Specific detection of Phytophthora cactorum in diseased strawberry plants using nested polymerase chain reaction

Validated protocols for DNA purification and PCR amplification are reported for detection of Phytophthora cactorum in diseased strawberry plants. To remove PCR inhibitors, necrotic strawberry tissues were soaked in 5% alconox solution for >12 h before DNA extraction, and the extracted genomic DNA was embedded in an agarose gel chamber and subjected to electrophoresis. The purified DNA was amplified reliably by PCR. Nested PCR was used to detect a portion of the rRNA gene of P. cactorum in samples. In the first round of PCR, primers ITS1 and ITS4 amplified fragments of varying sizes from total genomic DNA from diseased strawberry plants. In the second round of PCR, a 1:25 dilution of the first-round PCR products was used as template with two P. cactorum- specific primer pairs (BPhycacL87FRG and BPhycacR87RRG, which amplified a 340-bp fragment and a 480-bp fragment from the rRNA gene; and BPhycacL89FRG and BPhycacR176RRG, which amplified a 431-bp fragment). Validation tests using culture-based isolations as a standard for comparison indicated that the DNA purification and PCR primers and amplification protocols were reliable and specifically amplified a portion of the rRNA gene of P. cactorum from necrotic root, crown and petiole tissues of strawberry naturally infected by the pathogen.     

油茶叶枯病菌的生物学特性

近几年.油茶叶枯病已从油茶的次要病害上升为主要病害.为给此病害的防治提供依据,对该病菌的生物学特性进行了分析,分离纯化了油茶叶枯病原菌,并对影响其生长的温度、酸碱度、光照和培养基进行了试验研究.结果表明:病原菌在5～31℃之间均能生长,其中27℃的温度为菌丝生长的最适宜温度;病原菌在pH值为4～9之间均可以生长,当pH值为6时菌丝生长最快;光照对菌丝生长的影响不大;以蔗糖为碳源的培养基最适合该病菌的生长,以葡萄糖为碳源的培养基其生长最慢.     

Market Disease Pathogens Detection of Imported Fruits in Shanghai

A tremendous amount of imported fresh fruits has been delivered to Shanghai markets, increasing the risk of invasion by harmful plant pathogens. Therefore, it is important to establish an effective detection and supervision system to survey the outbreak of the market diseases of the imported fruits during marketing. The samples were regularly surveyed in different markets to examine varieties, prices, localities, selling conditions, and diseases of the imported fruits from 2004 to 2008. The survey showed that 58 species of 30 different fruits were imported to Shanghai from 16 countries with more expensive price. The larger products were bananas, grapes, apples, and oranges. During the investigation, we found that the imported fruits frequently brought about the relatively serious market diseases. On the basis of morphology and the nuclear internal transcribed spacer （ITS） of ribosomal DNA （rDNA） analysis, 151 isolates of 15 fungi genera, which shown to be pathogenic after the inoculation assay, were finally identified. Among the identified fungi, Alternaria was the most frequent one with the highest detection rate （47.68%）, followed by Penicillium （14.57%） and Fusarium （11.92%）, respectively. Additionally, Pestalotiopsis microspora （detected in grapes Red-Globe coming from the USA） and Botrytis sp. （detected in black-plums coming from the USA） were first reported in China market. The present study summarized the selling situation of the imported fruits in Shanghai markets and constructed a library of the pathogens detected in the imported fruits during the selling period. The results obtained are useful to offer technical parameters for Chinese quarantine in order to prevent an invasion of the foreign harmful micro-organisms.     

油茶白朽病菌ITS基因的克隆及序列分析

分离油茶白朽病的病原菌,并对该菌的ITS基因进行克隆和测序 将所得序列在Genbank中经B lastn搜索,构建分子系统发育树。结果表明：病原菌ITS序列与Genbank数据库中已有的序列同源性较低,最高仅为94%,系统发育树也显示该菌不与其他菌株聚类而是单独聚为1支。由于Genbank数据库有限,难以根据同源性来确定其属种关系。     

牛心朴子生物碱对奶牛乳房炎病原菌的体外抑菌试验

对4例临床型奶牛乳房炎的乳汁进行病原分离及鉴定,分离出了金黄色葡萄球菌、无乳链球菌和大肠杆菌。选择牛心朴子生物碱,采用试管2倍稀释法进行体外抑菌试验,测其最小抑菌浓度（MIC）和最小杀菌浓度（MBC）。结果显示：牛心朴子生物碱对3种菌都有不同程度的抑制作用,对金黄色葡萄球菌、无乳链球菌、大肠杆菌分离株的MIC及MBC分别是12.5mg/mL、1.561 25mg/mL、6.25mg/mL和6.25mg/mL、1.561 25mg/mL、3.125mg/mL。     

