Advances in Localization and Molecular Markers of Wheat Leaf Rust Resistance Genes

Genetic resistance is the most economical method of reducing yield losses caused by wheat leaf rust. To identify the leaf rust resistance genes in commonly used parental germplasm and released cultivars become very important for utilizing the genetic resistance to wheat leaf rust fully. Up to date, about 90 leaf rust resistance genes have been found, of which 51 genes have been located and mapped to special chromosomes, and 56 genes have been designated officially according to the standards set forth in the Catalogue of Gene Symbols for wheat. Twenty-four wheat leaf rust resistance genes have been developed for their molecular markers. It is very important to isolate, characterize, and map leaf rust resistance genes due to the resistance losses of the genes caused by the pathogen continuously.

Identification and Molecular Tagging of Leaf Rust Resistance Gene (Lr24) in Wheat

This research was aimed to develop AFLP markers co-segregated with gene Lr24 and validate the using for marker assisted selection （MAS）. An F2 population developed from the cross between the resistant line TcLr24 and the susceptible line Thatcher was tested for resistance to the Puccinia triticina races BGQQ and SHRT using for genetic analysis and molecular marker. A total of 224 AFLP primer combinations were used to test the resistant and susceptible parents, as well as the resistant bulk and the susceptible bulk. Four AFLP markers, P-AGA/M-CTT289 bp, P-AGC/M-CAC1ss bp, P-AGC/M- CAC162 bp, and P-ACG/M-CGC239 bp, were co-segregated with Lr24. The AFLP fragment from the primer combination P- ACG/M-CGC was cloned, sequenced and converted into a STS marker named as ASTS212. Thatcher backgrounded NILs and 115 varieties were examined by using this STS marker and the marker SCS13026oz developed by Gupta. 5R615, 5R616, IR13, and 1R17 were identified and validated to contain gene Lr24. The marker is dominant and may be useful in identification the resistance gene Lr24 in wheat and wheat breeding programs.

Postulation of seedling leaf rust resistance genes in 84 Chinese winter wheat cultivars

Wheat leaf rust（caused by Puccinia triticina） is one of the most important fungal diseases in China. There are tens of winter wheat cultivars which are approved to be released by the government at a national level and more than 100 wheat cultivars at the provincial level. But there is no information about leaf rust（Lr） genes in these cultivars, which makes it difficult for farmers and breeders to select which cultivars they should plant in their fields and use in their breeding programs. The objective of this paper was to identify the leaf rust resistant genes at seedling stage present in the 84 commercial wheat cultivars from China that have been released in the past few years. A set of 20 near isogenic lines with Thatcher background and 6 lines with known Lr genes were used to test the virulence of 12 races of P. triticina（Pt）. By comparing the infection types（ITs） produced on the 84 cultivars by the 12 Pt races with the ITs on the differential sets, the Lr genes were postulated. In addition, 8 molecular markers of Lr genes such as Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, Lr26 and Lr29, which are closely linked to or co-segregated with the Lr gene, were used for further validation of the genes in the 84 Chinese winter wheat cultivars. Twelve Lr genes, including Lr1, Lr3,（Lr3bg）,（Lr3ka）, Lr11, Lr13, Lr14 a, Lr16, Lr26, Lr27, Lr30 and Lr31 were postulated to be present either singly or in combinations in these Chinese wheat cultivars. Lr3 and Lr26 were detected most often in the tested cultivars, with frequencies of 51.2 and 38.1%, respectively. No wheat Lr genes were detected in 16 cultivars, and 4 cultivars may carry unknown Lr genes other than those used in this study. Lr9, Lr20, Lr21, Lr24, Lr25 and Lr29 were not present in any of the 84 tested accessions.

A SSR Marker for Leaf Rust Resistance Gene Lr19 in Wheat

Microsatellite was carded out in Thatcher, six near-isogenic lines and F2 progeny of TcLr19xThatcher to develop molecular markers for leaf rust resistance gene Lr19. Thirteen primer pairs were screened, of which one primer pair Xgwm44 displayed polymorphsim in the population of TcLr 19, Thatcher, and their F2 generations. One marker closed linked to Lr19 resistance trait was obtained, and was named Xgwm44139bp with the genetic distance 0.9 cM. The research shows that Lr19 has more potential in marker-assisted breeding programs in wheat and provides a step stone for mapping genetic map, physical map and the eventual cloning.

