磷肥施用方式及类型对冬小麦产量和磷素吸收的影响

采用田间微区试验研究不同磷肥施用方式和种类对冬小麦生长和当季磷素吸收的影响.磷肥种类为磷酸二氢钙(MCP)和磷酸氢二铵(DAP),施用方式包括表面撒施,种子正下方5 cm条施,种子下方5 cm、偏3 cm条施,种子下方5 cm、偏10 cm条施,种子正下方20%土体混施5种.结果表明: 种子正下方5 cm条施对小麦的增产效果最高,其中磷酸二氢钙的产量达到7.63 t·hm^-2,磷酸氢二铵的产量达到7.99 t·hm^-2,分别较农民习惯撒施方式增产10.3%和10.7%.在5种施磷方式中,偏10 cm条施的小麦产量最低(6.60～6.77 t·hm^-2).种子正下方5 cm条施和20%土体混施处理的小麦总吸磷量均处于较高水平(34.4～35.6 kg·hm^-2),偏10 cm条施在小麦各生长阶段的吸磷量均显著低于其他施磷方式,但磷酸氢二铵偏10 cm条施的小麦总吸磷量较磷酸二氢钙高11.9%.表明将磷肥近距离集中施用于种子附近为该地区较为合理的施磷方式,在偏远距离条施下磷酸氢二铵对小麦的磷素吸收利用效果优于磷酸二氢钙.     

The oxidative stress response in Bacillus subtilis

Abstract Bacillus subtilis undergoes a typical bacterial stress response when exposed to low concentrations (0.1 mM) of hydrogen peroxide. Protection is thereby induced against otherwise lethal, challenge concentrations (10 mM) of this oxidant and a number of proteins are induced including the scavenging enzymes, catalase and alkyl hydroperoxide reductase, and a putative DNA binding and protecting protein. Induced protection against higher concentrations (10–30 mM) of hydrogen peroxide is eliminated in a catalase-deficient mutant. Both RecA and Spo0A influence the basal but not the induced resistance to hydrogen peroxide. A regulatory mutation has been characterized that affects the inducible phenotype and is constitutively resistant to high concentrations of hydrogen peroxide. This mutant constitutively overexpresses the proteins induced by hydrogen peroxide in the wild-type. The resistance of spores to hydrogen peroxide is partly attributable to binding of small acid soluble proteins by the spore DNA and partly to a second step which coincides with the depletion of the NADH pool, which may inhibit the generation of hydroxyl radicals from hydrogen peroxide.     

Comparative genome analysis of Bacillus cereus group genomes with Bacillus subtilis

Genome features of the Bacillus cereus group genomes (representative strains of Bacillus cereus, Bacillus anthracis and Bacillus thuringiensis sub spp. israelensis) were analyzed and compared with the Bacillus subtilis genome. A core set of 1381 protein families among the four Bacillus genomes, with an additional set of 933 families common to the B. cereus group, was identified. Differences in signal transduction pathways, membrane transporters, cell surface structures, cell wall, and S-layer proteins suggesting differences in their phenotype were identified. The B. cereus group has signal transduction systems including a tyrosine kinase related to two-component system histidine kinases from B. subtilis. A model for regulation of the stress responsive sigma factor sigmaB in the B. cereus group different from the well studied regulation in B. subtilis has been proposed. Despite a high degree of chromosomal synteny among these genomes, significant differences in cell wall and spore coat proteins that contribute to the survival and adaptation in specific hosts has been identified.     

磷高效基因型小麦对缺磷胁迫的根际适应性反应

采用奶箱分隔栽培试验法,进行了磷高效与磷低效小麦基因型根际土壤ＰＨ与有效磷变化的研究,结果表明：小麦根际土壤ＰＨ和有效磷含量皆明显低于外围土壤,表现出明显的根际效应特征;磷高效基因型小麦的根际ＰＨ和有效磷含量明显低于磷低效基因型,ＰＨ变异范围和磷素亏缺区也表现明显较大。为了进一步验证磷高效小麦基因型这的这一根际特征,同时进行了施用水溶性,枸溶性磷肥的试验研究,结果表明,以水溶性磷肥对根际ＰＨ和有效     

