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Abstract Wheat leaf rust, caused by Puccinia triticina (Pt), remains a constant threat to wheat production worldwide. Deployment of race-specific leaf rust (Lr) resistance genes in wheat provides effective protection against leaf rust, but often leads to selective pressures that drive the rapid emergence of new virulent Pt isolates in nature. However, the molecular mechanisms underlying the evasion of Lr-delivered resistance by leaf rust remain largely unknown. Here, we identify an avirulence gene AvrLr21 in Pt that triggers Lr21-dependent immune responses. BSMV (Barley stripe mosaic virus)-mediated host-induced gene silencing assay shows that silencing AvrLr21 compromises Lr21-mediated immunity. AvrLr21 interacts directly with Lr21 protein to induce a hypersensitive response in tobacco leaves. The evolved Lr21-breaking Pt isolates can suppress Lr21-mediated immunity. Our data provide a basis for studying the molecular determinants in Pt-wheat incompatible interaction and monitoring natural Pt populations to prioritize the deployment of Lr resistance genes in the field.

BackgroundRadish seed is a functional food with many beneficial health effects. Glucosinolates are characteristic components in radish seed that can be transformed into bioactive isothiocyanates by gut microbiota.ObjectiveThe present study aims to assess anti-obesity efficacy of radish seed glucosinolates (RSGs) and explored the underlying mechanisms with a focus on gut microbiota and fecal metabolome.MethodsHigh-fat diet-induced obese mice were supplemented with different doses of RSGs extract for 8 weeks. Changes in body weight, serum lipid, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) levels; and pathological changes in the liver and adipose tissue were examined. Fecal metabolome and 16S rRNA gene sequencing were used to analyze alterations in fecal metabolite abundance and the gut microbiota, respectively.Results and conclusionResults showed that RSG extract prevented weight gain and decreased serum lipid, ALT, AST levels and lipid deposition in liver and epididymal adipocytes in obese mice. Treatment with RSG extract also increased gut microbiota diversity and altered the dominant bacteria genera in the gut microbiota, decreasing the abundance of Faecalibaculum and increasing the abundance of Allobaculum, Romboutsia, Turicibacter, and Akkermansia. Fecal metabolome results identified 570 differentially abundant metabolites, of which glucosinolate degradation products, such as sulforaphene and 7-methylsulfinylheptyl isothiocyanate, were significantly upregulated after RSG extract intervention. Furthermore, enrichment analysis of metabolic pathways showed that the anti-obesity effects of RSG extract may be mediated by alterations in bile secretion, fat digestion and absorption, and biosynthesis of plant secondary metabolites. Overall, RSG extract can inhibit the development of obesity, and the obesity-alleviating effects of RSG are related to alternative regulation of the gut microbiota and glucosinolate metabolites.

Wheat leaf rust, caused by Puccinia triticina, is one of the most common fungal diseases of wheat in China and occurs widely in various wheat-growing regions. To clarify the epidemic, spread rules, and population structure of P. triticina among different regions, 217 isolates of P. triticina collected from Hebei, Shandong, Sichuan, and Xinjiang in China were tested by 34 Thatcher near-isogenic lines and 21 pairs of EST-SSR primers. A total of 83 races were identified, and THTT, PHTT, THTS, and PHJT were the most predominant races in the four provinces in 2009. We found enriched virulence and genetic diversity in the four P. triticina populations and a significant correlation between genetic polymorphism and geographic regions. However, no significant correlation was found between virulence phenotypes and molecular genotypes. Moreover, a notable high level of gene flow (Nm = 2.82 > 1) among four P. triticina populations was detected. The genetic relationship among Hebei, Shandong, and Sichuan populations was close, possibly due to the spread of P. triticina from Sichuan to Shandong and then to Hebei. In contrast, the Xinjiang population was relatively independent. Genetic differentiation analysis showed some level of differentiation among or within populations of P. triticina in the four provinces, and the genetic variation within populations (74.97%) was higher than across populations (25.03%). Our study provides a basis for a better understanding of the regional migration, epidemic, and population structure of P. triticina in China.

Puccinia triticina (Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effector-mediated wheat susceptibility remains largely unexplored. In this study, an effector Pt_21 was identified to interact with the apoplast-localized wheat thaumatin-like protein TaTLP1 using a yeast two-hybrid assay and the Pt_21-TaTLP1 interaction was characterized. The interaction between Pt_21 and TaTLP1 was validated by in vivo co-immunoprecipitation assay. A TaTLP1 variant, TaTLP1C71A, that was identified by the site-directed mutagenesis failed to interact with Pt_21. Pt_21 was able to suppress Bax-mediated cell death in leaves of Nicotiana benthamiana and inhibit TaTLP1-mediated antifungal activity. Furthermore, infiltration of recombinant protein Pt_21 into leaves of transgenic wheat line overexpressing TaTLP1 enhanced the disease development of leaf rust compared to that in wild-type leaves. These findings demonstrate that Pt_21 suppresses host defense response by directly targeting wheat TaTLP1 and inhibiting its antifungal activity, which broadens our understanding of the molecular mechanisms underlying Pt effector-mediated susceptibility in wheat.

The binding capacity of 10 flavonoids with okra seed protein (OSP) was studied by fluorescence spectroscopy. The structure of flavonoids had an obvious impact on binding performance. The binding ability of flavanone was lower than that of flavone, isoflavone and dihydrochalcone. The binding capacity of flavonoid glycoside was superior to that of the corresponding flavonoid aglycone. The binding ability was positively correlated with the number of phenolic hydroxyl groups on the B ring. The steric field and electrostatic field model constructed by 3D-QSAR method could well explain the above interaction behavior. Thermodynamic analysis suggested that the quenching mechanism of OSP caused by flavonoids was static quenching, and the binding-site number was 1. In addition, hydrogen bonding and van der Waals force dominated this interaction. The 3D and synchronous fluorescence spectra showed that there was no significant change in the polarity of the environment around tryptophan and tyrosine residues during binding.

