Mapping Powdery Mildew Resistance Gene pmYBL on Chromosome 7B of Chinese Wheat (Triticum aestivum L.) Landrace Youbailan.

Chinese wheat landrace Youbailan has excellent resistance to powdery mildew caused by Blumeria graminis f. sp. tritici. In the present study, genetic analysis indicated that a recessive gene, tentatively designated pmYBL, was responsible for the powdery mildew resistance of Youbailan. pmYBL was located in the 695-to-715-Mb genomic region of chromosome 7BL, with 19 gene-linked single-nucleotide polymorphism (SNP) markers. It was flanked by SNP1-12 and SNP1-2 with genetic distances of 0.6 and 1.8 centimorgans, respectively. The disease reaction patterns of Youbailan and four cultivars (lines) carrying the powdery mildew (Pm) genes located on chromosome arm 7BL indicated that pmYBL may be allelic or closely linked to these genes. All of the SNP markers linked to pmYBL were diagnostic, indicating that these markers will be useful for pyramiding pmYBL using marker-assisted selection.

Correction Strategy of Mortars with Trajectory Correction Fuze Based on Image Sensor.

For a higher accuracy of projectiles, a novel trajectory correction fuze is proposed. In this design, the sensor and actuator were reduced to achieve a balance between performance and affordability. Following introduction of the fuze concept, the flight model was presented and the crossrange and downrange components of trajectory response under control were investigated. The relationship between the inertial coordinate system and the detector coordinate system was studied so that the imager feedback could be used to derive the actual miss distance. The deployment time of canards and roll angle of the forward fuze were derived and used as the inputs of the control system in this strategy. Example closed-loop simulations were implemented to verify the effectiveness of the strategy. The results illustrate that the accuracy increase is evident and the proposed correction concept is applicable for terminal correction of mortars.

Effects of Climate Change on Epidemics of Powdery Mildew in Winter Wheat in China.

Powdery mildew is a highly destructive winter wheat pathogen in China. Since the causative agent is sensitive to changing weather conditions, we analyzed climatic records from regions with previous wheat powdery mildew epidemics (1970 to 2012) and investigated the long-term effects of climate change on the percent acreage (PA) of the disease. Then, using PA and the pathogen's temperature requirements, we constructed a multiregression model to predict changes in epidemics during the 2020s, 2050s, and 2080s under representative concentration pathways RCP2.6, RCP4.5, and RCP8.5. Mean monthly air temperature increased from 1970 to 2012, whereas hours of sunshine and relative humidity decreased (P < 0.001). Year-to-year temperature changes were negatively associated with those of PA during oversummering and late spring periods of disease epidemics, whereas positive relationships were noted for other periods, and year-to-year changes in relative humidity were correlated with PA changes in the early spring period of disease epidemics (P < 0.001). Our models also predicted that PA would increase less under RCP2.6 (14.43%) than under RCP4.5 (14.51%) by the 2020s but would be higher by the 2050s and 2080s and would increase least under RCP8.5 (14.37% by the 2020s). Powdery mildew will, thus, pose an even greater threat to China's winter wheat production in the future.

Development of Weather- and Airborne Inoculum-Based Models to Describe Disease Severity of Wheat Powdery Mildew.

Disease severity of wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, was recorded weekly in fungicide-free field plots for three successive seasons from 2009 to 2012 in Langfang City, Hebei Province, China. Airborne conidia of B. graminis f. sp. tritici were trapped using a volumetric spore sampler, and meteorological data were collected using an automatic weather station. Cumulative logit models were used to relate the development of wheat powdery mildew to weather variables and airborne conidia density. Density of airborne conidia was the most important variate; further addition of weather variables, although statistically significant, increased model performance only slightly. A model based on variables derived from temperature and humidity had a generalized R2 of 72.4%. Although there were significant differences in model parameters among seasons, fine adjustment did not increase model performance significantly.

Fusarium pseudograminearum biomass and toxin accumulation in wheat tissues with and without Fusarium crown rot symptoms.

