红皮云杉外生菌根菌对苗木生长的影响

2000年8～10月自大兴安岭新林林业局和兴隆林业局采集各种林型下的伞菌担子果,进行分离培养、纯化,获得36个菌株。经对红皮云杉室内接种试验获得6个可与红皮云杉共生形成外生菌根的菌株。室内接种试验结果表明,在红皮云杉播种苗上接种菌根菌的时期应在出苗后30d左右。菌根形成的适宜温度为20℃左右。筛选得到的6个菌株对红皮云杉幼苗的生长有明显的促进作用。接种林地菇(Agaricus silvaticus)、031和L15菌株的苗木叶绿素a含量明显高于其他菌株和对照;接种009、004、林地菇和L15菌株的苗木叶绿素b含量也明显高于其他菌株和对照;接种林地菇、L15、031、009和025菌株的苗木与对照苗木在重量上差异显著;其中接种林地菇和L15菌株的苗木比对照苗木分别增加了19.23%和23.08%;接种6个菌株的苗木与对照苗木高度比较差异非常显著,接种林地菇和L15菌株的苗木与对照相比分别增加17.83%和16.37%。室外接种结果表明,接种林地菇的苗木高生长最好,接种70d高于对照9.24%;接种菌株L15的苗木地径生长最好,接种70d高于对照9.92%;接种菌株009的苗木侧根数最多,接种70d高于对照51.91%;接种菌株009的苗木主根最长,接种70d高于对照3.36%;而接种菌株031的苗木全株感染菌根率最高,接种70d高于对照13.90%;接种菌株031的苗木光合作用最强,高于对照20.02%。从几项主要的指标综合来看,菌株L15、菌株009和菌株031是有潜在研究和应用价值的优良菌株。     

Use of the Minitek system for characterizing lactobacilli.

Incubation of inoculated substrates of the Minitek system in anaerobic GasPak jars provided a method that produced results comparable to those of conventional tubed media for characterizing species of Lactobacillus. The use of sterile mineral oil to overlay inoculated substrate disks was responsible for erroneous results.     

Transmission of experimentally-induced rat virus infection

The duration of transmission of rat virus (RV) infection was determined using Sprague-Dawley rats inoculated oronasally as juveniles (4 weeks old) or as infants (2 days old). Contact transmission from rats inoculated as juveniles was detected for 3 weeks, whereas transmission from rats inoculated as infants occurred for 10 weeks. Transmission continued for at least 7 weeks after seroconversion occurred in rats inoculated as infants. Two of three rats that had ceased to transmit infection harbored infectious virus as detected by explantation of kidney. Intrauterine transmission occurred only after pregnant dams were inoculated with large doses of virus and was more efficient when virus was inoculated intravenously than by the oronasal route. Enzyme immunoassay antibody titers to RV in offspring of previously infected dams decreased steadily during the first 13 weeks of life and 27 of 29 offspring tested by immunofluorescence assay at 12 or 13 weeks of age were seronegative. These results indicate that RV was transmitted by rats inoculated as infants for long periods after seroconversion occurred. They also suggest that the offspring of previously-infected dams were not infected. In utero transmission of RV-Y is unlikely to occur after oronasal inoculation unless rats are exposed to large doses of virus.     

Experimental infection of white-tailed deer (Odocoileus virginianus) with Ehrlichia chaffeensis by different inoculation routes

The infection dynamics of the tick-transmitted organism Ehrlichia chaffeensis were investigated in white-tailed deer (Odocoileus virginianus) using different routes of inoculation. Six deer were each inoculated with 5.4 x 10(6) DH82 cells infected with E. chaffeensis (Arkansas strain) by three different routes: intravenous (n = 2), subcutaneous (n = 2), and intradermal (n = 2). Two control deer were inoculated with uninfected cells. Infections were monitored for 54 days and were continued in one deer from each E. chaffeensis inoculated group for an additional 31 days. All deer inoculated with E. chaffeensis seroconverted (> or = 1: 64) and became 16S rDNA polymerase chain reaction and/or cell culture positive by post-inoculation day 15. There was no apparent (difference in susceptibility to infection between deer inoculated by different routes for the first 50 days based on detection of E. chaffeensis infection by PCR assay of blood or culture isolation. These results demonstrate infection of (deer by intradermal and subcutaneous routes for the first time.     

