Development and molecular cytogenetic identification of a new wheat-rye 6RL ditelosomic addition and 1R(1B)substitution line with powderymildew resistance

Powdery mildew is a serious disease caused by Blumeria graminis f.sp.tritici(Bgt)that critically threatens the yield and quality of wheat(Triticum aestivum L.).Using effective powdery mildew resistance genes is the optimal method for controlling this disease.Against the background of high genetic homogeneity among the modern commercial cultivars that are mainly derived from conventional interbreeding,the resistance genes from wheat relatives have especially prominent advantages.Octoploid triticale,produced from common wheat and rye(Secale cereale L.)through distant hybridization,is a new synthetic species and valuable gene donor for wheat improvement.In this study,we developed the wheat-rye line YT5 through the hybridization of octaploid triticale and two wheat lines.YT5 was confirmed to be a 6RL ditelosomic addition and 1R(1B)substitution line using genomic in situ hybridization(GISH),multicolor fluorescence in situ hybridization(mc-FISH),multicolor GISH(mc-GISH)and molecular marker analysis.Genetic analysis showed that the powdery mildew resistance in YT5 was derived from the rye chromosome arm 6RL.After inoculation with different Bgt isolates at the seedling stage,YT5 had compound reaction patterns with both obvious spores and hypersensitivity,and it gradually became highly resistant until the adult-plant stage,thus showing a resistance response significantly different from the reported Pm genes from rye chromosome 6RL.YT5 also showed promising agronomic performance,so it is expected to be an elite resistance donor for wheat improvement.To promote the transfer of the chromosome arm 6RL of YT5 in marker-assisted selection(MAS)breeding,we selected and verified two 6RL-specific kompetitive allelespecific PCR(KASP)markers that can be applied to efficiently detect this chromosome arm in different wheat backgrounds.

Development and identification of two novel wheat-rye 6R derivative lines with adult-plant resistance to powdery mildew and high-yielding potential

Powdery mildew,caused by Blumeria graminis f.sp.tritici(Bgt),is a devastating disease that seriously threatens wheat yield and quality.To control this disease,host resistance is the most effective measure.Compared with the resistance genes from common wheat,alien resistance genes can better withstand infection of this highly variable pathogen.Development of elite alien germplasm resources with powdery mildew resistance and other key breeding traits is an attractive strategy in wheat breeding.In this study,three wheat-rye germplasm lines YT4-1,YT4-2,and YT4-3 were developed through hybridization between octoploid triticale and common wheat,out of which the lines YT4-1 and YT4-2 conferred adult-plant resistance(APR)to powdery mildew while the line YT4-3 was susceptible to powdery mildew during all of its growth stages.Using genomic in situ hybridization,multi-color fluorescence in situ hybridization,multi-color GISH,and molecular marker analysis,YT4-1,YT4-2,and YT4-3 were shown to be cytogenetically stable wheat-rye 6R addition and T1RS.1BL translocation line,6RL ditelosomic addition and T1RS.1BL translocation line,and T1RS.1BL translocation line,respectively.Compared with previously reported wheat-rye derivative lines carrying chromosome 6R,YT4-1 and YT4-2 showed stable APR without undesirable pleiotropic effects on agronomic traits.Therefore,these novel wheat-rye 6R derivative lines are expected to be promising bridge resources in wheat disease breeding.

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℃ with 0.5 mmol·L^(-1) 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.

Fine mapping of the powdery mildew resistance gene PmXQ-0508 in bread wheat

In a wheat breeding line XQ-0508 showing consistent resistance to powdery mildew disease,a recessive gene,designated PmXQ-0508,was identified and mapped to a distal region on chromosome arm 2BS.Of three resistance-associated genes in this region,one encoding a protein kinase was selected as the primary candidate for PmXQ-0508.Ten closely linked DNA markers developed in the study could be used for marker-assisted selection for powdery-mildew resistance in breeding programs.

Occurrence Regularity and Comprehensive Prevention and Control Techniques of Sunflower Downy Mildew

This paper examines the occurrence regularity and comprehensive prevention and control techniques for sunflower downy mildew.It provides a detailed discussion of the pathogens,symptoms,and associated risks,as well as the transmission pathways,underlying causes,and prevention and control techniques related to sunflower downy mildew.The aim is to offer valuable references and technical guidance for the effective management of this disease.

