Recombination-like sequences in the upstream region of the phage-type DNA polymerase in ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter asiaticus’

We determined and compared nucleotide sequences in the upstream region of the bacteriophage-type DNA polymerase in 15 Asian isolates of the citrus-greening bacterium ‘Candidatus Liberibacter asiaticus’ using the genomic information of American isolate Psy62. Some isolates had nucleotide changes and a large 156-bp deletion, which were concentrated in the putative bacteriophage genes ORF3 and ORF4 and corresponding intergenic region. Eight partial recombination-like patterns were observed in this variable region. Most were confirmed to have occurred between the original types 1 and 2, but a fragment, only several dozen nucleotides long and possibly derived from a third type, was also identified.     

Distribution of the double-spined spruce bark beetle <Emphasis Type="BoldItalic">Ips duplicatus</Emphasis> in the Czech Republic: spreading in 1997–2009

Ips duplicatus (Coleoptera: Curculionidae) was reported in the Euro-Siberian taiga from Sweden to Sakhalin and in the Alps at the beginning of the 20th Century. It has been recorded in spruce stands in the north-eastern part of the Czech Republic since the beginning of the 1970s; monitoring of I. duplicatus was conducted in the Czech Republic in 9 years between 1997 and 2009. Beetles of the overwintering generation were captured in Theysohn pheromone traps baited with a lure of ipsdienol and E-myrcenol, and after 2005 in ID Ecolure lures. Surprisingly, I. duplicatus was found throughout the Czech Republic already in 1997. In 2002, there was a nationwide decrease in their numbers. Particularly since 2005, there has been a spread from the main center of occurrence in the eastern areas to the south and west into the central part of Bohemia where, until that time, its population densities had been lower and had not caused economically significant damage. Moreover, it was observed that in the north-eastern parts of the Czech Republic up to 80% of spruce trees from sanitary felling had been infested by this pest in recent years. The recorded amount of spruce trees infested by I. duplicatus is correlated significantly with the amount of spruce trees recorded as damaged by Ips typographus, Ips amitinus and Pityogenes chalcographus infestation as well as with the numbers of individuals of I. duplicatus captured during the aforementioned monitoring.     

Is the current state of the art of weed monitoring suitable for site‐specific weed management in arable crops?

Weed monitoring is the first step in any site‐specific weed management programme. A relatively large variety of platforms, cameras, sensors and image analysis procedures are available to detect and map weed presence/abundance at various times and spatial scales. Remote sensing from satellites or aircraft can provide accurate weed maps when the images are obtained at late weed phenological stages. Cameras located on unmanned aerial vehicles (UAVs) have been shown to be adequate for early‐season weed detection in a variety of wide‐row crops, providing images with relatively high spatial resolutions. Alternatively, weed detection/mapping systems from ground‐based platforms can achieve even higher resolutions using a variety of non‐imaging and imaging technologies. These ground systems are suited, in some cases, for real‐time site‐specific weed management. Despite this rich arsenal of technologies, their commercial adoption is, apparently, low. In this study, we describe the state of the art of remotely sensed and ground‐based weed monitoring in arable crops and the current level of adoption of these technologies, exploring major constraints for adoption and trying to identify research gaps and bottlenecks.     

Phenological adaptation in weeds—an evolutionary response to the use of herbicides?

The possibility of phenological adaptation in weed species is discussed in terms of an evolved response to herbicide use. Weed populations often exhibit heritable variation in life history traits that may reflect phenological adaptations. Approaches to assessing ‘adaptedness’ are discussed. Selection for seed dormancy in a grass weed is considered through life history analysis. It is concluded that too little is known about both life histories and fitness of weed species in varying environments for conclusions to be drawn concerning phenological adaptation for use in herbicide resistance management. ©1997 SCI     

Elimination of in vitro bacterial contaminants in shoot cultures of ‘MRS 2/5’ plum hybrid by the use of <Emphasis Type="Italic">Melia azedarach</Emphasis> extracts

