Attempts to transferRubus andFragaria viruses into herbaceous hosts

Attempts were made to transfer several types of viruses affectingRubus andFragaria species to herbaceous hosts of low tannin content. Despite use of phosphate buffers, nicotine sulphate solutions, and various lyophilization techniques, all inoculations were unsuccessful.Samenvatting Er werden pogingen ondernomen, verschillende typen van mozaïekvirussen van framboos en van aardbei met sap op kruidachtige, weinig of geen looistoffen bevattende planten over te brengen. Er werd gebruik gemaakt van fosfaatbuffers, van oplossingen van nicotinesulfaat en van droogvries-methoden vooraf de looistoffen uit het materiaal te verwijderen. De resultaten van de inoculatieproeven waren echter alle negatief.

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Research carried on at the Instiuut voor Plantenziektenkundig Onderzoek, Wageningen, Nederland. Supported in part by a fellowship from the John Simon Guggenheim Memorial Foundation, New York.

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Technical Paper No. 944 of the Oregon Agricultural Experiment Station. Contribution of the Department of Botany and Plant Pathology of this Station, and of the Instituut voor Plantenziektenkundig Onderzoek (I.P.G.), Wageningen.     

A simple and effective inoculation method for Phytophthora and fungal species on woody plants

In inoculations carried out on buds, young shoots, and stems of woody plants in Norway, map pins contaminated with the pathogens to be tested proved to be convenient tools. The pins were inserted into the plant tissue and were left standing in the inoculation site throughout the experimental period. In the present investigation, the method was used successfully for one Phytophthora and four fungal species on different host plants. Inoculated plant parts developed clear symptoms, while no symptoms occurred where non‐contaminated pins were used. The advantages of this method are multiple: minimal physical damage to the tissue, inoculation points easily traced due to the relatively large top grip on the needles, and time saving as there is no need to cover with polyethylene or Parafilm after inoculation.     

Node inoculation: A quick and easy technique to screen pigeonpea for resistance to Phytophthora blight

The petiole on pigeonpea was removed for easy, precise inoculation of node with Phytophthora drechsleri f. sp. cajani. After node inoculation, 96.0% plants were infected compared with 89.0% after stem-cut inoculation. Among various nodes inoculated on 30-day-old plants, the 5th node had the greatest relative susceptibility (90.0%), followed by the 3rd node (78.0%). This technique was validated on different cultivars (ICP 7119, Bahar, MA 6 and MAL 13), and 586 lines were successfully screened in the field, confirming the rapidity and effectiveness of the technique for resistance screening.     

Detection of a broad range of potato viruses in a single assay by mechanical inoculation of herbaceous test plants

To optimize mechanical inoculation of test plants as part of the regulated post-entry quarantine testing of stolon- and tuber-forming Solanum spp. (including potato), nine test plant species have been screened for their ability to detect 22 potato-infecting viruses. These included all the common mechanically transmissible potato viruses and most of the viruses found in potato incidentally. The symptoms observed after mechanical inoculation are shown for each combination of test plant species and virus. Under given conditions, Nicotiana occidentalis -P1 and Nicotiana hesperis -67A were found to detect reliably 20 and 18 out of 22 viruses respectively. These and former results on the seed-transmissible viruses and Andean potato mottle virus demonstrate that these Nicotiana species are very suitable for post-entry quarantine testing. Addition of either Chenopodium amaranticolor or Chenopodium quinoa to these two species may slightly extend the range of viruses. Therefore, using C. amaranticolor / C. quinoa , N. hesperis -67A and N. occidentalis -P1 for the biological screening of imported Solanum spp. will improve both the efficiency and the quality of post-entry quarantine testing.     

