitochondrion, and a ribosome using in silico mapping. Then, we developed 19 pairs of primers based on these sequences and used them to determine the polymorphisms between the stay-green cultivars (CN12, CN17, and CN18) and the control cultivar MY11. Among the 19 pairs of primers, 5 pairs produced polymorphisms between the stay-green cultivar and the non-stay-green control. Further studies of Chinese Spring nullisomic-tetrasomics show that JK738991 is mapped to 3B, JK738983 is mapped to 5D, and JK738989 is mapped to 2A, 4A, and 3D. The other two ESTs, JK738994 and JK739003, were not assigned to a chromosome using the Chinese Spring nullisomic-tetrasomics, which indicates that these ESTs may be derived from rye DNA in the wide cross. In particular, the ESTs that produce polymorphisms are notably useful in identifying the stay-green cultivar using molecular marker-assisted selection. The results also suggest that the in silico mapping data, even from a comparison genomic analysis based on the homogeneous comparison, are useful at some points, but the data were not always reliable, which requires further investigation using experimental methods.
Aegilops sharonensis (Sharon goatgrass) is a valuable source of novel high molecular weight glutenin subunits, resistance to wheat rust, powdery mildew, and insect pests. In this study, we successfully hybridized Ae. sharonensis as the pollen parent to common wheat and obtained backcross derivatives. F1 intergeneric hybrids were verified using morphological observation and cytological and molecular analyses. The phenotypes of the hybrid plants were intermediate between Ae. sharonensis and common wheat. Observations of mitosis in root tip cells and meiosis in pollen mother cells revealed that the F1 hybrids possessed 28 chromosomes. Chromosome pairing at metaphase I of the pollen mother cells in the F1 hybrid plants was low, and the meiotic configuration was 25.94 I + 1.03 II (rod). Two pairs of primers were screened out from 150 simple sequence repeat markers, and primer WMC634 was used to identified the presence of the genome of Ae. sharonensis. Sequencing results showed that the F1 hybrids contained the Ssh genome of Ae. sharonensis. The sodium dodecyl sulfate polyacrylamide gel electrophoresis profile showed that the alien high molecular weight glutenin subunits of Ae. sharonensis were transferred into the F1 and backcross derivatives. The new wheat-Ae. sharonensis derivatives that we have produced will be valuable for increasing resistance to various diseases of wheat and for improving the quality of bread wheat.