Toxicity of fenpropimorph to fenarimol-resistant isolates of Penicillium italicum

Fenarimol-resistant isolates ofPenicillium italicum with cross-resistance to imidazole and triazole fungicides which inhibit ergosterol biosynthesis were tested for their sensitivity to fenpropimorph. Radial growth tests on PDA showed that the isolates (n=6) lacked cross-resistance to fenpropimorph or displayed enhanced sensitivity to the fungicide (negatively correlated cross-resistance). Control of blue mold decay of oranges incited by the wild-type isolate could be achieved by dipping fruits in suspensions of fenpropimorph at a concentration of 100 mg ml^–1. Decay of oranges incited by the fenarimol-resistant isolates was controlled at the same or at a lower concentration (30 mg ml^–1), thus showing that the normal or increased sensitivity to fenpropimorph is also expressed under in vivo conditions.Samenvatting Fenarimol-resistente isolaten vanPenicillium italicum met kruisresistentie tegen imidazool-en triazoolfungiciden die de ergosterolbiosynthese remmen, werden getoetst op hun gevoeligheid voor fenpropimorf. Radiale groeiproeven op PDA toonden aan dat de isolaten (n=6) geen kruisresistentie bezaten met fenpropimorf of een verhoogde gevoeligheid voor het middel vertoonden (negatief gecorreleerde kruisresistentie). Op sinaasappels konPenicillium-rot, veroorzaakt door het wild-type bestreden worden door middel van een dompelbehandeling met fenpropimorf bij een dosering van 100 g ml^–1). Bestrijding van rot veroorzaakt door fenarimil-resistente isolaten werd verkregen bij dezelfde of een lagere dosering (30 g ml^–1); aldus werd aangetoond dat de normale of verhoogde gevoeligheid voor fenpropimorf ook in vivo tot uiting komt.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

Characterisation of energy-dependent efflux of imazalil and fenarimol in isolates of Penicillium italicum with a low,medium and high degree of resistance to DMI-fungicides

Differential accumulation of [¹⁴C]imazalil and [¹⁴C]fenarimol by germlings of wild-type and DMI-resistant isolates ofPenicillium italicum was studied at various pH values. At pH 7 and 8 the low-resistant isolate E_300–3 accumulated 22% and 35%, respectively, less imazalil than the wild-type isolate W₅. Imazalil accumulation at pH 5 and 6 was similar. Isolate E_300–3 also accumulated less fenarimol as compared with the wild-type isolate. This difference was much more obvious than for imazalil and was observed at all pH values tested. Differences in accumulation of both imazalil and fenarimol between low (E_300–3), medium (H₁₇) and high resistant (I₃₃) isolates were not observed. These results suggest that decreased accumulation of DMIs is responsible for a low level of resistance only and that additional mechanisms of resistance might operate in isolates with a medium and high degree of resistance. With all isolates fenarimol accumulation was energy-dependent. This was not obvious for imazalil.The wild-type and DMI-resistant isolates had a similar plasma membrane potential as determined with the probe [¹⁴C]tetraphenylphosphonium bromide ([¹⁴C]TPP⁺). Various test compounds, among which ATPase inhibitors, ionophoric antibiotics and calmodulin antagonists, affected the accumulation of [¹⁴C]TPP⁺, [¹⁴C]imazalil and [¹⁴C]fenarimol. No obvious correlation between the effects of the test compounds on accumulation levels of the fungicides and [¹⁴C]TPP⁺ could be observed. These results indicate that the plasma membrane potential does not mediate the efflux of DMI fungicides byP. italicum.     

克瑞森无核葡萄的主要性状和栽培技术

克瑞森无核为晚熟无核葡萄优良品种。果穗圆锥形，平均重500g；果粒椭圆形，平均重4g，经赤霉素处理后可达6g；果皮红色，果肉黄色，脆硬味甜，口感极佳；10月中下旬成熟，耐贮运，冷藏案件下可存放5个月：抗病能力强于红地球。栽培时宜采用小棚架长梢修剪；合理灌水并严格控制生长后期氮肥的施用；采用化学疏花疏果技术，套袋果适时脱袋、及时采收。注意阴雨天不得进行抹芽、剪梢，以防白腐病侵染，     

Cyclooxygenase ‐2 Expression in Mammary Tumors in Dogs and its Correlation to Histologic and Biologic Behavior

