The influence of nitrogen fertilisation on bacterial activity in the rhizosphere of barley

The effect of nitrogen addition on the activity of rhizosphere bacteria was studied using barley seedlings. Three different nitrogen sources were added to the soil (nitrate, ammonium and ammonium+nitrate) at four different concentrations (0, 100, 300 and 500 mg N kg⁻¹ soil) and the plants were allowed to grow for 6 weeks. The bacterial activity was estimated by measuring thymidine and leucine incorporation into bacteria extracted using homogenisation-centrifugation. Bulk soil bacterial activity was low compared with that of rhizosphere bacteria. Nitrogen addition did not affect the activity of the bulk soil bacteria, indicating that the activity was not nitrogen limited. The thymidine and leucine incorporation rates of rhizosphere bacteria decreased when ammonium or ammonium+nitrate was applied compared with the non-amended controls. No effect on bacterial activity was found following nitrate addition. There was a significant positive correlation between rhizosphere bacterial activity and rhizosphere pH. Shoot length following ammonium treatment was significant lower than in the non-amended control, while nitrate and ammonium+nitrate addition had no effect. This indicates that the varying effects due to nitrogen sources on rhizosphere bacterial activity were not due to effects on plant growth.     

Survival of bacterial DNA and culturable bacteria in archived soils from the Rothamsted Broadbalk experiment

Dried soil samples from many sources have been stored in archives world-wide over the years, but there has been little research on their value for studying microbial populations. Samples collected since 1843 from the Broadbalk field experiment on crop nutrition at Rothamsted have been used to document changes in the structure and composition of soils as agricultural practices evolve, also offering an invaluable record of environmental changes from the pre- to post-industrial era in the UK. To date, the microbial communities of these soils have not been studied, in part due to the well-documented drop in bacterial culturability in dried soils. However, modern molecular methods based on PCR amplification of DNA extracted directly from soil do not require bacterial cells to be viable or intact and may allow investigations into the legacy of bacteria that were present at the time of sample collection.

In a preliminary study, to establish if dried soils can provide a historical record of bacterial communities, samples from the Broadbalk soil archive dating back to 1868 were investigated and plots treated with either farmyard manure (FYM) or inorganic fertilizer (NPK) were compared. As anticipated, the processes of air-drying and milling greatly reduced bacterial viability whilst DNA yields declined less and may be preserved by desiccation. A higher proportion of culturable bacteria survived the archiving process in the FYM soil, possibly protected by the increased soil organic matter. The majority of surviving bacteria were firmicutes, whether collected in 2003 or in 1914, but a wide range of genera was detected in DNA extracted from the samples using PCR and DGGE of 16S rRNA genes. Analysis of DGGE band profiles indicated that the two plots maintained divergent populations. Sequence analysis of bands excised from DGGE gels, from a sample collected in 1914, revealed DNA from - and β-proteobacteria as well as firmicutes. PCR using primers specific for ammonia oxidizing bacteria showed similar band profiles across the two treatments in recently collected samples, however older samples from the NPK plot showed greater divergence. Primers specific for the genus Pseudomonas were designed and used in real-time quantitative PCR to indicate that archived soil collected in 1868 contained 10-fold less pseudomonad DNA than fresh soil, representing around 10⁵ genomes g⁻¹ soil. Prior to milling, dramatically less pseudomonad DNA was extracted from recently collected air-dried soil from the NPK compared to the FYM plot; otherwise, the two plots followed similar trends. Overall bacterial abundance, diversity and survival during the archiving process differed in the two soils, possibly due to differences in clay and soil organic matter content. Nevertheless, the results demonstrate that air-dried soils can protect microbial DNA for more than 150 years and offer an invaluable resource for future research.     

冬枣溃疡病病原初步鉴定

2003~2004年在山东省滨州市沾化县、无棣县等冬枣产区调查发现,冬枣生长前期在枣吊和叶片上发生了一种细菌性新病害。通过对典型病害样本进行病原菌分离、纯化获得20个菌株。从中选出4个菌株进行致病性测定,同时对其细菌形态、培养性状、染色反应、生理生化测定及血清学反应等性状进行了较为系统研究。结果表明,该地区冬枣溃疡病是由油菜黄单胞Xanthom onas campestrisPamm e(Dowson,1939)侵染引起的。     

