Overexpression of a Broccoli Defensin Gene BoDFN Enhances Downy Mildew Resistance

Plant defensins are small, basic cysteine-rich peptides that play important roles in disease resistance. A gene, designated BoDFN, was isolated from Brassica oleracea var. italica. Gene sequence has been submitted to NCBI with an accession no. of HQ436486. Complete coding sequence of BoDFN is 243 bp in length encoding 80 amino acids. Sequence comparison results showed that BoDFN shared high homology with those of crucifer plants and there were only few DNA base differences. RT-PCR results indicated an increase of gene expression in Hyaloperonospora parasitica infected leaves and revealed a significant increase at 24 and 36 h of treatment. A recombinant plasmid, named pBI121-BoDFN, was constructed and introduced into Agrobacterium tumefacien LBA4404. PCR screening for 65 regenerated plantlets, 17 positive plantlets were obtained, PCR screening results revealed that 17 out of 65 regenerated plantlets were positive. Disease resistant identification results indicated that all positive plants showed an increase in resistance to H. parasitica.

Cloning and Expression Analysis of Downy Mildew Resistance-Related cDNA Sequences in Melon

Melon downy mildew caused by Pseudoperonospora cubensis leads to significant losses in melon yields worldwide. Reverse-transcription Polymerase Chain Reaction （RT-PCR） was performed using cDNAs as templates from melon-Huangdanzi induced with fungus Pseudoperonospora cubensis, and degenerate primers designed based on the conserved amino acid sequences of known plant disease-resistance genes. A polymorphic cDNA fragment which we named rap-19 was cloned and sequenced. The Open Reading Frame （ORF） of this product comprised of 510 base pairs which encodes DNA or RNA-binding protein with 170 amino acids. The putative amino acid sequence of mp-19 appeared highly homologous with those of NBS-type resistant-genes isolated from other plants. Southern blot indicated that the melon genome contained more than 3 copies of rap-19. The obvious expression differences detected by semi-quantitative RT- PCR could be observed between resistant-line Huangdanzi and susceptible-line Jiashi after Pseudoperonospora cubensis infection, which implied that mp-19 gene may be related to the resistance of downy mildew in melon.

Breeding melon(Cucumis melo)with resistance to powdery mildew and downy mildew

Melon(Cucumis melo L.)production is often restricted by a plethora of pests and diseases,including powdery mildew and downy mildew caused respectively by the fungal species Podosphaera xanthii/Golovinomyces orontii and oomycete species Pseudoperonospora cubensis.Many efforts have been directed on identification of resistant sources by screening(wild)melon germplasm.In the current review,we summarized such efforts from various publications of the last 50 plus years.Resistance to powdery mildew has been identified in 239 melon accessions and downy mildew resistance in 452 accessions of both C.melo and the wild relative species C.figarei.Among the resistance sources,C.melo var.cantalupensis accessions PMR 45,PMR 5,PMR 6,and WMR 29 as well as C.melo var.momordica accessions PI 124111,PI 124112,and PI 414723 have been considered as the most valuable germplasm because multiple resistance genes have been identified from these accessions and are widely used in melon resistance breeding.Further genetic mapping in a number of resistant sources has enabled identification of 25 dominant genes,two recessive genes and seven QTLs conferring powdery mildew resistance,as well as eight dominant genes and 11 QTLs for downy mildew resistances.Based on the reported sequences of associated markers,we anchored physically(many of)these genes and QTLs to chromosomes of the melon cv.DHL92 genome.In addition to presenting a comprehensive overview on powdery mildew and downy mildew resistance in(wild)melon germplasm,we suggest strategies aiming at breeding melon with durable and broad-spectrum resistance to pathogens and pests.

