Single-strand conformational polymorphism analysis of the ribosomal internal transcribed spacer 1 for rapid species identification within the genus Pythium

Single-strand conformational polymorphism (SSCP) of the ribosomal internal transcribed spacer 1 (ITS-1) was characterized for 58 isolates of Pythium, representing 41 species from the five groups of Plaats-Niterink. Thirty-one species each produced a distinct SSCP pattern. Three species produced more than one unique pattern, corresponding to morphological subgrouping. The remaining seven species produced three distinct patterns with two or three morphologically similar species sharing a pattern. A successful blind test with four samples and the identification of eight previously unknown isolates from irrigation water demonstrated the reliability of this technique for species identification. Each SSCP pattern was defined and described by the positions of the top and bottom bands and the number of bands in between, which allows laboratories to use this technique without need to access the type isolates of Pythium species.     

Genetic engineering of Streptococcus gordonii for the simultaneous display of two heterologous proteins at the bacterial surface

A new species Pythium canariense (CI-07), isolated from soil samples taken in the San Nicolas region of the Canary islands (Gran Canaria, Spain), is being described here. This species is characterised by its spherical to pyriform, intercalary to catenulate sporangia, smooth-walled terminal oogonia supplied with monoclinous and diclinous antheridia which at times are branched and wrap around the female gametangia. The fungus has an antagonistic effect on Botrytis cinerea, the grey mould fungus. Morphological features are being given here together with the sequence of the complete internal transcribed spacer region of the nuclear ribosomal DNA of the fungus, its comparison with related species, and some aspects of its antagonistic behaviour with B. cinerea.     

Influence of Sugar Beet Seed Treatment with Pseudomonas fluorescens and Low Fungicide Doses on Infection with Pythium and Plant Yield and Quality

The effect of sugar beet seed inoculation with the bacterium Pseudomonas fluorescens and treatment with the fungicides Thiram 42‐S and Dithane S‐60 with and without seed inoculation aiming to control the root decay agents Pythium ultimum and Pythium debarianum was studied during a 2‐year trial on two soil types (Mollic Gleysols and Eutric Cambisols). The influence of the treatments on parameters of sugar beet yield and quality such as root yield, sugar content, sugar in molasses, sugar yield as well as percentage of the infected and decayed plants as a consequence of parasitic oomycete infestation will be described.     

德里腐霉游动孢子诱发试验的研究

本试验研究了不同培养基、菌龄大小、温度高低、诱饵种类和预处理时间长短对德里腐霉(Pythium.deliense)游动孢子产生的影响。结果显示,CMA、WA、PDA、TSA和LBA培养基均适宜于培养P.deliense并使其产生较多量的游动孢子,其中尤以CMA和WA效果为好;幼龄培养物的产孢能力比老龄的强,并以菌龄2天为最好;此菌在8℃以下和35℃以上不产孢,15—25℃为其产孢适宜温度;以马唐叶和胡萝卜为诱饵的处理,其孢子始见期和产孢量均明显比对照组早和高,马唐叶的刺激效应较胡萝卜的为强;随着预处理时间的增加,产孢量随之相应增多,但龄期重迭现象则越严重。     

The role of strigolactones and ethylene in disease caused by Pythium irregulare

Plant hormones play key roles in defence against pathogen attack. Recent work has begun to extend this role to encompass not just the traditional disease/stress hormones, such as ethylene, but also growth‐promoting hormones. Strigolactones (SLs) are the most recently defined group of plant hormones with important roles in plant–microbe interactions, as well as aspects of plant growth and development, although the knowledge of their role in plant–pathogen interactions is extremely limited. The oomycete Pythium irregulare is a poorly controlled pathogen of many crops. Previous work has indicated an important role for ethylene in defence against this oomycete. We examined the role of ethylene and SLs in response to this pathogen in pea (Pisum sativum L.) at the molecular and whole‐plant levels using a set of well‐characterized hormone mutants, including an ethylene‐insensitive ein2 mutant and SL‐deficient and insensitive mutants. We identified a key role for ethylene signalling in specific cell types that reduces pathogen invasion, extending the work carried out in other species. However, we found no evidence that SL biosynthesis or response influences the interaction of pea with P. irregulare or that synthetic SL influences the growth or hyphal branching of the oomycete in vitro. Future work should seek to extend our understanding of the role of SLs in other plant interactions, including with other fungal, bacterial and viral pathogens, nematodes and insect pests.     

Pythium aphanidermatum Causing Pythium Rot on Lampranthus spectabilis in Korea

Pythium aphanidermatum was found to be associated with rotting of leaves and stems in Lampranthus spectabilis in the summers of 2013 and 2014. Infected plants were initially characterized by water‐soaked and discoloured tissue, which are soon covered with cottony aerial hyphae. Subsequently, infected tissues wilted, leaves and stems appeared desiccated, and infected plants died. Pathogenicity of a representative isolate was proved, fulfilling Koch's postulates. Sequence of the internal transcribed spacer region of ribosomal DNA from the isolate shared 100% identity with that of P. aphanidermatum. To our knowledge, this is the first report of P. aphanidermatum causing leaf and stem rot on L. spectabilis in Korea.     

Production and Application of Syringomycin E as an Organic Fungicide Seed Protectant against Pythium Damping‐off

Syringomycin E (SRE) is a cyclic lipodepsinonapeptide with potent antifungal activity and is produced by certain strains of Pseudomonas syringae pv. syringae. In this study, its potential as an organic‐compatible agrofungicide and vegetable seed treatment against the soilborne pathogen Pythium ultimum var. ultimum was examined. A variant of P. syringae pv. syringae strain B301D with enhanced SRE‐producing capabilities was isolated and grown in a bioreactor with SRE yields averaging 50 mg/l in 40 h. SRE was extracted and purified through a large‐scale chromatography system using organic‐compatible processes and reagents. The minimum concentrations of the purified product required to inhibit 50 and 90% of P. ultimum oospore germination were determined as 31.3 and 250 μg/ml, respectively. Drench treatment of cucumber seeds in P. ultimum‐infested potting medium (500 oospores/g) with 50 μg/ml SRE or water with no SRE resulted in 90.2 ± 4.5% and 65.7 ± 4.6% germination rates, respectively. Seed coating with 0.03% (w/w) SRE allowed 65.7 ± 4.6% seedlings to germinate on naturally infested soil while 100.0 ± 0.0% of non‐coated seeds were unable to germinate due to Pythium infection. Organic‐compatible and scalably produced SRE is potentially a novel organic fungicide seed protectant.