Intracellular reactive oxygen species mediate suppression of sporulation in Bacillus subtilis under shear stress

Sporulation is an important cellular response to stress that is also significant from a bioreactor operation viewpoint. While sporulating organisms are known to show an enhanced sporulation response under several stress situations, the sporulation response to shear stress has not been investigated thus far. Such a study could be of interest since shear stress, to a greater or lesser degree, is always present in bioreactor operation. In this article, we investigate the sporulation extents of the Gram-positive bacteria Bacillus subtilis at various defined shear levels. We show that, contrary to expectations, shear inhibits sporulation. We found an inverse correlation between the shear rate-dependent specific intracellular reactive oxygen species level (siROS), and the sporulation extent. A 10-fold increase in siROS resulted in about 17-fold decrease in sporulation extent. The involvement of reactive oxygen species (ROS) in sporulation was unknown thus far. Further, through experiments that specifically increased and reduced intracellular ROS (iROS), we established that siROS is responsible for the inhibition of sporulation under shear stress. In addition, we found that shear induced siROS regulated the expression levels of the general stress proteins Ctc and sigma(B). Based on the above, we hypothesize that siROS may regulate suppression of sporulation under high shear by altering sigma(B) and Ctc expression levels, and a model for the same is presented.     

旱地冬小麦吸收氮,磷的营养特征

通过田间试验,研究了不同施肥水平条件下冬小麦吸收氮、磷的营养特征。结果表明,不同施肥水平冬小麦返青前吸收氮、磷均很少,返青后迅速增多,至孕穗一灌浆初期达全生育期吸收的高峰（Ｎ０Ｐ０吸氮在拔节一孕穗阶段达高峰）,且峰值随施肥水平提高而增大。灌浆以后,氮磷营养不同,氮素积累减少２８．２７％－３１．１９％,减少绝对量随前期吸氮增大而增加。相对量则是施肥低于对照;而磷素于灌浆中期一成熟阶段还有１吸收高峰（     

脂肪酶基因在枯草芽孢杆菌中的表达及表达产物性质的研究

从环境中筛选到了脂肪酶高产菌株金黄色葡萄球菌JH,依据NCBI上发表的原核微生物脂肪酶基因序列的多序列比对,发现它具有很强的序列保守性。利用PCR从金黄色葡萄球菌JH基因组中扩增得到了脂肪酶基因,利用基因重组技术将其整合到质粒pC194中,并导入到枯草芽孢杆菌中进行表达。应用选择抗药性筛选重组子,利用硫酸铵沉淀法和离子交换层析分离纯化重组脂肪酶,并用DS-PAGE进行纯度鉴定,确定其相对分子量约为32000Da。通过对其酶学特性的研究发现,重组脂肪酶在反应温度为41℃, pH为8.0时具有最大活性,其Km和Vm各自为0.34mM和308μmol•mg-1min-1,Ca2+、K+、Mg2+能激活这种酶的活性鳩e2+、Cu2+、Co2+则抑制它的活性。     

枯草芽孢杆菌近缘种群鉴定方法研究进展

枯草芽孢杆菌近缘种群(简称枯草群)是一群表型相似的革兰氏阳性、产芽孢的杆菌,目前该群已包括10个有效发表种。该群细菌广泛应用于生物技术、工业和农业等领域,因此实现菌株的快速准确鉴别与鉴定,确保其使用的安全性和有效性是首要条件。本文在本研究室进行的枯草群鉴定研究基础上,总结提出了基于形态和生化特性测定的传统经典方法和以分子生物学技术为基础的基因序列分析技术、特异PCR技术,并在实践上进行了大量的验证,对指导该种群的准确鉴定与快速鉴别具有重要应用价值。     

Unraveling the molecular basis for ligand binding in truncated hemoglobins: The trHbO Bacillus subtilis case

