Changes in Hexokinase Activity in Echinochloa phyllopogon and Echinochloa crus-pavonis in Response to Abiotic Stress

Hexokinase (HXK; EC 2.7.1.1) regulates carbohydrate entry into glycolysis and is known to be a sensor for sugar-responsive gene expression. The effect of abiotic stresses on HXK activity was determined in seedlings of the flood-tolerant plant Echinochloa phyllopogon (Stev.) Koss and the flood-intolerant plant Echinochloa crus-pavonis (H.B.K.) Schult grown aerobically for 5 d before being subjected to anaerobic, chilling, heat, or salt stress. HXK activity was stimulated in shoots of E. phyllopogon only by anaerobic stress. HXK activity was only transiently elevated in E. crus-pavonis shoots during anaerobiosis. In roots of both species, anoxia and chilling stimulated HXK activity. Thus, HXK is not a general stress protein but is specifically induced by anoxia and chilling in E. phyllopogon and E. crus-pavonis. In both species HXK exhibited an optimum pH between 8.5 and 9.0, but the range was extended to pH 7.0 in air-grown E. phyllopogon to 6.5 in N₂-grown E. phyllopogon. At physiologically relevant pHs (6.8 and 7.3, N₂ and O₂ conditions, respectively), N₂-grown seedlings retained greater HXK activity at the lower pH. The pH response suggests that in N₂-grown seedlings HXK can function in a more acidic environment and that a specific isozyme may be important for regulating glycolytic activity during anaerobic metabolism in E. phyllopogon.     

Data mining of molecular dynamics trajectories of nucleic acids

Analysis, storage, and transfer of molecular dynamic trajectories are becoming the bottleneck of computer simulations. In this paper we discuss different approaches for data mining and data processing of huge trajectory files generated from molecular dynamic simulations of nucleic acids.     

Assessment of Epstein-Barr virus nucleic acids in gastric but not in breast cancer by next-generation sequencing of pooled Mexican samples

Gastric (GC) and breast (BrC) cancer are two of the most common and deadly tumours.Different lines of evidence suggest a possible causative role of viral infections forboth GC and BrC. Wide genome sequencing (WGS) technologies allow searching for viralagents in tissues of patients with cancer. These technologies have alreadycontributed to establish virus-cancer associations as well as to discovery new tumourviruses. The objective of this study was to document possible associations of viralinfection with GC and BrC in Mexican patients. In order to gain idea about costeffective conditions of experimental sequencing, we first carried out an insilico simulation of WGS. The next-generation-platform IlluminaGallx wasthen used to sequence GC and BrC tumour samples. While we did not find viralsequences in tissues from BrC patients, multiple reads matching Epstein-Barr virus(EBV) sequences were found in GC tissues. An end-point polymerase chain reactionconfirmed an enrichment of EBV sequences in one of the GC samples sequenced,validating the next-generation sequencing-bioinformatics pipeline.     

Population Dynamics of Plant Nematodes in Cultivated Soil: Length of Rotation in Newly Cleared and Old Agricultural Land

During a 6-year study of 1-, 2-, and 3-year crop rotations, population densities of Pratylenchus brachyurus, Trichodorus christiei, and Meloidogyne incognita were significantly affected by the choice of crops but not by length of crop rotation. The density of P. brachyurus and T. christiei increased rapidly on milo (Sorghum vulgate). In addition, populations of P. brachyurus increased significantly in cropping systems that involved crotalaria (C. rnucronata), millet (Setaria italica), and sudangrass (Sorghum sudanense). Lowest numbers of P. brachyurus occurred where okra (Hibiscus esculentus) was grown or where land was fallow. The largest increase in populations of T. christiei occurred in cropping systems that involved millet, sudangrass, and okra whereas the smallest increase occurred in cropping systems that involved crotalaria or fallow. A winter cover of rye (Secale cereale) had no distinguishable effect on population densities of P. brachyurus or T. christiei. Meloidogyne incognita was detected during the fourth year in both newly cleared and old agricultural land when okra was included in the cropping system. Detectable populations of M. incognita did not develop in any of the other cropping systems. Yields of tomato transplants were higher on the newly cleared land than on the old land. Highest yields were obtained when crotalaria was included in the cropping system. Lowest yields were obtained when milo, or fallow were included in the cropping system. Length of rotation had no distinguishable effect on yields of tomato transplants.     

Use of EDTA derivatization to characterize interactions between oligodeoxyribonucleoside methylphosphonates and nucleic acids

EDTA-derivatized oligonucleoside methylphosphonates were prepared and used to characterize hybridization between the oligomers and single-stranded DNA or RNA. The melting temperatures of duplexes formed between an oligodeoxyribonucleotide 35-mer and complementary methylphosphonate 12-mers were 4-12 degrees C higher than those of duplexes formed by oligodeoxyribonucleotide 12-mers as determined by spectrophotometric measurements. Derivatization of the methylphosphonate oligomers with EDTA reduced the melting temperature by 5 degrees C. Methylphosphonate oligomer-nucleic acid complexes were stabilized by base stacking interactions between the terminal bases of the two oligomers binding to adjacent binding sites on the target. In the presence of Fe2+ and DTT, the EDTA-derivatized oligomers produce hydroxyl radicals that cause degradation of the sugar-phosphate backbone of both targeted DNA and RNA. Degradation occurs specifically in the region of the oligomer binding site and is approximately 20-fold more efficient for single-stranded DNA than for RNA. In comparison to the presence of one oligomer, the extent of target degradation was increased considerably by additions of two oligomers that bind at adjacent sites on the target. For example, the extent of degradation of a single-stranded DNA 35-mer caused by two contiguously binding oligomers, one of which was derivatized by EDTA, was approximately 2 times greater than that caused by the EDTA-derivatized oligomer alone. Although EDTA-derivatized oligomers are stable for long periods of time in aqueous solution, they undergo rapid autodegradation in the presence of Fe2+ and DTT with half-lives of approximately 30 min. This autodegradation reaction renders the EDTA-derivatized oligomers unable to cause degradation of their complementary target nucleic acids.(ABSTRACT TRUNCATED AT 250 WORDS)     