Novel method for identifying wheat leaf disease images based on differential amplification convolutional neural network

In this study,a differential amplification convolutional neural network(DACNN)was proposed and used in the identification of wheat leaf disease images with ideal accuracy.The branches added between the deep convolutional layers can amplify small differences between the real output and the expected output,which made the weight updating more sensitive to the light errors return in the backpropagation pass and significantly improved the fitting capability.Firstly,since there is no large-scale wheat leaf disease images dataset at present,the wheat leaf disease dataset was constructed which included eight kinds of wheat leaf images,and five kinds of data augmentation methods were used to expand the dataset.Secondly,DACNN combined four classifiers:Softmax,support vector machine(SVM),K-nearest neighbor(KNN)and Random Forest to evaluate the wheat leaf disease dataset.Finally,the DACNN was compared with the models:LeNet-5,AlexNet,ZFNet and Inception V3.The extensive results demonstrate that DACNN is better than other models.The average recognition accuracy obtained on the wheat leaf disease dataset is 95.18%.

Molecular Markers for Leaf Rust Resistance Gene Lr45 in Wheat Based on AFLP Analysis

Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 progenies were used to develop markers for Lr45 gene. Sixty AFLP primer combinations were screened and most of them provided clear amplification products, 31 primer combinations displayed polymorphism of TcLr45 in 23 NILs. Two AFLP markers closely linked to the gene Lr45 were acquired： P-AGG/M-GAG261bp, which was found closely linked to the Lr45 locus at a distance of 0.6 cM on one side, and P-ACA/M-GGT105bp, which was found at a distance of 1.3 cM on the other side. The specific hands were cloned and subsequently sequenced. The 261-bp fragment produced by P-AGG/M-GAG showed 86% similarity with the sequence of Vulgate Hort I gene; the 105-bp fragment produced by P-ACA/M-GGT showed 96% similarity with the phosphatidylserine decarboxylase gene of the Triticum monococcum. Both included an open reading frame （ORF）.

Seedling and adult plant resistance to leaf rust in 46 Chinese bread wheat landraces and 39 wheat lines with known Lr genes

Wheat leaf rust,caused by Puccinia triticina(Pt),is an important foliar disease that has an important influence on wheat yield.The most economic,safe and effective way to control the disease is growing resistant cultivars.In the present study,a total of 46 wheat landraces and 34 wheat lines with known Lr(leaf rust resistance)genes were inoculated with 16Pt pathotypes for postulating seedling resistance gene(s)in the greenhouse.These cultivars and five wheat differential lines with adult plant resistance(APR)genes(Lr12,Lr22b,Lr34,Lr35 and Lr37)were also evaluated for identification of slow rusting resistance in the field trials in Baoding,Hebei Province of China in the 2014–2015 and 2015–2016 cropping seasons.Furthermore,10 functional molecular markers closely linked to 10 known Lr genes were used to detect all the wheat genotypes.Results showed that most of the landraces were susceptible to most of the Pt pathotypes at seedling stage.Nonetheless,Lr1 was detected only in Hongtangliangmai.The field experimental test of the two environments showed that 38 landraces showed slow rusting resistance.Seven cultivars possessed Lr34 but none of the landraces contained Lr37 and Lr46.Lr genes namely,Lr9,Lr19,Lr24,Lr28,Lr29,Lr47,Lr51 and Lr53 were effective at the whole plant stage.Lr18,Lr36 and Lr45 had lost resistance to part of pathotypes at the seedling stage but showed high resistance at the adult plant stage.Lr34 as a slowing rusting gene showed good resistance in the field.Four race-specific APR genes Lr12,Lr13,Lr35 and Lr37 conferred good resistance in the field experiments.Seven race-specific genes,Lr2b,Lr2c,Lr11,Lr16,Lr26,Lr33 and LrB had lost resistance.The 38 landraces showed slow rusting resistance to wheat leaf rust can be used as resistance resources for wheat resistance breeding in China.

