Evaluation of Borage (Borago officinalis L.) Genotypes for Nutraceutical Value Based on Leaves Fatty Acids Composition

Borage (Borago officinalis L.) is a traditional vegetable grown and consumed in some Spanish regions. The objective of this study was to determine the variability and evolution of fatty acid composition in a borage germplasm collection formed by wild types, breeding lines, commercial varieties, and landraces. Fatty acids were analysed in petioles, the commonly edible part of the leaves, and the leaf blades, the by-product of the borage industry, in two growth stages: at the optimal harvest period (120 days after sowing) and at the end of the harvest period (150 days after sowing). The results showed that for each of the eight fatty acids identified, there were significant differences among the twelve borage genotypes depending on the developmental plant stage at sampling date and the part of the leaf analysed, the interaction effect also being statistically significant. The main polyunsaturated fatty acids identified were: linoleic acid (18:2 n6, LA), α-linolenic acid (18:3 n3, ALA), γ-linolenic acid (18:3 n6, GLA), and stearidonic acid (SDA, 18:4, n-3), account for approximately 70% of polyunsaturated fatty acids. Blue-flowered genotypes differ from white-flowered genotypes by their high content of ALA and SDA, which can be exploited in borage breeding programs. Petioles from young plants present higher n6 fatty acids, while older plants produce a great amount of n3 fatty acids. Besides, the higher content of ALA in the leaf blades gives them a good dietary potential. All these fatty acids, with multiple health benefits, support the nutraceutical interest of borage leaves (both petioles and leaf blades) for human consumption, animal feeding, medicine, and pharmacy.

Classification Method of Rape Root Swelling Disease Based on Convolution Neural Network

Both the seedling stage and the adult plant stage of rape can be infected with root edema, and the damaged roots swell to form tumors of different sizes and shapes. The incidence of rape root swelling at the seedling stage reached 17%, and the average incidence at the adult plant stage was 15%, resulting in a 10.2% reduction in rape production. The average plant height, number of siliques, number of kernels per horn, 1000-seed weight and yield per plant of healthy plants were significantly higher than those of diseased plants. Grading root lesions can help trace the root causes of root lesions. However, the method of grading is often performed manually by professionals at present, which has the problems of low speed and low efficiency. In order to solve this problem, a method for grading rape root swelling based on deep convolutional neural network is proposed in this paper. Firstly, a rape root swelling model based on convolutional neural network and regional candidate network was established, and then implement it on the deep learning Tensorflow framework Model, and finally compare and analyze the results. The rape root swelling model uses the VGG16 network to extract the characteristics of the rape root swelling image. The regional candidate network generates the preliminary position candidate frame of the rape root swelling, and Fast-RCNN realizes the classification and positioning of the candidate frame. The results show that this method can achieve rapid and accurate detection of healthy, first-level tumors, second-level tumors, and third-level tumors of four-level rape root swelling, with an average accuracy rate of 84.12%. The experimental results show that the accuracy rate can reach more than 90%.

Deciphering resistance to Zymoseptoria tritici in the Tunisian durum wheat landrace accession ‘Agili39’

Background Septoria tritici blotch (STB), caused by Zymoseptoria tritici (Z. tritici), is an important biotic threat to durum wheat in the entire Mediterranean Basin. Although most durum wheat cultivars are susceptible to Z. tritici, research in STB resistance in durum wheat has been limited. Results In our study, we have identified resistance to a wide array of Z. tritici isolates in the Tunisian durum wheat landrace accession ‘Agili39’. Subsequently, a recombinant inbred population was developed and tested under greenhouse conditions at the seedling stage with eight Z. tritici isolates and for five years under field conditions with three Z. tritici isolates. Mapping of quantitative trait loci (QTL) resulted in the identification of two major QTL on chromosome 2B designated as Qstb2B_1 and Qstb2B_2. The Qstb2B_1 QTL was mapped at the seedling and the adult plant stage (highest LOD 33.9, explained variance 61.6 %), conferring an effective resistance against five Z. tritici isolates. The Qstb2B_2 conferred adult plant resistance (highest LOD 32.9, explained variance 42 %) and has been effective at the field trials against two Z. tritici isolates. The Qstb2B_1 QTL was mapped at the seedling and the adult plant stage (highest LOD 33.9, explained variance 61.6 %), conferring an effective resistance against five Z. tritici isolates. The Qstb2B_2 conferred adult plant resistance (highest LOD 32.9, explained variance 42 %) and has been effective at the field trials against two Z. tritici. The physical positions of the flanking markers linked to Qstb2B_1 and Qstb2B_2 indicate that these two QTL are 5Mb apart. In addition, we identified two minor QTL on chromosomes 1A (Qstb1A) and chromosome 7A (Qstb7A) (highest LODs 4.6 and 4.0, and explained variances of 16 % and 9%, respectively) that were specific to three and one Z. tritici isolates, respectively. All identified QTL were derived from the landrace accession Agili39 that represents a valuable source for STB resistance in durum wheat. Conclusion This study demonstrates that Z. tritici resistance in the ‘Agili39’ landrace accession is controlled by two minor and two major QTL acting in an additive mode.

