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Botrytis cinerea causes gray mold on thousands of plants, leading to huge losses in production. Anilinopyrimidine (AP) fungicides have been applied to control B. cinerea since the 1990s. Although resistance to AP fungicides was detected soon after their application, the mechanism of AP resistance remains to be elucidated. In this study, a sexual cross between resistant and sensitive isolates was performed, and the genomes of parental isolates and progenies were sequenced to identify resistance-related single nucleotide polymorphisms (SNPs). After screening and verification, mutation E407K in the Bcmdl1 gene was identified and confirmed to confer resistance to AP fungicides in B. cinerea . Bcmdl1 was predicted to encode a mitochondrial protein that belonged to a half-type ATP-binding cassette (ABC) transporter. Although Bcmdl1 was a transporter, it did not mediate resistance to multiple fungicides but mediated resistance specifically to AP fungicides. On the other hand, reductions in conidial germination and virulence were observed in Bcmdl1 knockout transformants compared to the parental isolate and complemented transformants, illustrating the biological functions of Bcmdl1 . Subcellular localization analysis indicated that Bcmdl1 was localized in mitochondria. Interestingly, the production of ATP was reduced after cyprodinil treatment in Bcmdl1 knockout transformants, suggesting that Bcmdl1 was involved in ATP synthesis. Since Mdl1 could interact with ATP synthase in yeast, we hypothesize that Bcmdl1 forms a complex with ATP synthase, which AP fungicides might target, thereby interfering with the metabolism of energy. IMPORTANCE Gray mold, caused by B. cinerea , causes huge losses in the production of many fruits and vegetables. AP fungicides have been largely adopted to control this disease since the 1990s, and the development of resistance to AP fungicides initiates new problems for disease control. Due to the unknown mode of action, information on the mechanism of AP resistance is also limited. Recently, mutations in mitochondrial genes were reported to be related to AP resistance. However, the mitochondrial process of these genes remains to be elucidated. In this study, we identified several AP resistance-related mutations by quantitative trait locus sequencing (QTL-seq) and confirmed that mutation E407K in Bcmdl1 conferred AP resistance. We further characterized the expression patterns, biological functions, subcellular localization, and mitochondrial processes of the Bcmdl1 gene. This study deepens our understanding of the mechanism of resistance to and mode of action of AP fungicides., Competing Interests: The authors declare no conflict of interest.

A systematic investigation on the isotopic and elemental signature, for both stable and radioactive elements, and mineral contents was performed to examine the characteristics of subsurface formations collected at different depths between 3.962 km and 4.115 km around deep-laying coal seams located under the Marmarica plateau in Egypt. Concentrations of major and minor oxides (Na 2 O, MgO, Al 2 O 3 , SiO 2 , SO 3 , K 2 O, CaO, TiO 2 , MnO, ΣFeO + Fe 2 O 3 , SrO, ZrO 2 , and BaO) were determined by X-ray fluorescence and dependencies among these concentrations revealed the type and sort of the formations. Organic contents were determined by Fourier Transform infrared spectroscopy to investigate the variation of the CO/CC bonding ratio with depth. Rare earth elements (REE), specifically Y, Sc, La, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, and Lu were determined by inductively coupled plasma mass spectrometry while actinoids were detected by the radioactive decay of its daughter nuclei. The results showed a high trapping of REE elements and actinoids in layers above the coal seams which indicates the occurrence of aqueous flow followed by possible sorption in these layers. The mobility of the fluid was investigated using the process radioactive decay series between Ra226 and Ac228 from one side and their daughters from the other side., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Parvalbumin-positive (PV + ) γ-aminobutyric acid (GABA) interneurons are critically involved in producing rapid network oscillations and cortical microcircuit computations, but the significance of PV + axon myelination to the temporal features of inhibition remains elusive. Here, using toxic and genetic mouse models of demyelination and dysmyelination, respectively, we find that loss of compact myelin reduces PV + interneuron presynaptic terminals and increases failures, and the weak phasic inhibition of pyramidal neurons abolishes optogenetically driven gamma oscillations in vivo. Strikingly, during behaviors of quiet wakefulness selectively theta rhythms are amplified and accompanied by highly synchronized interictal epileptic discharges. In support of a causal role of impaired PV-mediated inhibition, optogenetic activation of myelin-deficient PV + interneurons attenuated the power of slow theta rhythms and limited interictal spike occurrence. Thus, myelination of PV axons is required to consolidate fast inhibition of pyramidal neurons and enable behavioral state-dependent modulation of local circuit synchronization., Competing Interests: MD, MP, KH, DW, MH, SK, MK No competing interests declared, (© 2022, Dubey et al.)

