奶牛流产性疾病研究进展

随着中国奶牛养殖业的规模扩大,奶牛流产病例呈上升趋势.目前,奶牛流产已成为奶牛生产中危害较大的问题之一.流产可影响奶牛的泌乳性能、繁殖性能、牛群健康和使用年限,造成巨大的经济损失.某些引起奶牛流产的病原微生物是重要的人畜共患病原,严重威胁公共卫生安全,对公众和职业人群有巨大危害.因此,有效控制奶牛流产性疾病对奶牛生产和公共卫生安全都极其重要.     

拮抗菌对香蕉枯萎病菌的抑菌作用初步研究

从土壤中分离到一株对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)具有抑制作用的拮抗细菌--假单胞杆菌(Pseudomonas sp.),并对其抑菌效果、热稳定性以及作用机理进行了初步研究.结果表明,拮抗细菌发酵液及其无菌滤液对香蕉枯萎病菌的生长及其分生孢子的萌发具有显著的抑制作用(P＜0.05),并导致病菌菌丝和分生孢子芽管扭曲、膨大;拮抗细菌发酵液经121℃处理30min后,对病菌生长的抑制作用降低.     

蔬菜土传病原菌三重PCR检测体系的建立与应用

【目的】 针对生产中危害严重的3种蔬菜土传病原菌瓜果腐霉（Pythium aphanidermatum）、尖镰孢（Fusarium oxysporum）、大丽轮枝菌（Verticillium dahliae）,建立三重PCR（triplex PCR）检测体系,为蔬菜土传真菌病害的早期诊断和鉴定提供技术与方法。【方法】 筛选3种病原菌特异性引物组合,通过设定不同的PCR退火温度、引物浓度、循环次数以及延伸时间,探索影响三重PCR扩增的因素,建立蔬菜土传病原菌三重PCR检测体系,并对体系灵敏度进行检测。为了验证建立体系的稳定性,选择2×Taq Master PCR Mix（北京博迈德生物技术有限公司）和TaKaRa Taq酶（大连宝生物工程有限公司）2个不同公司的试剂,分别利用C1000 Touch ^TM型（赛默飞世尔科技有限公司）与Aeris ^TM型（新加坡艺思高科技有限公司）热循环仪进行三重PCR反应,比较检测结果的可重复性。对田间采集的35份病害样本和149份土壤样本,分别进行三重PCR和病原菌分离检测,以确定所建立得三重PCR检测体系的适用性。 【结果】 三重PCR反应体系中引物对AsAPH2B/AsPyF、FOF1/FOR1、VActF/VActR可分别扩增出瓜果腐霉、尖镰孢、大丽轮枝菌长度为163、328和530 bp的特异性目的片段。反应体系（25 μL）：0.12 μmol·L ^-1 AsAPH2B/AsPyF,0.16 μmol·L ^-1 FOF1/FOR1,0.24 μmol·L ^-1 VActF/VActR,2×Taq Master PCR Mix 12.5 μL,退火温度为60.8℃,35个循环。该体系对病原菌纯培养物的检测灵敏度为10 ^-1ng·μL ^-1,对土壤中大丽轮枝菌、尖镰孢和瓜果腐霉的检测灵敏度分别为10 ⁵、10 ⁶个孢子/g土壤和10 ^-2mg菌丝/g土壤。分别采用2×Taq Master PCR Mix和TaKaRa Taq酶,利用C1000 Touch ^TM型和Aeris ^TM型热循环仪进行三重PCR,扩增结果一致,说明三重PCR体系检测稳定性好。对田间采集的病害和土壤样本进行检测,25份病害样本和71份土壤样本中检测出带菌,三重PCR检测与病原菌分离培养结果一致。 【结论】 本研究建立的瓜果腐霉、尖镰孢、大丽轮枝菌三重PCR检测体系具有灵敏度高、稳定性和重复性好的特点,能够快速、准确地检测田间病株及其根围土壤中的瓜果腐霉、尖镰孢、大丽轮枝菌,为蔬菜土传病害的早期预防和流行监测提供了有效的技术手段。     