汾河太原段沉积物中细菌群落的分布特征

分析汾河太原段沉积物细菌群落结构,探讨细菌群落与典型环境因子的相关关系,丰富河流沉积物细菌群落资料,并为汾河太原段生态健康状况评价提供依据。于2020年10月枯水期,在汾河入城口至清徐段布设11个采样断面,采集汾河太原段表层沉积物,共采集到10份样本。采用Illumina高通量测序技术分析沉积物细菌丰度和群落结构,利用主成分分析（PCoA）、冗余分析（RDA）和相关性分析等方法探讨沉积物中细菌群落结构分布特征以及主要影响因素。结果表明,10个断面表层沉积物中5 422个细菌OTUs分属于61门140纲346目580科1 073属。细菌群落结构在断面fh6、fh7、fh8、fh9、fh10、fh11和fh4相似度较高,fh1和fh2断面相似度较高,而fh3断面与其他断面差异显著。除fh3,沉积物细菌优势菌门均为变形杆菌门（Proteobacteria）,次优势菌门为绿弯菌门（Chloroflexi）,优势菌纲为γ-变形菌纲（Gammaproteobacteria）。Pearson相关性分析表明,TN、OM和Pb与OTUs、Shannon指数和Simpson指数相关性显著。TN、OM和Pb...     

长期秸秆掩埋配施氮肥对土壤细菌群落特征的影响

为揭示典型潮土微生物群落及其生态网络对秸秆还田的响应机制,选择长期小麦-玉米轮作下不同秸秆还田处理的试验地为对象,应用高通量测序和生态网络等方法,阐明秸秆掩埋配施不同氮肥后土壤细菌群落组成、细菌网络共存关系以及与土壤养分的关联。结果表明：（1）与秸秆移除且不施氮肥处理相比,秸秆还田配合常规施肥处理显著降低土壤pH,提高了土壤全氮、有机碳、有效磷、速效钾和硝态氮含量（P < 0.05）。秸秆掩埋配施氮肥有利于增加土壤养分含量。（2）不同秸秆还田方式和氮肥施用量处理的细菌Alpha多样性无显著差异,而细菌群落结构差异显著。pH、SOC和TN等因子驱动了细菌群落结构变异。酸杆菌门、变形菌门、拟杆菌门和绿弯菌门是潮土细菌群落的主要优势菌门。（3）共现网络分析得到4个主要的生态集群,均与土壤养分有显著相关性。模块1中物种丰度与SOC、TN、TP、NO3- -N、AP和AK呈极显著负相关（P < 0.001）,与pH呈极显著正相关（P < 0.001）;模块2和模块3中物种丰度与大部分养分含量呈正相关,与pH呈负相关。因此秸秆还田配施氮肥可以通过调节细菌互作关系,从而调控土壤肥力。研究结果可为秸秆的资源化利用和土壤施肥管理提供科学依据。     

Resveratrol protects cortical neurons in infant rats with bacterial meningitis through miR-223-3p/NLRP3 pathway

AIM To investigate the protective effect of resveratrol (Res) on cortical neurons in rat bacterial meningitis (BM) model. METHODS Group B hemolytic Streptococcus was injected via the posterior cistern to establish a BM model. Resveratrol was administered intranasally and microRNA-223-3p (miR-223-3p) antagomir was administered by intracerebroventricular injection. HE staining was used to observe the pathological changes of the brain tissue. Loeffler scoring method was used to evaluate the neurobehavioral functions. TUNEL staining was used to detect neuronal apoptosis. The expression of interleukin-1β (IL-1β), IL-18, glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Iba1) was detected by immunofluorescence staining. The protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）, cleaved caspase-1, IL-1β and IL-18 were determined by Western blot. The expression level of miR-223-3p was detected by RT-qPCR. Online software TargetScan was used to search for the complementary nucleotide sequences between miR-223-3p and NLRP3 mRNA. RESULTS Compared with sham group, the thickness of meninges in BM model was increased, the neurological score was decreased (P<0.05), and the number of TUNEL positive neurons was increased significantly (P<0.05). Astrocytes and microglia were activated, the fluorescence intensity of IL-1β and IL-18 was increased (P<0.05), and the expression levels of NLRP3, cleaved caspase-1, IL-1β, IL-18 and miR-223-3p were increased (P<0.05). Compared with BM group, after treatment with resveratrol, the neurological score was increased (P<0.05), the number of TUNEL positive neurons was decreased significantly (P<0.05), and the inflammatory response of astrocytes and microglia was suppressed. The fluorescence intensity of IL-1β and IL-18 was decreased (P<0.05), the protein levels of NLRP3, cleaved caspase-1, IL-1β and IL-18 were decreased (P<0.05), and the expression level of miR-223-3p was increased (P<0.05). A nucleotide sequence in the 3'-UTR of NLRP3 mRNA might be targeted by miR-223-3p. In the brain of rat BM model, compared with antagomir control group, the expression of NLRP3 was increased in miR-223-3p antagomir group with resveratrol treatment (P<0.05). CONCLUSION Resveratrol may reduce the inflammatory death of cortical neurons in BM model of infant rats through miR-223-3p/NLRP3 pathway, thus playing a protective role for the neurons.     