The antimicrobial activity of leaf and callus extracts of Melia azedarach was tested on in vitro shoot cultures of the peach rootstoch ‘MRS 2/5’ (Prunus cerasifera × Prunus spinosa) that were heavily contaminated with Sphingomonas paucimobilis (Sp) and Bacillus circulans (Bc). The extracts were filter-sterilised and added at 0%, 1%, 5%, 10% and 20% to a modified Murashige and Skoog proliferation medium previously autoclave-sterilised. Up to about 17% shoots died with 10–20% extract, except for Sp-contaminated shoots, whose survival was reduced to 50% after treatment with 20% extract. No shoots died with 1% to 5% supplement. The undiluted leaf extract showed bactericidal activity on plated Sp and Bc isolates. The homogenates of shoots randomly collected from treated cultures were processed for bacterial colony counting. Thus the 10% supplement was the best treatment for ridding Bc-contaminated cultures of bacteria (although 5% had a similar bactericidal effect), and allowing shoot growth and proliferation comparable to controls at the fifth subculture on a standard medium, while 20% extract was needed to eliminate Sp, and could induce higher growth and proliferation rates in surviving shoots than in untreated cultures. Callus extract was ineffective. The bactericidal activity of the leaf extract seemed attributable to a synergistic effect of azadirachtin with other unidentified compounds present in the extract.     

Can poisons stimulate bees? Appreciating the potential of hormesis in bee–pesticide research

Hormesis, a biphasic dose response whereby exposure to low doses of a stressor can stimulate biological processes, has been reported in many organisms, including pest insects when they are exposed to low doses of a pesticide. However, awareness of the hormesis phenomenon seems to be limited among bee researchers, in spite of the increased emphasis of late on pollinator toxicology and risk assessment. In this commentary, we show that there are several examples in the literature of substances that are toxic to bees at high doses but stimulatory at low doses. Appreciation of the hormetic dose response by bee researchers will improve our fundamental understanding of how bees respond to low doses of chemical stressors, and may be useful in pollinator risk assessment. © 2015 Society of Chemical Industry     

Sex-specific probing behaviour of the carrot psyllid <Emphasis Type="Italic">Bactericera trigonica</Emphasis> and its implication in the transmission of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Liberibacter solanacearum’

‘Candidatus Liberibacter solanacearum (Lso)’ is a pathogen of Solanaceae but also causes serious physiological disorders in carrots and celery (Apiaceae). In carrots, this pathogen is transmitted by the psyllids Bactericera trigonica and Trioza apicalis. How vector sex influences Lso transmission has not been yet elucidated. Here we report the probing behaviours of male and female B. trigonica and their impact on Lso titre transmitted, percentage of transmission, and symptoms produced on carrots when Lso is transmitted by males or females of B. trigonica. Vector sex affected the inoculation of Lso; our results suggest that females might inoculate higher Lso titres than males. However, the percentage of transmission was not affected by vector sex at a density of one or eight psyllids per plant. The number of yellow leaves and root weight were not different when Lso was transmitted by males or females at either of the psyllid densities tested, except for groups of females whose Lso transmission resulted in a higher number of yellow leaves than did Lso transmitted by groups of males. Electrical penetration graphs (EPG) showed that the proportion of individuals who reached phloem tissues was similar for males and females. However, EPGs also showed that females probed more times, ingested longer from phloem sieve elements and reached phloem tissues more frequently than did males during an 8-h inoculation access period (IAP). Our study shows that differences in probing behaviours between males and females of B. trigonica could modulate how Lso is inoculated by psyllids. These results highlight the importance of taking sex into consideration in psyllid studies of probing behaviour and bacterial transmission.     