Truncated hemoglobins (trHbs) are heme proteins present in bacteria, unicellular eukaryotes, and higher plants. Their tertiary structure consists in a 2‐over‐2 helical sandwich, which display typically an inner tunnel/cavity system for ligand migration and/or storage. The microorganism Bacillus subtilis contains a peculiar trHb, which does not show an evident tunnel/cavity system connecting the protein active site with the solvent, and exhibits anyway a very high oxygen association rate. Moreover, resonant Raman results of CO bound protein, showed that a complex hydrogen bond network exists in the distal cavity, making it difficult to assign unambiguously the residues involved in the stabilization of the bound ligand. To understand these experimental results with atomistic detail, we performed classical molecular dynamics simulations of the oxy, carboxy, and deoxy proteins. The free energy profiles for ligand migration suggest that there is a key residue, GlnE11, that presents an alternate conformation, in which a wide ligand migration tunnel is formed, consistently with the kinetic data. This tunnel is topologically related to the one found in group I trHbs. On the other hand, the results for the CO and O₂ bound protein show that GlnE11 is directly involved in the stabilization of the cordinated ligand, playing a similar role as TyrB10 and TrpG8 in other trHbs. Our results not only reconcile the structural data with the kinetic information, but also provide additional insight into the general behaviour of trHbs. Proteins 2010. © 2009 Wiley‐Liss, Inc.     

化感物质对枯草芽孢杆菌(Bacillus subtilis)在厌氧条件下的生长及反硝化作用的影响

研究由秸秆腐解产生的化感物质 :阿魏酸 ( t-FA)、对羟基苯甲酸 ( p-HA)和苯甲酸 ( BA)在不同浓度下对厌氧培养的枯草芽孢杆菌 ( Bacillussubtilis)的生长及其反硝化活性的影响。结果表明 ,3种浓度的阿魏酸 ( 5.1 5、2 .58、0 .2 6mmol/L)均表现出对枯草芽孢杆菌的生长有抑制作用。对羟基苯甲酸 ( 0 .3 6、3 .62、7.2 4 mmol/L )对生长影响不明显。 8.1 9mmol/L和 4 .0 9mmol/L的苯甲酸有一定的刺激作用 ,而 0 .4 1 mmol/L的苯甲酸与对照无差别。实验表明枯草芽孢秆菌不仅能转化 NO- 3生成 NO- 2 ,而且还能转化 NO- 2 生成 N2 O。 3种化感物质对 NO- 3的转化均表现抑制作用 ,其抑制作用强弱依次为阿魏酸 >对羟基苯甲酸 >苯甲酸。高浓度的抑制作用强于低浓度。阿魏酸在 5.1 5mmol/L和 2 .58mmol/L浓度下 ,其抑制作用的差异显著性分别为 P<0 .0 1 ,P<0 .0 5。 NO- 2 的生成与 NO- 3的减少相互有关联 ,第 3天测定时 ,各处理中NO- 3急剧减少 ,而 NO- 2 急剧增加。在阿魏酸、苯甲酸处理中的 NO- 2 积累高峰在第 3天、第 4天 ,然后下降。而在对羟基苯甲酸的处理中 NO- 2 的积累一直上升 ,在第 6天的观察中仍未出现下降趋势。 3种化感物质均能抑制 N2 O的生成 ,至于在田间的抑制效果尚需进一步试验     

The effect of acid shock on sporulating Bacillus subtilis cells

AIMS: To study the effect of acid shock in sporulation on the production of acid-shock proteins, and on the heat resistance and germination characteristics of the spores formed subsequently. METHODS AND RESULTS: Bacillus subtilis wild-type (SASP-alpha+beta+) and mutant (SASP-alpha-beta-) cells in 2 x SG medium at 30 degrees C were acid-shocked with HCl (pH 4, 4.3, 5 and 6 against a control pH of 6.2) for 30 min, 1 h into sporulation. The D85-value of B. subtilis wild-type (but not mutant) spores formed from sporulating cells acid-shocked at pH 5 increased from 46.5 min to 78.8 min, and there was also an increase in the resistance of wild-type acid-shocked spores at both 90 degrees C and 95 degrees C. ALA- or AGFK-initiated germination of pH 5-shocked spores was the same as that of non-acid-shocked spores. Two-dimensional gel electrophoresis showed only one novel acid-shock protein, identified as a vegetative catalase 1 (KatA), which appeared 30 min after acid shock but was lost later in sporulation. CONCLUSIONS: Acid shock at pH 5 increased the heat resistance of spores subsequently formed in B. subtilis wild type. The catalase, KatA, was induced by acid shock early in sporulation, but since it was degraded later in sporulation, it appears to act to increase heat resistance by altering spore structure. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first proteomic study of acid shock in sporulating B. subtilis cells. The increasing spore heat resistance produced by acid shock may have significance for the heat resistance of spores formed in the food industry.     