A method to test wheat leaves for their reactions to inoculation with Septoria species

The construction and application is described of a polystyrol humidity box in which wheat leaves, while continuing to function as parts of living plants, can be tested for their reactions toSeptoria spp. in an atmosphere nearly saturated with water, as is required for successful infection. The method is simple, accurate ans inexpensive.Samenvatting Voor dit doel is een z.g. vochtdoos geconstrueerd (Fig. 1A). Het te toetsen blad wordt daar doorheen geleid en in de doos geïnoculeerd met een druppel conidiën-suspensie van de schimmel. Onder het blad staat wat water in de doos (Fig. 1B). Na afsluiting ontstaat in de doos de hoge luchtvochtigheid van bijna 100% die nodig is voor de infectie. Dit wordt op deze wijze eenvoudig en goedkoop gerealiseerd. De bladeren die zo worden getoetst blijven onbeschadigd functioneren aan de plant. Na enkele dagen kan de doos worden geopend en kan de symptoom-ontwikkeling worden afgewacht en gevolgd (Fig. 3A, 3B en 4). De methode leent zich voor nauwkeurig werk en vereist zeer weinig infectiemateriaal. De Technische en Fysische Dienst voor de Landbouw (TFDL), Wageningen, ontwierp en construeerde een statief voor het gebruik van de vochtdozen in serie (Fig. 2).     

A new method for inoculation of lettuce with Bremia lactucae

Samenvatting Het inwrijven van slabladeren en- bladschijfjes met carborundumpoeder vóór de bespuiting met een sporensuspensie vanB. lactucae heeft een opmerkelijk snellere infectie van de sla en een sterk vergrote sporulatie van de schimmel tot gevolg. De middelfijne poeders, graad 500–800 mesh, werken effectiever dan de grovere en de finjnere sorteringen. Voor het uitvoeren van proeven metB. lactucae en voor het toetsen van slarassen en- lijnen bij de resistentieveredeling tegen valse meeldauw is dit een belangrijke verbetering.He is much indebted to the Director, Dr J. G. ten Houten, Institute of Phytopathological Research (IPO), Wageningen, for putting laboratory facilities at his disposal and to the Netherlands Bureau of Technical Assistance, The Hague, for financial support.     

一种快速测定稻瘟病菌群体毒性的简便方法

选育和布局抗病品种是控制稻瘟病最经济有效的方法,但病菌群体的毒性类型频率逐步上升导致品种抗性丧失已成为抗性品种选育和推广的瓶颈问题。为寻找一种能够为基层单位所掌握且快速、准确的稻瘟病菌群体毒性监测方法,本文将不同来源的穗颈瘟标样分别接种于一套抗稻瘟病单基因系、杂交稻主栽品种及亲本和抗源上,30 d后调查最高病害级别和株发病率。结果表明,‘丽江新团黑谷’、标样来源品种及其所含抗瘟基因的单基因系均能稳定发病。4个来自四川盆地的病菌群体对四川主栽品种的毒性明显强于来自云南楚雄的群体,且这4个病菌群体均具备对Pi-2及‘宜香优2115’的毒性。来自万州甘宁镇‘F优498’的病菌群体对主栽品种的毒性谱比崇州‘宜香优2115’来源的病菌更广。该方法操作简单,能快速准确地监测稻瘟病菌群体毒性状态,有助于基层单位更有效地开展稻瘟病防控布局工作。     

A quick method of recording plant growth stages

The recording of the stage of growth of crops and weeds by directly photocopying the plants is described.     

糖类寄生螨快速分离检验方法

本文对糖类寄生螨的快速分离检验方法进行了改进。将糖类溶解后，经过500目/英寸标准检验筛进行过筛分离。此方法对各种可溶性糖类中的寄生螨类能取得100％的快速分离检验结果。     

A refined inoculation method to evaluate false smut resistance in rice

False smut, caused by Ustilaginoidea virens, is a serious disease of rice worldwide. To evaluate false smut resistance in rice, we developed a method combining the cultivation of the main culm of rice plants in the greenhouse and rapid preparation of a conidial suspension to inject into the leaf sheath. The method was used to evaluate false smut resistance in 18 varieties/lines of rice. For comparison, field trials were also carried out in 2007 and 2008. The results indicated that the greenhouse method was more reproducible than field trials: commercial varieties tested were resistant; almost all the forage varieties were highly susceptible; and blast-resistant varieties/lines were mostly resistant to false smut. Thus, this inoculation method will be useful for determining the level of false smut resistance in rice and for breeding resistant varieties.     