Polymerase chain reaction for detection of Clostridium botulinum types A, B and E in food, soil and infant faeces

The application of the polymerase chain reaction (PCR) for detection of Clostridium botulinum types A, B and E in foods, environmental and clinical samples was evaluated and compared to the mouse bioassay. Samples inoculated with 10, 100 and 1000 spores of Cl. botulinum types A and B included pasteurized milk, UHT milk, infant formula, infant faeces, meat juice, canned tuna, mushrooms, blood sausage and soil. Clostridium botulinum type E spores were inoculated into fish eggs, canned tuna, picked herring, raw fish and soil at similar levels. Spores were added to 2.5 g of each sample with the exception of soil which was inoculated in 10 g samples. The presence of Cl. botulinum in sample enrichments was determined by both PCR and the bioassay. An overall correlation of 95.6% was observed between PCR results and the mouse bioassay. Of the total of 114 samples tested there was disparity between the mouse bioassay and the PCR in three samples of soil inoculated with 100 type A or E spores and 10 type B spores per 10 g, respectively, and two samples of infant faeces inoculated with 10 type A or B spores per 2.5 g. All of these samples gave negative animal results and positive PCR results.     

Accumulation of Salicylic Acid and 4-Hydroxybenzoic Acid in Phloem Fluids of Cucumber during Systemic Acquired Resistance Is Preceded by a Transient Increase in Phenylalanine Ammonia-Lyase Activity in Petioles and Stems

Cucumber (Cucumis sativa) leaves infiltrated with Pseudomonas syringae pv. syringae cells produced a mobile signal for systemic acquired resistance between 3 and 6 h after inoculation. The production of a mobile signal by inoculated leaves was followed by a transient increase in phenylalanine ammonia-lyase (PAL) activity in the petioles of inoculated leaves and in stems above inoculated leaves; with peaks in activity at 9 and 12 h, respectively, after inoculation. In contrast, PAL activity in inoculated leaves continued to rise slowly for at least 18 h. No increases in PAL activity were detected in healthy leaves of inoculated plants. Two benzoic acid derivatives, salicylic acid (SA) and 4-hydroxybenzoic acid (4HBA), began to accumulate in phloem fluids at about the time PAL activity began to increase, reaching maximum concentrations 15 h after inoculation. The accumulation of SA and 4HBA in phloem fluids was unaffected by the removal of all leaves 6 h after inoculation, and seedlings excised from roots prior to inoculation still accumulated high levels of SA and 4HBA. These results suggest that SA and 4HBA are synthesized de novo in stems and petioles in response to a mobile signal from the inoculated leaf.     

Increase in Plasmodesmatal Permeability during Cell-to-Cell Spread of Tobacco Rattle Virus from Individually Inoculated Cells

A microinjection technique was devised for inoculation of single Nicotiana clevelandii leaf trichome cells with virus particles. By removing inoculated trichomes at various times after microinjection, it was shown that at least 4 hr were required for tobacco rattle virus (TRV; tobravirus group) to move out of primarily inoculated cells. Effects of the early stages of TRV infection on plasmodesmatal permeability were examined by microinjection of fluorochrome-labeled molecules. Fluorescein-labeled insulin A chain (Mr, 2921) and fluorescein-labeled dextran (Mr, 4400) were observed to pass out of individual N. clevelandii trichome cells that had been inoculated with TRV by microinjection 5 hr previously. By contrast, Lucifer Yellow CH-labeled dextran (Mr, 10,000) was restricted to the inoculated cell. None of these fluorescent probes were able to move out of uninoculated cells or out of cells that had been inoculated with TRV only 2 hr previously. The movement of macromolecules through plasmodesmata, therefore, coincided with and probably resulted from cell-to-cell movement of TRV. The results are discussed with reference to the interaction of viruses and plasmodesmata and mechanisms of intercellular virus movement.     

