纯化的玉米小斑病菌C小种毒素生物活性的研究

 纯化的HMC毒素对C细胞质玉米幼根生长的抑制,根电解质的渗漏,叶片和植株的伤害,超氧物歧化酶和过氧化物酶活性及原生质层透性的影响,与基因型相同的T、N细胞质相比,差异显著,进一步确定了HMC毒素对C细胞质的专化作用。     

玉米小斑病菌C小种毒素的分离、纯化及其植物病理反应

 魏建昆等证实在中国存在玉米小斑病C小种^[1]。本文通过凝胶柱层析(Sephadex G-25)和硅胶层析(Silica gel)将C毒素(HMC-toxin)分为四种亚毒素,毒素Ⅰ和毒素Ⅲ(占全部毒素管数的75%)主要侵染T、C、S、N四种同核异质玉米中的C胞质叶片,使之产生严重萎蔫,为专化性毒素,但少数管号内的毒素对T、S有轻微的伤害,而且毒素Ⅰ内少量对N也有影响,毒素Ⅱ(占10%)只轻微地侵染N细胞质,毒素Ⅳ(占15%)除了对S无影响外,对T、C、N三种细胞质均有一定的危害。由此可见,C毒素是一具有生物活性的混合体,其中以侵染C胞质的毒素为优势毒素,同时存在少量对T、S、N三种细胞质毒力较弱的毒素。对T、C、S、N四种同核体(Homocaryons)玉米人工接种C小种所出现的叶片抗性反应,从本实验获得了一定的解释。     

HMC-毒素对C型不育玉米根冠细胞原生质体及微丝分布的影响

 用玉米小斑病菌C小种毒素(HMC-毒素)处理C型不育玉米根冠细胞后,采用微丝的特异探针异硫氰酸荧光素-鬼笔环肽进行标记,荧光显微镜下发现HMC-毒素可引起根冠细胞原生质体收缩,并伴有原生质泄漏、原生质体荧光变暗、微丝分布异常等现象。HMC-毒素对根冠细胞的这种影响明显不同于细胞松弛素B (CB);并且与HMC-毒素处理后的同核保持系(N)玉米相比损坏程度要严重得多,N型玉米细胞原生质体只是轻度收缩,仍可见微丝网络分布。根冠细胞对HMC-毒素的这种反应与细胞质的抗病性相关。     

不同林龄胡杨林叶片与土壤的化学计量特征

为了解天然胡杨林生态系统的C、N、P、K含量及化学计量特征,在新疆轮台县轮南镇选取5种不同林龄胡杨林(幼龄林、中龄林、近熟林、成熟林和过熟林),研究不同林龄胡杨叶和林下土壤C、N、P、K含量及化学计量特征。结果表明:(1)不同林龄胡杨叶的C、N、P、K含量分别为437.77、13.67、1.87、5.26 g·kg-1,林龄对叶片C、N、P、K含量影响不显著。不同林龄胡杨叶片C∶N、C∶P、C∶K、N∶P、N∶K、P∶K为35.46、187.04、87.86、8.40、2.66、0.57,林龄对叶片的C∶P、C∶K影响显著,对C∶N、N∶P、N∶K、P∶K影响不显著。(2)土壤的C、N、P、K含量分别为6.89、0.62、0.57、18.69 g·kg~(-1),林龄对土壤C、N、P、K含量影响显著。不同林龄土壤C∶N、C∶P、C∶K、N∶P、N∶K、P∶K为11.48、12.34、0.36、1.12、0.032、0.032,林龄对土壤化学计量影响差异显著。(3)胡杨叶片N∶P比为8.40,远远小于阀值14,表明胡杨生长发育过程中严重受土壤氮素含量限制。     

安徽省玉米小斑病菌生理小种鉴定及对烯唑醇的敏感性

玉米小斑病是由玉蜀黍双极蠕孢(Bipolarismaydis)引起玉米叶部发病的病害之一,该病害广泛分布在世界玉米产区,严重影响玉米生产。玉米小斑病菌存在明显生理分化现象,目前已证实在中国存在T、C、S和O共4个小种。由于近年来不育细胞质的感病玉米自交系在玉米育     

