Detection of squash mosaic virus in seeds of melon (Cucumis melo) by enzyme-linked immunosorbent assay (ELISA)

Squash mosaic virus (SqMV, comovirus) is seed-transmitted in severalCucurbitaceae. Therefore, the use of virus-free seed is important to prevent establishment of this virus in the Netherlands and to avoid spread to other countries.This study was undertaken to develop an enzyme-linked immunosorbent assay (ELISA) for the detection of SqMV in melon seeds. An antiserum was produced to a serotype 1 isolate from melon. Two ELISA variants were investigated viz. an ELISA variant with simultaneous incubation of sample and enzyme conjugate (ELISA 1) and an ELISA variant with successive incubation of sample and enzyme conjugate (ELISA 2). The sensitivity of ELISA was tested by mixing fluor of ground infected and non-infected seeds in different proportions. SqMV was detected by both ELISA variants at dilutions of 1 160 (1 part of infected flour mixed with 159 parts of non-infected flour) or higher after a substrate incubation period of 4 h. However, ELISA 1 gave relatively higher absorbance values than ELISA 2 for nearly all dilutions. Since ELISA 1 is also faster than ELISA 2, ELISA 1 is advised for routine testing. In these test, using subsamples of 100 melon seeds SqMV is detected reliably. ELISA 1 is now used in the Netherlands for routine-indexing of melon seed lots for SqMV.Samenvatting Het pompoenemozaïekvirus gaat over met het zaad van verscheideneCucurbitaceae. Het gebruik van virusvrij zaad is belangrijk om te voorkomen dat het virus zijn intrede doet in Nederland en zich naar andere landen verspreidt.Een antiserum werd geproduceerd tegen een serotype 1 isolaat van meloen. Met behulp van dit antiserum werd een ELISA ontwikkeld om pompoenemozaïekvirus in zaden van meloen aan te tonen. Twee varianten van ELISA werden vergeleken, namelijk een variant waarbij monster en enzymconjugaat gelijktijdig geïncubeerd werden (ELISA 1) en een variant waarbij monster en enzymconjugaat na elkaar geïncubeerd werden (ELISA 2). De gevoeligheid van de ELISA varianten werd uitgetoetst door meel van zieke zaden in verschillende verhoudingen te mengen met meel van gezonde zaden. Het pompoenemozaïekvirus werd met beide ELISA varianten aangetoond in verdunningen van 1 160 (1 deel meel van zieke zaden gemengd met 159 delen meel van gezonde zaden) of hoger na 4 uur incubatie met substraat. ELISA 1 gaf doorgaans hogere extinctiewaarden dan ELISA 2 voor bijna alle verdunningen. Omdat ELISA 1 ook nog sneller is dan ELISA 2, wordt ELISA 1 aanbevolen voor routinematig gebruik. Wanneer voor routinematig gebruik 100 meloenezaden per submonster getoetst worden, kan het pompoenemozaïek virus betrouwbaar worden aangetoond. In Nederland worden momenteel per zaadpartij 20 submonsters van 100 zaden getoetst.     

Resistance to bifenthrin and resistance mechanisms of different strains of the two-spotted spider mite (Tetranychus urticae) from Turkey

Nine different strains of the two-spotted spider miteTetranychus urticae Koch (Acari: Tetranychidae) were collected on cotton from Adana, Antalya, Izmir, Manisa and Urfa in Turkey. Their responses to bifenthrin were investigated using conventional bioassay and biochemical assays. LC₅₀ and LC₉₀ values of bifenthrin were determined for all strains by using a residual bioassay with a petri dish-spray tower. Resistance ratios were determined by comparing the samples with a standard susceptible strain, GSS. The resistance ratios of the strains ranged from <1 to 669-fold (at LC₅₀). Of the investigated field strains, only three (two from Adana and one from Urfa) were resistant to bifenthrin. There was a correlation between esterase enzyme activity and bifenthrin resistance according to polyacrylamide gel electrophoresis and microtiter plate assays in the three resistant strains. http://www.phytoparasitica.org posting May 17, 2005.     

Age-related resistance to Pseudomonas syringae pv. tomato is associated with the transition to flowering in Arabidopsis and is effective against Peronospora parasitica

As plants mature it has been observed that some become more resistant to normally virulent pathogens. The ability to manifest the Age-Related Resistance (ARR) response in Arabidopsis to Pseudomonas syringae pathovars tomato (Pst) coincided with the transition to flowering in plants both delayed and accelerated in the transition to flowering. ARR was also associated with a change in PR-1 gene expression, such that young plants expressed PR-1 abundantly at 3 days post inoculation (dpi) while mature plants expressed much less. The Arabidopsis ARR response requires SA accumulation via isochorismate synthase (ICS1) [24]. ICS1 was expressed one dpi with virulent and avirulent Pst in both young and mature plants. The ARR response was also effective versus avirulent Pst providing an additional 4-fold limitation in bacterial growth. Arabidopsis ARR was found to be ineffective against two necrotrophs, Erwinia carotovora subspecies carotovora (bacterium) and Botrytis cinerea (fungus) and one obligate biotroph, Erysiphe cichoracearum (fungus). However, mature wild type, SA-deficient sid2 and NahG plants supported little growth of the obligate biotrophic oomycete, Peronospora parasitica. Therefore ARR to P. parasitica appears to be SA-independent, however the level of ARR resistance was somewhat reduced in these mutants in some experiments. Thus, there may be numerous defence pathways that contribute to adult plant resistance in Arabidopsis.     

