Quantitative detection of <Emphasis Type="Italic">Monosporascus cannonballus</Emphasis> in infected melon roots using real-time PCR

A method was developed for the specific detection, identification and quantification of Monosporascus cannonballus in infected melon roots based on real-time PCR (SYBR® Green chemistry) targeting the ITS1 region of the rDNA conserved between different strains of the pathogen. The specificity of the reaction was assessed using a number of fungi taxonomically and ecologically related to M. cannonballus. The method was highly sensitive and M. cannonballus was first detected in the roots of a susceptible Piel de Sapo cultivar 2 days after inoculation, before symptom appearance. Although conventional PCR methods could also provide such a specific and sensitive detection, real-time PCR was also able to produce reliable quantitative data over a range of 4 orders of magnitude (from 5 ng to 0.3 pg). The method allowed the quantitative monitoring of fungal growth from the very first stages of infection, and was successfully employed in the early screening of resistance. The assessment of disease progress and severity obtained with real-time PCR was more accurate than that obtained with the visual scoring of root lesions or root biomass losses. Therefore, there exists a great potential for its implementation in those steps of breeding programmes where high accuracy is required.     

Bacterial fruit blotch of melon: screens for disease tolerance and role of seed transmission in pathogenicity

Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax avenae subsp. citrulli, is a serious threat to the watermelon and melon industries. To date, there are no commercial cultivars of cucurbit crops resistant to the disease. Here we assessed the level of tolerance to bacterial fruit blotch of various commercial cultivars as well as breeding and wild lines of melon, using seed-transmission assays and seedling-inoculation experiments. Selected cultivars were also tested in a greenhouse experiment with mature plants. All tested cultivars/lines were found to be susceptible to the pathogen, and most of them showed different responses (relative tolerance vs. susceptibility) in the different assays; however, some consistent trends were found: cv. ADIR339 was relatively tolerant in all tested assays, and cv. 6407 and wild lines BLB-B and EAD-B were relatively tolerant in seed-transmission assays. We also provide evidence supporting a strong correlation between the level of susceptibility of a cultivar/line and the ability of the pathogen to adhere to or penetrate the seed. To the best of our knowledge, this is the first attempt to assess melon cultivars/lines for bacterial fruit blotch response.     

Ear Rot Susceptibility and Mycotoxin Contamination of Maize Hybrids Inoculated with Fusarium Species Under Field Conditions

The development of new maize hybrids with resistance to Fusarium infection is an effective means of minimizing the risk of mycotoxin contamination. Several maize hybrids have been investigated for Fusarium ear rot and accumulation of fumonisin B₁ (FB₁), fumonisin B₂ (FB₂), beauvericin (BEA) and fusaproliferin (FP) after artificial inoculation in the field with toxigenic strains of Fusarium verticillioides and Fusarium proliferatum. The year of inoculation had a significant influence on the disease severity and mycotoxin accumulation in maize kernels. Of all the hybrids tested, only Mona exhibited resistance to ear rot caused by F. verticillioides and produced low levels of fumonisins during three years of experiments. In Fusarium-damaged kernels (FDK), fumonisin B₁, fumonisin B₂, beauvericin and fusaproliferin were detected at concentrations much higher (up to 10–20 times) than in healthy-looking kernels (HLK). Animal and human exposure to these mycotoxins can be drastically reduced by removing mouldy and visibly damaged kernels from the commodity.     

The Use of a GUS Transformant of Trichoderma Harzianum, Strain T3a, to Study Metabolic Activity in the Spermosphere and Rhizosphere Related to Biocontrol of Pythium Damping-off And Root Rot

The activity of Trichoderma harzianum in the spermosphere and rhizosphere of different plant species was studied by use of a beta-glucuronidase (GUS) transformant (strain T3a). Hereby, direct observation of micro-habitats supporting metabolic activity of T. harzianum is reported. Germination of conidia and mycelial growth were not supported by exudates from healthy roots of various ages. Instead, growth and activity of T. harzianum depended on access to dead organic substrates such as seed coats, decaying roots, and wounds, including those caused by infecting pathogens. A correlation between the GUS activity of T. harzianum and the biomass of Pythium ultimum in infected roots was established. On the basis of our observations, we suggest that the biocontrol ability of T. harzianum involves competition with the pathogen for substrates including the seed coat, and wounded or infected root tissue.     

