Implication of peroxynitrite in defence responses of potato to Phytophthora infestans

In this study peroxynitrite (ONOO^?) is proposed as an important player in defence responses during the interaction of potato (Solanum tuberosum) and the oomycete pathogen Phytophthora infestans. The potato–avr P. infestans model system exhibited a transient programme of boosted ONOO^? formation correlated in time with the burst of nitric oxide (NO) and superoxide during the first 6 h post‐inoculation (hpi). The early ONOO^? over‐accumulation was not accompanied by TPx gene expression. In contrast, the compatible interaction revealed a 24 h delay of ONOO^? formation; however, an enhanced level of NO and superoxide correlated with TPx up‐regulation was recorded within the earlier stages of pathogen infection. Peroxynitrite over‐accumulation in the susceptible potato coincided with an enhanced level of protein tyrosine nitration starting from 24 hpi. Surprisingly, the nitroproteome profile of the resistant potato did not show any visible difference after inoculation, apart from one band containing subtilisin‐like protease‐like proteins, which appeared 48 h after pathogen attack. An additional pharmacological approach showed that treatment of the susceptible genotype with ONOO^? followed by inoculation with P. infestans contributed to slowing down of the colonization of host tissues by the pathogen via a faster and stronger up‐regulation of the key defence markers, including the PR‐1 gene. Taken together, the results obtained indicate that a precise control of emitted NO and superoxide in cooperation with thioredoxin‐dependent redox sensors in sites of pathogen ingress could generate a sufficient threshold of ONOO^?, triggering defence responses.     

A new classification of pre‐ovulatory oocyte maturation stages in pikeperch,Sander lucioperca (L.), and its application during artificial reproduction

This study aimed at improving the reproduction effectiveness and synchronization of ovulation in the pikeperch, Sander lucioperca (L.), during induced spawning, which is one of the main bottlenecks in the aquaculture of this species. For this purpose, a new categorization of maturation stages in pre‐ovulatory oocytes was applied. It is generally based on two morphological indicators: germinal vesicle migration or its breakdown (GVBD) and different oil droplet coalescence rates. This categorization covered seven stages (from I to VII) – from the end of vitellogenesis to ovulation. The categorization was verified by controlled reproduction with the use of hormonal stimulation (500 IU of hCG per kg of female body weight) and low spawning temperature (12 °C), which extended the latency time. In addition, some morphological indicators (pseudo‐gonadosomatic index, Fulton's condition coefficient) of females were calculated in order to determine their usability in determining the maturation stage. However, these indicators proved to be ineffective for this purpose, further highlighting the need to determine the maturational stages in pre‐ovulatory oocytes to synchronize ovulation in pikeperch. During the experiment, ovulation seemed to be synchronized among the experimental treatments. Statistical differences were found in terms of latency time between experimental groups at different maturity stages (II – 78–98 h; III – 57–78 h; IV – 48–58 h; V – 32–49 h; VI – 5–30 h) according to the proposed classification. This classification and the results presented in the study significantly improved the synchronization of ovulation, which may positively affect the effectiveness of pikeperch production under controlled conditions.     

Acute ammonia toxicity during early ontogeny of ide Leuciscus idus (Cyprinidae)

Acute ammonia toxicity was investigated in four developmental stages of the juvenile ide, Leuciscus idus: 1, 10, 20 and 30 days after the first feeding. Mean (±SD) total length of the larvae was 8.5 ± 0.3, 15.7 ± 0.7, 23.0 ± 2.0 and 29.7 ± 2.0 mm, and standard weight was 1.6 ± 0.3, 9.2 ± 5.5, 94.9 ± 31.0 and 196.0 ± 31.7 mg, respectively. The larvae used for toxicity tests were reared in the experimental, closed recirculating system. Groups of fishes (n from 7 to 10; in respect of fish size) were exposed to the ammonium chloride solutions in 1-L glass units. Water temperature was 25 ± 0.1 °C for both the rearing and the toxicity tests. pH was not adjusted and ranged between 8.4 and 8.7. The ammonium chloride solutions were renewed every 12 h. At the same time, dead larvae were counted and removed, and the pH and temperature measurements were taken. Each acute toxicity test duration was 96 h, and lethal concentration LC₁, LC₅₀ and LC₉₉ values were calculated for 24, 48, 72 and 96 h. The susceptibility of the ide larvae to ammonia decreased linearly with age up to 20th day and surprisingly increased during the next 10 days. The LC₅₀ (48 h) values ranged from 0.27 mg L^?1 of unionized ammonia nitrogen for 1 day after the first feeding (AFF) larvae to 1.42 mg L^?1 at day 20 after first feeding. The LC₅₀ (48 h) for 30 days AFF was as high as 0.67 mg L^?1. The critical level of the unionized ammonia nitrogen for ide larvae was suggested as 0.21 mg L^?1.     

Sperm quality and selected biochemical markers of seminal plasma at the beginning of the reproductive period of common carp, Cyprinus carpio L.