Experimental inoculation of conventional pigs with porcine reproductive and respiratory syndrome virus and porcine circovirus 2

Postweaning multisystemic wasting syndrome (PMWS) is a disease of nursery and fattening pigs characterized by growth retardation, paleness of the skin, dyspnea, and increased mortality rates. Porcine circovirus 2 (PCV2) has been demonstrated to be the cause of PMWS. However, other factors are needed for full development of the syndrome, and porcine reproductive and respiratory syndrome virus (PRRSV) infection has been suggested to be one of them. Twenty-four conventional 5-week-old pigs were distributed in four groups: control (n = 5), PRRSV inoculated (n = 5), PCV2 inoculated (n = 7), and PRRSV and PCV2 inoculated (n = 7). The two groups inoculated with PRRSV showed growth retardation. Pigs inoculated with both PRRSV and PCV2 had increased rectal temperature. One of these pigs developed wasting, had severe respiratory distress, and died. The most important microscopic lesion in pigs inoculated with PCV2 was lymphocyte depletion with histiocytic infiltration of the lymphoid organs, more severe and in a wider range of tissues in doubly inoculated pigs. Interstitial pneumonia was observed in the three inoculated groups. PCV2 nucleic acid was found by in situ hybridization in larger amounts and in a wider range of lymphoid tissues in PRRSV- and PCV2-inoculated than in PCV2-inoculated pigs. TaqMan PCR was performed to quantify the PCV2 loads in serum during the experiment. PCV2 loads were higher in doubly inoculated pigs than in pigs inoculated with PCV2 alone. These findings indicate that severe disease can be reproduced in conventional 5-week-old pigs by inoculation of PRRSV and PCV2. Moreover, these results support the hypothesis that PRRSV infection enhances PCV2 replication.     

Does Facilitation of Faunal Recruitment Benefit Ecosystem Restoration? An Experimental Study of Invertebrate Assemblages in Wetland Mesocosms

We used wetland mesocosms (1) to experimentally assess whether inoculating a restored wetland site with vegetation/sediment plugs from a natural wetland would alter the development of invertebrate communities relative to unaided controls and (2) to determine if stocking of a poor invertebrate colonizer could further modify community development beyond that due to simple inoculation. After filling mesocosms with soil from a drained and cultivated former wetland and restoring comparable hydrology, mesocosms were randomly assigned to one of three treatments: control (a reference for unaided community development), inoculated (received three vegetation/sediment cores from a natural wetland), and stocked + inoculated (received three cores and were stocked with a poorly dispersing invertebrate group—gastropods). All mesocosms were placed 100 m from a natural wetland and allowed to colonize for 82 days. Facilitation of invertebrate colonization led to communities in inoculated and stocked + inoculated treatments that contrasted strongly with those in the unaided control treatment. Control mesocosms had the highest taxa richness but the lowest diversity due to high densities and dominance of Tanytarsini (Diptera: Chironomidae). Community structure in inoculated and stocked + inoculated mesocosms was more similar to that of a nearby natural wetland, with abundance more evenly distributed among taxa, leading to diversity that was higher than in the control treatment. Inoculated and stocked + inoculated communities were dominated by non‐aerial invertebrates, whereas control mesocosms were dominated by aerial invertebrates. These results suggest that facilitation of invertebrate recruitment does indeed alter invertebrate community development and that facilitation may lead to a more natural community structure in less time under conditions simulating wetland restoration.     