Determination of critical nitrogen dilution curve based on leaf area index for winter wheat in the Guanzhong Plain, Northwest China

Excessive use of nitrogen (N) fertilizers in agricultural systems increases the cost of production and risk of environmental pollution. Therefore, determination of optimum N requirements for plant growth is necessary. Previous studies mostly established critical N dilution curves based on aboveground dry matter (DM) or leaf dry matter (LDM) and stem dry matter (SDM), to diagnose the N nutrition status of the whole plant. As these methods are time consuming, we investigated the more rapidly determined leaf area index (LAI) method to establish the critical nitrogen (Nc) dilution curve, and the curve was used to diagnose plant N status for winter wheat in Guanzhong Plain in Northwest China. Field experiments were conducted using four N fertilization levels (0, 105, 210 and 315 kg ha?1) applied to six wheat cultivars in the 2013–2014 and 2014–2015 growing seasons. LAI, DM, plant N concentration (PNC) and grain yield were determined. Data points from four cultivars were used for establishing the Nc curve and data points from the remaining two cultivars were used for validating the curve. The Nc dilution curve was validated for N-limiting and non-N-limiting growth conditions and there was good agreement between estimated and observed values. The N nutrition index (NNI) ranged from 0.41 to 1.25 and the accumulated plant N deficit (Nand) ranged from 60.38 to –17.92 kg ha?1 during the growing season. The relative grain yield was significantly affected by NNI and was adequately described with a parabolic function. The Nc curve based on LAI can be adopted as an alternative and more rapid approach to diagnose plant N status to support N fertilization decisions during the vegetative growth of winter wheat in Guanzhong Plain in Northwest China.

Slight shading after anthesis increases photosynthetic productivity and grain yield of winter wheat (Triticum aestivum L.) due to the delaying of leaf senescence

The solar radiation intensity and duration are continuously decreasing in the major wheat planting area of China. As a con- sequence, leaf senescence, photosynthesis, grain filling and thus wheat yield shall be affected by light deficiency. Therefore, two winter wheat （Triticum aestivum L.） cultivars, Tainong 18 （a large-spike cultivar） and Ji＇nan 17 （a multiple-spike cultivar）, were subjected to shading during anthesis and maturity under field condition in 2010-2011 and 2011-2012. Under the slight shading treatment （$1,88% of full sunshine）, leaf senescence was delayed, net photosynthesis rate （Po） and canopy apparent photosynthesis rate （CAP） were improved, and thus thousand-kernel weight （TKW） and grain yield were higher as compared with the control. However, mid and severe shading （S2 andS3, 67 and 35% of full sunshine, respectively） led to negative effects on these traits substantially. Moreover, superoxide dismutase （SOD）, peroxidase （POD） and cat- alase （CAT） activities in flag leaf were significantly greater under slight shading than those in other treatments, while the malondialdehyde （MDA） content was less than that under other treatments. In addition, the multiple-spike cultivar is more tolerant to shading than large-spike cultivar. In conclusion, slight shading after anthesis delayed leaf senescence, enhanced photosynthesis and grain filling, and thus resulted in higher grain yield.

Inverse Leaf Aging Sequence (ILAS) and Its Significance of Wheat

The implication of the revelation of the inverse leaf aging sequence（ILAS） of wheat needs to be probed in theory and practice.Since 2005,the comparison experiment of the ILAS and the conventional leaf aging sequence（CLAS） has been carried out to measure the canopy temperature and some important biological parameters of wheat.In nature,there existed the phenomenon that the wheat leaves aged in an sequence opposite to the conventional sequence and some of the leaves of ILAS wheat aged sequentially differed from those of CLAS wheat,i.e.,the penultimate leaves rather than the flag leaves aged last among the leaves at different leaf positions;in correspondence with the inverse leaf aging sequence,there appeared an unconventional leaf color structure at the late fruiting stage,which had yellow upper leaves and green lower leaves,opposite to the conventional leaf color structure that had green upper leaves and yellow lower leaves;the chlorophyll concentrations,soluble protein concentrations,transpiration rates,and net photosynthetic rates of the penultimate leaves of ILAS wheat unconventionally surpassed those of their flag leaves as their growth moved forward from one stage to another stage,and the ILAS wheat characteristically presented cold canopy temperature,i.e.,a cold tail canopy temperature at the late fruiting stage,or a cold canopy temperature at the whole fruiting stage;because ILAS wheat was unique in physiological process,its kernel weights were obviously higher than those of CLAS wheat,which closely related to the ＂relay＂ kernel-filling mode,which was different from the kernel-filling mode of CLAS wheat under which the flag leaves act as the main nutrient supply source at the whole fruiting stage.This study provided a new idea and approach for the theoretical exploration on wheat fruiting and aging,wheat yield further improvement,and cold type wheat and cold tail wheat breeding.