贺兰山青海云杉针叶C、N、P含量及其计量比随环境因子的变化特征

为了研究青海云杉叶片养分随环境的变化特征,通过测定贺兰山海拔梯度上的青海云杉针叶碳、氮、磷含量,研究不同海拔青海云杉叶片C、N、P的变异特征,同时探讨叶片C、N、P含量及其计量比与环境因子的关系,为进一步探讨植物对环境的响应提供科学依据。结果显示:(1)随着海拔升高、温度的下降以及降水的逐渐增加,青海云杉叶片C含量没有显著的变化,叶片N含量随海拔梯度显著下降。而叶片P随海拔梯度波动较大但并没有表现出明显的下降趋势,只是在海拔为2810m之后,P含量显著降低,C:N随着海拔的升高而升高,由于叶片P含量波动,使C:P和N:P的比值均表现先降低后迅速升高的趋势。(2)本研究青海云杉叶片C、N、P含量及C:N、C:P及N:P值的变异系数均小于30%,其大小排序为P(27.03%)>C:P(26.74%)>N:P(19.19%)>C:N(12.44%)>N(12.33)>C(1.07%)。(3)青海云杉叶片N、P含量及N:P比值相比于其他研究处于较低的水平。因此,在海拔梯度上,优势树种青海云杉叶片C含量受影响不大,而N、P含量随环境变化较大;较低的N:P值说明贺兰山青海云杉在生长中主要受N的限制。     

二斑叶螨侵染下菜豆叶片交替呼吸途径对光合作用、叶温和间接防御的影响

在害虫侵染下,植物能够通过气味吸引害虫天敌以防御害虫的侵染,这被称为间接防御反应.本文研究发现,二斑叶螨的侵染增加了叶片总呼吸、交替呼吸途径容量以及交替呼吸途径容量在总呼吸中比例的水平,并提升了叶片对二斑叶螨天敌的吸引作用.交替呼吸途径抑制剂(水杨基氧肟酸)的处理则降低了二斑叶螨侵染叶片对天敌的吸引作用.水杨基氧肟酸的处理并没有显著改变二斑叶螨侵染叶片的叶片温度,但降低了其光合速率;而二斑叶螨侵染叶片对天敌的吸引作用可能和其光合速率有关.基于以上结果,讨论了在害虫侵染下线粒体交替呼吸途径对于植物间接防御影响的可能机制.     

盐胁迫下交替呼吸对二斑叶螨侵染叶片及其间接防御的影响

为探索在遭受盐胁迫时二斑叶螨Tetranychus urticae Koch侵染叶片的交替呼吸途径的生理学功能,在实验室条件下研究了NaCl以及水杨基氧肟酸对二斑叶螨侵染叶片的间接防御反应,以及对过氧化氢含量和细胞死亡水平的影响。200 mmol/L NaCl胁迫没有明显影响二斑叶螨侵染叶片对智利小植绥螨Phytoseiulus persimilis的吸引作用,也没有提高二斑叶螨侵染叶片中过氧化氢含量和细胞死亡的水平,但显著提高了侵染叶片中交替呼吸途径的水平。200 mmol/L NaCl和1mmol/L水杨基氧肟酸的复合处理则显著降低了二斑叶螨侵染叶片对捕食螨的吸引作用,并提高了过氧化氢含量和细胞死亡的水平。研究表明,当二斑叶螨侵染叶片遭受盐胁迫时,交替呼吸途径不仅有助于维持植物的间接防御反应,而且缓解了植物体内的氧化压力和细胞死亡。     