Relationship Between Meloidogyne hapla Density and Damage to Carrots in Organic Soils

Field and growth chamber experiments were conducted to determine the effect of five initial densities (Pi - 20 - 240/100 cm³ soil) of Meloidogyne hapla on carrot development and yield of storage roots at maturity. Carrots growing in infested and noninfested organic soil were harvested after 15, 29, 44, 59, and 106 days of growth in controlled environment chambers and after l l 0 days in field plots. Nematodes affected weight of roots and foliage, weight and length of the storage portion of tap roots, and induced malformations (forking), galling, and hairiness of tap roots. In most cases the data could not be represented satisfactorily by the exponential model of Seinhorst: y = m + (l-m) Z^P-t. In growth chambers the weight of mature storage roots was not correlated to initial nematode density, but there was a significant negative correlation between weight of storage roots and initial nematode density in field plots. Tolerance levels were calculated as points where the regression lines reached the growth level on noninoculated plants. The tolerance levels of foliage were higher than those of roots, and increased with age of plants. The tolerance level of marketable weight in field plots, average crop value, and a hypothetical control cost function are used to discuss the possibility of optimizing chemical control of root-knot nematode in organic soils.     

蛋白质/核酸相互作用研究方法进展

蛋白质和核酸是构成生命体最为重要的两类生物大分子,蛋白质与核酸的相互作用是分子生物学研究的中心问题之一,它是许多生命活动的重要组成部分。研究蛋白质/核酸相互作用近期采用的新技术有:核酸适体技术、生物信息学方法、蛋白质芯片技术以及纳米技术等。本文就这些新的研究方法进行综述。     

The interactions of nucleic acids at elevated hydrostatic pressure

The application of elevated hydrostatic pressure on the order of a few thousand bar provides insight into the molecular forces responsible for stabilizing the conformations and non-covalent interactions of biological molecules in aqueous solution. In particular, the parameters derived from these studies have enabled researchers to glean information regarding the importance of hydration in the energetics and kinetics of these systems. This review presents data concerned with the application of hydrostatic pressure to study the thermodynamics, kinetics, and structure of nucleic acids and the interactions between nucleic acids and proteins, enzymes, and drugs. These complexes often form extremely stable non-covalent complexes in which electrostatic interactions play an important role. The sensitivity of these interactions to pressure offers a valuable experimental tool for investigating the energetics of the complexes.     

Dimethylsulfoniopropionate Biosynthesis in Spartina alterniflora : Evidence That S-Methylmethionine and Dimethylsulfoniopropylamine Are Intermediates

The osmoprotectant 3-dimethylsulfoniopropionate (DMSP) occurs in Gramineae and Compositae, but its synthesis has been studied only in the latter. The DMSP synthesis pathway was therefore investigated in the salt marsh grass Spartina alterniflora Loisel. Leaf tissue metabolized supplied [³⁵S]methionine (Met) to S-methyl-l-Met (SMM), 3-dimethylsulfoniopropylamine (DMSP-amine), and DMSP. The ³⁵S-labeling kinetics of SMM and DMSP-amine indicated that they were intermediates and, consistent with this, the dimethylsulfonium moiety of SMM was shown by stable isotope labeling to be incorporated as a unit into DMSP. The identity of DMSP-amine, a novel natural product, was confirmed by both chemical and mass-spectral methods. S. alterniflora readily converted supplied [³⁵S]SMM to DMSP-amine and DMSP, and also readily converted supplied [³⁵S]DMSP-amine to DMSP; grasses that lack DMSP did neither. A small amount of label was detected in 3-dimethylsulfoniopropionaldehyde (DMSP-ald) when [³⁵S]SMM or [³⁵S]DMSP-amine was given. These results are consistent with the operation of the pathway Met → SMM → DMSP-amine → DMSP-ald → DMSP, which differs from that found in Compositae by the presence of a free DMSP-amine intermediate. This dissimilarity suggests that DMSP synthesis evolved independently in Gramineae and Compositae.     

Structural diversity and isomorphism of hydrogen-bonded base interactions in nucleic acids

The wide structural diversity of RNA results in part from the diversity of non-Watson-Crick interactions between bases. To examine the repertoire of possible hydrogen bond interactions among bases, we computed databases of base-pairs and base-triples by systematically matching all possible hydrogen-bond donors and acceptors between bases and evaluating the geometries of each planar configuration. For base-pairs, we find 53 arrangements having at least two hydrogen bonds, including 23 pairs with protonated bases that have not previously been modeled. A comparison with experimentally observed base-pairs reveals an unexpected G:U pair recently observed in the ribosome. For base-triples, we find 840 arrangements in which the three bases are constrained by a total of at least three hydrogen bonds. Base-triples in particular exhibit a wide range of structural diversity, suggesting how compact or elongated nucleic acid structures may be constructed using different hydrogen-bonding patterns. Base-pair and base-triple conformations were systematically compared to identify structurally isomorphic combinations, and the experimentally observed arrangements within double and triple helices are among the most isomorphic. Unexpectedly, however, other combinations in the database are even more isomorphic, including several in which all-purine arrangements overlap with all-pyrimidine arrangements. These studies highlight some of the combinatoric and geometric versatility of base interactions and help provide a framework for analyzing and modeling isomorphic interactions and potentially for designing novel nucleic acid structures.