Interactions between cytokinin and nitrogen contribute to grain mass in wheat cultivars by regulating the flag leaf senescence process

Premature senescence after anthesis reduces crop yields.Delaying leaf senescence could maintain photosynthetic activity for a longer period and lead to a higher photosynthetic rate.Recent studies have provided some insights into the interaction between cytokinin and nitrogen(N)in the regulation of plant development.In the present study,foliar application of exogenous 6-benzylaminopurine(6-BA)and lovastatin,an inhibitor of cytokinin synthesis,was combined with three N rates[0 kg ha^(-1)(low nitrogen,LN),240 kg ha^(-1)(normal nitrogen,NN),and 360 kg ha^(-1)(high nitrogen,HN)]in two wheat cultivars,Wennong 6(with a staygreen phenotype)and Jimai 20(with a non-staygreen phenotype).Flag leaf senescence was assessed using a Gompertz growth curve.Grain mass,dry matter accumulation and distribution,total N of flag leaf,and concentrations of zeatin riboside(ZR)and abscisic acid(ABA)were also used to evaluate the functional characteristics of flag leaves.Grain mass was negatively correlated with initial senescence rate(r_0)and duration of rapid chlorophyll loss(Chl_(loss)),whereas it was positively correlated with maximum senescence rate(r_(max)),average senescence rate(r_(aver)),persistence phase(Chl_(per)),total duration of flag leaf(Chl_(total))and inflection point cumulative temperature(M).Compared to Jimai 20,Wennong 6 had larger r_(aver),Chl_(per),and Chl_(total).The concentration of ZR was highest under the 6-BA×NN treatment,followed by the 6-BA×HN and 6-BA×LN treatments.However,the concentration of ABA showed the opposite trend.It was concluded that the staygreen phenotype Wennong 6 was associated with greater grain mass and altered cytokinin metabolism and could be classified as a functional staygreen type.Foliar application of 6-BA interacting with N at the NN level(240 kg ha^(-1))may be a beneficial strategy for improving grain yield of wheat by regulating endogenous hormones and the flag leaf senescence process.Increasing endogenous cytokinin promoted the transport of dry matter to grain.

The Stimulating Effects of Rewatering on Leaf Area of Winter Wheat Suffering Water Stress

After water stress at various levels and durations at different growth stages, rewatering could greatly stimulate the leaf area development of winter wheat. The results showed that the stimulation effect changed with water stress time, degree and duration. Rewatering under earlier stress had greater stimulation effect on leaf area than that under later stress. Higher stimulation effect was observed under severe water stress than that under moderate stress. Longer duration of stress resulted in low stimulation effect. In spite of the greater stimulation effect under severe and longer stress, the final leaf area in these situations was lower than that under moderate stress and shorter duration. Whenever the stress occurred, the stimulating effect was due to the increase of the leaf area of the tillers. Once the leaf on the main stem emerged during stress period, rewatering had no effect on its size, and consequently its leaf area. The stimulation of rewateirng on leaf area contributed to the final grain yield by 45% under moderate stress, and 67% under severe stress. Although the stimulation partly compensated for the loss during stress, the final leaf area and the grain yield could not reach the level without water stress.

