Bulking generated considerable bias in detection of amplified fragment length polymorphism variations in oat,fringed brome and smooth bromegrass

Amplified fragment length polymorphism (AFLP) variations in bulk and plant‐by‐plant (PBP) samples of five oat (Avena sativa L.) accessions, one fringed brome (Bromus ciliatus L.) accession and two smooth bromegrass (B. inermis Leyss.) accessions were compared. The proportions of AFLP bands detected in PBP, but lost in bulk, samples of oat, fringed brome, and smooth bromegrass ranged from 19 to 31%, 40 to 44%, and 22 to 33% of the total bands scored, respectively. These lost bands had occurred at frequencies ranging from 0.1 to 1 in the PBP samples. These findings demonstrate bulking can generate substantial bias in detection of AFLP variations.     

Stimulatory effect of polyethylene glycol (PEG) on gene expression in mouse liver following hydrodynamics-based transfection

BACKGROUND: Rapid intravenous injection of a large volume of plasmid DNA (pDNA), i.e. a transfection procedure based on hydrodynamics, is known to be an efficient and liver-specific method of in vivo gene delivery. However, the gene expression is transient. METHODS: We investigated the effect of addition of polyethylene glycol (PEG) to a solution of naked pDNA (luciferase) on the expression of the gene in mouse liver following transfection by the hydrodynamics-based technique. In addition, the mechanism leading to the enhancement of the gene expression was studied. RESULTS: The addition of 1% (w/v) PEG2000 to the pDNA solution enhanced the resulting gene expression in the liver. Increasing the PEG2000 concentration to more than 1 and up to 10% (w/v) rather diminished the gene expression level. By contrast, increasing the molecular weight of PEG to over 2000 up to 10 000 did not affect the level of gene expression. Histopathological and serum-chemistry examinations indicated that hydrostatic or osmotic pressure increased tissue and hepatocellular damage in a PEG-concentration-dependent manner, and resulted in a decrease in gene expression. Quantitative evaluation showed that the enhanced gene expression resulted from stabilization of the pDNA introduced into the hepatocytes and an enhancement of the transport of intact pDNA to the nucleus. CONCLUSIONS: For most gene therapy applications and gene function studies, sustained expression of the introduced gene(s) is necessary. This simple method to achieve enhanced gene expression in liver may have a great potential for a wide variety of laboratory studies in molecular and cellular biology as well as possibly for future clinical applications in humans.     

Selective grazing of methanotrophs by protozoa in a rice field soil

Biological methane oxidation is a key process in the methane cycle of wetland ecosystems. The methanotrophic biomass may be grazed by protozoa, thus linking the methane cycle to the soil microbial food web. In the present study, the edibility of different methanotrophs for soil protozoa was compared. The number of methanotroph-feeding protozoa in a rice field soil was estimated by determining the most-probable number (MPN) using methanotrophs as food bacteria; naked amoebae and flagellates were the dominant protozoa. Among ten methanotrophic strains examined as a food source, seven yielded a number of protozoa comparable with the yield with Escherichia coli [10⁴ MPN (g soil dry weight)⁻¹], and three out of four Methylocystis spp. yielded significantly fewer numbers [10²–10³ MPN (g soil dry weight)⁻¹]. The lower edibility of the Methylocystis spp. was not explained either by their growth phase or by harmful effects on protozoa. Incubation of the soil under methane resulted in a higher number of protozoa actively grazing on methanotrophs, especially on the less-edible group. Protozoa isolated from the soil demonstrated a grazing preference on the different methanotrophs consistent with the results of MPN counts. The results indicate that selective grazing by protozoa may be a biological factor affecting the methanotrophic community in a wetland soil.     