Altogether, 192 Botrytis cinerea isolates collected from tomato greenhouses at different locations in Hubei Province were evaluated for their sensitivity to fungicides procymidone and zoxamide. The mean effective concentration to cause 50% growth inhibition (EC 50 ) values of procymidone for sensitive and resistant isolates were 0.25 and 3.60 μg/ml, respectively. The frequency of procymidone-resistant (Pro R ) isolates was 18%, and the highest frequency was recorded in Jingmen. Positive cross-resistance was observed for Pro R isolates to other dicarboximide fungicides but not to phenylpyrroles. Significant differences were observed for fitness parameters (i.e., mycelial growth, osmotic sensitivity, and virulence between sensitive and resistant isolates). Amino acid sequence of the Bos1 gene revealed that Pro R isolates carried either point mutations at codon 365 (I365S) or a pair of point mutations at codons 369 (Q369P) and 373 (N373S). For zoxamide, the mean EC 50 values for sensitive and resistant isolates were 0.22 and 5.32 μg/ml, respectively. Approximately 14% of the isolates were found to be resistant to zoxamide, and the highest frequency of resistance was also observed in Jingmen. There was positive cross-resistance for zoxamide-resistant (Zox R ) isolates to carbendazim. No significant differences were observed for fitness parameters between zoxamide-sensitive and Zox R isolates. Sequence analysis of the β-tubulin gene of Botrytis cinerea revealed two previously reported point mutations (E198A and E198K) and one new point mutation (T351I). This new mutation was detected in only those isolates which possessed the E198K but not E198A substitution. This study allows for a better understanding of the resistance development profile in Hubei Province. Results will be useful for the improvement of fungicide resistance management strategies.

Two hundred and forty isolates of Botrytis cinerea were collected during the early summer of 2012 and 2013 from strawberry greenhouses in 10 locations in Hubei Province and examined for sensitivity to five fungicides, most of which were commonly used to control this fungus. High frequency of resistance to carbendazim (Car, 63.63%) and cyprodinil (Cyp, 42.42%) was detected. Boscalid-resistant (Bos R ) isolates were detected for the first time in China, whereas no fludioxonil-resistant isolates were identified. Dual resistance to carbendazim and diethofencarb (Die) was also detected. There were six phenotypes of resistance profile (i.e., Car R Die S Bos S Cyp S , Car R Die R Bos S Cyp S , Car R Die S Bos S Cyp R , Car R Die S Bos R Cyp S , Car R Die R Bos S Cyp R , and Car R Die S Bos R Cyp R ). Car R Die S Bos S Cyp S and Car R Die S Bos S Cyp R were the most common phenotypes, occurring at eight and seven locations, respectively. After 10 successive transfers on fungicide-free potato dextrose agar, tested resistant isolates retained levels of resistance similar to or comparative with the initial generation, indicating the stability of these resistances. Fitness evaluations based on investigation of mycelial growth, osmotic sensitivity, sporulation in vitro and in vivo, and virulence revealed the uncompromising fitness in resistant isolates, except that decreased virulence was observed in Bos R isolates. The molecular basis of carbendazim, diethofencarb, and boscalid resistance was investigated. Results showed that all 13 sequenced carbendazim-resistant isolates harbored the mutation E198V or E198A in the β-tubulin gene and the five isolates with dual resistance to carbendazim and diethofencarb showed the mutation E198K in the same gene. Bos R isolates possessed the H272R mutation in succinate dehydrogenase subunit B gene. The results achieved in this study challenge the current management strategies for B. cinerea, which largely depend on applications of these fungicides.