Olive leaves are rich in phenolic compounds. This study explored the chemical profiles and contents of free phenolics (FPs) and bound phenolics (BPs) in olive leaves, and further investigated and compared the antioxidant properties of FPs and BPs using chemical assays, cellular antioxidant evaluation systems, and in vivo mouse models. The results showed that FPs and BPs have different phenolic profiles; 24 free and 14 bound phenolics were identified in FPs and BPs, respectively. Higher levels of phenolic acid (i.e., sinapinic acid, 4-coumaric acid, ferulic acid, and caffeic acid) and hydroxytyrosol were detected in the BPs, while flavonoids, triterpenoid acids, and iridoids were more concentrated in the free form. FPs showed a significantly higher total flavonoid content (TFC), total phenolic content (TPC), and chemical antioxidant properties than those of BPs (p < 0.05). Within the range of doses (20–250 μg/mL), both FPs and BPs protected HepG2 cells from H2O2-induced oxidative stress injury, and there was no significant difference in cellular antioxidant activity between FPs and BPs. The in vivo experiments suggested that FP and BP treatment inhibited malondialdehyde (MDA) levels in a D-galactose-induced oxidation model in mice, and significantly increased antioxidant enzyme activity of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT), and the total antioxidant capacity (T-AOC). Mechanistically, FPs and BPs exert their antioxidant activity in distinct ways; FPs ameliorated D-galactose-induced oxidative stress injury partly via the activation of nuclear factor erythroid-2-related factor 2 (Nrf2) signaling pathway, while the BP mechanisms need further study.

Wheat leaf rust, caused by Puccinia triticina, is one of the most important fungal diseases of wheat in China. However, little is known about the dynamic changes of population structure and genetic diversity of P. triticina during a period of time. In this study, 247 isolates of P. triticina collected from Hebei Province from 2001 to 2010 were tested on 36 Thatcher near-isogenic lines for virulence diversity and detected by 21 pairs of Expressed Sequence Tag derived Simple Sequence Repeat (EST-SSR) primers for genetic diversity. A total of 204 isolates were successfully identified as 164 races, and THTT, THST, PHRT, THTS, and PHTT were the most common races in Hebei Province from 2001 to 2010. The cluster analysis based on virulence showed that P. triticina has a rich virulence polymorphism, which had a certain correlation with the years, while the cluster analysis based on EST-SSR showed that the genetic diversity of the P. triticina population was significantly different between years in Hebei Province from 2001 to 2010. In addition, the population structure of P. triticina may have changed greatly in 2007 and 2009, which was significantly different from that of 2001–2006 on either virulence or genetic characteristics. The variation frequency of the population structure had an increasing trend during this period. From 2001 to 2010, there was a certain degree of gene flow among the P. triticina populations. No significant correlation was found between virulence and molecular polymorphism. The genetic differentiation analysis of the 10 tested populations (each year as a population) showed that the coefficient of genetic differentiation (Gst) was 0.27, indicating that there was a certain genetic differentiation among or within populations of P. triticina in Hebei Province. The genetic variation within populations (73.08%) was higher than that among populations (26.92%), which indicated that the genetic variations were mainly found within populations. Our study provides the foundation for a better understanding of the population structure change and genetic diversity of P. triticina over a period in Hebei Province of China.

IntroductionWheat leaf rust caused by Puccinia triticina (Pt) remains one of the most destructive diseases of common wheat worldwide. Understanding the pathogenicity mechanisms of Pt is important to control wheat leaf rust.MethodsThe urediniospores of Pt race PHNT (wheat leaf rust resistance gene Lr19-avirulent isolate) were mutagenized with ethyl methanesulfonate (EMS), and two Lr19-virulent mutants named M1 and M2 were isolated. RNA sequencing was performed on samples collected from wheat cultivars Chinese Spring and TcLr19 infected with wild-type (WT) PHNT, M1, and M2 isolates at 14 days post-inoculation (dpi), respectively. Screening AvrLr19 candidates by quantitative reverse transcription PCR (qPCR) and Agrobacterium-mediated transient assays in Nicotiana benthamiana.Results560 genes with single nucleotide polymorphisms (SNPs) and insertions or deletions (Indels) from non-differentially expressed genes were identified. Among them, 10 secreted proteins were screened based on their fragments per kilobase of exon model per million mapped reads (FPKM) values in the database. qPCR results showed that the expression profiles of 7 secreted proteins including PTTG_27471, PTTG_12441, PTTG_28324, PTTG_26499, PTTG_06910, PTTG_26516, and PTTG_03570 among 10 secreted proteins in mutants were significantly different with that in wild-type isolate after infection wheat TcLr19 and might be related to the recognition between Lr19 and AvrLr19. In addition, a total of 216 differentially expressed genes (DEGs) were obtained from three different sample comparisons including M1-vs-WT, M2-vs-WT, and M1-vs-M2. Among 216 DEGs, 15 were predicted to be secreted proteins. One secreted protein named PTTG_04779 could inhibit programmed progress of cell death (PCD) induced by apoptosis-controlling genes B-cell lymphoma-2 associated X protein (BAX) on Nicotiana benthamiana, indicating that it might play a virulence function in plant. Taken together, total 8 secreted proteins, PTTG_04779, PTTG_27471, PTTG_12441, PTTG_28324, PTTG_26499, PTTG_06910, PTTG_26516, PTTG_03570 are identified as AvrLr19 candidates.DiscussionOur results showed that a large number of genes participate in the interaction between Pt and TcLr19, which will provide valuable resources for the identification of AvrLr19 candidates and pathogenesis-related genes.

