水稻白叶枯病菌GX1329基因组文库的构建及含编码TAL效应物基因的克隆的分离

由革兰氏阴性细菌水稻白叶枯病菌引起的水稻白叶枯病是亚洲、北美以及非洲部分地区最严重的水稻病害之一,水稻白叶枯病可使水稻减产高达50％以上。研究表明水稻白叶枯病菌的毒力主要依靠三型分泌系统所分泌的效应物。为了解水稻白叶枯病菌广西菌株GX1329中含有avrBs3／pthA家族基因的情况,本研究应用Alu I部分酶切其基因组DNA,构建了含有736个克隆的菌株GX1329的基因组文库。BamHI酶切分析随机挑取的15个文库克隆表明,克隆的外源DNA随机性良好,克隆的最小片段为27．7kb,最大为58．5kb,平均大小为39．9kb,文库克隆容量约为2．8×10^3Mb,该文库中包含基因组中任一个基因的概率为99．4％。利用来自水稻白叶枯病菌菲律宾菌株PX086的无毒基因avrXa10的第252位～第486位核苷酸序列作为探针,通过菌落原位杂交从GX1329基因组文库中筛选到37个含avrBs3／pthA家族基因的克隆。再通过Southern杂交分析,得到了17个独立克隆。这17个克隆中至少含有13个不同的avrBs3／pthA家族基因。这些基因在GX1329基因组中有的单独存在,有的两个或两个以上串联存在。本工作基本上明确了菌株GX1329基因组中avrBs3／pthA家族基因的数量,为进一步研究菌株GX1329中avrBs3／pthA家族基因的功能奠定了基础。     

Effects of Cd and Pb pollution on soil enzymatic activities and soil microbiota

Based on a representative sampling method and pot experiment with different concentrations of Cd and Pd, the enzymatic activities (urease, phosphatase, catalase, invertase), population of bacteria, fungus and actinomycete in the soil, the Cd and Pd pollution status of soil samples (from the wastewater-irrigated area of Baoding suburb) were appraised. Unitary linear and nonlinear curve-fitting optimization models were applied in the research, and the relationship between Pb and Cd causing pollution and enzymatic activities of the tested soils were discussed. The research may provide a theoretical basis for protecting the environment in the region of Baiyangdian Lake, Hebei province, prevent soil pollution, and ascertain biochemical indexes, which reflect soil heavy metal pollution levels. The research results indicated that: (1) there was obvious accumulation of Pb and Cd in the wastewater-irrigated area, also the accumulation in wastewater-irrigated soil is more than that in fresh water-irrigated soil, and accumulation on surface layer was more than that in the lower layer. Pb and Cd contents in the tested soils exceeded the standards of soil background values for some major cities at home and abroad and the world soil Cd and Pb contents range. This means that the tested soil had reached a lightly polluted level; (2) there existed an obvious negative correlation between soil enzymatic activities and Pb and Cd contents in wastewater-irrigated soil, where the soil urease and catalase activities decreased obviously with the increase of Pb and Cd contents in soil. Therefore, the urease and catalase can be considered as biochemical indexes that reflect the degree of soil Pb and Cd pollution; (3) the pot experiments indicated that the influence of Cd on soil enzymatic activities was greater than that of Pb. Generally, the effect of Cd on soil phosphatase, urease, catalase is more obvious than that on invertase, while Pb has a more obvious effect on invertase than Cd; (4) pot experiments of triple cropping showed that, Cd and Pb had an obvious inhibiting effect on microbes. The influence sensitivity of Cd and Pb on different microbes was ordered as bacteria > actinomycete > fungus. Cd had a greater inhibiting effect on microbes than Pb, the influence of Cd and Pb on actinomycete and fungus was relatively smaller, therefore, bacteria can be chosen as an important index that reflects the effect of Cd and Pb on microbe sensitivity.     

