Diversity and mycotoxin production by Fusarium temperatum and Fusarium subglutinans as causal agents of pre-harvest Fusarium maize ear rot in Poland.

Maize ear rot is a common disease found worldwide, caused by several toxigenic Fusarium species. Maize ears and kernels infected by Fusarium subglutinans contained significant amounts of beauvericin, fusaproliferin, moniliformin, and enniatins. In 2011, F. subglutinans sensu lato has been divided into two species: Fusarium temperatum sp. nov. and F. subglutinans sensu stricto, showing different phylogeny and beauvericin production within the populations of maize pathogens in Belgium. Isolates of the new species-F. temperatum-were also identified and characterized in Spain, Argentina, Poland, France, and China as one of the most important pathogens of maize. Moreover, F. temperatum was proved to be pathogenic to maize seedlings and stalks. We identified Fusarium isolates obtained from diseased maize ears collected between 2013 and 2016 in Poland (321 isolates). Based on morphological analyses, six Fusarium species were identified. Molecular identification performed on the set of selected isolates (42 isolates) revealed 34 isolates to be F. temperatum and only five to be F. subglutinans. Interestingly, the phylogenetic analysis showed that the population of F. temperatum infecting maize in Poland remained quite uniform for over 30 years with only a few exceptions. For the first time, a single isolate of Fusarium ramigenum was detected from the area of Poland. Significant amounts of BEA were found in Fusarium-damaged kernels. The same kernel samples contained also enniatins A1, A, B1, and B. The results clearly demonstrate the occurrence of F. temperatum as maize pathogen in Poland for over the last three decades.

Fusarium temperatum as a New Species Causing Ear Rot on Maize in Poland.

Diseases caused by Fusarium spp. are probably the most important factors affecting the yield and grain quality of maize (Zea mays L.) in Poland. While Fusarium stalk rot problems were reported to be stable in all regions, ear rot tends to increase. This tendency probably results from warmer climate and conservation tillage techniques combined with maize- and wheat-dominated rotation systems increasingly practiced in these regions. Maize kernel samples were harvested from trials conducted with hybrids moderately resistant and susceptible to ear rot in three locations in 2011 (12 samples) and in seven locations in 2012 (20 samples) from regions of Poland where maize for seed production is grown for many years. For each sample, 56 kernels (1,792 total) were subjected to the investigation of fungal colonization as following: surface-disinfected in alcohol (15 sec), rinsed with distilled water, dried, placed on the water agar in petri plates, incubated 7 to 12 days at 22°C, and sub-cultured using the single spore technique on SNA. Pure cultures were grown on SNA at 22°C for 10 days to produce macroconidia of uniform size and form, as well as on PDA (1). The presence of Fusarium spp. was determined based on the SCAR-PCR markers and the translation elongation factor (tef-1α) sequence analyses (4). All samples were contaminated by Fusarium spp. at level ranging from 17.3 to 40.5% of kernels (444 strains were isolated). F. verticillioides was the most frequent species. The frequencies of F. temperatum (a recently described species closely related to F. subglutinans [3]) ranged from 2.3 to 54.3%, F. subglutinans from 0 to 37.9%, F. verticillioides 41.0 to 90.0%, and F. proliferatum from 0 to 11.5% of the total Fusarium strain number. After extraction and purification, Fusarium mycotoxins were detected using HPLC method. The majority of maize kernel samples contained fumonisins B1 to B3. Zearalenone and deoxynivalenol were not detected or their levels were below limit detections. Aggressiveness of 10 representative isolates (3 each of F. temperatum, F. verticillioides, and F. subglutinans, and 1 F. proliferatum) was evaluated under field conditions after kernel inoculation. Susceptible and moderate resistant inbreed lines were used. The experimental design consisted of a factorial arrangement with two factors and three replicates (7 plants per replicate). To produce inoculum, isolates were grown on a liquid SNA medium. After 2 weeks, cultures were filtered through cheesecloth and conidial concentrations were adjusted to ~106 spores ml-1. Inoculation of each ear was conducted 10 to 12 days after silking time using 1.5 ml of spore suspension. Control plants were inoculated with distilled water. Visual evaluation of ear rot severity was scored during harvesting time as the percentage of the ear area covering by mycelium (2). Differences of aggressiveness between genotypes, Fusarium spp., and isolates were statistically significant; ear area covered by mycelium ranged from 3.57 to 26.95% for F. temperatum, from 10.10 to 41.39% for F. subglutinans, from 5.90 to 14.33% for F. verticillioides, and from 9.57 to 17.14% for F. proliferatum. To our knowledge, this is the first report of F. temperatum causing ear rot of maize in Poland. References: (1) J. F. Leslie et al. Page 388 in: The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, 2006. (2) L. M. Reid et al. Can. J. Plant Pathol. 19:185, 1997. (3) J. Scauflaire et al. Mycologia 103:586, 2011. (4) L. Stepien et al. J. Appl. Genet. 54:367, 2013.