Proximate, Chemical and Functional Properties of Wheat, Soy and Moringa Leaf Composite Flours

The study presents the effect of utilizing wheat, soy and moringa leaf flour and quality analysis of the flour. The composite flour was prepared using refined wheat flour, soy flour and moringa leaf flour. Four composite flour, compositions were formulated such as 100% wheat flour (control) designated as sample A. Sample B consisted of 75% wheat, 20% soybean and 5% moringa leaf. Sample C consisted of 70% wheat, 20% soybean and 10% moringa leaf. Sample D was 65% wheat, 20% soybean and 15% moringa leaf. Sample E was 60% wheat, 20% soybean and 20% moringa leaf, respectively. Proximate, chemical, and functional properties of wheat, soy and moringa leaf flours were studied in composite flour variation and preparations. The present study highlighted the nutrients enrichment of flour on incorporation of soy and moringa leaf. Relevant statistical tests were done to analyse the significance of means for all tested parameters. Composite flour composition with 20% soybean was identified to produce optimal nutrient, mineral quality and yield. The addition of soybean and moringa flour in baked products has been shown in this study to improve the nutrition and health benefits of the body. It also serves as a good cut on the cost of wheat importation in communities with supply challenges.

PCR-based molecular markers linked to the leaf rust resistance gene Lr19 in different bread wheat cultivars

61 varieties of wheat collected in the gene fund of the Research Institute of Crop Husbandry were screened using SCAR-markers associated with the gene of resistance to brown leaf rust, Lr19. As a result of PCR analysis using SCS123 marker the 737 bp locus was detected in 48 genotypes. The expected fragment of the 688 bp was detected in 53 genotypes using the SCS253 marker. The results obtained using both markers indicate that the Lr19 gene is present on 7D chromosomes of 45 genotypes. The existence of the Lr19 gene has not been proven only for 5 from the 61 analyzed wheat genotypes.

The Relationship Between Developmental Accumulation of Leaf Soluble Proteins and Vernalization Response of Wheat(Triticum aestivum L.em. Thell)

The relationship between vernalization requirement and quantitative and qualitative changes in total leaf soluble proteins were determined in one spring （cv. Kohdasht） and two winter （cvs. Sardari and Norstar） cultivars of wheat （Triticum aestivum L.） exposed to 4℃. Plants were sampled on days 2, 14, 21 and 35 of exposure to 4℃. The final leaf number （FLN） was determined throughout the vernalization periods （0, 7, 14, 24, and 35 d） at 4℃. The final leaf number decreased until days 24 and 35 in Sardari and Norstar eultivars, respectively, indicating the vernalization saturation at these times. No clear changes were detected in the final leaf number of Kohdash cultivar, verifying no vernalization requirement for this spring wheat cultivar. Comparing with control, clear cold-induced 2-fold increases in proteins quantity occurred after 48 h following the 4℃-treatment in the leaves of the both winter wheat cultivars but, such response was not detected in the spring cultivar. However, the electrophoretic protein patterns showed between-cultivar and between-temperature treatment differences. With increasing exposure time to 4℃, the winter cultivars tended to produce more HMW polypeptides than the spring cultivar. Similar proteins were induced in both Sardari and Norstar winter wheat cultivars, however, the long vernalization requirement in Norstar resulted in high level and longer duration of expression of cold-induced proteins compared to Sardari with a short vernalization requirement. These observations indicate that vernalization response regulates the expression of low temperature （LT） tolerance proteins and determines the duration of expression of LT- induced proteins.

Identification of AFLP Markers Linked to Leaf Rust Resistance Genes Using Near Isogenic Lines of Wheat

The present investigation was undertaken to find molecular markers linked to leaf rust resistance genes, Lr9 and Kharchia local mutant KLM4-3B. Preliminary AFLP analysis was carried out with different stocks, a survey of primer combinations with different selective nucleotide indicated that for each primer combination, the number of scorable loci ranged from 34 to 123. Only a limited primer combination used in the set of parental and near isogenic lines showed a high level of polymorphism for AFLP marker. Putative AFLP marker were found to be linked to Lr9, Lr19 and KLM4-3B. The alien genes were readily identified.

Common Spectral Bands and Optimum Vegetation Indices for Monitoring Leaf Nitrogen Accumulation in Rice and Wheat