Study on the electro-transformation conditions of improving transformation efficiency for Bacillus subtilis

Aims: To optimize the transformation conditions and improve the transformation efficiency of Bacillus subtilis WB800 and DB104. Methods and Results: Trehalose, which could decrease the damage of electric shock to the cells, was added to the electroporation medium containing sorbitol and mannitol. The factors affecting the transformation efficiency, such as the growth phase of bacteria, cell concentration, electric field strength and plasmid variety, were examined and improved. The new method increased the transformation efficiency of B. subtilis by nearly 100‐fold compared with the conventional one. Conclusions: With the optimized method, the transformation efficiency came up to 3·64 × 10⁵ transformants μg^?1 DNA for WB800, and 2·10 × 10⁵ transformants μg^?1 DNA for DB104. Significance and Impact of the Study: This improvement in transformation efficiency will be largely attributed to the research of expression of exogenous genes in B. subtilis, gene library construction for directed evolution and transformation of wild‐type B. subtilis strains.     

蛋白激发子PeaT1在枯草芽胞杆菌中的分泌表达及重组菌株提高小麦抗旱和促生的作用

PeaT1是从极细链格孢菌Alternaria tenuissima中分离的一种蛋白激发子,具有促进植物生长和诱导植物产生系统获得抗性的功能,为了实现peaT1基因在枯草芽胞杆菌Bacillus subtilis中的分泌表达,增加其应用途径,从枯草芽胞杆菌基因组DNA中分别扩增获得P43启动子和nprB基因的信号肽序列,并用SOE (Splicing by over lapping extension) 方法与peaT1基因连接,将连接产物克隆到大肠杆菌-枯草芽胞杆菌穿梭表达载体pHY300-PLK上,构建了重组表达载体pHY43N-peaT1。将重组载体转化枯草芽胞杆菌WB800菌株,SDS-PAGE和Western blotting分析证实,在NprB信号肽的引导下,枯草芽胞杆菌成功分泌表达了PeaT1蛋白。构建的重组菌株能够显著增强幼苗抗旱性,提高小麦株高。     

Combined effect of chemical fertilisers and rhizosphere-competent Bacillus subtilis BSK17 on yield of Cicer arietinum

A chemical fertiliser-adaptive variant, Bacillus subtilis BSK17, showed induction in growth at 0.32?M of Urea, 0.05?M of DAP, 0.04?M of MoP and 0.08 of gypsum. In addition, B. subtilis BSK17 produced various plant growth-promoting substances and showed higher colony growth inhibition of Fusarium oxysporum that increased with increase in incubation time and reached the maximum by 78% at 120?h. In field, antibiotic-resistant marker strains of B. subtilis BSK17^ery+ and B. subtilis BSK17^tet+ showed more improvement in seed yield (90% than the control and 24% than full dose of chemical fertilisers) of Cicer arietinum when applied with half dose of chemical fertilisers (N₅₊₅P₁₅₊₁₅K₁₅S_10+10+10). Root length, shoot length, fresh and dry weight of root and shoot of plants were enhanced after 120?days in comparison to control; all values were significant at 1% CD. The strain significantly colonised the rhizosphere of C. arietinum by 6.64 log cfu after 120?days.     