原子吸收次灵敏线法直接测定土壤中Fe、Mn

用原子吸收次灵敏线法塞曼效应扣除背景测定土壤中Fe、Mn,实验结果表明:此方法有较好的准确度,精密度,操作简单,适合一般土壤中Fe、Mn的测定。     

土著B[a]P降解菌群的富集及最佳降解条件研究

为了获得能够降解苯并[a]芘(BaP)的微生物菌群,为生物修复多环芳烃污染的土壤提供菌种资源,本试验从长期石油污染的土壤中,富集了一个能够降解BaP的微生物菌群,并研究了其最佳降解条件,通过高通量测序研究了其群落结构。该菌群可以在15 d内将30 mg·L^-1的BaP降解33.34%。高通量测序的结果表明,该菌群主要由Bacillus、Zobellella、Gordonia和Rheinheimera组成,其中Bacillus是主要的降解菌。试验结果表明,该菌群的最佳降解条件为1%盐度,酵母浓度为80 mg·L^-1,pH值7.0。该菌群对芘、菲、荧蒽等多环芳烃也有一定的降解效果,其中荧蒽的降解效果最好,降解率达到99%。试验结果表明,该菌群在实际应用中具有很大的潜力。     

叶面喷施微生物合成纳米硒对水稻硒镉吸收的影响

为探讨喷施微生物合成纳米Se(B)对水稻Se、Cd吸收的影响与籽粒富Se降Cd的效果,本研究通过盆栽试验,研究微生物合成SeNPs喷施5(B5)、10(B10)、20(B20) mg·L^-1对水稻Se、Cd吸收、转运效应,以及与无机Se(S)、有机Se(M)喷施对比,比较水稻籽粒富Se降Cd效果。结果表明,与不喷Se(CK)、S、M处理相比,B5、B10、B20处理提高籽粒Se含量分别为227.52%~366.46%、3.38%~23.31%、30.54%~42.57%,降低籽粒Cd含量分别为10.14%~55.07%、4.08%~36.37%、9.03%~40.77%,籽粒富Se降Cd效果表现为B>S>M>CK,B处理中,籽粒富Se降Cd效果以喷施20 mg·L^-1浓度最佳,高Se与低Se水稻品种间对Se、Cd吸收差异不显著。与CK处理相比,B处理促进了水稻叶片中Cd在细胞壁、液泡组分中的积累,减少了叶绿体、线粒体组分中的积累,促进了叶片中Se在细胞壁、叶绿体组分中的积累,减少了线粒体、液泡组分中的积累。主成分分析和Pearson相关矩阵表明,水稻各部位Se与Cd含量呈负相关关系(P ≤ 0.01)。与CK、M处理相比,B处理对水稻Se生物富集系数分别提高20.63%~264.70%、20.37%~32.86%,表现为S>B>M>CK,B5、B10、B20处理对水稻Cd富集系数分别降低9.10%~25.10%、5.70%~9.65%、10.64%~22.10%,表现为CK>M>S>B。供试水稻Se转移系数表现为根-茎<茎-籽粒<茎-叶,Cd转移系数表现为根-茎<茎-叶<茎-籽粒,与CK、S、M处理相比,B处理均促进了Se向籽粒的转移,减少了Cd向籽粒的富集。与CK、S、M处理相比,B处理均显著提高了水稻叶片SOD、CAT、POD、APX、PAL、Pro、GSH含量,降低了MDA、H₂O₂的含量,高Se水稻品种(泰丰优208)对提高水稻抗氧化酶活性与抗氧化性物质含量效果高于低Se水稻品种(桂野丰)。因此,叶面喷施微生物合成SeNPs在调控水稻Se、Cd吸收、转运与籽粒富Se降Cd方面有着更显著的优势。     

基于激光诱导击穿光谱的地下水中Fe和Mn的检测

采用激光诱导击穿光谱(LIBS)技术并结合在锌基板上液固转换的样品处理方法,对标准水溶液样品中Fe、Mn进行检测,并根据标准曲线法建立Fe、Mn的定量模型;再利用所建立的LIBS定量模型,对洞庭湖区地下水实际水样中Fe、Mn含量进行检测,并与电感耦合等离子体质谱法(ICP–MS)的检测结果进行比较。结果表明：得到Fe、Mn的LIBS定标曲线的决定系数均大于0.98,相对标准偏差均为4.3%,留一交叉验证均方根误差分别为0.024、0.007 mg/L,Fe、Mn的检出限分别为0.028、0.008 mg/L;采用LIBS对洞庭湖地下水实际水样中Fe、Mn含量检测的结果与ICP–MS的检测结果基本一致,平均相对误差小于10%。     