Multiple facets of response to fungicides – the influence of azole treatment on expression of key mycotoxin biosynthetic genes and candidate resistance factors in the control of resistant <Emphasis Type="Italic">Fusarium</Emphasis> strains

The spread of fungicide resistant and/or tolerant phytopathogenic fungi is an important factor affecting crop protection. However, the mechanisms of fungal response to fungicide application are not entirely characterised. In particular, the contribution of previously known resistance factors and the final influence of fungicide treatments on metabolism of surviving mycelia (e.g. mycotoxin increased release and biosynthesis potentially causing contamination of the crops) merit investigation, in order to improve future molecular diagnostics of fungicide resistant strains. The performed experiments have shown distinct expression changes for homologs of a known fungicide resistance factor Flr1 (yeast; DHA1 family of major facilitator superfamily transporters) after azole application in cultured fusaria. Two distantly related homologs of that gene were selected, based on the unsupervised clustering and phylogenetic analysis of transporter sequences. One of these (FGSG_02865), was found to occur across the Fusarium sambucinum complex (F. graminearum, F. culmorum, F. cerealis) and was upregulated starting 24 h after fungicide treatments. This delayed response may point to possible involvement of DHA1 antiporters in a generalised response to stress resulting from fungicide treatment. Additional expression profiling was conducted for the mycotoxin biosynthetic genes (trichothecene and zearalenone gene clusters) in strains of Fusarium sambucinum complex cereal pathogens. The expression changes, when subjected to treatment with the fungicides (flusilazole, carbendazim), show that even an effective treatment (in this study, the benzimidazole fungicide carbendazim) applied to the grown mycelium, can result in enhanced activation of mycotoxin biosynthetic genes in fungal cells which survive the treatment. Our results suggest that increased mycotoxin contamination can be strongly influenced not only by the amount or the type of antifungal compound, but also the timing of fungicide exposition (stage of infection).     

Involvement of the β-cinnamomin elicitin in infection and colonisation of cork oak roots by <Emphasis Type="Italic">Phytophthora cinnamomi</Emphasis>

The virulence of two wild type (PA45 and PA37) and two genetically modified (13C: hygromycin resistant; FATSS: hygromycin resistant and β-cin knock-down) Phytophthora cinnamomi strains towards cork oak (Quercus suber) was assessed via a quantitative evaluation of disease symptoms arising from a soil infestation assay, and by a histological analysis of root colonization. Comparison of virulence, as expressed by symptom severity, resulted in the following ranking: highly virulent (wild type strains), medium virulence (strain 13C) and weakly virulent (FATSS). Both transgenic strains were compromised in their virulence, as expressed by symptom severity, but strain 13C was much less affected than FATSS. Microscopic observation showed that the FATSS strain was unable to effectively invade the root, while 13C and the two wild type strains were all able to rapidly colonize the whole root, including the vascular tissue. These results strengthen the notion that elicitins are associated, either directly or indirectly, with the infection process of Phytophthora.     

Analysis of the relationship between parameters of resistance to <Emphasis Type="Italic">Fusarium</Emphasis> head blight and <Emphasis Type="Italic">in vitro</Emphasis> tolerance to deoxynivalenol of the winter wheat cultivar WEK0609 <Superscript>®</Superscript>

Fusarium head blight (FHB) symptom development, relative spikelet weight (RSW), fungal DNA (FDNA) and deoxynivalenol (DON) content of grain was assessed in the FHB resistant winter wheat cv. WEK0609 and the FHB susceptible cv. Hobbit sib, and among doubled haploid progeny lines (DHLs) developed from a cross between these cultivars. In addition, the relationship between FHB resistance traits and germination on DON-containing medium (in vitro DON tolerance (IVDT)) was also investigated to assess the possibility of using this test as in vitro method of screening for FHB resistance in this cultivar. Analysis indicated that WEK0609 resistance significantly reduced symptom development, yield loss and the FDNA and DON content of grain relative to Hobbit sib. Although both the DON and FDNA content were greater in susceptible than in resistant progeny lines, the ratio of DON to FDNA decreased with increasing susceptibility. The resistance derived from WEK0609 appears to have a greater effect on colonisation of the grain by the fungus than on the accumulation of DON within the grain. In vitro tolerance to DON does not appear to relate to FHB resistance in WEK0609 and thus does not provide a means of selecting for FHB resistance derived from this cultivar.     