小麦磷素利用效率的品种差异

通过土培盆栽试验,研究了小麦不同品种在低磷水平下生物量、磷浓度、磷素干物质生产效率及磷素籽粒生产效率的差异,以筛选磷高效利用小麦品种.结果表明：分蘖期、拔节期、扬花期和成熟期,供试小麦品种单株生物量的变幅分别为0.46～1.09、0.85～2.10、3.00～7.00和3.85～12.88 g,磷浓度变幅分别为2.21～4.26、2.38～4.42、2.44～4.96和1.30～5.09 mg·g^-1.随生育时期的推进,小麦磷素累积量、磷素干物质生产效率对生物量形成的影响程度呈减小趋势.分蘖期（CV=16.3%）、拔节期（CV=15.0%）、扬花期磷素干物质生产效率（CV=13.3%）和成熟期磷素籽粒生产效率（CV=20.5%）的品种差异较大.CD1158-7和省A3宜03-4具有较高的磷素干物质生产效率和磷素籽粒生产效率,而渝02321较低;不同生育时期高效品种磷浓度极显著低于低效品种,而高效品种CD1158-7和省A3宜03-4的籽粒产量分别是低效品种渝02321的1.98和1.78倍.     

小麦不同生育时期Cd、Cr、Pb污染监测指标体系

以小麦为供试材料,分别采用发芽试验、溶液培养、土柱栽培等毒理试验方法,研究了小麦萌芽期、幼苗期及成株期受重金属污染毒害的指标体系。结果表明,①小麦根伸长抑制率可作为萌芽期重金属污染评价的一项生物指标;Cd、Cr、Pb对小麦根生长的7d半效应浓度(EC50)值分别为1.39、0.20mmo.lL-1和2.75mmol.L-1,据此得到3种重金属对小麦同一性状的毒性次序为CrCdPb;此外各性状抑制率与胁迫浓度的关系可用双曲线模型y=x/(a+x/100)或指数曲线模型y=a(1-exp(-bx))较好模拟。②Cd、Cr、Pb单一污染胁迫对小麦幼苗性状的毒性次序均以影响叶面积和冠部干重为主,表明叶面积和冠部干重为幼苗期污染监测的敏感指标;重金属对除根干重外所有性状的毒性次序:CrCdPb,该毒性次序是根据引起50%抑制的临界浓度即EC50值确定的;各性状抑制率与胁迫浓度的关系可用直线模型、或双曲线模型、或指数曲线模型较好模拟。③重金属单一污染胁迫造成小麦籽粒产量下降幅度是CdCrPb;二元互作对产量影响的重要性次序为CdCrCdPbCrPb;在Cd、Cr、Pb复合污染胁迫条件下,穗数可以作为成株期重金属污染监测的首选指标。以上指标可为农田小麦重金属污染的监测和综合治理提供一些理论依据。     

不同基因型小麦磷素营养阈值的研究

利用温室盆栽试验,对不同磷效率小麦基因型进行了磷素营养阈值的研究。结果表明,磷低效基因对缺磷反应敏感,在不施或施磷量较低时,籽粒产量及生物量均较低,随着施磷量增加,产量缓慢增加,表现出对磷肥效应较迟钝的钝的现象,而磷高效小麦基因型在不同施磷肥或施磷量较低条件下的产量与生物量相对较高,表现出明显的耐低磷特性及对土壤难溶性磷的高效活化、吸收能力。且随产丰施磷量增加,磷高效基因型的产量急剧增加,达到一定     

小麦-大豆间作中小麦对大豆磷吸收的促进作用

采用塑料膜,２０μｍ尼龙网分司以及不分隔两种俄根系的盆栽装置研究了小麦,大豆间作种间磷吸收的促进作用。结果表明,两种作物根系用尼龙网分隔后,即种间磷竞争作用基本消除后,小麦对大豆磷吸收具有明显的促进和,表现为大豆生物学产量明显提高,是塑料膜分隔的２．５倍,是无分隔４．６倍,尼龙网分隔,大豆分隔,大豆的根系活力高于塑料膜分隔４５．２％,达显著水平,此外,尼龙网分隔形成小麦根面,大豆的根系呈偏向小麦根     

A review of phosphorus efficiency in wheat

More efficient utilization of phosphorus by wheat plants is needed to extend the useful life of the phosphate reserves in the world, to reduce the cost of producing crops, and to improve the value of the grain and the straw produced. In this paper definitions of efficient use of phosphorus by wheat are reviewed, genotypic variation in phosphorus efficiency is reported, some consequences of breeding for greater efficiency are discussed, and ways to select more efficient genotypes are suggested.