In cortical pyramidal neurons the presynaptic terminals controlling transmitter release are located along unmyelinated axon collaterals, far from the original action potential (AP) initiation site, the axon initial segment (AIS). Once initiated, APs will need to reliably propagate over long distances and regions of geometrical inhomogeneity like branch points (BPs) to rapidly depolarize the presynaptic terminals and confer temporally precise synaptic transmission. While axon pathologies such as demyelinating diseases are well established to impede the fidelity of AP propagation along internodes, to which extent myelin loss affects propagation along BPs and axon collaterals is not well understood. Here, using the cuprizone demyelination model, we performed optical voltage-sensitive dye (VSD) imaging from control and demyelinated layer 5 pyramidal neuron axons. In the main axon, we find that myelin loss switches the modality of AP propagation from rapid saltation towards a slow continuous wave. The duration of single AP waveforms at BPs or nodes was, however, only slightly briefer. In contrast, by using two-photon microscopy-guided loose-seal patch recordings from axon collaterals we revealed a presynaptic AP broadening in combination with a reduced velocity and frequency-dependent failure. Finally, internodal myelin loss was also associated with de novo sprouting of axon collaterals starting from the primary (demyelinated) axon. Thus, the loss of oligodendrocytes and myelin sheaths bears functional consequences beyond the main axon, impeding the temporal fidelity of presynaptic APs and affecting the functional and structural organization of synaptic connectivity within the neocortex.

In mammalian neurons, the axon initial segment (AIS) electrically connects the somatodendritic compartment with the axon and converts the incoming synaptic voltage changes into a temporally precise action potential (AP) output code. Although axons often emanate directly from the soma, they may also originate more distally from a dendrite, the implications of which are not well-understood. Here, we show that one-third of the thick-tufted layer 5 pyramidal neurons have an axon originating from a dendrite and are characterized by a reduced dendritic complexity and thinner main apical dendrite. Unexpectedly, the rising phase of somatic APs is electrically indistinguishable between neurons with a somatic or a dendritic axon origin. Cable analysis of the neurons indicated that the axonal axial current is inversely proportional to the AIS distance, denoting the path length between the soma and the start of the AIS, and to produce invariant somatic APs, it must scale with the local somatodendritic capacitance. In agreement, AIS distance inversely correlates with the apical dendrite diameter, and model simulations confirmed that the covariation suffices to normalize the somatic AP waveform. Therefore, in pyramidal neurons, the AIS location is finely tuned with the somatodendritic capacitive load, serving as a homeostatic regulation of the somatic AP in the face of diverse neuronal morphologies., Competing Interests: The authors declare no conflict of interest.

Myelination and voltage-gated ion channel clustering at the nodes of Ranvier are essential for the rapid saltatory conduction of action potentials. Whether myelination influences the structural organization of the axon initial segment (AIS) and action potential initiation is poorly understood. Using the cuprizone mouse model, we combined electrophysiological recordings with immunofluorescence of the voltage-gated Nav1.6 and Kv7.3 subunits and anchoring proteins to analyze the functional and structural properties of single demyelinated neocortical L5 axons. Whole-cell recordings demonstrated that neurons with demyelinated axons were intrinsically more excitable, characterized by increased spontaneous suprathreshold depolarizations as well as antidromically propagating action potentials ectopically generated in distal parts of the axon. Immunofluorescence examination of demyelinated axons showed that βIV-spectrin, Nav1.6, and the Kv7.3 channels in nodes of Ranvier either dissolved or extended into the paranodal domains. In contrast, while the AIS in demyelinated axons started more closely to the soma, ankyrin G, βIV-spectrin, and the ion channel expression were maintained. Structure-function analysis and computational modeling, constrained by the AIS location and realistic dendritic and axonal morphologies, confirmed that a more proximal onset of the AIS slightly reduced the efficacy of action potential generation, suggesting a compensatory role. These results suggest that oligodendroglial myelination is not only important for maximizing conduction velocity, but also for limiting hyperexcitability of pyramidal neurons., (Copyright © 2015 Hamada and Kole.)