Occurrence of boscalid resistance in cucumber powdery mildew in Japan and molecular characterization of the iron–sulfur protein of succinate dehydrogenase of the causal fungus

A total of 74 mass isolates of cucumber powdery mildew fungus (Podosphaera xanthii) were collected from commercial greenhouses with a history of boscalid use in Ibaraki Prefecture, Japan, and tested in a leaf disk assay for their sensitivity to boscalid. The mildew development of 40 of 74 isolates and five sensitive reference isolates on the disks was completely suppressed at 5 μg boscalid/ml. The minimum inhibitory concentrations (MIC) for the remaining 34 isolates were 50 μg/ml or higher, and 21 of these isolates also grew well at 500 μg/ml. Six single-spore isolates were resistant to boscalid with MIC values higher than 500 μg/ml; four of these were moderately resistant (MR), and two were very highly resistant (VHR) isolates. The growth of MR isolates was almost completely suppressed at 500 μg/ml, whereas two VHR isolates grew vigorously at 500 μg/ml. In foliar inoculation tests of potted cucumber plants, the efficacy of boscalid (500 μg/ml) against both MR and VHR isolates was very low. Partial DNA fragment of the iron–sulphur protein subunit (SdhB) gene of succinate dehydrogenase was PCR-amplified from five sensitive and five resistant isolates and directly sequenced, revealing that VHR isolates possess a substitution from a highly conserved histidine (CAT) to tyrosine (TAT) in a third cysteine-rich center of a putative SdhB, whereas MR isolates so far have not been found to have such substitution in SdhB.     

‘<Emphasis Type="Italic">Candidatus</Emphasis> Phlomobacter fragariae’ and the proteobacterium associated with the low sugar content syndrome of sugar beet are related to bacteria of the arsenophonus clade detected in hemipteran insects

The phylogeny of ‘Candidatus Phlomobacter fragariae’(Ca. P. fragariae), the agent of the strawberry marginal chlorosis (SMC), and the proteobacterium associated with the low sugar content syndrome of sugar beet (SBRp) is not well understood. The spoT-spoU-recG genetic locus initially characterised by genome walking from a ‘Ca. P. fragariae’ partial spoT sequence was used to determine relatedness of ‘Ca. P. fragariae’ and SBRp with bacteria detected in hemipteran insects. Both plant pathogenic bacteria belong to the same phylogenetic group as bacteria of the arsenophonus clade detected in hemipteran insects. The SBRp is closely related to arsenophonus-like proteobacteria from cixiids and more distantly related to psyllid and delphacid secondary endosymbionts, whereas the relatives of ‘Ca. P. fragariae’ remain to be discovered. No genetic variability was found among isolates of ‘Ca. P. fragariae’ or SBRp. Implications for explaining the emergence of both ‘Ca. P. fragariae’ and SBRp as epidemic plant pathogens are discussed.     

On the apple proliferation symptom display and the canopy colonization pattern of “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali” in apple trees

Notwithstanding the availability of several different real time PCR protocols for “Candidatus Phytoplasma mali”, it is still unclear how informative is the estimation of the concentration of phytoplasma cells in the leaves of apple proliferation infected trees, and how the reliability of the estimations may be affected by an erratic and uneven distribution of the pathogen in the host. Here we investigated these issues systematically and showed that phytoplasma concentration varies significantly among seasons, but not between two cultivars that appeared to have different degree of susceptibility on the basis of the symptoms displayed. In fully symptomatic trees sampled at the end of the season the phytoplasmas were detectable in most leaves, but in more than half of the leaves at low concentrations. Both the pattern of colonization of the canopy and the amount of phytoplasmas varied greatly in trees that show symptom remission, although a direct relation between symptom severity and colonization could not be established. The sampling of the apple canopy for the purpose of evaluation of concentration of “Candidatus Phytoplasma mali” should take into consideration the complex pattern of colonization and seasonal variation.     