快速检测单头柑橘木虱体内黄龙病病原

本研究通过采用简易的柑橘木虱制样方法,利用PCR检测技术在单头柑橘木虱体内检测到黄龙病病原,并利用XbaⅠ限制性内切酶将从柑橘木虱体内扩增的黄龙病病原16SrDNA酶切成520bp和640bp两个片段,证实了木虱体内黄龙病病原为亚洲种。本研究成功地建立了一套快速检测柑橘木虱体内黄龙病病原的方法,可为检疫工作者和相关研究人员提供重要参考。     

甘蓝抗软腐病离体鉴定方法探究

通过分析不同因素对甘蓝软腐病离体接种后发病效率的影响,建立了一套甘蓝软腐病抗性的快速鉴定方法。结果发现,用浓度为6.4×10~8 cfu/mL的软腐病液体培养基悬浮液,采用针刺法接种苗龄为5～7叶期植株的离体叶片,接种后空气湿度为90%以上是最有利于软腐病发病的方法。利用该方法鉴定27份栽培甘蓝,发现高抗材料3份,抗病材料8份,耐病材料8份,感病材料8份。     

Responses of white yam (Dioscorea rotundata) cultivars to inoculation with three viruses

The responses of 24 white yam ( Dioscorea rotundata ) cultivars to mechanical and vector transmission with each of three viruses infecting yams were assessed through enzyme-linked immunosorbent assay (ELISA) and symptom development. The viruses were Dioscorea alata virus (DAV), genus Potyvirus ; Dioscorea alata bacilliform virus (DaBV), genus Badnavirus ; and Cucumber mosaic virus (CMV), genus Cucumovirus . Only TDr 95-128, a landrace cultivar from Nigeria, developed symptoms of infection with CMV and DaBV following mechanical and vector transmission, respectively. PAS-ELISA showed that nine genotypes remained uninfected by DAV and 11 were uninfected by CMV or DaBV. Genotypes TDr 747 and TDr 1640 both showed resistance to all three viruses.     

Molecular characterization of a phytoplasma causing Phyllody in Clover and other herbaceous hosts in Northern Italy

Red clover (Trifolium pratense) and Ladino clover (Trifolium repens) plants showing phytoplasma-associated symptoms (yellowing/reddening, virescence and phyllody) have been recovered in Friuli-Venezia Giulia, Italy. Using AluI RFLP analysis of PCR amplified 16S rDNA we showed that the disease can be caused independently by two phylogenetically distinct phytoplasmas. One of them showed the very typical 16S rDNA RFLP pattern of the agent of Clover Phyllody in Canada (CCPh). The 16S rDNA of the other phytoplasma (Italian Clover Phyllody phytoplasma, ICPhp) has been PCR amplified, cloned and sequenced. The sequence revealed high similarity (>98%) with phytoplasmas belonging to the X disease cluster, which includes organisms not reported to cause phyllody on their hosts. The analysis by AluI RFLP of the PCR amplified pathogen 16S rDNA from other herbaceous plants (Crepis biennis, Taraxacum officinale, Leucanthemum vulgare) collected nearby with phytoplasma-associated symptoms showed similar patterns. Southern blot hybridization of their EcoRI digested total DNA revealed identical RFLP patterns, suggesting that the causative agent may be the same organism.Abbreviations PCR Polymerase Chain Reaction - rDNA gene for the small subunit ribosomal RNA - RFLP Restriction Fragment Length Polymorphism     