Monitoring physiological races of<Emphasis Type="Italic">Podosphaera xanthii</Emphasis> (syn.<Emphasis Type="Italic">Sphaerotheca fuliginea</Emphasis>), the causal agent of powdery mildew in cucurbits: Factors affecting race identification and the importance for research and commerce

Identification of the physiological races ofPodosphaera xanthii (syn.Sphaerotheca fuliginea), the causal agent of powdery mildew in cucurbits, is based upon the differing responses of various melon cultigens to the pathogen. Eight races of the pathogen have been identified to date in the USA, Africa, Europe and around the Mediterranean Sea, and four new races were reported from greenhouse melons in the major growing area of Japan. Plant responses to powdery mildew may be affected by environmental factors such as light intensity, temperature and humidity, as well as by age and nutritional status of the plants. The same factors affect the accuracy and reliability of race identification. In an attempt to overcome those obstacles, the genetic diversity ofP. xanthii was studied using molecular markers. Unfortunately, no correlation was found between DNA polymorphism and the race of the pathogen as identified by biological tests. The usefulness of race identification as a guide for the grower in selecting appropriate cultivars is limited because changes or shifts in the pathogen population are common. Such changes may be found among growing seasons, geographic regions and hosts, and also within a single greenhouse during a single season. On the other hand, race identification is important for basic research and is especially important for the commercial seed industry, which requires accuracy in declaring the type and level of resistance to powdery mildew in its products. http://www.phytoparasitica.org posting March 2, 2004. Contribution from the Agricultural Research Organization. No. 501/04.     

西瓜品种资源抗细菌性果斑病苗期鉴定

为筛选高抗瓜类细菌性果斑病的西瓜资源,以24份西瓜品种资源为试材,采用苗期喷雾接种法分别接种分离自甜瓜上的瓜类嗜酸菌Acidovorax citrulli菌株pslb96和ZZ-1,鉴定各品种资源对瓜类细菌性果斑病的苗期抗性。结果表明,24份西瓜品种资源中未发现有对2株菌株表现免疫的材料,有7份资源对菌株pslb96表现高抗,12份资源对菌株ZZ-1表现高抗;9份资源对菌株pslb96表现中感或感病,7份资源对菌株ZZ-1表现中感和感病;对2株菌株均表现高抗的品种资源有野生型种质资源A9及商品种华欣、申蜜968、申选958和申抗988,占总品种资源的20.83%。部分品种资源A4、A13和申蜜7号对菌株pslb96和ZZ-1的抗性表现出明显的差异,表明相同寄主来源的2株不同菌株致病力存在差异。     

豌豆蚜防控的关键RNAi靶标基因筛选与分析

为筛选防控豌豆蚜Acyrthosiphon pisum的关键RNA干扰(RNA interference,RNAi)靶标基因,通过构建精氨酸激酶(arginine kinase,AK)、超氧化物歧化酶(superoxide dismutase,SOD)和几丁质合成酶(chitin synthase,CHS)这3个与豌豆蚜生长发育相关基因的dsRNA,于室内检测ds AK、ds SOD和ds CHS处理后对其致死效果,并比较4种dsRNA递送方式对其致死效果的影响,筛选获得防治效果明显的dsRNA及其递送方式。结果显示,处理48～168 h后,不同基因dsRNA处理组豌豆蚜的死亡率比阴性对照组高2.19倍～4.39倍,其中ds AK处理组豌豆蚜的死亡率为27.7%～91.1%,显著高于ds SOD、ds CHS处理组及阴性对照组。使用ds AK通过人工饲喂法和叶片注射饲喂法处理豌豆蚜,48 h即可出现显著致死效果。ds AK通过人工饲喂方法处理72 h后,豌豆蚜AK基因表达量比空白对照组降低43.2%,同时可抑制豌豆蚜的生长发育,使其体长比阴性对照组减少了5.9%。表明ds AK对豌豆蚜...     

基于MaxEnt模型的突颊侧琵甲新疆潜在地理分布预测和主要影响因子分析

为有效预警和防控突颊侧琵甲Prosodes(Lioprosodes)dilaticollis Motschulsky发生,基于该物种分布数据、1950―2000年环境下生物气候数据和典型受害区土壤因子数据、气象数据等,采用MaxEnt模型对其在新疆的潜在地理分布及其主要影响因子进行预测和分析验证。结果表明:受试者工作特征曲线下面积(area under the ROC curve,AUC)都大于0.990,预测结果可靠。突颊侧琵甲的潜在分布区位于新疆天山一带,高、中度适生区分布在伊犁河谷草原和昌吉州玛纳斯县、呼图壁县草原及塔城地区沙湾县等地;刀切法得出降水量是影响其潜在地理分布的主要因素,特别是5、6、11、12月降水量的影响最大。该虫典型受害区的降水数据验证了分布概率响应曲线的准确性;土壤湿度在15%~30%范围时,30 cm处土壤湿度与虫口密度呈极显著弱正相关。表明降水作为影响草原土壤湿度、土壤含氧量的主要和直接因素,对不同生长期突颊侧琵甲生存起到重要作用。