Precipitation changes in the mid-latitudes of the Chinese mainland during 1960–2014

Based on daily precipitation data from 163 meteorological stations, this study investigated precipitation changes in the mid-latitudes of the Chinese mainland(MCM) during 1960–2014 using the climatic trend coefficient, least-squared regression analysis, and a non-parametric Mann-Kendall test.According to the effects of the East Asian summer monsoon on the MCM and the climatic trend coefficient of annual precipitation during 1960–2014, we divided the MCM into the western MCM and eastern MCM. The western MCM was further divided into the western MCM1 and western MCM2 in terms of the effects of the East Asian summer monsoon. The main results were as follows:(1) During the last four decades of the 20~(th) century, the area-averaged annual precipitation presented a significant increasing trend in the western MCM, but there was a slight decreasing trend in the eastern MCM, where a seesaw pattern was apparent. However, in the 21~(st) century, the area-averaged annual precipitation displayed a significant increasing trend in both the western and eastern MCM.(2) The trend in area-averaged seasonal precipitation during 1960–2014 in the western MCM was consistent with that in the eastern MCM in winter and spring. However, the trend in area-averaged summer precipitation during1960–2014 displayed a seesaw pattern between the western and eastern MCM.(3) On an annual basis,both the trend in rainstorms and heavy rain displayed a seesaw pattern between the western and eastern MCM.(4) The precipitation intensity in rainstorms, heavy rain, and moderate rain made a greater contribution to changes in the total precipitation than precipitation frequency. The results of this study will improve our understanding of the trends and differences in precipitation changes in different areas of the MCM. This is not only useful for the management and mitigation of flood disasters, but is also beneficial to the protection of water resources across the MCM.     

Variations of precipitation characteristics during the period 1960–2014 in the Source Region of the Yellow River,China

Precipitation, a natural feature of weather systems in the Earth, is vitally important for the environment of any region. Under global climate change condition, the characteristics of precipitation have changed as a consequence of enhanced global hydrological cycle. The source region of the Yellow River(SRYR), locating within the Qinghai-Tibet Plateau, is sensitive to the global climate change due to its complex orography and fragile ecosystem. To understand the precipitation characteristics and its impacts on the environment in the region, we studied the characteristics of rainy days and precipitation amount of different precipitation classes, such as light(0–5 and 5–10 mm), moderate(10–15, 15–20 and 20–25 mm) and heavy(≥25 mm) rains by analyzing the precipitation data of typical meteorological stations in the SRYR during the period 1961–2014, as well as the trends of persistent rainfall events and drought events. Results showed that annual average precipitation in this area had a non-significant(P>0.05) increasing trend, and 82.5% of the precipitation occurred from May to September. Rainy days of the 0–5 mm precipitation class significantly decreased, whereas the rainy days of 5–10, 10–15, and 20–25 mm precipitation classes increased and that of ≥25 mm precipitation class decreased insignificantly. The persistent rainfall events of 1-or 2-day and more than 2-day showed an increasing trend, with the 1-or 2-day events being more frequent. Meanwhile, the number of short drought periods(≤10 days) increased while long drought periods(>10 days) decreased. Since the 0–5 mm precipitation class had a huge impact on the grasslands productivity; the 5–10, 10–15, and 20–25 mm precipitation classes had positive effects on vegetation which rely on the deep soil water through moving nutrients and water into the root zone of these vegetation or through the plant-microbe interactions; the ≥25 mm precipitation class contributed to the floods; and more persistent rainfall events and fewer long drought events inferred positive effects on agriculture. Thus, these results indicate grassland degradation, less risk of floods, and the upgrading impact of climate change on agriculture. This study may provide scientific knowledge for policymakers to sustain the eco-environmental resources in the SYSR.     