Comparative proteomic profiles of resistant/susceptible cucumber leaves in response to downy mildew infection

Downy mildew is a serious disease in cucumber production worldwide,which is caused by Pseudoperonospora cubensis(Berk.&Curt.)Rostov.Underlying the mechanism of cucumber response to downy mildew infection is important for breeding improvement and production;however,the research remains largely elusive.A comparative proteomic approach was used to reveal the differential accumulation of the proteomes from leaves of cucumber(susceptible line and resistant line)that were inoculated with P.cubensis or not by two-dimensional electrophoresis and MALDI-TOF/TOF MS.A total of 76 protein spots were successfully identified with significant changes in abundance(>2-fold,P<0.05)in downy mildew infected or not leaves for the susceptible line and resistant line.By the functional annotation,these proteins were classified into 8 groups including photosynthesis(29%),energy and metabolism(29%),cell rescue and defense(17%),and protein biosynthesis,folding and degradation(13%),unclassified(7%),nucleotide metabolism(3%),signal transduction(1%)and cellular process(1%).Among the 17 differentially expressed proteins between the resistant and susceptible cucumber line,most of the protein spots were concentrated in cell rescue and defense(4)and energy and metabolism(4).Moreover,a schematic diagram containing majority of the metabolic pathways of cucumber leaves in response to downy mildew was proposed.This network revealed the positive effect of several functional components in cucumber seedlings’resistant to downy mildew such as accumulation of energy supply and resistance-related proteins,hastened protein metabolism and photorespiratory,inhibited photosynthesis,and triggered photosystem repair and programmed cell death.Taken together,these results have advanced a further understanding of the key metabolic pathways of cucumber resistance to downy mildew and pathogen control in the proteomic level.

Identification and Characterization of Simple Sequence Repeats (SSR) Markers Associated with Downy Mildew Resistance Locus “<i>RPV1</i>” in Grapes

The cultivation of grapes is severely impacted by the emergence of downy mildew (DM) disease which negatively affects quality and yield possibly resulting in heavy losses. Due to certain shortcomings in the usage of fungicides and the development of new cultivars by plant breeding, marker assisted selection (MAS) will be an efficient alternative method to introduce desired genes into the cultivated varieties in a short time period. The Simple sequence repeats (SSR) markers seem to be the most popular genetic marker of choice for MAS. In the present study, we identified 14 new SSR markers in RPV1 locus that are associated with downy mildew resistance in grapes. The characterization of the identified markers was carried out on the basis of various parameters such as types of repeat motifs, number of repeats, different classes and structure of microsatellites. Additionally, SSR genotyping in 56 different grape accessions was done to determine the susceptibility or resistance of these accessions to DM.

Identification and Molecular Mapping of the Rs Dm R Locus Conferring Resistance to Downy Mildew at Seedling Stage in Radish(Raphanus sativus L.)

Downy mildew （DM）, caused by the fungus Peronospora parasitica, is a destructive disease of radish （Raphanus sativus L.） worldwide. Host resistance has been considered as an attractive and environmentally friendly approach to control the disease. However, the genetic mechanisms of resistance in radish to the pathogen remain unknown. To determine the inheritance of resistance to DM, F1, F2 and BC1F1 populations derived from reciprocal crosses between a resistant line NAU-dhp08 and a susceptible line NAU-qtbjq-06 were evaluated for their responses to DM at seedling stage. All F1 hybrid plants showed high resistance to DM and maternal effect was not detected. The segregation for resistant to susceptible individuals statistically iftted a 3：1 ratio in two F2 populations （F2（SR） and F2（RS））, and 1：1 ratio in two BC1F1 populations, indicating that resistance to DM at seedling stage in radish was controlled by a single dominant locus designated as RsDmR. A total of 1 972 primer pairs （1 036 SRAP, 628 RAPD, 126 RGA, 110 EST-SSR and 72 ISSR） were screened, and 36 were polymorphic between the resistant and susceptible bulks, and consequently used for genotyping individuals in the F2 population. Three markers （Em9/ga24370, NAUISSR826700 and Me7/em10400） linked to the RsDmR locus within a 10.0 cM distance were identiifed using bulked segregant analysis （BSA）. The SRAP marker Em9/ga24370 was the most tightly linked one with a distance of 2.3 cM to RsDmR. These markers tightly linked to the RsDmR locus would facilitate marker-assisted selection and resistance gene pyramiding in radish breeding programs.