Candidate Genes Underlying QTL for Powdery Mildew Resistance in Triticale (x Triticosecale Wittm.)

Combining to tolerance to biotic and abiotic stresses is important target for modern triticale breeding. Cultivation of varieties resistant to fungal pathogens is economically and environmentally important and may lead to reducing of the use of fungicides. Molecular markers are necessary for accumulation of advantageous alleles in the best genotypes by means of marker-assisted and genomic selection approaches. In present research, QTL regions associated with the powdery mildew resistance at adult plant stage were evaluated in order to provide the effective selection tools. Testing of DH population in multiple environments under natural infestation revealed 20 QTL on wheat (4A, 3B, 4B) and rye (2R, 4R, 5R, 6R) chromosomes. Regions explained 8.1% - 29.3% of phenotypic variation depending of the trait, localization and year of the experiment. Main QTL with effect exceeding 15% were found on chromosomes 3B, 4B, 2R, 5R and 6R. QTL and candidate genes located on chromosomes 4B, 2R, 5R and 6R are so far reported for the first time as regions associated with PM resistance in the adult triticale plants. Additionally, within all QTL, 21 candidate genes associated with the PM resistance were revealed. Predicted function of protein encoded by these genes include triggering a defense system which restricts the pathogen growth, enzyme activity, regulation of hormone-activated pathways, transcriptional corepressor complex and cell wall construction. Availability of QTL, molecular markers together with candidate genes linked with the powdery mildew resistance can be validated on triticale lines and varieties and then, used in MAS to improve modern breeding.

Evaluations of Genomic Prediction and Identification of New Loci for Resistance to Stripe Rust Disease in Wheat (Triticum aestivum L.)

Stripe rust is one of the most destructive diseases of wheat (Triticum aestivum L.), caused by Puccinia striiformis f. sp. tritici (Pst), and responsible for significant yield losses worldwide. Single-nucleotide polymorphism (SNP) diagnostic markers were used to identify new sources of resistance at adult plant stage to wheat stripe rust (YR) in 141 CIMMYT advanced bread wheat lines over 3 years in replicated trials at Borlaug Institute for South Asia (BISA), Ludhiana. We performed a genome-wide association study and genomic prediction to aid the genetic gain by accumulating disease resistance alleles. The responses to YR in 141 advanced wheat breeding lines at adult plant stage were used to generate G × E (genotype × environment)-dependent rust scores for prediction and genome-wide association study (GWAS), eliminating variation due to climate and disease pressure changes. The lowest mean prediction accuracies were 0.59 for genomic best linear unbiased prediction (GBLUP) and ridge-regression BLUP (RRBLUP), while the highest mean was 0.63 for extended GBLUP (EGBLUP) and random forest (RF), using 14,563 SNPs and the G × E rust score results. RF and EGBLUP predicted higher accuracies (∼3%) than did GBLUP and RRBLUP. Promising genomic prediction demonstrates the viability and efficacy of improving quantitative rust tolerance. The resistance to YR in these lines was attributed to eight quantitative trait loci (QTLs) using the FarmCPU algorithm. Four (Q.Yr.bisa-2A.1, Q.Yr.bisa-2D, Q.Yr.bisa-5B.2, and Q.Yr.bisa-7A) of eight QTLs linked to the diagnostic markers were mapped at unique loci (previously unidentified for Pst resistance) and possibly new loci. The statistical evidence of effectiveness and distribution of the new diagnostic markers for the resistance loci would help to develop new stripe rust resistance sources. These diagnostic markers along with previously established markers would be used to create novel DNA biosensor-based microarrays for rapid detection of the resistance loci on large panels upon functional validation of the candidate genes identified in the present study to aid in rapid genetic gain in the future breeding programs.