科尔沁沙地小叶锦鸡儿人工林防风固沙及改良土壤效应研究

为探讨小叶锦鸡儿防风固沙、改良土壤养分状况和生物活性的能力,选取6年和11年生小叶锦鸡儿人工固沙林为对象,以流动沙丘为对照,研究不同年龄固沙林降低风速和风沙流结构特征、土壤有机质和N,P含量、土壤酶的活性以及微生物生物量C,N,P含量的变化特征.结果表明:在小叶锦鸡儿灌木林内,各个高度风速和输沙量均显著低于流动沙丘,防风固沙效果明显.随着小叶锦鸡儿固沙群落发育时间的增长,其综合防风性能提高,土壤有机质、N,P,K含量和生物活性逐渐得到改善.O～30 cm土层中磷酸单酯酶、蛋白酶、脲酶、蔗糖酶、硝酸还原酶、多酚氧化酶、脱氢酶的活性和土壤微生物生物量C,N和P含量均大幅度提高,其中O～10 cm土层增幅最大.土壤酶中蔗糖酶的活性增加最为迅速,6年和11年生固沙林O～10 cm 土层分别是流动沙丘的28.58倍和55.21倍.小叶锦鸡儿不仅具有较好的防风固沙性能,而且表现出强大的改善土壤养分和生物活性的能力,可作为优良的固沙植物材料在科尔沁沙地大面积推广应用.     

河西走廊藜麦C、N、P生态化学计量学特征对物候期的响应

藜麦（Chenopodium quinoa）是南美洲传统作物,具有极高的营养价值和较强的环境适应能力,但其在我国西北干旱区的适应性还有待深入研究。生态化学计量学是对有机体的元素组成（主要是C、N、P）及其关系进行研究的科学,能一定程度地反映有机体的特征及其与环境的关系。在河西走廊大田栽培条件下,本试验研究了藜麦主要物候期的碳（C）、氮（N）、磷（P）含量及其生态化学计量比的变化。结果表明：随物候期的变化,藜麦的有机碳（organic carbon,OC）含量变化不显著,而全氮（total nitrogen,TN）和全磷（total phosphorus,TP）含量则显著降低;各器官间的OC含量较稳定,而TN和TP含量差异显著且叶和穗较高。藜麦的C∶N、C∶P随物候期的变化呈升高趋势,N∶P则呈先降低后显著升高趋势;根和茎C∶N、C∶P较高而叶N∶P较高。物候期和器官显著影响藜麦的C、N、P含量和计量比,后者与藜麦的生长和物质积累速度显著相关。     

小麦赤霉病菌毒素对小麦抗病突变体及其亲本细胞超微结构的影响

 在小麦赤霉病菌毒素的作用下,小麦抗赤霉病突变体92k809及其亲本新克旱九内颖细胞超微结构发生了变化。随着处理时间的增长,细胞器和膜系统破坏程度加剧,但在相同处理时间内,突变体细胞的超微结构破坏程度比其亲本要轻。对赤霉病菌毒素最敏感的是线粒体,其次是叶绿体、细胞质膜和细胞核膜。本文还探讨了赤霉病菌毒素在致病过程中的作用及其致病机理。     

Effects of tebuconazole on morphology,structure, cell wall components and trichothecene production of Fusarium culmorum in vitro

The effects of tebuconazole, a systemic fungicide, on the morphology, structure, cell wall components and toxin production of Fusarium culmorum were investigated in vitro. Treatment was by application of four filter paper strips (0.75 cm × 5.0 cm) soaked in 20 µg ml ^?1 fungicide placed around a point inoculum in Petri dishes. Mycelial growth was strongly inhibited by fungicide treatment. Scanning electron microscopic observations showed that the fungicide caused irregular swelling and excessive branching of hyphae. The morphological changes induced by the fungicide at the ultrastructural level included considerable thickening of the hyphal cell walls, excessive septation, the formation of the incomplete septa, extensive vacuolisation, accumulation of lipid bodies and progressing necrosis or degeneration of the hyphal cytoplasm. Non‐membrane inclusion bodies were often detected in the hyphal cytoplasm. Furthermore, the formation of new hyphae (daughter hyphae) inside collapsed hyphal cells was common following treatment. The daughter hyphae also displayed severe alterations such as irregular thickening of the cell walls and necrosis of the cytoplasm. Using cytochemical techniques, the labelling densities of chitin and β‐1,3‐glucan in the cell walls of the fungicide‐treated hyphae were more pronounced than in those of the control hyphae. Moreover, immunogold labelling with antiserum against deoxynivalenol (DON) revealed that Fusarium toxin DON was localized in the cell walls, cytoplasm, mitochondria and vacuoles of the hyphae from the control and the fungicide treatment, but the labelling density in the fungicide‐treated hyphae decreased dramatically compared with the control hyphae, indicating that tebuconazole reduced Fusarium toxin production of the fungus. © 2001 Society of Chemical Industry     