Fusarium crown rot (FCR) is an important and devastating disease of wheat (Triticum aestivum) caused by the fungus Fusarium pseudograminearum and related pathogens. Using two distinct susceptible cultivars, we investigated the isolation frequencies of F. pseudograminearum and quantified its biomass accumulation and the levels of the associated toxins deoxynivalenol (DON) and DON-3-glucoside (D3G) in inoculated field-grown wheat plants. We detected F. pseudograminearum in stem, peduncle, rachis, and husk tissues, but not in grains, whereas DON and D3G accumulated in stem, rachis, husk, and grain tissues. Disease severity was positively correlated with the frequency of pathogen isolation, F. pseudograminearum biomass, and mycotoxin levels. The amount of F. pseudograminearum biomass and mycotoxin contents in asymptomatic tissue of diseased plants were associated with the distance of the tissue from the diseased internode and the disease severity of the plant. Thus, apparently healthy tissue may harbor F. pseudograminearum and contain associated mycotoxins. This research helps clarify the relationship between F. pseudograminearum occurrence, F. pseudograminearum biomass, and mycotoxin accumulation in tissues of susceptible wheat cultivars with or without disease symptoms, providing information that can lead to more effective control measures.

Characterization of the sterol 14α-demethylases of Fusarium graminearum identifies a novel genus-specific CYP51 function.

CYP51 encodes the cytochrome P450 sterol 14α-demethylase, an enzyme essential for sterol biosynthesis and the target of azole fungicides. In Fusarium species, including pathogens of humans and plants, three CYP51 paralogues have been identified with one unique to the genus. Currently, the functions of these three genes and the rationale for their conservation within the genus Fusarium are unknown. Three Fusarium graminearum CYP51s (FgCYP51s) were heterologously expressed in Saccharomyces cerevisiae. Single and double FgCYP51 deletion mutants were generated and the functions of the FgCYP51s were characterized in vitro and in planta. FgCYP51A and FgCYP51B can complement yeast CYP51 function, whereas FgCYP51C cannot. FgCYP51A deletion increases the sensitivity of F. graminearum to the tested azoles. In ΔFgCYP51B and ΔFgCYP51BC mutants, ascospore formation is blocked, and eburicol and two additional 14-methylated sterols accumulate. FgCYP51C deletion reduces virulence on host wheat ears. FgCYP51B encodes the enzyme primarily responsible for sterol 14α-demethylation, and plays an essential role in ascospore formation. FgCYP51A encodes an additional sterol 14α-demethylase, induced on ergosterol depletion and responsible for the intrinsic variation in azole sensitivity. FgCYP51C does not encode a sterol 14α-demethylase, but is required for full virulence on host wheat ears. This is the first example of the functional diversification of a fungal CYP51.

The Quantitative Analyses for the Effects of Two Wheat Varieties With Different Resistance Levels on the Fungicide Control Efficacies to Powdery Mildew.

Effective strategies to reduce the occurrence of wheat powdery mildew include the use of resistant varieties and application of fungicides. However, most studies rarely focus on the quantitative value of fungicide reduction using resistant varieties. To explore how the fungicides performed on different resistant wheat varieties to powdery mildew, field experiments were conducted during wheat growing seasons in 2018/19 and 2019/20 to investigate the control efficacies of enostroburin⋅epoxiconazole 18% SC and triadimefon 20% EC to wheat powdery mildew on a highly resistant wheat variety ("Baofeng104") and a highly susceptible wheat variety ("Jingshuang16"). The analyses of variance on control efficacies showed that the control efficacies of enostroburin⋅epoxiconazole 18% SC to wheat powdery mildew were mostly significantly higher than triadimefon 20% EC under the same conditions (i.e., varieties, dosages). However, both fungicide and variety resistance made variabilities in the mildew disease index and played a significant role in mildew management. Particularly, the variety resistance made the greatest contribution in mildew-reducing, and the disease index could significantly be reduced on the highly resistant variety even in the absence of fungicide treatment. The control efficacies to mildew on the highly susceptible variety mainly depended on the high efficiency of fungicides whereas the highly resistant variety were mainly by virtue of variety resistance through the comparative analyses of linear regression models. Furthermore, the random-coefficient regression models and quantile models quantificationally expounded that the relationships between active ingredient dosage and disease index or control efficacy varied from the effects of variety, fungicide, and year, particular from variety. Thus, a dosage reference table of enostroburin⋅epoxiconazole 18% SC or triadimefon 20% EC for different resistant wheat varieties were provided; it would be helpful for users to formulate an appropriate dosage of fungicide on mildew management in the field and avoid overusing or superfluous application. Further study needs to consider the effects of fungicide reduction on wheat yields, only then the maximum-economic benefits on mildew management can be determined.