Powdery mildew resistance genes in wheat: verification of STS markers.

Five accessions of Aegilops speltoides and 67 European wheat cultivars (winter and spring) originating from the Czech Republic, Germany, Poland, Russia, Slovakia, United Kingdom, and 4 non-European wheat cultivars from Brazil and the USA were examined with molecular Sequence Tagged Site (STS) markers for resistance genes to powdery mildew: Pm 1, Pm 2, Pm 3 and Pm 13. All markers gave clear, repeatable results, although three of them (Pm 1, Pm 2 and Pm 3) appeared as not specific for resistance genes. Comparison of STS analysis results with Pm genes, postulated as the reaction type after inoculation with differential isolates of Erysiphe graminis f.sp. tritici (Blumeria graminis), revealed a high number of disparities. The marker for Pm 13 was not detected in any examined cultivar but was present in five accessions of Aegilops speltoides.

Molecular markers for leaf rust resistance genes in wheat.

Over 100 genes of resistance to rust fungi: Puccinia recondita f. sp. tritici, (47 Lr - leaf rust genes), P. striiformis (18 Yr - yellow rust genes) and P. graminis f. sp. tritici (41 Sr - stripe rust genes) have been identified in wheat (Triticum aestivum L.) and its wild relatives according to recent papers. Sixteen Lr resistance genes have been mapped using restriction fragments length polymorphism (RFLP) markers on wheat chromosomes. More than ten Lr genes can be identified in breeding materials by sequence tagged site (STS) specific markers. Gene Lrk 10, closely linked to gene Lr 10, has been cloned and its function recognized. Available markers are presented in this review. The STS, cleaved amplified polymorphic sequence (CAPS) and sequence characterized amplified regions (SCAR) markers found in the literature should be verified using Triticum spp. with different genetic background. Simple sequence repeats (SSR) markers for Lr resistance genes are now also available.

Function of dog's auditory cortex in tests involving auditory location cues and directional instrumental response.

Twenty one dogs, distributed into four groups, were trained pre-operatively in differentiation of auditory location or frequency cues. In each group instrumental responses, reinforced by food consisted in placing by the animal its right paw on the side levers. The first differentiation, go-left, go-right task with two location cues, required the animal to place its paw on the lever opposite to the source of the cue. The second differentiation task with the same location cues required placing the paw on the lever located close to the cue. The third task, involving 700 Hz vs. 1,000 Hz tone, required responding to one lever to the presentation of one tone and responding to the opposite lever to the presentation of the other. The last task was a symmetrically reinforced go, no-go differentiation with, again, auditory location cues: the animals were trained to place the paw on a lever to one location cue and to withhold this response to the other location cue. Bilateral ablation of the primary auditory cortex produced a considerable impairment of the performance of the two go-left, go-right tasks involving location cues. The go-left, go-right task employing frequency cues, and the symmetrically reinforced go, no-go task with location cues, were only slightly disturbed by this lesion.

[Case of severe epilepsy treated by 2-stage hemispherectomy]. / Przypadek ciezkiej postaci padaczki leczonej za pomoca hemisferektomii wykonanej w dwóch etapach operacyjnych.

The authors describe a 7-year-old boy with severe epilepsy cured by means of two-stage hemispherectomy. Epileptic seizures occurred in the left extremities and recurred several tens of times daily despite intensive pharmacological treatment. EEG investigations demonstrated seizure activity in the right motor-parietal-temporal area. Stereotaxic destruction of Forel's field and thalamus on the right side gave only brief improvement lasting several days. In view of this the frontal, temporal and a part of the parietal lobe were removed according to electrocorticographic investigations. During 6 months after the operation epileptic attacks were absent, than they recurred with increasing frequency. After 10 months full hemispherectomy was done. The child tolerated the operation very well, and control examination after 2 years showed disappearance of epileptic attacks and considerable compensation of motor power loss and sensory disturbances in the left extremities. The authors believe that hemispherectomy carried out in two stages at an interval of several months is better tolerated by the patients and makes possible better compensation of functional disturbances than classic hemispherectomy. In the discussion the authors stress the importance of extrapyramidal pathways for compensation of motor disturbances.