The Relationship of Methyl Jasmonate Enhanced Powdery Mildew Resistance in Wheat and the Expressions of 9 Disease Resistance Related Genes

[Objective] This study was carried out to determine the induction effect of jasmonic acid（JA）on powdery mildew resistance in wheat,the activation effect on the expressions of plant disease resistance related genes,and to investigate the relationship between the induced resistance and the gene expression patterns.[Method] Three powdery mildew susceptible cultivars of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂ typically representing different phenotypes in the field were employed.The powdery mildew was assessed by detached leaf assay,and real time quantitative RT-PCR was used to determine the expression patterns of 9 disease resistance related genes of PR1（PR1.1）,PR2（β,1-3 glucanase）,PR3（chitinase）,PR4（wheatwin1）,PR5（thaumatin-like protein）,PR9（TaPERO,peroxidase）,PR10,TaGLP2a（germin-like）and Ta-JA2（jasmonate-induced protein）in leaf of the three cultivars.[Result] MeJA application enhanced the powdery mildew resistances of ＂Chinese Spring＂,＂Pumai 9＂ and ＂Zhoumai 18＂.The induced powdery mildew resistance could be detected from 12 h to 96 h after MeJA treatment,and the peak value was at 24 h.Though there were differences between the three cultivars,MeJA significantly effect on the expressions of the 8 disease resistance related genes except TaGLP2a,and the peak values were at 12 h,24 h or 48 h after treatments.The strongest activation of MeJA was on PR9 and PR1 that their expressions could reach more than 100 times of the untreated samples.MeJA strongly activated PR2、PR4、PR5、PR3、PR10 and Ta-JA2,their expression could reach 10 to 70 times,and there was almost no activation effect on TaGLP2a.The induced powdery mildew resistance positively correlated with the induced expressions of the 8 disease related genes.[Conclusion] The induced powdery mildew resistance positively correlated with the induced expressions of the disease related genes.Jasmonate signalling plays a role in defence against Blumeria graminis f.sp.tritici.and future manipulation of this pathway may improve powdery mildew resistance in wheat.

Field Efficacy of Six Kinds of Fungicides for Control and Prevention of Apple Powdery Mildew(Podosphaera leucotricha)

[Objective] This study aimed to screen effective fungicide for prevention and control of apple powdery mildew. [Method] Field efficacy of six kinds of fungi- cides with 18 concentrations against apple powdery mildew was investigated. [Re- sult] 4 000-fold triflumizole WP exhibited the highest control efficacy of 82.96%, fol- lowed by 3 000-fold triflumizole WP with control efficacy of 77.75%, while 800-fold and 1 000-fold 70% thiophanate methyl WP exhibited relatively low control efficacy, which was both lower than 60%. [Conclusion] In actual production, 4 000-fold triflu- mizole WP and 3 000-fold triflumizole WP should be used alternately to control and prevent apple powdery mildew.

Toxicity Test and Field Control Effects of 4 Different Fungicides on Grape Downy Mildew

[Objective] This study was conducted to investigate the biocontrol activity of Streptomyces corchorusii strain NF0919 and Bacillus subtilis D J-6 WP to grape downy mildew. [Methed] We determined the indoor toxicity of the supernatant of S. corchorusii strain NF0919, 1.0×1011 cfu/g B. subtilis D J-6 WP, mancozeb and dimethomorph on Plasmopara viticola by the leaf disc method, respectively, and a field efficacy trial was conducted. [Result] The results showed that the ECso values of the supernatant of strain NF0919, 1.0×1011cfu/g B. subtilis D J-6 WP, mancozeb and dimethomorph were 96.285 9, 86.603 8, 69.947 2 and 7.263 6 μg/ml, respec- tively. The values of field efficacy in preventive experiments for grape downy mildew on the 7th day after 2 times of spraying 20 times diluent of the supernatant of strain NF0919 and 1 000 times diluent of 1.0×1011 cfu/g B. subtilis D J-6 WP were 71.55% and 70.71%, respectively, and the values of field efficacy on the 14th day after the 2 times of fungicide application were 67.54% and 68.19%, respectively. The values of field efficacy in curative experiments on the 7th day after 2 times of spraying 20 times diluent of the supematant of strain NF0919 and 1 000 times diluent of 1.0×1011 cfu/g B. subtilis D J-6 WP were 59.72% and 56.07%, respectively, and the val- ues of field efficacy on the 14th day after the 2 times of fungicide application were 56.88% and 57.46%, respectively. The field efficacy values of the 2 tested biocon- trol agents were equivalent. The protective effect showed no significant difference between each of tested biocontrol agents and 300 times diluent of 50% mancozeb WP, but there was a significant difference in the efficacy between each of tested biocontrol agents and 200 times diluent of 40% dimethomorph SC. [Conclusion] The S. corchorusii strain NF0919 and B. subtilis D J-6 WP had certain biocontrol poten- tial to grape downy mildew and development value.