In vivo DNA electrotransfer into muscle

Naked plasmid DNA injected into skeletal muscle is taken up by muscle cells and the genes in the plasmid are expressed. Among the non-viral techniques for gene transfer in vivo , this method is especially simple, inexpensive, and safe. However, the relatively low expression levels attained by this method have limited its applications for uses other than as a DNA vaccine. We and other groups investigated the applicability of in vivo electroporation for gene transfer into muscle, using plasmid DNA vector. The results demonstrated that gene transfer into muscle by in vivo electroporation is far more efficient than simple intramuscular DNA injection and provides a potential approach to systemically delivering cytokines, growth factors, and other serum proteins for basic research and human gene therapy.     

Incidence of crown rust and virulence of puccinia coronata cda. f.sp. avenae eriks. and the effectiveness of pc genes for resistance in europe,middle east and north africa: Inzidenz von kronenrost und virulenz von puccinia coronata cda. f.sp. avenae eriks. und die wirksamkeit von pc-genen für resistenz in europa,nahost und nordafrika

Crown rust on oat incited by Puccinia coronata Cda f.sp. avenae is a wide-spread disease in Europe, Middle East (Israel) and North Africa (Morocco). High natural levels of the disease were recorded in Austria, Bulgaria, Czech Republic, Estonia, Israel, Italy, Poland, Russia and the former Yugoslavia. The severity of the disease in the European and Mediterranean Oat Disease Nursery (EMODN) between 1995 and 1999 showed that it occurred at damaging levels in a number of countries. Considerable differences in a disease resistance index (DRI) to crown rust among 67 oat lines in the period 1995?–?2000 were found. The values of the DRI ranged from18 (KR 8122) to over 290 (Pc 59, Pc 68). Detailed studies of the virulence combinations of P.coronata f.sp. avenae in Europe, Middle East and North Africa were carried out between 1995?–?2001. There were considerable differences in the average number of virulence among countries. Virulence to Pc 39 and 68 was found for the first time and is a significant finding. Nevertheless the major genes Pc 68, Pc 39, Pc 50-2, Pc Pc 59, Pc 60, Pc 61 proved to be highly effective.     

The family 6 carbohydrate-binding module (CtCBM6B) of Clostridium thermocellum alpha-L-arabinofuranosidase binds xylans and thermally stabilized by Ca2+ ions

Abstract

The gene encoding CtCBM6B of Clostridium thermocellum α-L-arabinofuranosidase (Ct43Araf) was cloned in pET-21a(+) vector, over-expressed using Escherichia coli BL-21(DE3) cells and purified by immobilized metal-ion affinity chromatography (IMAC). The recombinant CtCBM6B showed a molecular size close to 15 kDa by SDS-PAGE analysis, which was close to the expected size of 14.74 kDa. The ligand-binding affinity of CtCBM6B was assessed against ligands for which the catalytic enzyme, Ct43Araf showed maximum activity. The affinity-gel electrophoresis of CtCBM6B with rye arabinoxylan showed lower equilibrium association constant (K_a, 4.0% C^{? 1}), whereas, it exhibited higher affinity (K_a, 19.6% C^{? 1}) with oat spelt xylan. The ligand-binding analysis of CtCBM6B by fluorescence spectroscopy also revealed similar results with low K_a (3.26% C^{? 1}) with rye arabinoxylan and higher affinity for oat spelt xylan (K_a, 17.9% C^{? 1}) which was corroborated by greater blue-shift in case of oat spelt xylan binding. The CtCBM6B binding with insoluble wheat arabinoxylan by adsorption isotherm analysis showed significant binding affinity as reflected by the equilibrium association constant (K_a), 9.4 × 10³ M^{? 1}. The qualitative analysis by SDS-PAGE also corroborated the CtCBM6B binding with insoluble wheat arabinoxylan. The protein-melting curve of CtCBM6B displayed the peak shift from 53°C to 59°C in the presence of Ca²⁺ ions indicating that Ca²⁺ ions impart thermal stability to the CtCBM6B structure.     