Key infection stages defending heat stress in high-temperature-resistant Blumeria graminis f. sp. tritici isolates.

With the increase of temperature in the winter wheat-growing regions in China, the high-temperature-resistant Blumeria graminis f. sp. tritici (Bgt) isolates developed in the fields. To clarify the key infection stages and the roles of heat shock protein (HSP) genes of high-temperature-resistant Bgt isolates defending high temperature, 3 high-temperature-resistant and 3 sensitive Bgt isolates were selected from 55 isolates after determination of temperature sensitivity. And then they were used to investigate the infection stages and the expression levels of HSP genes, including Bgthsp60, Bgthsp70, Bgthsp90, and Bgthsp104, at 18°C and 25°C. The formation frequency of abnormal appressoria and inhibition rate of haustoria formation of high-temperature-resistant isolates at 25°C were lower than those of high-temperature-sensitive isolates, while major axis of microcolonies of high-temperature-resistant isolates was higher than those of high-temperature-sensitive isolates at 25°C. The results indicated that haustoria formation and hyphal expansion were the key infection stages of defense against heat stress in high-temperature-resistant isolates. Further analyses of HSP genes found the expression levels of Bgthsp60 and Bgthsp70c were upregulated at 24 and 72 h post-inoculation in high-temperature-resistant isolates, while no significant difference was observed for Bgthsp90 and Bgthsp104 genes. Taken together, the basis of high-temperature-resistant Bgt isolates is associated with induced expression of Bgthsp60 and Bgthsp70c response to heat stress in haustoria formation and hyphal expansion stages.

Involvement of NO and Ca2+ in the enhancement of cold tolerance induced by melatonin in winter turnip rape (Brassica rapa L.).

As a multifunctional phytohormone, melatonin (Mel) plays pivotal roles in plant responses to multiple stresses. However, its mechanism of action remains elusive. In the present study, we evaluated the role of NO and Ca2+ signaling in Mel enhanced cold tolerance in winter turnip rape. The results showed that the NO content and concentration of intracellular free Ca2+ ([Ca2+]cyt) increased by 35.42% and 30.87%, respectively, in the leaves of rape seedlings exposed to cold stress. Compared with those of the seedlings in cold stress alone, the NO content and concentration of [Ca2+]cyt in rape seedlings pretreated with Mel increased further. In addition, the Mel-mediated improvement of cold tolerance was inhibited by L-NAME (a NO synthase inhibitor), tungstate (a nitrate reductase inhibitor), LaCl3 (a Ca2+ channel blocker), and EGTA (a Ca2+ chelator), and this finding was mainly reflected in the increase in ROS content and the decrease in osmoregulatory capacity, photosynthetic efficiency and antioxidant enzyme activities, and expression levels of antioxidant enzyme genes. These findings suggest that NO and Ca2+ are necessary for Mel to improve cold tolerance and function synergistically downstream of Mel. Notably, the co-treatment of Mel with L-NAME, tungstate, LaCl3, or EGTA also inhibited the Mel-induced expression of MAPK3/6 under cold stress. In conclusion, NO and Ca2+ are involved in the enhancement of cold tolerance induced by Mel through activating the MAPK cascades in rape seedlings, and a crosstalk may exist between NO and Ca2+ signaling.

Molecular Cytogenetic Identification of a New Wheat-Rye 6R Addition Line and Physical Localization of Its Powdery Mildew Resistance Gene.