Real-time monitoring of nitrogen status in rice and wheat plant is of significant importance for nitrogen diagnosis, fertilization recommendation, and productivity prediction. With 11 field experiments involving different cultivars, nitrogen rates, and water regimes, time-course measurements were taken of canopy hyperspeetral reflectance between 350-2 500 nm and leaf nitrogen accumulation （LNA） in rice and wheat. A new spectral analysis method through the consideration of characteristics of canopy components and plant growth status varied with phenological growth stages was designed to explore the common central bands in rice and wheat. Comprehensive analyses were made on the quantitative relationships of LNA to soil adjusted vegetation index （SAVI） and ratio vegetation index （RVI） composed of any two bands between 350-2 500 nm in rice and wheat. The results showed that the ranges of indicative spectral reflectance were largely located in 770-913 and 729-742 nm in both rice and wheat. The optimum spectral vegetation index for estimating LNA was SAVI （R822, R738） during the early-mid period （from jointing to booting）, and it was RVI （Rs22, R73s） during the mid-late period （from heading to filling） with the common central bands of 822 and 738 nm in rice and wheat. Comparison of the present spectral vegetation indices with previously reported vegetation indices gave a satisfactory performance in estimating LNA. It is concluded that the spectral bands of 822 and 738 nm can be used as common reflectance indicators for monitoring leaf nitrogen accumulation in rice and wheat.

The Chromosomal Control of Leaf Characteristics of Early-Stage Plants in Wheat (Triticum aestivum L.)

Rapid expansion of leaves of early-stage plants in wheat produced by chromosomal control of characteristics related to rapid expansion of the first six leaves of wheat were investigated using a set of single chromosome substitution lines under two different temperature regimes（TRs）.Results from this study indicated that several chromosomes could be responsible for each of the four characteristics studied（including leaf width,leaf length,leaf elongation rate and leaf appearance index） and that different chromosomes may be responsible for the same characteristics at different TRs.Only a small number of substitutions that showed the most significant effects on leaf length were found among those that showed the most significant effects on leaf width,providing further evidence that these two characteristics are likely controlled by different sets of genes.Substitutions 4B,4D,and 5A are among those that were repeatedly detected to having significant effects on different leaf characteristics at the lower-TR and genes residing on these three chromosomes could be homologous.This possibility and its implication are discussed.

Molecular mapping of leaf rust resistance genes in the wheat line Yu 356-9

The Chinese wheat line Yu 356-9 exhibits a high level of resistance to leaf rust. In order to decipher the genetic base of resistance in Yu 356-9, gene postulation, inheritance analyses, and chromosome linkage mapping were carried out. Gene postulation completed using 15 leaf rust pathotypes and 36 isogenic lines indicated that Yu 356-9 was resistant to all pathotypes tested. F1 and F2 plants from the cross Yu 356-9 （resistant）/Zhengzhou 5389 （susceptible） were tested with leaf rust pathotype ＂FHNQ＂ in the greenhouse. Results indicated a 3：1 segregation ratio, indicative of the presence of a single dominant leaf rust resistance gene in Yu 356-9 which was temporarily designated as LrYu. Bulk segregant analysis and molecular marker assays were used to map LrYu. Five simple sequence repeat （SSR） markers on chromosome 2BS were found closely linked to LrYu. Among these markers, Xwmc770 is the most closely linked, with a genetic distance of 5.7 cM.

Mapping Spatial and Temporal Variations of Leaf Area Index for Winter Wheat in North China

Leaf area index （LAI） is an important parameter in a number of models related to ecosystem functioning, carbon budgets, climate, hydrology, and crop growth simulation. Mapping and monitoring the spatial and temporal variations of LAI are necessary for understanding crop growth and development at regional level. In this study, the relationships between LAI of winter wheat and Landsat TM spectral vegetation indices （SVIs） were analyzed by using the curve estimation procedure in North China Plain. The series of LAI maps retrieved by the best regression model were used to assess the spatial and temporal variations of winter wheat LAI. The results indicated that the general relationships between LAI and SVIs were curvilinear, and that the exponential model gave a better fit than the linear model or other nonlinear models for most SVIs. The best regression model was constructed using an exponential model between surface-reflectance-derived difference vegetation index （DVI） and LAI, with the adjusted R2 （0.82） and the RMSE （0.77）. The TM LAI maps retrieved from DVILAI model showed the significant spatial and temporal variations. The mean TM LAI value （30 m） for winter wheat of the study area increased from 1.29 （March 7, 2004） to 3.43 （April 8, 2004）, with standard deviations of 0.22 and 1.17, respectively. In conclusion, spectral vegetation indices from multi-temporal Landsat TM images can be used to produce fine-resolution LAI maps for winter wheat in North China Plain.