Origin and circulation of saline springs in the Kuqa Basin of the Tarim Basin,Northwest China

It is widely accepted that hydrogeochemistry of saline springs is extremely important to understand the water circulation and evolution of saline basins and to evaluate the potential of potassium-rich evaporites.The Kuqa Basin,located in the northern part of the Tarim Basin in Northwest China,is a saline basin regarded as the most potential potash-seeking area.However,the origin and water circulation processes of saline springs have yet to be fully characterized in this saline basin.In this study,a total of 30 saline spring samples and 11 river water samples were collected from the Qiulitage Structural Belt(QSB)of the Kuqa Basin.They were analyzed for major(K^+,Ca2^+,Na^+,Mg2^+,SO42-,Cl-and HCO3-)and trace(Sr2^+and Br-)ion concentrations,stable H-O-Sr isotopes and tritium concentrations in combination with previously published hydrogeochemical and isotopic(H-O)data in the same area.It is found that the water chemical type of saline springs in the study area belonged to the Na-Cl type,and that of river water belonged to the Ca-Mg-HCO3-SO4 type.The total dissolved solid(TDS)of saline springs in the QSB ranged from 117.77 to 314.92 g/L,reaching the brine level.On the basis of the general chemical compositions and the characteristics of the stable H-O-Sr isotopes of saline springs,we infer that those saline springs mainly originated from precipitation following river water recharging.In addition,we found that saline springs were not formed by evapo-concentration because it is unlikely that the high chloride concentration of saline springs resulted in evapo-concentration and high salinity.Therefore,we conclude that saline spring water may have experienced intense evapo-concentration before dissolving the salty minerals or after returning to the surface.The results show that the origin of salinity was mainly dominated by dissolving salty minerals due to the river water and/or precipitation that passed through the halite-rich stratum.Moreover,there are two possible origins of saline springs in the QSB:one is the infiltration of the meteoric water(river water),which then circulates deep into the earth,wherein it dissolves salty minerals,travels along the fault and returns to the surface;another is the mixture of formation water,or the mixture of seawater or marine evaporate sources and its subsequent discharge to the surface under fault conditions.Our findings provide new insight into the possible saltwater circulation and evolution of saline basins in the Tarim Basin.     

Identification of a novel point mutation in the <Emphasis Type="Italic">β</Emphasis>-tubulin gene of <Emphasis Type="Italic">Botrytis cinerea</Emphasis> and detection of benzimidazole resistance by a diagnostic PCR-RFLP assay

The molecular basis of resistance to benzimidazole fungicides with laboratory and field mutant isolates of Botrytis cinerea was investigated. After chemical mutagenesis with N-methyl-N-nitrosogouanidine (NMNG) two different benzimidazole-resistant phenotypes were isolated on media containing carbendazim or a mixture of carbendazim and diethofencarb. The mutant isolates from the fungicide-mixture-containing medium were moderately resistant to carbendazim with wild-type tolerance to diethofencarb while mutant isolates from carbendazim-containing medium were highly resistant to carbendazim but sensitive to diethofencarb. The studied field isolates were highly resistant to benzimidazoles and sensitive to diethofencarb. Study of fitness characteristics of benzimidazole highly-resistant isolates showed that the resistance mutation(s) had no apparent effect on fitness-determining parameters. Contrary to this, the moderately benzimidazole-resistant strains, with no increased diethofencarb sensitivity, had a significant reduction in certain ecological fitness-determining characteristics. Analysis of the sequence of the β-tubulin gene revealed two amino acid replacements in the highly benzimidazole-resistant mutants compared to that of the wild-type parent strain. One was the glutamic acid (GAG) to alanine (GCG) change at position 198 (E198A), identified in both laboratory and field highly benzimidazole-resistant isolates, a mutation previously implicated in benzimidazole resistance. The second was a novel benzimidazole resistance mutation of glutamic acid (GAG) to glycine (GGG) substitution at the same position 198 (E198G), identified in a highly benzimidazole-resistant laboratory mutant strain. Molecular analysis of the moderately benzimidazole-resistant strains revealed no mutations at the β-tubulin gene. A novel diagnostic PCR-RFLP assay utilising a BsaI restriction site present in the benzimidazole-sensitive (E198) but absent in both resistant genotypes (E198G and E198A) was developed for the detection of both amino acid replacements at the β-tubulin gene.