Molecular Detection of Resistance Genes to Stripe Rust and Powdery Mildew in Common Wheat Cultivar Yunmai52

Yunmai52, developed by crossing with common wheat-Haynaldia villosa6AL/6VS translocation line 92R149 as a resistant parent in 1992, was a common wheat cultivar approved and released in 2007 in Yunnan Province, China, which is characterized by high resistance to powdery mildew and stripe rust. In this study,an F_2 population derived from a cross K78S/Yunmai52 was constructed to investigate the resistance genes, where K78 S is a wheat male sterile line susceptible to powdery mildew and stripe rust. Phenotypic identification of the parents, F_1 and F_2 populations and chi-square analyses showed that F_1 population was immune to stripe rust and powdery mildew; the segregation ratio of resistance and susceptibility to powdery mildew（χ~2=1.10χ~2_（1,0.05）=3.84） and stripe rust（χ~2=0.15χ~2_（1,0.05）=3.84） fit to a 3：1 ratio in F_2 population, indicating that Yunmai52 harbors a dominant stripe rust resistance gene and a dominant powdery mildew resistance gene. The individuals were further detected with a marker co-segregated with Pm21（SCAR_（1400）） and two markers closely linked with Yr26（XWe173 and Xbarc181）. The results showed that polymorphic bands could be amplified between the parents and between resistance and susceptibility gene pools at the same locus. Randomly 96 individuals of F_2 population were selected for verification. The results showed that the phenotype was significantly correlated with the genotype. The detection accuracy of markers SCAR_（1400）, XWe173 and Xbarc181 was 100%, 97.91% and 92.70%, respectively.Yunmai52 harbored powdery mildew resistance gene Pm21 and stripe rust resistance gene Yr26, which were both derived from 6AL/6VS translocation line 92R149.In addition, the results also demonstrate that Pm21 and Yr26 are two genes conferring durable resistance to powdery mildew and stripe rust in wheat.

Irradiation Dose for the Insect Disinfestation and Mildew Control of Peanut

The bulk peanuts were used as the materials. After ）,-ray irradiation of different doses, the peanuts were stored up at room temperature, and the degree of pest infection and gradient of mould infection were observed regularly. The results showed that insects began to appear in the unirradiated peanuts in 2 months, while after 4 months, the pest infection rate increased with the prolonged storage time, and the pest infection rate reached up to 58%-100% after 6 months of storage. However, the irradiated peanuts did not suffer from the pest infection throughout the storage. The unirradiated peanuts were contaminated by moulds after three months of storage, while the peanuts with an irradiation dose of 0.3-0.5 kGy were mildewed after 6 months of storage, while those with an irradiation dose of 1.0, 4.0 kGy had no mould after 12 months of storage. Combined with relevant international and national standards as well as the findings of scholars at home and abroad, the minimum effective irradiation dose for insect disinfestations of peanuts was 0,3 kGy, and the minimum effective irradiation dose for mildew control was 1.0 kGy, while the maximum tolerance dose was 4.0 kGy.

Studies on Photosynthetic Characteristics of Tomato Leaves after Inoculation with Tomato Powdery Mildew (Oidium neolycopersici)

[ Objective ] The paper was to explore the pathogenic mechanism of tomato powdery mildew, and to study the effects of the disease on photosynthetic characteristics of tomato. [ Method ] With four tomato varieties as materials, the pathogen of tomato powdery mildew was artificially inoculated. After the varieties were infected, the parameters including net photosynthetic rate, stomatal conductance and transpiration rate of tomato leaf were measured by Li-6400 portable photo- synthesis detector under natural lighting conditions. [ Result] The net photosynthetic rate, stomatal conductance and transpiration rate of four tomato varieties all decreased after infection. However, the decrease extent of these parameters of four varieties was different. The parameters of seriously damaged Jinyangdajuxdng （ No. 4） and Xinsheng No. 1 （ No. 5 ） decreased greatly, while the parameters of slightly damaged Lujia （ No. 13 ） and improved 98-6 decreased lightly. [ Condu- sion] The results could provide theoretical basis for the study on pathogenic mechanism, new prevention way and resistance breeding of tomato powdery mildew.