含双病原物诱导启动子植物安全表达载体的构建

 本研究用来自烟草的具有高度病原物特异性的两个诱导启动子EAS4和hsr203J,串联驱动GUS基因的表达,同时考虑到转基因植物的安全性,引入双边界序列,构建了含双病原物诱导启动子的植物安全表达载体。将表达载体转化烟草获得转基因植株。分析显示,在正常生长情况下转基因烟草检测不到GUS活性,或活性极低;而受疫霉激发子parasiticein、Phytophthora nicotianea[0]的孢子悬浮液和Ralstonia solanacarum的菌悬液诱导后,转基因烟草叶片中可检测到明显的GUS活性。结果表明,构建的植物表达载体所含双病原物诱导启动子均具有良好的诱导活性,可用于植物遗传转化。     

Genotypic differences in storage losses of sugar beet – causes and indirect criteria for selection

To improve the storability of sugar beets, this study aimed at determining reasons for genotypic variability in sugar losses and invert sugar accumulation during storage, and at identifying indirect criteria to select for varieties with low storage losses prior to storage. In 2011 and 2012, 18 genotypes, and in 2012 and 2013, six genotypes cultivated at two locations were stored for 8 and 12 weeks at 8°C under controlled conditions. The same 18 genotypes were grown under stress conditions in Spain in 2012/2013. Sugar losses were closely correlated with the invert sugar accumulation after storage. Genotypic differences in storage losses were primarily caused by differences in the level of infestation with microorganisms. The invert sugar accumulation was lower for genotypes with high marc concentration before storage, pointing to a non‐specific resistance. Additionally, the sugar concentration in dry matter before storage, and the invert sugar concentration after cultivation under stress conditions correlated with the invert sugar concentration after storage. These parameters are therefore suggested as criteria to select for improved storability of sugar beet genotypes.     

管芯片技术通量检测杨树溃疡病病菌的应用

正基因芯片技术因能对生物样本进行快速、高通量的定性和定量分析,而在植物病害诊断中也得到普遍的应用[1]。树木溃疡病是指引起木本植物树皮上出现溃疡腐烂等症状的病害[2]。引起杨树溃疡病的真菌种类较多,加之其无性型和有性型多样,所以,关于杨树溃疡病病原菌的报道多有不同。另外,受地理位置、寄主植物关系及分类体系的限制,迫切需要发展不依赖于常规分离培养的病原鉴定新技术。     

Suppression of the fungal pathogen Magnaporthe grisea by Stenotrophomonas maltophilia,a seed-borne rice (Oryza sativa L.) endophytic bacterium

The present work was carried out to study the potential of bacteria isolated from the seeds of rice plant for the biocontrol of five rice pathogenic fungi. Eleven endophytic bacteria isolated from rice seeds were evaluated for their antagonistic potential. Of five pathogens studied, only the growth of Magnaporthe grisea was inhibited by one bacterial isolate in an in vitro dual culture assay. Based on 16S rDNA sequence analysis, and biochemical and morphological characteristics, this strain was closely related to Stenotrophomonas maltophilia. We named this new isolate to be S. maltophilia SEN₁ (seed endophyte). This isolate was further tested for the production of volatile and diffusible antibiotics against M. grisea, for plant growth-promoting (PGP) traits and colonization of some rice cultivars. In addition, S. maltophilia SEN₁ was tested for its ability to promote plant growth and reduce the incidence of rice blast disease under greenhouse conditions. When applied to the soil, this isolate increased seedling growth and suppressed blast disease in plants of three studied cultivars. This study also showed this isolate could colonize the root interior of other rice cultivars. This study indicates that the S. maltophilia isolate studied has an excellent potential to be used as biocontrol agents of M. grisea or biofertilizer under in vitro and in vivo conditions.     

食源性病原及其毒素检测技术研究进展

食源性病原包括细菌、真菌、病毒、朊病毒和原虫,它们在生产加工及储运环节污染食物,同时不断分泌包含毒素在内的各种成分到细胞外直至自身死亡和裂解,其中一些病原菌及其毒素能耐受复杂的食物加工,直至进入人体干扰细胞生理过程并引发疾病。详细介绍了如何利用各种"组"学(包括蛋白质组、肽组、代谢组和食品组学)技术检测和寻找食源性病原微生物的生物标志物,深入了解其产毒机理、毒力和致病机理,及对胁迫的适应能力。这对于追踪食源性病原微生物内、外毒素的来源,保证食品安全并控制食源性疾病爆发具有重要意义。