Rye (Secale cereale L.), a naturally cross-pollinating relative of wheat, is a tertiary gene donor and of substantial value in wheat improvement. Wheat powdery mildew is caused by Blumeria graminis f. sp. tritici (Bgt), which seriously affects yield and quality worldwide. Identifying and transferring new, effective resistance genes against powdery mildew from rye is important for wheat breeding. The current study developed a wheat-rye line YT2 resistant to powdery mildew by crossing, backcrossing, and self-pollination for multiple generations between octoploid triticale 09R2-100 and common wheat cultivar Shixin 616. YT2 was confirmed to be a 6R disomic addition and T1RS⋅1BL translocation line by genomic in situ hybridization (GISH), multicolor fluorescence in situ hybridization (mc-FISH), multicolor-GISH (mc-GISH), and molecular marker analyses. Disease responses to different Bgt isolates and genetic analysis showed that the powdery mildew resistance gene of YT2 was derived from the rye chromosome 6R of 09R2-100, which differed from the previously reported Pm genes from rye including Pm20 on 6RL. Resistance phenotype of different translocation lines and deletion lines derived from YT2 combined with newly developed 6RL-specific markers analysis suggested that the powdery mildew resistance gene of YT2 was localized to the region in chromosome 6RL: 890.09-967.51 Mb and flanked by markers XM189 and X4M19, corresponding to the reference genome of Weining rye. Therefore, YT2 could be used as a promising bridging parent for wheat disease resistance improvement.

PM2b, a CC-NBS-LRR protein, interacts with TaWRKY76-D to regulate powdery mildew resistance in common wheat.

Powdery mildew caused by Blumeria graminis f. sp. tritici (Bgt) is a destructive disease of wheat throughout the world. Host resistance is considered the most sustainable way to control this disease. Powdery mildew resistance gene Pm2b was mapped to the same genetic interval with Pm2a and PmCH1357 cloned previously, but showed different resistance spectra from them, indicating that they might be caused by different resistance genes or alleles. In this study, Pm2b was delimited to a 1.64 Mb physical interval using a large segregating population containing 4,354 F2:3 families of resistant parent KM2939 and susceptible cultivar Shimai 15. In this interval, TraesCS5D03G0111700 encoding the coiled-coil nucleotide-binding site leucine-rich repeat protein (CC-NBS-LRR) was determined as the candidate gene of Pm2b. Silencing by barley stripe mosaic virus-induced gene silencing (BSMV-VIGS) technology and two independent mutants analysis in KM2939 confirmed the candidate gene TraesCS5D03G0111700 was Pm2b. The sequence of Pm2b was consistent with Pm2a/PmCH1357. Subcellular localization showed Pm2b was located on the cell nucleus and plasma membrane. Pm2b had the highest expression level in leaves and was rapidly up-regulated after inoculating with Bgt isolate E09. The yeast two-hybrid (Y2H) and luciferase complementation imaging assays (LCI) showed that PM2b could self-associate through the NB domain. Notably, we identified PM2b interacting with the transcription factor TaWRKY76-D, which depended on the NB domain of PM2b and WRKY domain of TaWRKY76-D. TaWRKY76-D negatively regulated the resistance to powdery mildew in wheat. The specific KASP marker K529 could take the advantage of high-throughput and high-efficiency for detecting Pm2b and be useful in molecular marker assisted-selection breeding. In conclusion, cloning and disease resistance mechanism analysis of Pm2b provided an example to emphasize a need of the molecular isolation of resistance genes, which has implications in marker assisted wheat breeding.

Characterization of a new splicing variant of powdery mildew resistance gene Pm4 in synthetic hexaploid wheat YAV249.

Powdery mildew, caused by Blumeria graminis f. sp. tritici (Bgt), is a destructive fungal disease of wheat throughout the world. Utilization of effective powdery mildew resistance genes and cultivars is considered as the most economic, efficient, and environmental-friendly method to control this disease. Synthetic hexaploid wheat (SHW), which was developed through hybridization of diploid Aegilops and tetraploid wheat, is a valuable genetic resource for resistance to powdery mildew. SHW line YAV249 showed high levels of resistance to powdery mildew at both the seedling and adult stages. Genetic analysis indicated that the resistance was controlled by a single dominant gene, temporarily designated PmYAV. Bulked segregant analysis with wheat 660K single nucleotide polymorphism (SNP) array scanning and marker analysis showed that PmYAV was located on chromosome 2AL and flanked by markers Xgdm93 and Xwgrc763, respectively, with genetic distances of 0.8 cM and 1.2 cM corresponding to a physic interval of 1.89 Mb on the Chinese Spring reference genome sequence v1.0. Sequence alignment analysis demonstrated that the sequence of PmYAV was consistent with that of Pm4a but generated an extra splicing event. When inoculated with different Bgt isolates, PmYAV showed a significantly different spectrum from Pm4a, hence it might be a new resistant resource for improvement of powdery mildew resistance. The flanked markers GDM93 and WGRC763, and the co-segregated markers BCD1231 and JS717/JS718 were confirmed to be easily performed in marker-assisted selection (MAS) of PmYAV. Using MAS strategy, PmYAV was transferred into the commercial cultivar Kenong 199 (KN199) and a wheat line YK13 was derived at generation BC3F3 from the population of YAV249/4*KN199 due to its excellent agronomic traits and resistance to powdery mildew. In conclusion, an alternative splicing variant of Pm4 was identified in this study, which informed the regulation of Pm4 gene function.