Assessment and modeling using machine learning of resistance to scald (Rhynchosporium commune) in two specific barley genetic resources subsets

Barley production worldwide is limited by several abiotic and biotic stresses and breeding of highly productive and adapted varieties is key to overcome these challenges. Leaf scald, caused by Rhynchosporium commune is a major disease of barley that requires the identification of novel sources of resistance. In this study two subsets of genebank accessions were used: one extracted from the Reference set developed within the Generation Challenge Program (GCP) with 191 accessions, and the other with 101 accessions selected using the filtering approach of the Focused Identification of Germplasm Strategy (FIGS). These subsets were evaluated for resistance to scald at the seedling stage under controlled conditions using two Moroccan isolates, and at the adult plant stage in Ethiopia and Morocco. The results showed that both GCP and FIGS subsets were able to identify sources of resistance to leaf scald at both plant growth stages. In addition, the test of independence and goodness of fit showed that FIGS filtering approach was able to capture higher percentages of resistant accessions compared to GCP subset at the seedling stage against two Moroccan scald isolates, and at the adult plant stage against four field populations of Morocco and Ethiopia, with the exception of Holetta nursery 2017. Furthermore, four machine learning models were tuned on training sets to predict scald reactions on the test sets based on diverse metrics (accuracy, specificity, and Kappa). All models efficiently identified resistant accessions with specificities higher than 0.88 but showed different performances between isolates at the seedling and to field populations at the adult plant stage. The findings of our study will help in fine-tuning FIGS approach using machine learning for the selection of best-bet subsets for resistance to scald disease from the large number of genebank accessions.

Molecular and Phenotypic Analysis of Bread Wheat Varieties in Relation to Durable Rust Resistance

Global wheat production is constantly threatened by rust diseases. Identifying resistant genes is a useful tactic to control wheat rust pathogen. Twenty-six wheat varieties were screened with twelve Simple Sequence Repeats (SSR) markers to detect rust resistant genes and the efficacy of genes was validated through field testing. The alleles Lr32, Lr39, Lr50, SrCad and SrWeb were not amplified in the varieties included in this study. The SSR markers indicated that the varieties viz., Chakwal-97, Bakhar-2002 and Lasani-2008 had a combination of 02 slow rusting alleles (Lr46/Yr29 and Yr18/Lr34). The adult plant resistance (APR) allele Yr17 was less prevalent and found only in BWL-97. However, Noshera-96 had a slow rusting combination of Lr67/Yr46 and Lr46/Yr46 alleles. The Lr46/Yr29 identified in 50% of the varieties, Yr18/Lr34 in 19.23%, Lr32 in 11.54%, and multiple APR alleles in 19.32%. Their resistance was validated through a field trap nursery for 3 consecutive seasons. The slow rusting combination of Lr46/Yr29 and Yr18/Lr34 was comparatively more effective than Lr67/Yr46 and Lr46/Yr29 alleles under field conditions. The varieties Yecora-70, Lylpure-73 and Tandojam-83 showed highly susceptible phenotype. The varieties Chakwal-86, Pirsabak-2005, Fareed-2006, and Sehar-2006 showed resistant to moderately resistant phenotype at high-temperature adult-plant stage. The cluster diagram divided the varieties into two distinct clades. The clade II depicted the abundance of APR allele Lr46/Yr29. The varieties contain valuable sources of durable rust resistant alleles that can be exploited to deploy rust resistance in future wheat cultivars. It has been observed that the varieties approved for commercial cultivation after 1990s and onwards contain APR alleles. © 2021 Friends Science Publishers

Control of stripe rust of wheat using indigenous endophytic bacteria at seedling and adult plant stage