Control of Phytophthora cryptogea in the hydroponic forcing of witloof chicory with the rhamnolipid-based biosurfactant formulation PRO1

Rhamnolipids, extracellular metabolites of Pseudomonas aeruginosa with surfactant properties, proved to be very effective in controlling the spread of brown root rot disease caused by Phytophthora cryptogea in the hydroponic forcing system of witloof chicory ( Cichorium intybus var. foliosum ). The biosurfactant was applied as the product PRO1, a formulation of 25% rhamnolipids in oil. Both an in vitro screening and in vivo experiments in a mini-hydroponic system demonstrated the ability of PRO1 to control brown root rot. A 25  µ g mL⁻¹ rhamnolipids nutrient solution was enough to obtain good control of an artificial infection with a zoospore suspension of P. cryptogea . The biosurfactant PRO1 performed well in a semicommercial system under growers' conditions. A treatment of 25  µ g mL⁻¹ rhamnolipids (100  µ g mL⁻¹ PRO1) reduced the disease incidence significantly in two independent experiments. However, PRO1 was not effective when a mycelial suspension was used as inoculum. Rhamnolipids have good potential to limit the spread of P. cryptogea in the hydroponic forcing system of witloof chicory, and can be used as a preventive measure against brown root rot.     

Epidemiology of beet necrotic yellow vein virus in sugar beet at different initial inoculum levels in the presence or absence of irrigation: Dynamics of inoculum

Using field plots where rhizomania had not previously been detected, different inoculum levels of beet necrotic yellow vein virus (BNYVV) were created by application of infested soil. A susceptible sugar beet cultivar (cv. Regina) was grown for two consecutive years (1988 and 1989), in the presence or absence of drip irrigation. In soil samples taken in spring 1989, the different initial inoculum levels of 1988 could be distinguished using a quantitative bioassay estimating most probable numbers (MPNs) of infective units per 100 g dry soil. The first sugar beet crop resulted in a tenthousandfold multiplication of inoculum of BNYVV (viruliferousPolymyxa betae). Mean MPNs of BNYVV ranged from 0.6 and 7 per 100 g soil for the lowest inoculum level to 630 and 1100 per 100 g for the highest level, in plots without and with irrigation, respectively. In spring 1990, MPNs had again increased. In both years, the initial inoculum level of 1988 had a significant linear effect on log-transformed MPNs of BNYVV determined. Log-transformed MPNs for 1990 and 1989 showed a positive linear correlation, despite a decreasing multiplication ratio at higher inoculum levels. Drip irrigation during one or two years enhanced the increase in MPN of BNYVV, which was reflected by the enhancement of multiplication ratios at all inoculum levels. The totalP. betae population was also higher after growing two irrigated crops than after growing two non-irrigated ones.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

Production,survival and evaluation of solid-substrate inocula ofConiothyrium minitans againstSclerotinia sclerotiorum

Coniothyrium minitans grew on all ten solid-substrates (barley, barley-rye-sunflower, bran-vermiculite, bran-sand, maizemed-perlite, millet, oats, peat-bran, rice and wheat) tested, producing high numbers of germinable pycnidiospores (1.9–9.3×10⁸ g^–1 air dry inocula). All solid substrate inocula survived better in the laboratory at 5 and 15 °C than at 30 °C for at least 64 weeks.In pot bioassays carried out in the glasshouse and field, soil incorporations of each inoculum almost completely inhibited carpogenic germination ofS. sclerotiorum. In the field bioassay, no sclerotia were recovered after 38 weeks fromC. minitans-treated pots compared to 56% from control pots. In the glasshouse bioassay, 9–30% of sclerotia were recovered after 20 weeks fromC. minitans-treated pots, but 88–100% of these were infected by the antagonist. The antagonist also spread to infect sclerotia in control pots.In larger scale glasshouse trials, single preplanting soil-incorporations of five inocula (barley-ryesunflower, maizemeal-perlite, peat-bran, rice and wheat) controlled Sclerotinia disease in a sequence of lettuce crops, with only small differences between the types of inocula tested. At harvest,C. minitans reduced sclerotial populations on the soil surface and over 74% of sclerotia recovered fromC. minitans-treated plots were infected by the antagonist.C. minitans survived in soil in all solid-substrate inocula-treated plots for at least 39 weeks at levels of 10⁴–10⁵ colony forming units cm^–3 soil and spread to infect over 36% of sclerotia recovered from control plots.     