Wheat leaf rust, caused by Puccinia triticina Eriks. (Pt), is a global wheat disease threatening wheat production. Dissecting how Pt effector proteins interact with wheat has great significance in understanding the pathogenicity mechanisms of Pt. In the study, the cDNA of Pt 13-5-72 interacting with susceptible cultivar Thatcher was used as template to amplify Pt13024 gene. The expression pattern and structure of Pt13024 were analyzed by qRT-PCR and online softwares. The secretion function of Pt13024 signal peptide was verified by the yeast system. Subcellular localization of Pt13024 was analyzed using transient expression on Nicotiana benthamiana. The verification that Pt13024 inhibited programmed cell death (PCD) was conducted on N. benthamiana and wheat. The deletion mutation of Pt13024 was used to identify the virulence function motif. The transient transformation of wheat mediated by the type III secretion system (TTSS) was used to analyze the activity of regulating the host defense response of Pt13024. Pt13024 gene silencing was performed by host-induced gene silencing (HIGS). The results showed that Pt13024 was identified as an effector and localized in the cytoplasm and nucleus on the N. benthamiana. It can inhibit PCD induced by the Bcl-2-associated X protein (BAX) from mice and infestans 1 (INF1) from Phytophthora infestans on N. benthamiana, and it can also inhibit PCD induced by DC3000 on wheat. The amino acids 22 to 41 at N-terminal of the Pt13024 are essential for the inhibition of programmed cell death (PCD) induced by BAX. The accumulation of reactive oxygen species and deposition of callose in near-isogenic line TcLr30, which is in Thatcher background with Lr30, induced by Pt13024 was higher than that in 41 wheat leaf rust-resistant near-isogenic lines (monogenic lines) with different resistance genes and Thatcher. Silencing of Pt13024 reduced the leaf rust resistance of Lr30 during the interaction between Pt and TcLr30. We can conclude that Pt13024 is avirulent to TcLr30 when Pt interacts with TcLr30. These findings lay the foundation for further investigations into the role of Pt effector proteins in pathogenesis and their regulatory mechanisms.

Vegetables from the Brassica species are excellent sources of glucosinolates (GLSs), the precursors of health-promoting isothiocyanates (ITCs). Fermentation enhances the biotransformation of GLSs into potential bioactive ITCs. To explore the biotransformation of GLSs during Brassica fermentation, the changes in GLSs during the fermentation of two Brassica species (i.e., cauliflower and broccoli); the formation of corresponding breakdown products; and the shifts in physicochemical parameters, bacterial communities, and myrosinase activities involved in GLSs degradation were systematically investigated. Nine aliphatic, three indolic, and two benzenic GLSs were identified in fermented cauliflower (FC) and fermented broccoli (FB). Aliphatic glucoiberin and glucoraphanin were the major forms of GLS in FC and FB, respectively; indolic glucobrassicin was also abundant in both FC and FB. The total GLS content decreased by 85.29% and 65.48% after 3 d of fermentation in FC and FB, respectively. After 2 d of fermentation, a significant increase in bioactive GLS degradation products (P

The impact of fermentation by Levilactobacillus brevis YSJ3 on sleep-promoting components (SPCs) of carrot juice was evaluated. The contents of acetic acid, isovaleric acid, butyric acid, and γ-aminobutyric acid (GABA) significantly increased after fermentation. The beneficial effects of fermented carrot juice (FCJ) on sleep were evaluated in animal experiments. Behavioral test reveal SPCs-enriched FCJ could effectively relieve anxiety. The sleep duration in the FCJ group were extended compared to the control (NC) group and the unfermented carrot juice (UCJ) group. Moreover, the relative abundances of Ruminiclostridium and Akkermansia in the FCJ group and PC group, respectively, increased significantly, compared to the NC group the UCJ group. The contents of gut short-chain fatty acids in the FCJ group were significantly higher than that in the NC group and the UCJ group. The levels of GABA and 5-hydroxytryptamine in the brain for the FCJ group also increased significantly, compared to the NC group and the UCJ group. It indicated that SPCs-enriched FCJ effectively improved sleep in mice, which might be related to the fermentation of carrot juice and the compounds produced during the fermentation.

Puccinia triticina, which is the causative agent of wheat leaf rust, is widely spread in China and most other wheat-planting countries around the globe. Cultivating resistant wheat cultivars is the most economical, effective, and environmentally friendly method for controlling leaf rust-caused yield damage. Exploring the source of resistance is very important in wheat resistance breeding programs. In order to explore more effective resistance sources for wheat leaf rust, the resistance of 112 wheat accessions introduced from the U.S. National Plant Germplasm System were identified using a mixture of pathogenic isolates of THTT, THTS, PHTT, THJT and THJS which are the most predominant races in China. As a result, all of these accessions showed high resistance at seedling stage, of which, ninety-nine accessions exhibited resistance at adult plant stage. Eleven molecular markers of eight effective leaf rust resistance genes in China were used to screen the 112 accessions. Seven effective leaf rust resistance genes Lr9, Lr19, Lr24, Lr28, Lr29, Lr38 and Lr45 were detected, except Lr47. Twenty-three accessions had only one of those seven effective leaf rust resistance gene. Eleven accessions carried Lr24+Lr38, and 7 accessions carried Lr9+Lr24+Lr38, Lr24+Lr38+Lr45, Lr24+Lr29+Lr38 and Lr19+Lr38+Lr45 respectively. The remaining seventy-one accessions had none of those eight effective leaf rust resistance genes. This study will provide theoretical guidance for rational utilization of these introduted wheat accessions directly or for breeding the resistant wheat cultivars.