采后ASM诱导处理对鸭梨果实黑霉病的控制

 Acibenzolar-S-methyl (ASM) 是重要的植物抗病基因诱导剂。鸭梨果实经过负压渗透ASM诱导处理后接种黑霉病(Alternaria alternata) 。结果表明, 0.5 mmol·L ^{- 1} ASM能有效抑制鸭梨果实损伤接种病害和自然侵染病害的发生。ASM处理果实具有较高的苯丙氨酸解氨酶( PAL) 和几丁质酶(CHT) 活性, 积累了较高水平的H₂O₂ 和酚类物质。离体试验表明, ASM不对病原菌生长产生抑制作用。ASM处理对果实病害的抑制作用可能与其增强了果实抗性系统有关。     

Campylobacter spp., Salmonella spp., Verocytotoxic Escherichia coli,and Antibiotic Resistance in Indicator Organisms in Wild Cervids

Faecal samples were collected, as part of the National Health Surveillance Program for Cervids (HOP) in Norway, from wild red deer, roe deer, moose and reindeer during ordinary hunting seasons from 2001 to 2003. Samples from a total of 618 animals were examined for verocytotoxic E. coli (VTEC); 611 animals for Salmonella and 324 animals for Campylobacter. A total of 50 samples were cultivated from each cervid species in order to isolate the indicator bacterial species E. coli and Enterococcus faecalis/E. faecium for antibiotic resistance pattern studies. Salmonella and the potentially human pathogenic verocytotoxic E. coli were not isolated, while Campylobacter jejuni jejuni was found in one roe deer sample only. Antibiotic resistance was found in 13 (7.3%) of the 179 E. coli isolates tested, eight of these being resistant against one type of antibiotic only. The proportion of resistant E. coli isolates was higher in wild reindeer (24%) than in the other cervids (2.2%). E. faecalis or E. faecium were isolated from 19 of the samples, none of these being reindeer. All the strains isolated were resistant against one (84%) or more (16%) antibiotics. A total of 14 E. faecalis-strains were resistant to virginiamycin only. The results indicate that the cervid species studied do not constitute an important infectious reservoir for either the human pathogens or the antibiotic resistant microorganisms included in the study.     

以白菜为例的转基因植物环境安全性评价研究

以转除草剂抗性基因的白菜为例,通过对具体事实的展现和分析,介绍了关于转基因植物环境安全性评价的方法,以及目前国际上关于转基因作物的一些比较一致的结论。针对我国的农业生态环境,结合试验结果,讨论了获得的转基因白菜的环境安全性。     

柑橘抗CTV转基因与分子标记研究进展

综述了柑橘抗衰退病基因工程中两方面的研究进展。介绍多种来源于柑橘衰退病毒(Citrustriztezavirus,CTV)核酸序列的转基因柑橘和抗性种质资源中抗性基因的分子标记,以及所涉及的方法和遇到的问题。目前研究表明,虽然已成功实现对病毒衣壳蛋白(CP)等基因的转化和Ctv等抗性基因的标记,但尚未获得对CTV有高度抗性的转基因柑橘,而抗性基因亦不能实现定点克隆和转化。因此上述两方面研究还有待深入。     

不同砧木嫁接网纹甜瓜试验

为预防网纹甜瓜枯萎病等土传病害,以新土佐、灰甘瓜、全能铁甲南瓜,甬砧3号甜瓜为砧木,以当地主栽的拿玻里网纹甜瓜为接穗,进行嫁接试验。结果表明:拿玻里网纹甜瓜嫁接后,植株的抗病性有了显著提高,对枯萎病的防效高达100%,对白粉病的防效也有所提高,综合抗性增强。其中甬砧3号在4个参试砧木品种中,与接穗拿玻里网纹甜瓜的亲和性最好,抗病性提高明显,品质也较自根苗好。     

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。     

园林布局中必然性和偶然性要素的辩识与运用

阐释了园林构成要素中的必然性要素和偶然性要素及其相互关系与运用意义；以昆明世博园中“齐鲁园”为例，辩识了“齐鲁园”中必然性与偶然性园林要素，分析了诸要素在特定园林环境布局中的运用。