Relation between cropping frequency of peas and other legumes and foot and root rot in peas

The relation between the frequency of legume crops in a rotation and the root rot severity in pea was examined in a field survey. Additionally, greenhouse experiments were performed with soil samples from legume rotation trials or from farmers' fields. The frequency of pea crops in current rotations proved to be much less than the recommended value of one in six years. The correlation between pea root rot and the number of years that pea or other legumes were not grown on the field under consideration (called crop interval) was weak. Root rot severity correlated better with the frequency of peas or legumes in general over a period of 18 years, but the frequency still explained only a minor fraction of the variation in disease index. Some experimental data pointed to the occurrence of a highly specific pathogen microflora with continuous cropping of only one legume species, but this phenomenon probably does not occur in farmers' fields. In field samples, root disease index for pea correlated well with that for field bean. The survival of resting structures of pathogens such asAphanomyces euteiches probably explains why the frequency of legume cropping has a higher impact than crop interval on root disease incidence. Pea-free periods and legume frequencies have a poor predictive value for crop management purposes.     

黄瓜根际细菌J352对大豆根腐病菌的作用

J352是从黄瓜根际筛选到的一株对病原真菌具有拮抗作用的细菌。室内培养结果表明,该菌对大豆根腐镰刀病菌有较强的抑制效果。J352发酵液对2种病原真菌孢子萌发有较高的抑制率;细菌与病原菌共培养后发现真菌菌丝和孢子发生畸变,菌丝和孢子内部均发生原生质体凝聚并出现空洞。与其它供试病原真菌的对峙培养发现,对烟草赤星病菌抑制作用最明显,抑菌率达85.5%。     

Relation between Soil Health, Wave-like Fluctuations in Microbial Populations, and Soil-borne Plant Disease Management

A healthy soil is often defined as a stable soil system with high levels of biological diversity and activity, internal nutrient cycling, and resilience to disturbance. This implies that microbial fluctuations after a disturbance would dampen more quickly in a healthy than in a chronically damaged and biologically impoverished soil. Soil could be disturbed by various processes, for example addition of a nutrient source, tillage, or drying-rewetting. As a result of any disturbance, the numbers of heterotrophic bacteria and of individual species start to oscillate, both in time and space. The oscillations appear as moving waves along the path of a moving nutrient source such as a root tip. The phase and period for different trophic groups and species of bacteria may be shifted indicating that succession occurs. DGGE, Biolog and FAME analysis of subsequent populations in oscillation have confirmed that there is a cyclic succession in microbial communities. Microbial diversity oscillates in opposite direction from oscillations in microbial populations. In a healthy soil, the amplitudes of these oscillations will be small, but the background levels of microbial diversity and activity are high, so that soil-borne diseases will face more competitors and antagonists. However, soil-borne pathogens and antagonists alike will fluctuate in time and space as a result of growing plant roots and other disturbances, and the periods and phases of the oscillations may vary. As a consequence, biological control by members of a single trophic group or species may never be complete, as pathogens will encounter varying populations of the biocontrol agent on the root surface. A mixture of different trophic groups may provide more complete biological control because peaks of different trophic groups occur at subsequent locations along a root. Alternatively, regular addition of soil organic matter may increase background levels of microbial activity, increase nutrient cycling, lower the concentrations of easily available nutrient sources, increase microbial diversity, and enhance natural disease suppression.