Wheat stripe rust, caused by Puccinia striiformis Westend. f.sp. tritici Eriks (Pst), is one of the most important diseases of bread wheat worldwide. Breeding resistant wheat cultivars is the most economical, effective and environmentally friendly way for controlling wheat stripe rust in China. The Romanian wheat line Fundulea 900 showed good resistance to wheat stripe rust at the adult stage. The present study aimed to map the quantitative trait loci (QTLs) for stripe rust resistance in 176 F2:6 recombinant inbred lines (RIL) derived from the cross of Fundulea 900 × Thatcher. The RIL population was phenotyped for stripe rust (YR) severity at Mianyang in the Sichuan province and Baoding in the Hebei province in the 2016/2017 and 2017/2018 cropping seasons. SSR markers combined with a preferred screened group (PSG) analysis were used to identify the QTLs for stripe rust in the population. Three QTLs for stripe rust resistance were mapped on chromosomes 1AL, 7BL and 7DS, respectively. All three QTLs originated from Fundulea 900 and were detected in all the environments. The QTL on 7DS was provided by the known resistance gene Yr18/Lr34. The two QTLs on chromosomes 1AL and 7BL were explained by 9.2 to 21.5% and 5.1 to 10.1% of the phenotypic variance, respectively and might be new QTLs. The QTLs identified in the study and their closely linked markers can be used for marker-assisted selection (MAS) in wheat breeding programmes.

Pathogenesis-related (PR) proteins play important roles in plant defense response and systemic acquired resistance (SAR). PR1 has antifungal activity against many plant pathogens. In our previous study, RNA sequencing (RNA-seq) was conducted on resistant wheat line TcLr19 and sensitive wheat cultivar Chinese Spring inoculated with Puccinia triticina (Pt) race PHNT. In this study, seven salicylic acid (SA)-induced TaPR1 genes involved in plant disease resistance were found in the RNA-seq library. Quantitative PCR (qPCR) results showed that TaPR1-4 was most induced by Pt among these seven TaPR1 genes in the incompatible interaction. Yeast two-hybrid (Y2H) results showed that TaPR1-4 interacted with TaTLP1 via the αIV helix. Protein-mediated phenotyping assays in vivo and antifungal activity in vitro demonstrated that wheat leaves infiltrated with pure TaPR1-4 protein developed significantly less disease compared to control leaves. This effect was correlated with a strong increase in defense gene expression, and resistance activity was dependent on the CAPE1 motif located in the C-terminal region of TaPR1-4. These findings increase current knowledge regarding the interaction of TaPR1 and TaTLP1 and provide new insights on the role of TaPR1 protein in the resistance of wheat to Pt.

Abstract NPR1 is a key regulator of systemic acquired resistance (SAR) in plant species. In our previous study, we identified a conserved fungal effector PNPi from Puccinia striiformis f. sp. tritici (Pst) that can suppress acquired resistance in local leaf by directly targeting the wheat NPR1 protein. In this investigation, we identified and validated a novel protein interaction between PNPi and wheat pathogenesis-related TaPR1a in the apoplastic space. TaPR1a-overexpressing wheat lines exhibited enhanced resistance to both Pst and Puccinia triticina (Pt). We further determined that exogenous expression of PNPi RNA in transgenic wheat lines reduced the degree of acquired resistance to Magnaporthe oryzae isolate P131 in the region adjacent to Pseudomonas syringae pv. tomato DC3000 infection area. Additionally, when PNPi was overexpressed, the expression levels of two plant defense responsive genes were suppressed upon P. syringae DC3000 infection in the local infiltration region. These findings established the mechanism of a single rust effector that can suppress multiple defense responses in wheat plants by targeting different components.

Stripe or yellow rust (YR) and leaf rust (LR) cause large losses in wheat production worldwide. Resistant cultivars curtail the levels of losses. The present study aimed to identify quantitative trait loci (QTL) for YR and LR resistance in 147 F2:6 recombinant inbred lines (RIL) derived from the cross Fuyu 3/Zhengzhou 5389. The RIL population and parents were genotyped with the Wheat55K single nucleotide polymorphism (SNP) array and simple sequence repeat (SSR) markers. All materials were also phenotyped for YR severity at Mianyang in Sichuan province and Baoding in Hebei province in the 2015/2016, 2016/2017, and 2017/2018 cropping seasons, and LR severity at Zhoukou in Henan province and at Baoding in 2017/2018. Eleven QTL for YR resistance and five for LR resistance were detected using inclusive composite interval mapping (IciMapping). Four of these QTL on chromosomes 1BL, 2BS, 3AL, and 5AL conferred resistance to both YR and LR. The QTL on 1BL was Lr46/Yr29, and that on 7BL might be Lr68. The QTL on chromosome 2BS was detected at a similar position to previously detected loci. QYr.hebau-3AL/QLr.hebau-3AL, QYr.hebau-5AL/QLr.hebau-5AL, QYr.hebau-7DL, QYr.hebau-4BS, QYr.hebau-6DL, and QYr.hebau-2AS are likely to be new. An SSR marker for QYr.hebau-7DL was developed and validated in a diverse wheat panel from China, suggesting effectiveness in different genetic backgrounds. These QTL with closely linked SNP and SSR markers could be useful for marker-assisted selection in wheat breeding programs targeting durable resistance to both diseases.