光周期对燕麦生育时期和穗分化的影响

盆栽试验研究了不同光周期对不同熟期燕麦品种的穗分化进程和生育时期影响及生理生态机制。试验材料包括早熟品种白燕8号,中熟品种白燕2号以及晚熟品种坝莜3号,跟踪测定了叶片保护酶活性、膜脂过氧化作用和质膜透性等生理指标的动态变化。结果表明,晚熟品种坝莜3号对光周期反应敏感,短日照(8h)条件下,其穗分化只能到二棱期,未能正常抽穗;早熟品种白燕8号在短日照条件下可完成穗分化、抽穗开花,但3个品种生育时期和穗分化时间均延长。随着光周期延长,各燕麦品种单株小穗数和穗重均增加。光周期对穗分化进程的影响机制可从叶片保护酶活性、膜脂过氧化作用和质膜透性等指标中得到证明,白燕8号在8h短日照处理下SOD,POD活性均高于白燕2号和坝莜3号,而MDA含量和相对电导率低于另两个品种。MDA含量和相对电导率与光周期呈显著正相关。燕麦早熟性与光周期不敏感性具有一定相关性,且燕麦光周期适应性调控机理可能与保护酶活性、膜脂过氧化作用和质膜透性等生理变化密切相关。     

燕麦基因组学研究进展

燕麦具有较高的营养价值和保健功能,是一种可用于均衡营养、科学饮食的健康食品,正逐渐受到人们的青睐和认可。基因组学研究有助于燕麦重要农艺性状的定位和克隆,对开发利用燕麦优质种质资源具有重要意义。本文从以下几个方面对燕麦基因组学研究进展进行综述:(1)燕麦属基因组类型、大小及染色体倍性研究;(2)基于多种分子标记手段构建燕麦基因组遗传图谱进展;(3)二倍体、六倍体燕麦基因组测序进展;(4)基于数量性状基因座定位和全基因组关联性分析手段对燕麦基因组功能基因的注释研究;(5)燕麦群体基因组/泛基因组学研究。同时对燕麦基因组学研究方向进行了探讨,以期为今后燕麦遗传育种提供参考信息。     

硫化氢对盐碱胁迫下裸燕麦叶片抗坏血酸-谷胱甘肽循环的调控效应

盐碱胁迫是植物遭受的常见非生物胁迫之一,气体信号硫化氢(H₂S)在植物响应盐碱胁迫中发挥着重要作用。为探讨H₂S对盐碱胁迫下裸燕麦抗坏血酸(AsA)-谷胱甘肽(GSH)循环的调控效应,以品种‘定莜9号’为材料,研究了喷施H₂S供体硫氢化钠(NaHS)或H₂S合成抑制剂羟胺(HA)对盐碱混合胁迫下植株生长、叶片活性氧、膜脂过氧化和AsA-GSH循环中抗氧化物质和关键酶的影响。结果表明: 喷施50 μmol·L^-1 NaHS可缓解50 mmol·L^-1盐碱混合胁迫对裸燕麦生长的抑制,降低超氧阴离子、H₂O₂、丙二醛、氧化型抗坏血酸(DHA)、还原型谷胱甘肽(GSH)和氧化型谷胱甘肽(GSSG)含量,提高AsA/DHA和GSH/GSSG,而对还原型抗坏血酸(AsA)含量无显著影响。喷施NaHS还提高了盐碱混合胁迫下裸燕麦叶片AsA合成关键酶L-半乳糖脱氢酶(GalDH)和L-半乳糖-1,4-内酯脱氢酶(GalLDH)及AsA-GSH循环中单脱氢抗坏血酸还原酶(MDHAR)活性,降低了抗坏血酸过氧化物酶(APX)和脱氢抗坏血酸还原酶(DHAR)活性,而对抗坏血酸氧化酶(AO)和谷胱甘肽还原酶(GR)活性的影响不大。增添HA后部分或完全解除了喷施NaHS的上述作用。这说明H₂S可通过促进AsA合成和增强MDHAR活性提高AsA-GSH循环效率,降低盐碱胁迫对裸燕麦的氧化伤害。