Using Pyroxasulfone for Downy Brome (<i>Bromus tectorum</i>L.) Control in Winter Wheat

Downy brome is one of the most troublesome weeds in no-till wheat production systems of the US Great Plains. Pyroxasulfone is a relatively new, soil-applied residual herbicide (root/shoot growth inhibitor) labeled for use in wheat. Multiple field experiments were conducted near Huntley, MT from 2012 through 2016 to determine the efficacy of pyroxasulfone to control downy brome in imidazolinone (IMI)-tolerant (Clearfield&#8482) winter wheat. Pyroxasulfone did not cause any injury to wheat in any of the three studies. Downy brome injury with pyroxasulfone preemergence (PRE) only program did not differ between 89 or 178 g&middot;ai (active ingredient)&middot;ha-1 rates, and averaged 82% and 84% in 2 separate studies. In a preplant (PP) burndown program, the addition of pyroxasulfone (178 g&middot;ai&middot;ha-1) to glyphosate improved downy brome end-season injury from 15% to 74%. In a separate study, the end-season injury with pyroxasulfone was greater when applied PRE (84%) compared to the delayed PRE (DPRE) timing (74%). In addition, the water dispersible granule (WDG) formulation of pyroxasulfone performed slightly better than the suspension concentrate (SC) formulation for downy brome injury. Pyroxasulfone applied PRE in the fall at a rate of 89 g&middot;ai&middot;ha-1 followed by (fb) imazamox (44 g&middot;ai&middot;ha-1 rate) applied postemergence (POST) in the spring effectively controlled downy brome (99% end-season injury). Furthermore, the injury was consistent with the standard program comprising of propoxycarbazone (29 g&middot;ai&middot;ha-1) PRE fb imazamox POST in IMI-tolerant winter wheat. In conclusion, pyroxasulfone applied PRE in the fall can be effectively utilized in conjunction with a standard acetolactate synthase (ALS)-inhibitor-based POST herbicide program for a season-long downy brome management in winter wheat.

Cucumber Downy Mildew: Research Progress and Registered Fungicides

Downy mildew is an important disease of cucumber,seriously threatening the production of cucumber.This article briefly described the symptoms and pathogen characteristics of cucumber downy mildew,the pathogenesis of the pathogen,plant disease resistance,pathogen drug resistance and integrated prevention and control techniques.In addition,officially registered fungicides,which are used against cucumber downy mildew,were summarized and categorized.We emphatically analyzed the active ingredients and formula combinations of the fungicides for controlling cucumber downy mildew,and the problems of the registered fungicides.The characteristics and control techniques of the newly registered fungicides for controlling cucumber downy mildew in the past two years are introduced.The problems existing in the research of cucumber downy mildew are discussed.Finally,the future research direction is put forward.

Effects of Microbial Pesticides on Prevention and Control of Cucumber Downy Mildew

This experiment was carried out to explore the effects of microbial pesticides on prevention and control of cucumber downy mildew.Three kinds of microbial pesticides( Recharge,Dynamic F,and 3. 0% amino-oligosaccharin AS) were used and clean water was taken as the control( CK). The experiment showed that Recharge and 3. 0% amino-oligosaccharin AS had better control effect on cucumber downy mildew.After the first application,the average control effect of Recharge and 3. 0% amino-oligosaccharin AS was higher than 50%. After the third application,the average control effect of Recharge and 3. 0% amino-oligosaccharin AS was higher than 20%. The control effect of Dynamic F on cucumber downy mildew was relatively poor,and the average control effect after the first application was 38. 34%,and the average control effect after the third application was only 15. 64%. According to the experimental results,the average control effect of three pesticides after application showed a gradual decline trend.