Regulation and function of AtNRAMP4 metal transporter protein

The NRAMP gene family encodes integral membrane proteins mediating the transport of a broad range of transition metals in bacteria, fungi, plants, and animals. We studied the regulation of AtNRAMP4 in Arabidopsis. In a previous study, we showed that AtNRAMP3 and AtNRAMP4 transport manganese (Mn), iron (Fe), and cadmium (Cd). In this study, we show that, in contrast to AtNRAMP3, AtNRAMP4 complements the growth phenotype of the zrt1zrt2 Zn uptake deficient yeast mutant. In a previous study, we have shown that, under Fe starvation, AtNRAMP4 mRNA levels are up-regulated in Arabidopsis. To analyze the regulation of AtNRAMP4 at the protein level, we generated specific antibodies against AtNRAMP4 protein. The antiserum was able to recognize a tagged version of AtNRAMP4 expressed in yeast. The antibody did not reveal any change in AtNRAMP4 protein level upon Fe starvation in Arabidopsis thaliana ecotype Columbia plants. In AtNRAMP4 overexpressing plants, high levels of AtNRAMP4 protein could be detected. AtNRAMP4 overexpressing plants display cadmium hypersensitivity in a medium containing 50 μm FeEDTA as Fe source. However, despite the constitutive accumulation of AtNRAMP4 protein, AtNRAMP4 over-expressing plants did not display Cd hypersensitivity under high Fe supply (100 μm FeHBED). AtNRAMP4 over-expressing lines displayed the same sensitivity to Zn as controls under all conditions tested. Our results suggest a translational level for the regulation of AtNRAMP4. Over-expression of AtNRAMP4 in Arabidopsis thaliana confers a slight hypersensitivity to Cd but not to Zn.     

深松铲入土力学分析与振动减阻机理研究

对深松铲入土时与土壤的力学作用进行分析,得出土壤的变形阻力是影响牵引阻力的主要因素,从而提出振动减阻机理,即利用振动使土壤从无结构转变为有结构,达到减小耕作阻力、提高作业质量、降低能源消耗、提高生产效率的效果。     

Direct-fed microbes: A tool for improving the utilization of low quality roughages in ruminants

For many years, ruminant nutritionists and microbiologists have been interested in manipulating the microbial ecosystem of the rumen to improve production efficiency of different ruminant species. Removal and restriction of antibiotics subtherapeutic uses from ruminant diets has amplified interest in improving nutrient utilization and animal performance and search for more safe alternatives. Some bacterial and fungal microorganisms as a direct-fed microbial(DFM) can be the most suitable solutions. Microorganisms that are commonly used in DFM for ruminants may be classified mainly as lactic acid producing bacteria(LAB), lactic acid utilizing bacteria(LUB), or other microorganism's species like Lactobacillus, Bifidobacterium, Enterococcus, Streptococcus, Bacillus, Propionibacterium, Megasphaera elsdenii and Prevotellabryantii, in addition to some fungal species of yeast such as Saccharomyces and Aspergillus. A definitive mode of action for bacterial or fungal DFM has not been established; although a variety of mechanisms have been suggested. Bacterial DFM potentially moderate rumen conditions, and improve weight gain and feed efficiency. Fungal DFM may reduce harmful oxygen from the rumen, prevent excess lactate production, increase feed digestibility, and alter rumen fermentation patterns. DFM may also compete with and inhibit the growth of pathogens, immune system modulation, and modulate microbial balance in the gastrointestinal tract. Improved dry matter intake, milk yield, fat corrected milk yield and milk fat content were obtained with DFM administration. However, the response to DFM is not constant; depending on dosages, feeding times and frequencies, and strains of DFM. Nonetheless, recent studies have supported the positive effects of DFM on ruminant performance.     

基于减量化目标的农村宅基地整理分区及模式优选

农村宅基地整理是推进低效建设用地减量复垦的有效举措。该文以上海市奉贤区8个建制镇约374个自然村为例,综合运用GIS空间分析技术与层次分析法,融合自上而下宅基地整理潜能的镇域分区和自下而上宅基地整理效能的自然村分类,统筹农村建设用地减量化范围与目标,将奉贤区宅基地整理划分为5类区域3种模式。结果显示:异地迁移和减量储备区可通过复垦还耕推进基本农田集中区建设;调整归并和组团保留区可培育中心村以新增建设用地指标并优化用地布局;腾退并点区可强化新市镇建设以提升镇域用地效益。该研究拟在时序上形成"整理"、"储备"、"保留"渐进式的整理过程,在空间上推进节约集约的减量化实践。