Abstract Background Wheat leaf rust is an important disease worldwide. Understanding the pathogenic molecular mechanism of Puccinia triticina Eriks. (Pt) and the inconstant toxic region is critical for managing the disease. The present study aimed to analyze the pathogenic divergence between Pt isolates. Results Total RNA was extracted from the wheat cultivar Thatcher infected by two Pt isolates, Tc361_1 (THTT) and Tc284_2 (THTS), at 144 h post inoculation (hpi). The mRNA was then sequenced, and a total of 2784 differentially expressed genes (DEGs) were detected. Forty-five genes were specifically expressed in THTT; these genes included transcription initiation factors and genes with transmembrane transporter activity and other genes. Twenty-six genes were specifically expressed in THTS, including genes with GTPase activity, ABC transporters and other genes. Fifty-four differentially expressed candidate effectors were screened from the two isolates. Two candidate effectors were chosen and validated on tobacco, and the results showed that they could inhibit necrosis induced by BAX. qRT-PCR of 12 significant DEGs was carried out to validate that the results are similar to those of RNA-seq at 144 hpi, to show the expression levels of these DEGs in the early stage and to elucidate the differences in expression between the two Pt pathotypes. Conclusion The results obtained in this study showed that although the two pathotypes of THTT and THTS contribute similar virulence to wheat, there are a large number of genes participate in the interaction with the susceptible wheat cultivar Thatcher, and revealed the pathogenicity of rust is very complicated.

In Arabidopsis, both pathogen invasion and benzothiadiazole (BTH) treatment activate the nonexpresser of pathogenesis-related genes 1 (NPR1)-mediated systemic acquired resistance, which provides broad-spectrum disease resistance to secondary pathogen infection. However, the BTH-induced resistance in Triticeae crops of wheat and barley seems to be accomplished through an NPR1-independent pathway. In the current investigation, we applied transcriptome analysis on barley transgenic lines overexpressing wheat wNPR1 (wNPR1-OE) and knocking down barley HvNPR1 (HvNPR1-Kd) to reveal the role of NPR1 during the BTH-induced resistance. Most of the previously designated barley chemical-induced (BCI) genes were upregulated in an NPR1-independent manner, whereas the expression levels of several pathogenesis-related (PR) genes were elevated upon BTH treatment only in wNPR1-OE. Two barley WRKY transcription factors, HvWRKY6 and HvWRKY70, were predicted and further validated as key regulators shared by the BTH-induced resistance and the NPR1-mediated acquired resistance. Wheat transgenic lines overexpressing HvWRKY6 and HvWRKY70 showed different degrees of enhanced resistance to Puccinia striiformis f. sp. tritici pathotype CYR32 and Blumeria graminis f. sp. tritici pathotype E20. In conclusion, the transcriptional changes of BTH-induced resistance in barley were initially profiled, and the identified key regulators would be valuable resources for the genetic improvement of broad-spectrum disease resistance in wheat.[Graphic: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Olive leaves, which are the most abundant byproducts of the olive industry, offer multiple health benefits. The investigation of the phytochemical profiles and relevant biological activities is an essential step toward transforming these low-value byproducts into value-added ones. This study systematically investigated the phytochemical profiles, antioxidant capacity, and inhibition rates of olive leaves from four cultivars on the α-glucosidase, α-amylase, and angiotensin-converting enzyme (ACE). The leaves were prepared using two common drying methods, namely, hot air-drying and freeze-drying. A total of 33 bioactive compounds were identified in the olive leaves, namely, 19 flavonoids, 2 phenylethanoids, 2 coumarins, 2 hydroxycinnamic acids, 2 iridoids, and 6 triterpenic acids. Quantification of the bioactive compounds revealed high amounts of polyphenols, especially flavonoids [2,027–8,055 mg/kg dry weight (DW)], iridoids (566–22,096 mg/kg DW), and triterpenic acids (13,824–19,056 mg/kg DW) in the olive leaves. The hot air-dried leaves showed significantly (P < 0.05) higher iridoid (oleuropein and secoxyloganin) content than the fresh leaves, while freeze-drying resulted in significantly (P < 0.05) higher flavonoid aglycone and hydroxytyrosol content. Additionally, freeze-drying led to samples with the highest radical scavenging, α-amylase, α-glucosidase, and ACE inhibition abilities. The flavonoid (e.g., quercetin, luteolin, eriodictyol, kaempferol-7-O-glucoside, and luteolin-7-O-glucoside), hydroxytyrosol, and oleanolic acid contents in the olive leaves were positively correlated (P < 0.05) with their bioactive potentials.

Wheat bran supplementation in cereal food processing improves the nutritional value and quality of the final products. However, whether wheat bran has the potential as a biofortifier to enhance nutritional and flavor of fermented vegetables remains unknown. The study aimed to evaluate the potential of wheat bran supplementation for nutrition and flavor fortification during radish fermentation, and to explore the role of microorganisms in nutritional and flavor development. Using high-throughput sequencing coupled with high-performance liquid chromatography and headspace solid-phase microextraction-gas chromatography-mass spectrometry, the microbial community profiles and nutritional and flavor changes of wheat bran-treated samples were analyzed and compared with control samples. Correlation analysis between bacteria taxa with metabolites were also performed. The results showed that wheat bran treatment increased the content of most free amino acids (FAAs), α-linolenate, thiamine, and riboflavin in the samples (p < 0.05). In addition, the increased consumption of reducing sugar and glutamate in the wheat bran-treated samples was due to the production of secondary metabolites such as lactic acid, ethanol, acetic acid, and GABA (p < 0.05). Moreover, compared with control samples, the flavor of the wheat bran-treated pickled radish was preferable. Wheat bran increased the amount of alcohol, ester, acid, and ketones compounds but reduced the number of sulfides, which increased the aroma but decreased the pungent flavor. Additionally, the correlation analysis suggested that Lactobacillus, the most dominant genus, was boosted by wheat bran and was positively associated with most of FAAs, GABA, and lactate, while negatively associated with most sulfides. Therefore, compared with the control, wheat bran treatment could improve the nutritional values and sensorial properties of radish pickles. New areas of research should explore the co-fermentation of other vegetables with wheat bran, and the potential of this processing technique to provide consumers with products of high nutritional quality.