转Xa21基因杂交水稻选育和评价

通过基因枪转基因方法和双质粒共转化体系将Xa21基因转入优良恢复系明恢63,得到转基因系M12和M22,并进一步做田间试验、新品种选育和食用安全性评价。结果显示:M12和M22对中国的所有白叶枯病小种都表现出高度抗性,但与原受体品种明恢63相比,农艺性状上有许多变异,主要表现在:结实率、千粒重等农艺性状变差,与珍汕97A的配合力显著降低;但与温敏不育系(X07S、056S)配组的F1有较好的田间表现。通过多代回交和分子标记辅助选择,成功地将Xa21基因从M12株系转到保持系80-4B和不育系80-4A中,得到抗白叶枯病的皖21B和皖21A。并利用皖21A不育系选育出优良杂交组合抗优97(皖21A×R-18),该组合在两年区域试验和一年生产试验中产量表现突出,米质优良。对不同世代和不同遗传背景的转基因品系进行白叶枯病鉴定和Southern分析表明:Xa21基因都能稳定遗传和正常表达,连续16代的种植并没使其白叶枯病的抗性减弱或丧失,而且不论Xa21基因是纯合的还是杂合的都有相同的抗性表现。对大鼠和小鼠的饲喂试验表明:转基因大米实质上等同于非转基因大米,是安全无害的。     

旱田除草剂对玉米内根圈土壤微生物的影响

研究了3种旱田除草剂(氟乐灵、乙草胺和阿特拉津)对玉米品种瓦试13(抗)和瓦试15(感)内根圈土壤微生物(真菌、细菌、放线菌等)种群和数量的影响。结果表明：随着玉米的生长，不同抗性品种内根圈土壤中的真菌、细菌呈增长趋势，而放线菌则在幼苗初期数量最多。从抗感玉米内根圈的分离获得的细菌数量远远高于真菌和放线菌的数量。经常规鉴定明确玉米内根圈土壤有真菌6个属，细菌6个菌落形态，放线菌3个菌落形态。通过对立枯丝核菌的拮抗试验表明，拮抗菌中细菌数量最多，真菌和放线菌对立枯丝核菌的拮抗作用不明显。     

长白赤松和红松土壤微生物活性对高浓度CO_2的响应(英文)

研究了2003 年夏季长白赤松和红松土壤微生物活性对高浓度CO2的响应规律。结果表明,长白赤松和红松土壤细菌数量受高浓度CO2影响显著(p < 0.05)减少;与对照箱(350 μmol ·mol-1 CO2)和裸地(350 μmol ·mol-1 CO2)相比,红松土壤淀粉酶和转化酶活性降低,而长白赤松土壤淀粉酶和转化酶活性却表现为增加;同时发现受700 μmol ·mol-1 CO2处理的红松和长白赤松土壤微生物生物量碳均表现为显著降低。DGGE 结果表明:受高浓度CO2 的影响,长白赤松和红松土壤细菌群落结构发生了明显的变化。研究结果表明土壤微生物对高浓度CO2的响应规律与所研究的的树种有关。图2 表2 参29。     

种子带菌作为玉米弯孢菌叶斑病初侵染源的研究

[目的]对种子带菌作为玉米弯孢菌叶斑病的初侵染源进行研究,为生产上防治该病提供理论依据。[方法]采用玻片萌发法和组织分离法测定种子外部带菌、内部带菌的存活力;采用沙培法在室内检测种子带菌对幼苗的侵染;应用田间培养法测定种子带菌对玉米的侵染。[结果]玉米种子所带弯孢菌孢子和菌丝可以越冬成活,种子内部带菌影响种子萌发,种子外部带菌不影响种子萌发;种子内外带菌均能侵染玉米幼苗的胚根和胚芽。[结论]玉米种子带弯孢菌孢子和菌丝可以越冬并侵染幼苗,使幼苗长势衰弱。