To develop an effective biocontrol strategy for management of Fusarium head blight on wheat caused by Fusarium graminearum, the bacterial biocontrol agent Pcho10 was selected from more than 1,476 wheat-head-associated bacterial strains according to its antagonistic activity in vitro. This strain was subsequently characterized as Pseudomonas chlororaphis subsp. aurantiaca based on 16S ribosomal DNA sequence analysis, assays of the BIOLOG microbial identification system, and unique pigment production. The major antifungal metabolite produced by Pcho10 was further identified as phenazine-1-carboxamide (PCN) on the basis of nuclear magnetic resonance data. The core PCN biosynthesis gene cluster in Pcho10 was cloned and sequenced. PCN showed strong inhibitory activity against F. graminearum conidial germination, mycelial growth, and deoxynivalenol production. Tests both under growth chamber conditions and in field trials showed that Pcho10 well colonized on the wheat head and effectively controlled the disease caused by F. graminearum. Results of this study indicate that P. chlororaphis subsp. aurantiaca Pcho10 has high potential to be developed as a biocontrol agent against F. graminearum. To our knowledge, this is the first report of the use of P. chlororaphis for the management of Fusarium head blight.

Background: Carbendazim has been used in the control of Fusarium head blight (FHB) for more than 30 years in China. Thus, carbendazim-resistant (Car(R) ) populations of Gibberella zeae have developed in some areas. In this study, 9341 G. zeae isolates were collected from the ten main wheat-producing regions of China in the period from 2008 to 2012, and sensitivity to carbendazim was detected., Results: A high frequency of Car(R) isolates was observed in Zhejiang and Jiangsu provinces. Car(R) isolates were recovered from Anhui and Henan provinces in 2009 and 2012, respectively, but were not detected in the other six regions. Available (F167Y, E198Q and F200Y) and newly developed (E198L and E198K) allele-specific PCR assays were used to genotype field Car(R) isolates. The β-tubulin variants harbouring point mutation F167Y or E198Q accounted for >95% in Car(R) populations. Quantitative allele-specific real-time PCR assays were developed to determine the frequencies of five different β-tubulin variants present in populations of perithecia sampled from rice stubble., Conclusion: Car(R) populations of G. zeae develop rapidly under the selection pressure of carbendazim. Real-time PCR assays detecting the resistance frequencies in populations of perithecia would provide useful information for FHB control and management of resistance., (© 2013 Society of Chemical Industry.)

Estimation of computer model parameters using field data is sometimes attempted in the presence of model bias. In this paper, using simulated field data, a vector-valued model bias is fit simultaneously with a scalar model calibration parameter. Our main finding is that simultaneous estimation of a bias vector and a scalar calibration parameter can be sensitive to assumptions made prior to data collection. Possible implications of this finding are considered in two examples of process monitoring for nuclear safeguards., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Rhizoctonia cerealis, the causal agent of sharp eyespot on wheat, was not considered to be an important pathogen for many years. Recently, the disease has become endemic in many countries except for South America. The disease has created a new threat to world wheat production because the damage of wheat sharp eyespot has become increasingly severe. In this paper, previous studies on this pathogen, including the disease geographical distribution, pathogen identification, life cycle, symptoms, favourable environmental conditions, effects on wheat yield and control strategy, are reviewed. Such information will be helpful in management of sharp eyespot., (Copyright © 2011 Society of Chemical Industry.)