Cyst nematodes are one of the most important pathogens worldwide. Most cyst nematode species have been reported recently in China. From 2016 to 2020, an extensive survey of cyst nematodes was conducted in the Bashang region of Hebei Province. A total of 158 soil samples were collected, and cyst-forming nematodes were isolated from five soil samples. Morphological and molecular characterization showed that four of the cyst-forming nematode populations were Heterodera glycines (SCN), named populations of ZM, KM, CB and FN, respectively. These SCN populations were collected from Zhangbei County, Kangbao County, Chabei Management Area of Zhangjiakou and Fengning Manchu Autonomous County of Chengde, respectively, where the corresponding cyst densities were 57, 41, 103 and 31 cysts/200 cc soil. Furthermore, the populations of ZM, KM and CB were identified as race 4, whereas the FN population was identified as race 3. The cyst-forming nematode population was collected from Zhangbei County of Zhangjiakou, which was confirmed to be Heterodera schachtii (SBCN), named population ZZ, and the cyst density was 94 cysts/200 cc soil. It is a new disease of Chinese cabbage caused by SBCN based on Koch’s postulates. Fourteen cultivars from five plant families were evaluated as hosts for SBCN. Chinese cabbage (cv. Linglonghuang012) and cabbage (cv. Chunwang) were suitable hosts, while celery (cv. Yuhuang), potato (cv. Helan 15) and eggplant (cv. Junlang) were nonhosts. The obtained results regarding the occurrence, distribution, races of SCN and hosts of SBCN in the Bashang region in this study provide a reference for SCN and SBCN management.

Differences in flavor compounds and bacterial communities of Xuecai by traditional and modern fermentation are poorly understood. Allyl isothiocyanate (E9), ethyl acetate (E1), 3-butenenitrile (N1), phenol (P1), ethanol (A1), and 3-(2,6,6-trimethyl-1-cyclohexen-1-yl) acrylaldehyde (L11) were the main flavor compounds that differed between Xuecai produced by traditional and modern fermentation. Among these compounds, the contents of N1 and E9 were higher in modern fermentation Xuecai. Traditional fermentation Xuecai possessed higher contents of A1, P1, E1, and L11. High-throughput sequencing showed that Lactobacillus-related genera was the most abundant genus (50%) in modern fermentation Xuecai. However, in traditional fermentation Xuecai, Halanaerobium (29.06%) and Halomonas (12.96%) were the dominant genera. Halophilic bacteria (HB) positively contribute to the flavor of Xuecai. Carbohydrate metabolism and amino acid metabolism were the most abundant pathways associated with the bacterial communities of the Xuecai. This indicated that Xuecai flavor formation is mainly dependent on protein and carbohydrate degradation. This study provides a novel insight that HB may be important for flavor formation of Xuecai.

NAC transcription factors are one of the largest transcription factor families having functions in a variety of stress responses. Few NACs have been reported for interactions between wheat and the wheat rust fungus Puccinia triticina (Pt). In this study, based on analysis of RNA-seq data from wheat line TcLr19 inoculated by Pt, the NAC transcription factor TaNAC069 was cloned from wheat, and its transcriptional activity and homologous dimer formation were verified. Quantitative real-time PCR analysis showed that the expression of TaNAC069 was induced by Pt and associated signaling molecules. To further characterize the function of the TaNAC069 gene in wheat resistance to Pt, virus-induced gene silencing (VIGS) was utilized, and it revealed that Pt resistance in TaNAC069-silenced plants was significantly reduced. Potential interaction targets of TaNAC069 from wheat and Pt were screened and identified by yeast two-hybrid technology. Eukaryotic elongation factor eEF1A, CBSX3 protein, and cold acclimation protein WCOR410c were screened by yeast one-hybrid technology. The results indicate that the TaNAC069 gene plays a positive regulatory role in wheat resistance to Pt, laying a good foundation to analyze the molecular mechanisms of TaNAC069 and its functional role in wheat resistance to Pt.

Olive (Olea europaea L.) is a world-famous woody oil tree and popular for redundant unsaturated fatty acids. Fatty acid desaturase (FAD) genes are responsible for fatty acid desaturation and stress regulation but have not yet been identified in olive at the whole genome level. This study identified 40 and 27 FAD genes in the cultivated olive O. europaea cv. Farga and the wild olive O. europaea var. Sylvestris, respectively. Phylogenetic analysis showed that all the FAD genes could be classified into the soluble FAB2/SAD clade and membrane-bound clade, including ADS/FAD5, DES, FAD4, SLD, ω-6 and ω-3, with the high consistency of subcellular localization, motif composition and exon-intron organization in each group. FAD genes in olive showed the diverse functional differentiation in morphology of different tissues, fruit development and stress responses. Among them, OeFAB2.8 and OeFAD2.3 were up-regulated and OeADS.1, OeFAD4.1 and OeFAD8.2 were down-regulated under the wound, Verticillium dahliae and cold stresses. This study presents a comprehensive analysis of the FAD genes at the whole-genome level in olives and will provide guidance for the improvement of oil quality or stress tolerance of olive trees.