拟澳洲赤眼蜂和短管赤眼蜂在米蛾卵上的种间竞争

By means of single and mixed inoculation, this paper studied the interspecific competition between T. confusum and T. pretiosum on the factitious eggs of Corcyra cephaloica under different parasitoid densities, host densities and inoculated spaces. The results showed that for both singly and mixed inoculated groups, the parasitism increased with parasitoid density but decreased with host density, whereas the percentage of female progeny dropped with parasitoid density but enhanced with host density. No significant effect was observed on adult emergence for all parasitoid and host density treatments. In mixed inoculated group, the proportion of T. pretiosum in the progeny decreased with parasitoid density but increased with host density, and was more than 50% in all treatments, indicating that T. pretiosum had a stronger competitive ability than T. confusum. In the spaces ranging from 4cm^3 to 102cm^3, the parasitism decreased gradually in both singly and mixed inoculated groups. The percentage of female progeny and adult emergence had no significant difference among different inoculated spaces. In mixed inoculated group, the proportion of T. pretiosum in the progeny was more than 50% in all treatments except space 102 cm3, but decreased with space, which suggested that T. confusum could improve their competitive ability through increasing their search areas and looking for more hosts.     

接种乳黄粘盖牛肝菌和荧光假单胞菌对油松苗生长及猝倒病的影响

在盆栽条件下研究了单接种外生菌根真菌乳黄粘盖牛肝菌(SL)、单接种荧光假单胞菌(PF)、双接种(SL+PF)和不接种菌剂(CK)4个处理对油松苗生长、盆栽土壤指标、油松苗生理生化指标及猝倒病发生的影响.结果显示:(1)不同接种处理的油松苗成活率、生长状况及根系活力均高于对照,且SL+PF促生效果最好,其成活率、苗高、地径、地上干重、总干重及根系活力分别比对照提高22.10%、36.16%、44.86%、124.14%、161.54%和100%.(2)双接种可同时有效提高土壤酶活性和土壤养分含量,土壤脲酶、碱性磷酸酶和蔗糖酶活性均与速效氮、速效磷和有机碳呈显著或极显著正相关.(3)接种病原菌前后油松苗的MDA含量表现为CK>PF>SL>SL+PF,SOD、CAT和PAL活性均为SL+PF>SL>PF>CK,但接种病原菌前后不同处理酶活性增长幅度不同,CAT和PAL活性均以SL+PF增幅最大,分别比对照增长76.18%和112.64%.(4)接种处理的油松苗猝倒病发病率及病情指数均低于对照,且以SL+PF抗病效果最好,分别比对照降低60.94%和70.55%.研究表明,接种菌根真菌可有效提高土壤酶活性和土壤养分含量,增强植株体内的抗氧化酶活性,从而促进油松苗生长,抑制猝倒病的发生,菌根促生菌荧光假单胞菌增强了菌根真菌的作用.     

Rhizostabilization of metals in soils using Lupinus luteus inoculated with the metal resistant rhizobacterium Serratia sp. MSMC541

The aim of this work was to test Lupinus luteus plants, inoculated with metal resistant rhizobacteria, in order to phytostabilise metals in contaminated soils. The resistance to heavy metals of strains isolated from nodules of Lupinus plants was evaluated. The strain MSMC541 showed multi-resistance to several metals (up to 13.3 mM As, 2.2 mM Cd, 2.3 mM Cu, 9 mM Pb and 30 mM Zn), and it was selected for further characterization. Furthermore, this strain was able to biosorb great amounts of metals in cell biomass. 16S rDNA sequencing positioned this strain within the genus Serratia. The presence of arsenic resistance genes was confirmed by southern blot and PCR amplification. A rhizoremediation pot experiment was conducted using Lupinus luteus grown on sand supplemented with heavy metals and inoculated with MSMC541. Plant growth parameters and metal accumulation were determined in inoculated vs. non-inoculated Lupinus luteus plants. The results showed that inoculation with MSMC541 improved the plant tolerance to metals. At the same time, metal translocation to the shoot was significantly reduced upon inoculation. These results suggest that Lupinus luteus plants, inoculated with the metal resistant strain Serratia sp. MSMC541, have a great potential for phytostabilization of metal contaminated soils.     