Construction of Cucumber Powdery Mildew Early Warning System in Solar Greenhouse Based on Internet of Things

ln order to explore the design and construction of cucumber powdery mildew warning system in solar greenhouse, internet of things technology was used to conduct the real-time dynamic monitoring of the incidence of cucumber powdery mildew and cucumber growth environment in solar greenhouse. The growth environ-ment included temperature and humidity of air and soil. Logistic regression model was used to construct cucumber powdery mildew warning model. The results showed that humidity characteristic variable （maximum air humidity） and temperature characteristic variable （maximum air temperature） had significant effects on the inci-dence probability of cucumber powdery mildew in solar greenhouse. And it was fea-sible to construct cucumber powdery mildew warning system in solar greenhouse with internet of things.

Comparison of hydroxyl radical-scavenging abilities between white rot fungus Lentinula edodes and five mildews on lignocellulose substance

The hydroxyl radical（-OH）-scavenging ability of culture filtrates from submerged culture of Lentinula edodes AX3 and five mildews on lignocellulose substance was analysed. Only L. edodes AX3 showed significant -OH-scavenging ability which reached 52.2% at about the 48th hour. All mildews could hardly scavenge -OH under the experimental conditions. -OH-scavenging ability is considered related to the mode and ability of lignocellulose degradation of a strain. The degradation or bioconversion products might be the substance base for.this effect.

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.

Control Effect of Vegetable Oil on Cucumber Powdery Mildew

[Objective] The study discussed the indoor activity and field control effect of vegetable oil on cucumber powdery mildew.[Method] The cucumber seedlings of Changchunmici were adopted as the material,which were inoculated with the fungus pathogen of cucumber powdery mildew naturally occurred in the field.The indoor protective test and field test were performed respectively to observe the control effects of different plant oils EC on the cucumber powdery mildew.[Result] When six plant oils EC including cottonseed oil,soybean oil,canola oil,corn oil,sesame oil and sunflower oil was diluted into 10 and 5 ml/L,they had better control effect on cucumber powdery mildew.As for the indoor protective effect,the protective effect of the vegetable oil EC was equivalent to the control agent triadimefon EC,the field control effect of vegetable oil EC was in the range of 60%-75%,which was consistent with 69%-70% control effect of contrast agent with 1 ml/L triadimefon EC dilution.[Conclusion] All vegetable oils EC had significant control effect on cucumber powdery mildew in the test.

Prevention of 25% Azoxystrobin Suspending Agent to Cucumber Downy Mildew

[Objective] The aim was to research prevention effects of 25％ azoxys trobin suspending agent to cucumber downy mildew.[Method] The test,including control group and four treatment groups with 450,600,and 750 g/hm2 25％ azoxystrobin suspending agents,and 80％ mancozeb wettable powder,was carried out in 2012 and 2013 consecutively to test the prevention effects of 25％ azoxystrobin suspending agent to cucumber downy mildew.[Result] The prevention effects were significant of 25％ azoxystrobin suspending agent to cucumber downy mildew in the range of 70.0％-83.6％,and it is recommended 25％ azoxystrobin suspending agent at 600 g/hm2 be spread before plants diseased or in initial stage.In the whole stage,it was spread once every 7 d,totaling 3 times.[Conclusion] The research provides references for wide application of 25％ azoxystrobin suspending agent in production.

Genetics,resistance mechanism,and breeding of powdery mildew resistance in cucumbers(Cucumis sativus L.)

Cucumber is an important vegetable worldwide,and powdery mildew(PM)is a common and serious disease of cucumbers.Breeding disease-resistant cucumber varieties is the most advantageous strategy to control this disease.In recent years,exploration and identification of cucumber PM resistance genes have achieved great advancement,and many genes have been cloned and verified using different methods.However,the resistance mechanism of cucumber PM is still unclear,and many ambiguities need to be elucidated urgently.In this review,we summarized the research advances in PM resistance in cucumbers,including genetic analysis,quantitative trait locus mapping,map-based cloning,transcriptomics,mlo-mediated PM resistance,and mining of noncoding RNAs involved in resistance.Finally,the research directions and the problems that need to be solved in the future were discussed.