F-box proteins play critical roles in plant responses to biotic/abiotic stresses. In the present study, a total of 68 wheat F-box/Kelch (TaFBK) genes, unevenly distributed across 21 chromosomes and encoding 74 proteins, were identified in EnsemblPlants. Protein sequences were compared with those of Arabidopsis and three cereal species by phylogenetic and domain analyses, where the wheat sequences were resolved into 6 clades. In silico analysis of a digital PCR dataset revealed that TaFBKs were expressed at multiple developmental stages and tissues, and in response to drought and/or heat stresses. The TaFBK19 gene, a homolog of the Attenuated Far-Red Response (AFR) genes in other plant species, and hence named TaAFR, was selected for further analysis. Reverse-transcription quantitative real-time PCR (RT-qPCR) was carried out to determine tissue-specific, hormone and stress (abiotic/biotic) responsive expression patterns. Of interest, TaAFR was expressed most abundantly in the leaves, and its expression in response to leaf rust variants suggests a potential role in compatible vs incompatible rust responses. The protein was predicted to localize in cytosol, but it was shown experimentally to localize in both the cytosol and the nucleus of tobacco. A series of protein interaction studies, starting with a yeast-2-hybrid (Y2H) library screen (wheat leaf infected with incompatible leaf rust pathogens), led to the identification of three TaAFR interacting proteins. Skp1/ASK1-like protein (Skp1) was found to interact with the F-box domain of TaAFR, while ADP-ribosylation factor 2-like isoform X1 (ARL2) and phenylalanine ammonia-lyase (PAL) were shown to interact with its Kelch domain. The data presented herein provides a solid foundation from which the function and metabolic network of TaAFR and other wheat FBKs can be further explored.

Wheat leaf rust caused by Puccinia triticina is one of the most common and serious diseases in wheat production. The constantly changing pathogens overcome the plant resistance to P. triticina. Plant pathogens secrete effector proteins that alter the structure of the host cell, interfere plant defenses, or modify the physiology of plant cells. Therefore, the identification of effector proteins is critical to reveal the pathogenic mechanism. We used SignalP v4.1, TargetP v1.1, TMHMM v2.0, and EffectorP v2.0 to screen the candidate effector proteins in P. triticina isolates – KHTT, JHKT, and THSN. As a result, a total of 635 candidate effector proteins were obtained. Structural analysis showed that effector proteins were small in size (50AA to 422AA) and of diverse sequences, and the conserved sequential elements or clear common elements were not involved, regardless of their secretion from the pathogen to the host. There were 427 candidate effector proteins that contain more than or equal to 4 cysteine residues, and 339 candidate effector proteins contained the known motifs. Sixteen families, 9 domains, and 53 other known functional types were found in 186 candidate effector proteins using the Pfam search. Three novel motifs were found by MEME. Heterogeneous expression system was performed to verify the functions of 30 candidate effectors by inhibiting the programmed cell death (PCD) induced by BAX (the mouse-apoptotic gene elicitor) on Nicotiana benthamiana. Hypersensitive response (HR) can be induced by the six effectors in the wheat leaf rust resistance near isogenic lines, and this would be shown by the method of transient expression through Agrobacterium tumefaciens infiltration. The quantitative reverse transcription PCR (qRT-PCR) analysis of 14 candidate effector proteins secreted after P. triticina inoculation showed that the tested effectors displayed different expression patterns in different stages, suggesting that they may be involved in the wheat–P. triticina interaction. The results showed that the prediction of P. triticina effector proteins based on transcriptomic analysis and multiple bioinformatics software is effective and more accurate, laying the foundation of revealing the pathogenic mechanism of Pt and controlling disease.

Thaumatin-like proteins (TLPs), which are defined as pathogenesis-related protein family 5 (PR5) members, are common plant proteins involved in defense responses and confer antifungal activity against many plant pathogens. Our earlier studies have reported that the TaTLP1 gene was isolated from wheat and proved to be involved in wheat defense in response to leaf rust attack. The present study aims to identify the interacting proteins of TaTLP1 and characterize the role of the interaction between wheat and Puccinia triticina (Pt). Pull-down experiments designed to isolate the molecular target of TaTLP1 in tobacco resulted in the identification of TaPR1, a pathogenesis-related protein of family 1, and the interaction between TaTLP1 and TaPR1 was confirmed by yeast two-hybrid experiments (Y2H), bimolecular fluorescence complementation (BiFC), and co-immunoprecipitation (Co-IP). In vitro, TaTLP1 and TaPR1 together increased antifungal activity against Pt. In vivo, the disease resistance phenotype, histological observations of fungal growth and host responses, and accumulation of H2O2 in TaTLP1-TaPR1 in co-silenced plants indicated that co-silencing significantly enhanced wheat susceptibility compared to single knockdown TaTLP1 or TaPR1 plants. The accumulation of reactive oxygen species (ROS) was significantly reduced in co-silenced plants compared to controls during Pt infection, which suggested that the TaTLP1-TaPR1 interaction positively modulates wheat resistance to Pt in an ROS-dependent manner. Our findings provide new insights for understanding the roles of two different PRs, TaTLP1 and TaPR1, in wheat resistance to leaf rust.