Immunocytochemical localization of the Bacillus sphaericus binary toxin components in Culex quinquefasciatus (Diptera: Culicidae) larvae midgut

This study describes the immunocytochemical localization of the Bacillus sphaericus 2362 binary toxin components, BinA and BinB, in Culex quinquefasciatus larvae that had been intoxicated with this entomopathogen. Ultrathin sections of C. quinquefasciatus larvae midgut embedded in the hydrophilic resin L.R. White were incubated with the antibodies anti-BinA or anti-BinB and then revealed with goat anti-rabbit IgG coupled to gold particles. Immunocytochemical detection demonstrated the presence of specific labeling in ultra-thin sections that had been incubated with the BinA antiserum. Gold particles were detected on the apical areas of cell membranes and inside the epithelial cell cytoplasm, particularly the mitochondria, of cells from the gastric caeca and posterior stomach in larvae exposed during 2 or 24 h to the entomopathogen. A similar labeling pattern was observed in ultrathin sections from both midgut regions when incubated with BinB antiserum.     

小麦穗组织中脱氧镰刀菌烯醇毒素的免疫细胞化学定位

 采用免疫细胞化学技术对禾谷镰刀菌(Fusarium graminearum)在侵染小麦穗部过程中产生的脱氧镰刀菌烯醇毒素(deoxynivalenol,DON)进行了定位分析。在接种后24h,当菌丝在外稃、内稃的内侧表面扩展而尚未侵入寄主细胞前,病菌已分泌DON,并且DON已扩散到寄主组织内。在菌丝细胞内,DON主要被定位于细胞质、线粒体及细胞壁上;在寄主细胞中DON主要分布于细胞壁、叶绿体、细胞质和内质网上。在侵染初期(接种后2 d),菌丝仅能在寄主细胞间隙扩展,随寄主组织中DON浓度的升高,寄主细胞相应发生了一系列病理变化。随寄主细胞坏死(接种后3~4d),病菌进入坏死的寄主细胞。上述结果表明,DON在禾谷镰刀菌的侵染、致病和定殖过程中起着重要的作用。毒素标记结果表明病菌产生的毒素可通过穗轴微管束组织从侵染部位向上、向下转输,毒素向上的转输量明显高于向下转输     

Alternaria brassicae produces a host-specific protein toxin from germinating spores on host leaves

Spore suspensions of Alternaria brassicae, the causal agent of gray leaf spot in Brassica plants, were incubated on the leaves of cabbage (B. oleracea) and spore germination fluid (SGF) was collected after 48 h. A high molecular weight (HMW) fraction (>10 kDa) was separated from the SGF by ultrafiltration. In a detached leaf assay, the HMW fraction induced visible symptoms only on host leaves and the toxicity was lost by treatment with proteinase K or heat at 60 degrees C for 15 min, indicating the presence of host-specific protein toxin(s). A protein toxin in the HMW fraction was purified by several chromatography steps. The toxin induced water-soaked symptoms followed by chlorosis at concentrations of 0.5 to 1 microg/ml on host leaves, but not on nonhost leaves even at 50 microg/ml. The toxin also had infection-inducing activity when added to spore suspension of a nonpathogenic isolate of A. alternata, causing symptoms similar to the infection of A. brassicae only on host leaves. These results indicate that a new host-specific protein toxin named ABR-toxin is released from germinating spores of A. brassicae on host leaves. ABR-toxin migrated as a protein of 27.5 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of ABR-toxin was estimated to be approximately 7.0 and 21 N-terminal amino acid residues were sequenced.     