Spectroscopic Leaf Level Detection of Powdery Mildew for Winter Wheat Using Continuous Wavelet Analysis

Powdery mildew （Blumeria graminis） is one of the most destructive crop diseases infecting winter wheat plants, and has devastated millions of hectares of farmlands in China. The objective of this study is to detect the disease damage of powdery mildew on leaf level by means of the hyperspectral measurements, particularly using the continuous wavelet analysis. In May 2010, the reflectance spectra and the biochemical properties were measured for 114 leaf samples with various disease severity degrees. A hyperspectral imaging system was also employed for obtaining detailed hyperspectral information of the normal and the pustule areas within one diseased leaf. Based on these spectra data, a continuous wavelet analysis （CWA） was carried out in conjunction with a correlation analysis, which generated a so-called correlation scalogram that summarizes the correlations between disease severity and the wavelet power at different wavelengths and decomposition scales. By using a thresholding approach, seven wavelet features were isolated for developing models in determining disease severity. In addition, 22 conventional spectral features （SFs） were also tested and compared with wavelet features for their efficiency in estimating disease severity. The multivariate linear regression （MLR） analysis and the partial least square regression （PLSR） analysis were adopted as training methods in model mildew on leaf level were found to be closely related with the development. The spectral characteristics of the powdery spectral characteristics of the pustule area and the content of chlorophyll. The wavelet features performed better than the conventional SFs in capturing this spectral change. Moreover, the regression model composed by seven wavelet features outperformed （R2=0.77, relative root mean square error RRMSE=0.28） the model composed by 14 optimal conventional SFs （R2---0.69, RRMSE--0.32） in estimating the disease severity. The PLSR method yielded a higher accuracy than the MLR method. A combination of CWA and PLSR was found to be promising in providing relatively accurate estimates of disease severity of powdery mildew on leaf level.

Identification of Co-Segregating RAPD Marker Linked to Powdery Mildew Resistance Gene Pm18 in Wheat

The Pm18 gene of wheat confers resistance to the powdery mildew which is one of the mostserious diseases in many regions of the world. In this study, bulked segregant analysis(BSA) was used to develop randomly amplified polymorphic DNA (RAPD) markers linked toPm18 gene. Three hundred and twenty decamer primers were screened and one of them wasidentified as RAPD marker (S411600) linked to Pm18. Using the F2 mapping population fromthe cross Pm18Chancellor, the marker S411600 was shown to co-segregate with the genePm18. This marker can be conveniently used for marker-assisted selection in wheatbreeding programs for the identification or pyramiding of Pm18 with other resistancegenes.

Inheritance of Powdery Mildew Resistance in Cucumber (Cucumis sativus L.) and Development of an AFLP Marker for Resistance Detection

Cucumber powdery mildew is one of the most destructive diseases of cucumber throughout the world. In the present study, inheritance of powdery mildew resistance in three crosses, and linkage of resistance with amplified fragment length polymorphism （AFLP） markers are studied to formulate efficient strategies for breeding cultivars resistant to powdery mildew. The joint analysis of multiple generations and AFLP technique has been applied in this study. The best model is the one with two major genes, additive, dominant, and epistatic effects, plus polygenes with additive, dominant, and epistatic effects （E-l-0 model）. The heritabilities of the major genes varied from 64.26% to 97.82%, and susceptibility was incompletely dominant for the two major genes in the three crosses studied. The additive effects of the two major genes and the dominant effect of the second major gene were high, and the epistatic effect of the additive-dominant between the two major genes was the highest in cross I . In cross II, the absolute value of the additive effect, dominant effect, and potential ratio of the first major gene were far higher than those of the second major gene, and the epistatic effect of the additive-additive was the highest. The genetic parameters of the two major genes in cross III were similar to those in cross II. Correlation and regression analyses showed that marker E25/M63-103 was linked to a susceptible gene controlling powdery mildew resistance. The marker could account for 19.98% of the phenotypic variation. When the marker was tested on a diverse set of 29 cucumber lines, the correlation between phenotype and genotype was not significant, which suggested cultivar specialty of gene expression or different methods of resistance to powdery mildew. The target DNA fragment was 103 bp in length, and only a small part was found to be homologous to DNA in the other species evaluated, which indicated that it was unique to the cucumber genome.