Changes in semen quality and selected biochemical markers were analyzed during a week of spawning season of common carp Cyprinus carpio L. Semen was obtained twice, on May 30 and on June 7, and each time it was collected 24 h after hormonal stimulation using Ovopel [(d-Ala⁶, Pro⁹ NEt)-mGnRH + metoclopramide] in 1 pellet kg^?1. The total volume of semen (ml), volume of semen per kg of body weight (ml kg^?1 b.w.), sperm concentration (×10⁹ ml^?1), total number of sperm per kg of body weight (×10⁹ kg^?1 b.w.), pH of semen, pH of seminal plasma, seminal plasma osmotic pressure (mOsm kg^?1) and the total protein content in seminal plasma (mg ml^?1) were determined. A 10 mM Tris buffer containing 100 mM NaCl with 0.5 % BSA (pH 9.0, osmolality 200 mOsm kg^?1) was used to activate sperm. The following computer-assisted sperm analysis (CASA) parameters were determined: percentage of motile sperm (MOT, %), progressively motile sperm (PRG, %), curvilinear velocity (VCL, μm s^?1), straight-line velocity (VSL, μm s^?1), movement linearity (LIN, %), wobbling index (WOB, %), amplitude of lateral head displacement (ALH, μm) and beat cross-frequency (BCF, Hz). The volume of semen per kg of BW, total number of sperm per kg of BW and semen pH were significantly lower at the second semen sampling compared to the first semen sampling. Volume of semen at the second sampling correlated positively with CASA parameters. A lack of differences among CASA parameters between both collection periods indicates good quality of carp sperm hormonally stimulated with Ovopel twice at a 1-week interval.     

Early detection of sugar beet pathogen Ramularia beticola in leaf and air samples using qPCR

A quantitative PCR method (qPCR) was developed for the detection and quantification of Ramularia beticola causing Ramularia leaf spot in sugar beet. R. beticola specific primers were designed based on the internal transcribed spacer region 2 (ITS2). The assay was applied on DNA extracted from spores trapped on tape from Burkard spore traps placed in an artificially inoculated sugar beet field trial and in two sugar beet fields with natural infections. R. beticola DNA was detected at variable amounts in the air samples 14 to 16 days prior to first visible symptoms. R. beticola DNA was detected in air samples from fields with natural infection at significant and increasing levels from development of the first symptoms, indicating that spore production within the crop plays a major role in the epidemic development of the disease. Sugar beet leaves sampled from the inoculated field trial were also tested with the qPCR assay. It was possible to detect the presence of R. beticola in the leaves pre-symptomatic at least 10 days before the occurrence of the visible symptoms of Ramularia leaf spot. This is the first report of a molecular assay, which allows screening for the presence of R. beticola in plant material and in air samples prior to the appearance of visible symptoms. An early detection has potential as a tool, which can be part of a warning system predicting the onset of the disease in the sugar beet crop and helping to optimise fungicide application.     

Risk assessment posed by diseases in context of integrated management of wheat

Journal of Plant Diseases and Protection - Three strict field experiments (2010–2013) were conducted at the Experimental Station of Cultivar Testing in Chrząstowo in Poland...     

Within-Field Variability of Winter Wheat Yield and Grain Quality versus Soil Properties

Within-field variability in wheat grain yield and its quality always exists in production fields and depends, among other factors, mainly on various soil properties related to nutrients and water availability. The aim of the research was to examine the relationships between selected soil properties such as texture; pH; content of the available phosphorus (P), potassium (K), magnesium (Mg), and organic carbon; and winter wheat grain yield and quality under rainfed conditions. Six crop fields with winter wheat (Triticum aestivum, L.) in three sites located in different regions of Poland were examined during two seasons. The grain yield was mainly determined by the soil texture, and the majority of the chemical soil properties did not have a significant effect on grain yield. The grain quality traits were determined by the examined soil properties to a smaller degree than grain yield. The relationships were not consistent across sites and years.     

Rearing river lamprey <Emphasis Type="Italic">Lampetra fluviatilis</Emphasis> (L.) larvae under controlled conditions as a tool for restitution of endangered populations

A method for actively protecting river lamprey and increasing the number of river lamprey populations is presented as protocol of rearing stocking material. The objective of the study was to accommodate lamprey larvae during the most critical life period under controlled conditions until they reach a sufficient size, which will increase their chances to survive in the natural environment. The eggs obtained from wild breeders were incubated in Weiss’s jars at the water temperature of 12 °C. The larvae were reared in tanks set up in closed recirculating aquaculture systems. During the rearing period, the water temperature was 20 °C and the density of larvae was 150 individuals per 1 dm² of bottom. In the initial rearing phase, the main focus was on the type of feeds and the nourishment method. It was determined that the most suitable feed type was Artemia nauplii combined with Hikari dry food. Additionally, the type of substrate was tested under controlled conditions. In this case, the optimum one was composed of sand of the grain size between 0.3 and 0.5 mm. The experiment proved that it is possible to successfully conduct river lamprey larvae rearing under controlled conditions. The growth rates are comparable to or better than those achieved under natural conditions. After 30 days of the rearing period, the lamprey larvae were 25 mm long. The results create a possibility to develop a technology for the restitution of river lamprey and other lamprey species, which are endangered throughout the world.