Detecting Wheat Powdery Mildew and Predicting Grain Yield Using Unmanned Aerial Photography.

High-resolution aerial imaging with an unmanned aerial vehicle (UAV) was used to quantify wheat powdery mildew and estimate grain yield. Aerial digital images were acquired at Feekes growth stage (GS) 10.5.4 from flight altitudes of 200, 300, and 400 m during the 2009-10 and 2010-11 seasons; and 50, 100, 200, and 300 m during the 2011-12, 2012-13, and 2013-14 seasons. The image parameter lgR was consistently correlated positively with wheat powdery mildew severity and negatively with wheat grain yield for all combinations of flight altitude and year. Fitting the data with random coefficient regression models showed that the exact relationship of lgR with disease severity and grain yield varied considerably from year to year and to a lesser extent with flight altitude within the same year. The present results raise an important question about the consistency of using remote imaging information to estimate disease severity and grain yield. Further research is needed to understand the nature of interyear variability in the relationship of remote imaging data with disease or grain yield. Only then can we determine how the remote imaging tool can be used in commercial agriculture.

Detection of powdery mildew in two winter wheat plant densities and prediction of grain yield using canopy hyperspectral reflectance.

To determine the influence of plant density and powdery mildew infection of winter wheat and to predict grain yield, hyperspectral canopy reflectance of winter wheat was measured for two plant densities at Feekes growth stage (GS) 10.5.3, 10.5.4, and 11.1 in the 2009-2010 and 2010-2011 seasons. Reflectance in near infrared (NIR) regions was significantly correlated with disease index at GS 10.5.3, 10.5.4, and 11.1 at two plant densities in both seasons. For the two plant densities, the area of the red edge peak (Σdr680-760 nm), difference vegetation index (DVI), and triangular vegetation index (TVI) were significantly correlated negatively with disease index at three GSs in two seasons. Compared with other parameters Σdr680-760 nm was the most sensitive parameter for detecting powdery mildew. Linear regression models relating mildew severity to Σdr680-760 nm were constructed at three GSs in two seasons for the two plant densities, demonstrating no significant difference in the slope estimates between the two plant densities at three GSs. Σdr680-760 nm was correlated with grain yield at three GSs in two seasons. The accuracies of partial least square regression (PLSR) models were consistently higher than those of models based on Σdr680760 nm for disease index and grain yield. PLSR can, therefore, provide more accurate estimation of disease index of wheat powdery mildew and grain yield using canopy reflectance.

Simultaneous real-time PCR detection of Fusarium asiaticum, F. ussurianum and F. vorosii, representing the Asian clade of the F. graminearum species complex.

Due to the repeated discovery of new members of the Fusarium graminearum species complex (FGSC), some of the F. graminearum sensu stricto (s.s.)-specific qPCR assays developed to date have since been shown to be non-specific. In this study, a probe-based qPCR method was developed, targeting a sterol 14-alpha demethylase (CYP51) paralogue, CYP51C unique to the genus Fusarium, for the simultaneous detection of F. asiaticum, F. ussurianum and F. vorosii. Specificity of the assay was demonstrated for a wide range of Fusarium species, including all tested FGSC members (n=6), originating from different hosts and geographic regions. Alongside a previously published assay for detection of F. graminearum, we were able to show that members of the Asian clade of the FGSC (i.e. F. asiaticum, F. ussurianum and F. vorosii) were the primary etiological agent in wheat seeds samples originating from Central-East China. The grain samples from the UK tested negative for the presence of the FGSC's Asian clade and positive for presence of F. graminearum. It is likely that only F. graminearum s.s. is present in the UK, but the presence of other FGSC members cannot be ruled out and need further investigation.