Introduction:  Cyclooxygenase‐2 (Cox‐2) is the inducible form and the rate‐limiting enzyme, for conversion of arachidonic acid to prostaglandins. Cox‐2 overexpression, common in carcinomas, is associated with increased growth rate, resistance to apoptosis, angiogenesis, and overall, both local and distant aggressive behavior. Cox‐2 overexpression has been detected in human and canine mammary tumors (MTs). Histopathology of canine MT is not always predictive of biologic behavior, and anecdotally, only 50% of the malignant MTs are expected to metastasize. We hypothesize that Cox‐2 expression correlates with aggressive behavior.
Methods:  This retrospective study evaluated 48 bitches, presented for excision of MT between 2000 and 2003 at FMVZ de Botucatu‐UNESP, Brasil. Follow‐up varied from 18 months to 24 months and included physical examination and thoracic radiographs. Histopathologic examination was performed in all tumors, as well as in metastatic lesions when detected in the follow‐up period. Immunohistochemistry was used to detect expression of Cox‐2 in paraffin blocks (Rabbit polyclonal anti‐PGHS‐2. Oxford Biomedical). 10 adenomas, 10 carcinomas, 10 benign mixed tumors, 10 malignant mixed tumors and 8 cases of primary carcinomas and their metastatic lesions.
Results:  Expression of Cox‐2 varied among groups. Adenomas (32.1%), mixed benign tumors (38%), carcinomas (60.3%), malignant mixed tumors (65.8%), and metastatic carcinomas (81.25%) and their metastatic lesions (84.35%). Statistically significant differences (p < 0.05) were observed between the benign and malignant counterparts and between carcinomas and metastatic carcinomas.
Conclusions:  Cox‐2 expression correlates with both histologic and biologic behavior in mammary carcinomas, and may serve as a predictor of metastatic potential.     

压力蒸汽灭菌器对液体灭菌时常见故障浅析

针对压力蒸汽灭菌器对液体进行灭菌时造成事故的现象，分析了事故发生的主要原因是由于限压阀瞬间降压造成的。指出应慎用于液体的灭菌处理。     

西瓜中类胡萝卜素的高效液相色谱法分析

通过高效液相色谱法对西瓜中的类胡萝卜素进行分析,采用Vydac201TP54C18色谱柱,比较2种不同流动相的检测结果,表明乙腈:甲醇:二氯甲烷=75:20:5(V:V:V)的分离效果较好;初步定性分析确定西瓜中含有7种以上类胡萝卜素。     

西瓜核型雄性不育两用系POD同工酶研究

用PAGE(聚丙烯酰胺凝胶)电泳技术,研究了西瓜核型雄性不育两用系不同器官的POD(过氧化物酶)同工酶酶谱差异。试验结果表明,不育株与可育株主要差异在雄花蕾,表现为不育株较可育株多2条活性强的酶带;其他器官差异小,仅表现为酶谱带强弱不同。并对POD同工酶与雄性不育性的相关性进行了研究。     

披碱草和野大麦及其杂种苗期抗旱性研究

在室内人工模拟干旱条件,测定了干旱胁迫下披碱草和野大麦及其杂种幼苗的相对含水量、细胞膜透性、游离脯氨酸含量的变化。结果表明,干旱胁迫下亲本披碱草幼苗的组织相对含水量下降的幅度最低,细胞膜受害最轻,游离脯氨酸积累的最多,相对生长率最高,表现出很强的抗旱性。亲本野大麦幼苗的组织相对含水量下降的幅度最大,细胞膜受害最重,游离脯氨酸积累的最少,相对生长率最低,抗旱性弱。杂种F1幼苗的保水能力、细胞膜受害程度和游离脯氨酸积累能力、相对生长率均介于双亲之间,亦即其抗旱能力大于野大麦而小于披碱草,表现为双亲的中间型。正、反交对杂种F1的抗旱能力没有明显影响。     

剡溪山水文化旅游资源开发设想

文化底蕴越是丰厚的旅游资源，开发价值就越高。正是众多文人贤士游剡访古，使浙东剡溪的历史、文化与自然山水完美地结合在一起，形成了绚丽多彩的剡溪山水文化。文化被认为是旅游业的灵魂，是一种潜在的旅游产品。要提高山水旅游产品的文化品位，必须将山水文化的潜在价值转化为旅游产品。本文对剡溪山水文化旅游资源的开发提出了以下设想：再现“唐诗之路”山水旅游美景，并着重于按唐诗及有关文献、传说等增添新景点，沿江、沿山、于水中进行山水、文化全面开发，形成立体式、全景式的剡中山水文化游道；可开发出文化生态、修学、科考、宗教、节庆等剡溪山水文化旅游产品。