Late results of operation in temporal lobe epilepsy in adults and children.

In our clinic at the Medical Academy in Warsaw 282 patients with temporal lobe epilepsy were treated surgically. The causes of the illness were trauma in 43.2%, including birth trauma in 19.5%, infections diseases in early life in 19.8%, and microtumours in 6.8%. In 30% the aetiology was unknown. Fifty per cent had fits before 10 years and 75% before 20 years of age. The period of conservative treatment was on average nine years. Serial and stereo-EEG recordings including activation by ketamine, as well as intracarotid sodium amytal tests were performed routinely. The interictal EEG epileptiform abnormalities were found most frequently in both temporal lobes (154 cases). The operation was carried out according to Penfield's technique, with electrocorticography and resection of the temporal lobe extending for 6 to 10 cm in the nondominant hemisphere and for 4 to 5 cm in the dominant hemisphere using suction technique. In 75% microscopical changes in the hippocampus were found, in 20% so-called hamartomas and in 6.8% small gliomas. Two patients died and in 13 patients there was a hemiparesis which was transient in 10. The authors present the analysis of late results of 262 cases with a follow-up from 2 to 22 years after operation. Very good results were obtained in 127 cases (48.5%) - no attacks since leaving the clinic. Good results were observed in 42 patients (16%) - not more than 1-2 attacks a year. Thus, the operation resulted in freedom from attacks, or nearly so, in 169 cases (64.5%). In an additional 47 patients (18%) there was a significant reduction (at least 50%) in seizures without complete freedom from attacks, and in 44 cases (16.8%) no improvement was observed. The analysis of our series suggests that the best results may be obtained in patients with unilateral temporal EEG changes. The existence of an additional focus in parts of the other temporal lobe does not impair the operative results when the dominant epileptic focus has been removed. Among 24 cases with equally pronounced bitemporal EEG abnormalities the stereo-EEG studies allowed detection of the epileptic focus in 13 patients (54%). Detailed analysis of the results, obtained in 51 children below 15 years of age, led to the conclusion that temporal lobe epilepsy should be operated upon even in young children, provided that the epileptic focus can be clearly identified.

[Relationship between blood serum luminal and diphenylhydantoin level and the results of treatment and other clinical data in drug-resistant epilepsy]. / Stosunek miedzy poziomem luminalu i dwufenylhydantoiny u chorych z lekooporna padaczka a wynikami leczenia i innymi danymi klinicznymi.

The authors report a statistical analysis of the levels of phenobarbital (PH) and diphenylhydantoin (DPH) in a group of 100 patients with different types of epilepsy mostly resistant to pharmacological treatment. The mean values of PH and DPH were compared with the types of epilepsy, duration of the disease, additional treatment, EEG changes and therapeutic results. The following conclusions have been reached: a) therapeutic ranges of the PH and DPH levels in this group of patients were greater than those reported by others. b) no significant correlation is found between the levels of these drugs and the type of epilepsy, disease duration and EEG changes, with the exception of a positive correlation with PH level. c) Clonazepam, diazepam and mysodin raised the serum PH level in the serum, while only amizepine raised the DPH level. d) higher levels of PH and DPH were found in patients with poor therapeutic results.

Prefrontal cortex and manipulatory go left-go right differentiation to acoustic directional cues in dogs.

In 20 dogs the manipulatory go left – go right differentiation to acoustic directional cues was elaborated. All dogs received total prefrontal, or dorsolateral (total or partial) or medial (total or partial) cortical ablations. All total ablations markedly affected performance of the task, whereas the partial removals produced moderate or no impairment. Thus, both the dorsolateral and medial prefrontal cortex are involved in this type of differentiation.

The effects of the removal of the somatosensory areas I and II on left leg-right leg differentiation to tactile stimuli in dogs.

Dogs were trained to lift the right forelegs to a rhythmic tactile stimulus applied to the right side of the trunk, and to lift the left forelegs to a tactile stimulus applied to the left side of the trunk. After training was completed various ablations of sensory I area and sensory II area were made. It was found that the task was impaired after unilateral or bilateral SII lesions, but was virtually preserved after SI lesions.

Effects of prefrontal lesions on left leg-right leg differentiation in dogs.

Twenty five animals trained in the left leg-right leg differentiation were given orbital, proreal or medial precruciate lesions. Pure cortical ablations did not affect the task whereas these deep lesions involving the bundle of fibers inside the prefrontal region caused severe deficit. The medial lesions (superficial and deep) produced disinhibition of the intertrial reactions. The nature of the deficit in the differentiation and the relation between this deficit and the disinhibition is discussed.