接种两种固氮菌增强小麦幼苗抗渗透胁迫及生长能力

苗期是小麦(Triticum aestivum)物质和能量积累的关键时期, 苗期干旱影响小麦的后期群体建成。利用田菁茎瘤固氮根瘤菌(Azorhizobium caulinodans) ‘ORS571’与巴西固氮螺菌(Azospirillum brasilense) ‘Yu62’浸种侵染小麦和聚乙二醇(PEG)模拟渗透胁迫, 研究渗透胁迫下接菌小麦种子的发芽状况; 利用固氮菌涂抹小麦幼苗叶部, 测定PEG模拟渗透胁迫下小麦幼苗根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量, 探究固氮菌增强小麦幼苗抗渗透胁迫的能力。结果表明, 接种混合固氮菌后在渗透胁迫下小麦种子的发芽率明显提高; 在渗透胁迫下叶部涂抹固氮菌小麦的根体积、叶片相对含水量、脯氨酸含量及可溶性蛋白含量明显升高, 表明接种固氮菌可提高小麦幼苗的抗渗透胁迫能力, 且混合固氮菌对小麦幼苗叶片的增强效果优于单一固氮菌。     

接种量对花绒寄甲繁殖的影响

释放花绒寄甲防治天牛已成为重要手段,而繁殖花绒寄甲时,接种花绒寄甲幼虫到大麦虫蛹体时,控制合适的接种量是提高花绒寄甲繁殖数量和质量的关键技术。本研究通过在替代寄主上人工接种不同数量的花绒寄甲幼虫,观察其发育情况及其子代数量、质量等指标,明确最佳接种量。结果表明：随着接种量的增加,花绒寄甲幼虫历期和蛹历期明显缩短,其中接种4头/个和6头/个,幼虫历期为13 d,蛹历期为33 d,而接种14头/个和16头/个时,幼虫历期短于12 d,蛹历期明显缩短为28~29 d。花绒寄甲结茧数随接种量增加而增加,接种数为16头/个时,结茧数最多,接近7个。花绒寄甲结茧率随着接种量增多而降低,4头/个时,结茧率最高为72.3%。接种量对花绒寄甲子代个体数量和大小均有显著影响,其中接种8头/个时,羽化数平均为4.3头,显著高于接种4头/个（羽化数平均为2.8头）,明显低于接种16头/个(羽化数平均为6.9头)。接种量为4头/个和6头/个时,子代成虫个体最大,单头重平均每头可达0.035 g,接种量为8头/个时,成虫单头重平均每头0.032 g左右,接种量达到16头/个时,单头重最轻,为0.023 g。对花绒寄甲羽化率无显著影响,7个处理下子代羽化率均较高,平均在94.4%~100%。接种量越少,更利于花绒寄甲的生长发育,当接种量为4头/个时,花绒寄甲成虫发育最好,其子代个体最大,但子代数较少。因此,利用大麦虫蛹繁育花绒寄甲种虫时,最佳接种量为4头/个,而需要规模化繁育花绒寄甲作为天敌使用时,综合考虑子代数量和质量以及经济成本,最佳接种量为8头/个。     