Mitochondrial changes during storage of untreated or CIPC-treated potatoes

During storage of potato tubers (Solanum tuberosum L., var. Bintje), important changes appear which affect respiratory control, $\text{ADP}\text{O}$, intensity of O₂ consumption in the presence of different substrates, and NAD⁺ dependence, In mitochondria extracted under strictly similar conditions, from patato tubers stored at 4°C, the respiratory control (RC) maintains a value near 4 for 4 to 5 months. It then declines progressively to low values. At 20°C, a stable RC of 4 can be observed for several months, which then decreases at the end of dormancy. Then, the RC increases sharply; at this stage, $\text{ADP}\text{O}$ are abnormally low, and, some time later, NAD⁺ dependence disappears. Mitochondria treated with 250 μM chlorpropham show a 50% inhibition of the electron transfer with exogenous NADH as substrate. After tuber treatment with 1% chlorpropham, sprouting is inhibited for several months. The activities of mitochondria extracted from such tubers remain unaffected by the treatment. The use of this phenylcarbamate for potato tuber treatment permits obtaining functional mitochondria from tubers after a slightly longer period of storage.     

Optimization of in vitro growth conditions of Pyrenophora teres for production of the phytotoxin aspergillomarasmine A

In liquid cultures of Pyrenophora teres, three phytotoxins may be found: L, L - N -(2-amino-2-carboxyethyl) aspartic acid (toxin A), anhydroaspergillomarasmine A (toxin B) and aspergillomarasmine A (toxin C). In particular, toxins A and C cause chlorotic and necrotic symptoms in detached barley leaves, toxin C being the most damaging, whereas toxin B is only weakly phytotoxic. When P. teres is grown in liquid modified Fries medium, toxin B is the main toxin accumulated, possibly due to a ring closure of toxin C at the low pH value of the medium. The amount of toxin B produced by 11 isolates of P. teres was compared in modified Fries medium. Generally, the most virulent isolates of P. teres produced higher amounts of toxin B than the less virulent isolates. During growth, the pH of the media decreased from 6.7 to about 3.0–3.5, followed by a slight increase to about 3.5–4.0. All isolates, except one, produced toxin B, whereas only two isolates produced toxin C and toxin A. Maintaining the pH at about 6.5 by sterile titration with 1 M NaOH resulted in a shift in toxin accumulation from toxin B to toxin C. The addition of tris or phosphate buffer to the media resulted in higher pH during the growth period, an increase in the total amount of toxins produced, and a shift in toxin accumulation from toxin B to toxin C. The higher pH value probably prevented the conversion of toxin C into toxin B. No toxins were produced in two routinely used media, potato glucose broth and grass broth. Toxin B and toxin C were purified by ion exchange chromatography and precipitation with HCl.     

苯醚甲环唑在茶叶中的残留量及其在茶汤中的浸出动态研究

采用气相色谱-电子捕获检测器(GC-ECD)对田间采集的茶叶样品中的苯醚甲环唑残留量进行了测定,同时对泡茶后茶汤中苯醚甲环唑的浸出动态进行了研究。结果表明:苯醚甲环唑10%水分散粒剂分别按有效成分100和150 mg/kg于茶叶炭疽病发生初期施药2~3次,距末次施药后间隔5 d采样,成茶中苯醚甲环唑的残留量低于最大残留限量(MRL)值10.0 mg/kg;茶汤中苯醚甲环唑残留量及浸出率随冲泡次数的增加逐渐下降,4次冲泡的总浸出率为19.6% ~21.8%,人体摄入的苯醚甲环唑残留量占每日允许摄入量(ADI)的9.0%。