Stripe rust (caused by Puccinia striiformis tritici) is one of the most devastating diseases of wheat. The most effective ways to control stripe rust are the use of resistant cultivars and the timely use of an appropriate dose of fungicide. However, the changing nature of rust pathogen outwits the use of resistant cultivars, and the use of a fungicide is associated with environmental problems. To control the disease without sacrificing the environment, we screened 16 endophytic bacteria, which were isolated from stripe rust-resistant wheat cultivars in our previous study, for their biocontrol potential. A total of 5 bacterial strains Serratia marcescens 3A, Bacillus megaterium 6A, Paneibacillus xylanexedens 7A, Bacillus subtilis 11A, and Staphyloccus agentis 15A showed significant inhibition of Puccinia striiformis f. sp. tritici (Pst) urediniospores germination. Two formulations i.e., fermented liquid with bacterial cell (FLBC) and fermented liquid without bacterial cells (FL) of each bacterial strain, were evaluated against the urediniospores germination. Formulations of five selected endophytic bacteria strains significantly inhibited the uredinioospores germination in the lab experiments. It was further confirmed on seedlings of Pakistani susceptible wheat cultivar Inqilab-91 in the greenhouse, as well as in semi-field conditions. FLBC and FL formulations applied 24 h before Pst inoculation (hbi) displayed a protective mode. The efficacy of FLBC was between 34.45 and 87.77%, while the efficacy of FL was between 39.27 and 85.16% when applied 24 hbi. The inoculated wheat cultivar Inqilab-91 was also tested under semi-field conditions during the 2017–2018 cropping season at the adult plant stage. The strains Bacillus megaterium 6A and Paneibacillus xylanexedens 7A alone significantly reduced the disease severity of stripe rust with the efficacy of 65.16% and 61.11% for the FLBC in protective effect, while 46.07% and 44.47% in curative effect, respectively. Inoculated seedlings of Inqilab-91 showed higher activities of antioxidant enzymes, superoxide dismutase (SOD), peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia-lyase (PAL). The treated seedlings also showed higher expressions of pathogenesis-related (PR) protein genes, antifungal protein (PR-1), β-1,3-endoglucanases (PR-2), endochitinases (PR-4), peroxidase (PR-9), and ribonuclease-like proteins (PR-10). These results indicated that endophytic bacteria have the biocontrol potential, which can be used to manage stripe rust disease. High production antioxidant enzymes, as well as high expression of PR protein genes, might be crucial in triggering the host defense mechanism against Pst.

Identification of Resistant Germplasm and Detection of Genes for Resistance to Powdery Mildew and Leaf Rust from 2,978 Wheat Accessions

Powdery mildew and leaf rust, caused by Blumeria graminis f. sp. tritici (Bgt) and Puccinia triticina (Pt), respectively, are widespread diseases of wheat worldwide. Utilizing resistant cultivars is considered as the most economical, environmental-friendly, and effective method to control these diseases. In the present study, a collection of 2,978 wheat accessions consisting of 1,394 advanced breeding lines, 1,078 Chinese cultivars, 291 introduced cultivars, 132 lines containing alien chromosomes, and 83 landraces was tested for reactions to powdery mildew and leaf rust. The results indicated that 659 (22.1%) wheat accessions were highly resistant to a widely prevalent Bgt isolate, E09, at the seedling stage, and 390 were consistently resistant to the mixture of Bgt isolates at the adult-plant stage. Meanwhile, 63 (2.1%) accessions were highly resistant to leaf rust at the adult-plant stage, of which 54 were resistant to a predominant and highly virulent Pt race, THTT, at the seedling stage. Notably, 17 accessions were resistant to both powdery mildew and leaf rust. To detect known genes for resistance to powdery mildew and leaf rust, these accessions were tested with gene-specific or tightly linked markers for seven Pm genes (Pm2, Pm4, Pm5, Pm6, Pm8, Pm21, and Pm24) and ten Lr genes (Lr1, Lr9, Lr10, Lr19, Lr20, Lr24, Lr26, Lr34, Lr37, and Lr46). Of the 659 powdery mildew-resistant accessions, 328 might carry single Pm genes and 191 carry combined Pm genes. Pm2 was detected at the highest frequency of 59.6%, followed by Pm8, Pm6, Pm21, Pm4, and Pm5, while Pm24 was not detected. Besides, 139 accessions might contain unknown Pm genes different from those tested in this study. In the 63 accessions resistant leaf rust, four Lr genes (Lr1, Lr10, Lr26, and Lr34) were detected in 41 accessions either singly or in combination, while six genes (Lr9, Lr19, Lr20, Lr24, Lr37, and Lr46) were not detected. Twenty-two accessions might contain unknown Lr genes different from those tested in this study. This study not only provided important information for rationally distributing resistance genes in wheat breeding programs, but also identified resistant germplasm that might have novel genes to enrich the diversity of resistance sources.