Effects of CO2 Enrichment on Yield,Soluble Solid Content and Downy Mildew of Greenhouse Grape

With‘Victoria'grape as a material,the effects of increasing 1200 mg/L of CO_2 on yield,soluble solid content and downy mildew of grape were studied in greenhouse.The results showed that when CO_2 was increased at a rate of 1200 mg/L in greenhouse,‘Victoria'fruit was harvested 8 d in advance,and compared with no increase of CO_2,the transverse diameter,longitudinal diameter,average single grain weight,commodity fruit yield at early stage and total yield of commodity fruit increased by 35.07%,7.37%,17.39%,24.88%,46.14% and 20.61%,respectively,with very significant differences.The incidence and disease index of grape downy mildew decreased by more than 50%,indicating a significant control effect.Therefore,increasing CO_2 in greenhouse could bring forward grape harvest,significantly improve yield and soluble solids content of grape and reduce the incidence of grape downy mildew.

Preliminary Studies on Induced Resistance of Cucumber to Downy Mildew

1 mL of sporangial suspensions (5 x 10 5spporangia per milliliter) of Pseudoperonospora cubensis was droplet-inoculated on the surface of the second leaf of the plant grown in greenhouse ( inducing inoculatoin), then the lower surfaces of the third, the forth and the fifth leaves were uniformly sprayed with inoculum of the same fungi (5 x 10 4sporangia per milliliter, about 5 mL per plant) every 3 days interval (challenge inoculation). Plants were moistened at 18- 22 C for 18 h, then kept at room temperature (24 - 28 C) and supplemented with cool-white fluorescent lights. All three challenge leaves were collected after 7 days of challenge to measure the amount of sporulation and area of necrosis. Plants prior inoculated with P. cubensis were protected 38% (based on the area of necrosis) against disaesc caused by subsequent foliar challenge with the pathogen. Protective action was about 12% after 3 days, and maintained this level until 9 days, suddenly reached 34% after 12 days, and came to a maximum after 15 days, then dropped down slowly.

Identif ication of a hypersensitive response core peptide of HrpZ and its role in increasing grape downy mildew resistance

Harpins play a key role in inducing disease resistance in crops,and identifying their core functional regions and establishing a system for their efficient expression would be very valuable.In this study,large amounts of soluble fusion proteins of harpin HrpZ and its subpeptides were obtained via the optimized induction conditions(28
C with 0.5 mmol$L1 IPTG for 6 h)in Escherichia coli BL21(DE3).Hypersensitive response(HR)assays demonstrated that the C-terminal 66 aa of HrpZ(HrpZ_C_2_2)elicited a strong HR in tobacco(Nicotiana benthamiana)and grape(Flame Seedless)leaves.Additionally,treatment with HrpZ,and particularly HrpZ_C_2_2,significantly reduced the disease incidence and severity index of field vine leaves and those inoculated with downy mildew.The determination of the physiological parameters indicated that HrpZ,and especially HrpZ_C_2_2,improved the photosynthesis-and chlorophyll fluorescence-related parameters,enhanced the activity of defense-related enzymes,including SOD,POD,CAT and PAL,and increased the H_(2)O_(2)level.Collectively,we efficiently expressed a core peptide of HrpZ and elucidated its strong ability to elicit a HR and resistance to downy mildew.This research provides insight into understanding the structure and function of HrpZ and will advance the application of HrpZ_C_2_2 to increase the resistance of grapevine to downy mildew.