Pathogenicity of aseptic Bursaphelenchus xylophilus

Pine wilt is a disease of pine (Pinus spp.) caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus. However, the pathogenic mechanism of pine wilt disease (PWD) remains unclear. Although the PWN was thought to be the only pathogenic agent associated with this disease, a potential role for bacterial symbionts in the disease process was recently proposed. Studies have indicated that aseptic PWNs do not cause PWD in aseptic pine trees, while PWNs associated with bacteria cause wilting symptoms. To investigate the pathogenicity of the PWN and its associated bacteria, 3-month-old microcuttings derived from certain clones of Pinus densiflora Siebold & Zucc. produced in vitro were inoculated under aseptic conditions with aseptic PWNs, non-aseptic PWNs and bacteria isolated from the nematodes. Six-month-old aseptic P. densiflora microcuttings and 7-month-old P. massoniana seedlings were also inoculated under aseptic conditions with aseptic PWNs and non-aseptic PWNs. The results showed that the aseptic microcuttings and seedlings inoculated with aseptic PWNs or non-aseptic PWNs wilted, while those inoculated with bacterial isolates did not wilt. Nematodes were recovered from wilted microcuttings and seedlings inoculated with aseptic PWNs and non-aseptic PWNs, and the asepsis of nematodes recovered from aseptic PWN-inoculated microcuttings and seedlings was reconfirmed by culturing them in NB liquid medium at 30°C for more than 7 days. Taken together, the results indicate that the asepsis of PWN did not cause the loss of pathogenicity.     

Replication of cauliflower mosaic virus DNA in leaves and suspension culture protoplasts of cotton

Cauliflower mosaic virus (CaMV) replicated in protoplasts and in inoculated leaves of the non-host, cotton (Gossypium hirsutum, L.). Protoplasts prepared from suspension-cultured cotton cells were infected by incubation with liposome-encapsulated CaMV virions. During a 1-week culture period the amount of CaMV nucleic acid as detected by nucleic acid hybridization in the protoplasts increased significantly regardless of whether or not the protoplasts contained vacuoles. In leaves inoculated with CaMV virions or CaMV DNA, viral DNA sequences were found by leaf skeleton hybridization to be located in small circular areas. DNA extracted from ultracentrifugal pellets of homogenates of inoculated leaves contained circular, gapped CaMV DNA only when inocula contained CaMV virions, CaMV DNA, or partial nested dimer CaMV plasmid DNA. When plants had been heavily watered, the CaMV DNA recovered contained degraded CaMV DNA. The results suggest that the host range limitation for CaMV is not due to an inability to replicate or spread locally in inoculated leaves.     

马传染性贫血病毒非编码区LTR嵌合克隆的生物学特性

将已构建的马传染性贫血病毒LTR强弱毒嵌合克隆衍生毒vLGFD9-12体内接种健康试验马,在150d观察期内,各组试验动物体症均未见异常。血液学分析发现,vLGFD9-12嵌合克隆衍生病毒与亲本弱毒疫苗株的白细胞与血红蛋白含量总体上没有明显的规律性的变化。在动物外周血中均检测到一定的病毒RNA拷贝数,但拷贝数较低。二者在诱导EIAV特异性淋巴细胞增殖功能和特异性细胞毒性杀伤反应中,亦具有相似的变化趋势和效应。本项研究为进一步确定我国马传贫弱毒疫苗株毒力致弱及免疫保护的分子机制奠定了重要的分子生物学基础。     

Scrapie I. Transmission and Pathogenesis

Homogenized brain, extracts, and residues of brain from normal and scrapie sheep were inoculated into 116 sheep. Of 72 sheep inoculated with scrapie material 27 developed the disease, whereas four of 44 inoculated with ;normal' brain material showed symptoms similar to those of scrapie. The scrapie agent survived extraction for 18 hours with diethyl ether followed by water for 24 hours. Similarly the agent survived extraction in a Soxhlet apparatus with 95 per cent ethanol containing 2.0 per cent 4 M HC1 for 24 hours. There was no evidence of demyelination in sections of brain and spinal cord supporting the view that allergic encephalomyelitis is not responsible for the scrapie symptoms. Liver function, as measured by the bromsulfalein test, remained normal until just before death. These results preclude that liver dysfunction contributes to the disease but do not exclude the possibilities of other metabolic derangements.     

Some pea cultivars as local lesion hosts for potato virus X

SixteenPisum sativum cultivars were mechanically inoculated with two isolates of PVX. Distinct necrotic local lesions on the inoculated leaves were found in six cultivars 5 to 6 days after inoculation. The best results were obtained with the Meteor and Raman cultivars, 17 to 23 days old.