Linear-motion tattoo machine and prefabricated needle sets for the delivery of plant viruses by vascular puncture inoculation

Vascular puncture inoculation (VPI) of plant viruses previously has been conducted either manually or by use of a commercial engraving tool and laboratory-fabricated needle arrays. In an effort to improve this technique, a linear-motion tattoo machine driving industry-standard needle arrays was tested as a means of delivering plant viruses into maize and small grain seed embryos. The new method was applied in the successful transmission of maize rayado fino virus (MRFV), the type member of the genus Marafivirus, from an archived sample to maize. Subsequent transfer of MRFV from the sap of an infected plant using the method produced an average infection rate in maize of 70% (range 39–93%). Maize, oat, and triticale were successfully infected with oat blue dwarf virus (OBDV) using the method; similar infection rates were observed between maize seeds inoculated with the tattoo machine and those inoculated with the engraving machine when using prefabricated needle arrays. No infection was obtained in repeated tests with barley yellow dwarf virus (BYDV-PAV) or cereal yellow dwarf virus (CYDV-RPV) using either sap or RNA from infectious cloned cDNA. Replacement of the original engraving-tool with a linear-motion tattoo machine in VPI provides greater flexibility and convenience in a quiet, readily-available instrument, while improving reproducibility through the use of prefabricated needle arrays.     

Genetic diversity, presence of the syrB gene, host preference and virulence of Pseudomonas syringae pv. syringae strains from woody and herbaceous host plants

A total of 101 Pseudomonas syringae pv. syringae strains, obtained from international culture collections or isolated from diseased tissues of herbaceous and woody plant species, were assessed by repetitive PCR using the BOX primer, and for the presence of the syrB gene. Representative strains were also tested for pathogenicity to lilac, pear, peach, corn and bean, as well as for virulence to lemon and zucchini fruits. The unweighted pair-group method using arithmethic averages analysis (UPGMA) of genomic fingerprints revealed 17 different patterns which grouped into three major clusters, A, B and C. Most of the strains (52·4%) were included in patterns 1–4 of group A. These patterns comprised strains obtained from either herbaceous or woody species, and showed four fragments of similar mobility. Genetic variability was ascertained for strains isolated from apple, pear, apricot, Citrus spp. and cereals. No clear relationship was observed between host plant and bacterial genomic fingerprint. Variability was also observed in pathogenicity and virulence tests. The inoculation of pear leaves discriminated strains isolated from pear as well as the very aggressive strains, whereas inoculation of lilac, peach and corn did not discriminate the host plant from which the strains were originally isolated. Lemon fruit inoculation proved very effective for P. syringae pv. syringae virulence assessment. The syrB gene was present in almost all strains.     

应用稻苗离体叶片法筛选水稻纹枯病新农药活性的研究

采用水稻离体叶片法测定了农药对水稻纹枯病的活性。结果表明,该法与“植株法”比较有很好的相关性,具有占用空间小、温湿度容易控制、快速、准确等优点,适合于大量化合物的筛选试验。     

甘蔗花叶病抗病性鉴定接种新技术研究

研究了甘蔗花叶病抗病性鉴定的一种新技术—甘蔗生长期切茎接种法。此法和传统苗期手指摩擦接种法相比,接种后发病率极显著地高于手指摩擦接种法,且发病均匀、结果稳定,可以有效地区分不同甘蔗品种(材料)之间对病害的抗感性。比较11个甘蔗品种材料生长期切茎接种法和田间自然感病法对SrMV-HH1的抗性鉴定,结果表明:两种方法发病率呈正相关,且相关程度极密切,相关系数达0.9997,鉴定所得的抗病性和抗病等级结果完全一致,说明切茎接种法鉴定结果能真实反映甘蔗品种材料的自然抗感性;此外该方法简便实用、可操作性强、接种工效高,因此可以对大批量材料进行鉴定和筛选。