Downy feather-like para-aramid fibers and nonwovens with enhanced absorbency,air filtration and thermal insulation performances

Fiber morphology with off-standing branches,as found in nature,e.g.,in goose downy feather,provides exquisite functions that can be barely achieved by man-made fiber systems.In this work,we develop a simple and scalable method for generating downy feather-like para-aramid fibers and assemblies.Through treating commercial para-aramid microfibers with mild alkaline solution(low concentration of NaOH),a synergistic effect of chemical hydrolysis and physical shearing is successfully triggered to generate abundant nanofiber branches on the surface of para-aramid fibers.When compared with conventional monotonous structures,nonwovens composed of downy feather-like fibers exhibit a typical multiscale fiber morphology,larger specific surface area and smaller pore size,thus showing enhanced particles adsorption capacity(over twice of the pristine nonwoven),excellent oil absorption capacity(increased by~50%),improved air filtration performances(doubled the filtration efficiency)and effective thermal insulation(thermal conductivity=26.1 mW·m^(−1)·K^(−1)).More attractively,the intrinsic flame-retardant nature of para-aramid is well inherited by the downy feather-like fibers,and the fabrication process requires neither sophisticated equipment,nor tedious procedures,making us believe the strong competitiveness of these fibers and assemblies.

TiO_2 photocatalytic antifungal technique for crops diseases control

TiO2 and 1% Ce^3＋-TiO2 prepared by hydrothermal method were characterized by means of XRD and UV-visible diffusive reflectance spectra（DRS）. The results of DRS analysis indicated that the 1% Ce^3＋-TiO2 catalyst had significant optical absorption in the visible region between 400-450 nm because electrons could be excited from the valence band of TiO2 or cerium oxides to Ce 4f level. To investigate the photocatalytic activity of different catalysts for crop fungal disease control, a series of Quine tests were carried out for cucumber powdery mildew and litchi downy blight control in home-made photocatalytic reactor. The results showed that TiO2 photocatalysis technique should be effective to inhibit the growth of fungal diseases to some extent and P-25 had a higher activity for antifungal control than home-made TiO2 catalysts. TiO2-A prepared with Ti（SO4）2 is more active to contro 1 antifungal disease than TiO2-B prepared with TiOSO4 as precursor because the crystalline of TiO2-A was higher than that of TiO2-B. The antifungal index litchi downy blight control was greatly enhanced by doping 1% cerium ion. The antifungal index of 1.0%Ce3^＋-TiO2 was（47.0 ± 4.7）%, （82.2 ± 3.5）%, （100 ±0.0）% under indoor weaker light, solar light and black light, respectively. The results of field experiments showed that the antifungal index of 1.0%Ce^3＋-TiO2 was more than that of P-25. The antifungal index of 1.0%Ce^3＋-TiO2 was （81.7 ± 6.5）%, （67.5 ±4.7）%, （38.6±1.9）% for litchi downy blight, maize southern leaf spot, and rice blast, respectively. It was concluded that TiO2 photocatalysis technique should be an effective way for litchi fungal disease control in practice.

Development of real-time onion disease monitoring system using image acquisition

In this study, real-time disease monitoring was conducted on onion which is the most representative crop in Republic of Korea, using an image acquisition system newly developed for the mobile measurement of phenotype. The purpose of this study was to improve the accuracy of prediction of disease and state variables by processing images acquired from monitoring. The image acquisition system was consisted of two parts, a motorized driving system and a PTZ(pan, tilt and zoom) camera to take images of the plants. The acquired images were processed as follows. Noise was removed through an image filter and RGB(red, green and blue) colors were converted to HSV(hue, saturation and value), which enabled thresholding of areas with different colors and properties for image binarization by comparing the color of onion leaf with ambient areas. Four objects with the most significant browning in the onion leaf to the naked eye were selected as the samples for data acquired. The thresholding method with image processing was found to be superior to the naked eye in identifying accurate disease areas. In addition, it was found that the incidence of disease was different in each disease area ratio. As a result, the use of image acquisition system in image processing analysis will enable more prompt detection of any changes in the onion and monitoring of disease outbreaks during the crop lifecycle.