Rust fungi secrete various specialized effectors into host cells to manipulate the plant defense response. Conserved motifs, including RXLR, LFLAK-HVLVxxP (CRN), Y/F/WxC, CFEM, LysM, EAR, [SG]-P-C-[KR]-P, DPBB_1 (PNPi), and ToxA, have been identified in various oomycete and fungal effectors and are reported to be crucial for effector translocation or function. However, little is known about potential effectors containing any of these conserved motifs in the wheat leaf rust fungus (Puccinia triticina, Pt). In this study, sequencing was performed on RNA samples collected from the germ tubes (GT) of uredospores of an epidemic Pt pathotype PHTT(P) and Pt-infected leaves of a susceptible wheat cultivar “Chinese Spring” at 4, 6, and 8 days post-inoculation (dpi). The assembled transcriptome data were compared to the reference genome of “Pt 1-1 BBBD Race 1.” A total of 17,976 genes, including 2,284 “novel” transcripts, were annotated. Among all these genes, we identified 3,149 upregulated genes upon Pt infection at all time points compared to GT, whereas 1,613 genes were more highly expressed in GT. A total of 464 secreted proteins were encoded by those upregulated genes, with 79 of them also predicted as possible effectors by EffectorP. Using hmmsearch and Regex, we identified 719 RXLR-like, 19 PNPi-like, 19 CRN-like, 138 Y/F/WxC, and 9 CFEM effector candidates from the deduced protein database including data based on the “Pt 1-1 BBBD Race 1” genome and the transcriptome data collected here. Four of the PNPi-like effector candidates with DPBB_1 conserved domain showed physical interactions with wheat NPR1 protein in yeast two-hybrid assay. Nine Y/F/WxC and seven CFEM effector candidates were transiently expressed in Nicotiana benthamiana. None of these effector candidates showed induction or suppression of cell death triggered by BAX protein, but the expression of one CFEM effector candidate, PTTG_08198, accelerated the progress of cell death and promoted the accumulation of reactive oxygen species (ROS). In conclusion, we profiled genes associated with the infection process of the Pt pathotype PHTT(P). The identified effector candidates with conserved motifs will help guide the investigation of virulent mechanisms of leaf rust fungus.

Wheat cultivar Een1, 34 near isogenic lines (NILs), and two cultivars were used as plant materials to evaluate the resistance of Een1 to leaf rust disease. Infection type identification and gene postulation were carried out by inoculation of 12 Chinese Puccinia triticina (Pt) pathotypes. Based on the unique phenotype of Een1, we speculated that Een1 might carry Lr gene(s) different from the tested ones. The chromosomal locations for resistance gene to leaf rust disease was employed using SSR primers mapping the populations derived from the cross between Een1 and susceptible Thatcher. A total of 285 plants in the F2 population were tested by inoculating Pt pathotype FHNQ during the seedling stage. Results from the segregation analysis fits a ratio of 3:1 ( ${\chi }_{3:1}^{2}=2.37$ χ 3 : 1 2 = 2 . 37 , P = 0.12), indicating the presence of a single dominant gene in Een1 conferring resistance to FHNQ. A total of 1,255 simple sequence repeat (SSR) primers were first used to identify the likely linked markers based on bulk segregation analysis (BSA), and then those likely linked markers were further genotyped in the F2 population for linkage analysis. Our linkage analysis found that the resistance gene (LrE1) was distal to seven SSR loci on the long arm of chromosome 7B, with distances from 2.6 cM (Xgwm344) to 27.1 cM (Xgwm131). The closest marker Xgwm344 was further verified with F3 lines.

Olives (Olea europaea L.) are a significant part of the agroindustry in China. Olive leaves, the most abundant by-products of the olive and olive oil industry, contain bioactive compounds that are beneficial to human health. The purpose of this study was to evaluate the phytochemical profiles and antioxidant capacities of olive leaves from 32 cultivars grown in China. A total of 32 phytochemical compounds were identified using high-performance liquid chromatography–electrospray ionization–tandem mass spectrometry, including 17 flavonoids, five iridoids, two hydroxycinnamic acids, six triterpenic acids, one simple phenol, and one coumarin. Specifically, olive leaves were found to be excellent sources of flavonoids (4.92–18.29 mg/g dw), iridoids (5.75–33.73 mg/g dw), and triterpenic acids (15.72–35.75 mg/g dw), and considerable variations in phytochemical content were detected among the different cultivars. All tested cultivars were classified into three categories according to their oil contents for further comparative phytochemicals assessment. Principal component analysis indicated that the investigated olive cultivars could be distinguished based upon their phytochemical profiles and antioxidant capacities. The olive leaves obtained from the low-oil-content (20%) cultivars contained mainly triterpenic acids in their compositions. Correspondingly, the low-oil-content cultivars (OL3, Frantoio selection and OL14, Huaou 5) exhibited the highest ABTS antioxidant activities (758.01 ± 16.54 and 710.64 ± 14.58 mg TE/g dw, respectively), and OL9 (Olea europaea subsp. Cuspidata isolate Yunnan) and OL3 exhibited the highest ferric reducing/antioxidant power assay values (1228.29 ± 23.95 mg TE/g dw and 1099.99 ± 14.30 mg TE/g dw, respectively). The results from this study may be beneficial to the comprehensive evaluation and utilization of bioactive compounds in olive leaves.

Abstract Background Plants have evolved multifaceted defence mechanisms to resist pathogen infection. Production of the pathogenesis-related (PR) proteins in response to pathogen attack has been implicated in plant disease resistance specialized in systemic-acquired resistance (SAR). Our earlier studies have reported that a full length TaLr35PR5 gene, encoding a protein exhibiting amino acid and structural similarity to a sweet protein thaumatin, was isolated from wheat near-isogenic line TcLr35. The present study aims to understand the function of TaLr35PR5 gene in Lr35-mediated adult resistance to Puccinia triticina. Results We determined that the TaLr35PR5 protein contained a functional secretion peptide by utilizing the yeast signal sequence trap system. Using a heterologous expression assay on onion epidermal cells we found that TaLr35PR5 protein was secreted into the apoplast of onion cell. Expression of TaLr35PR5 was significantly reduced in BSMV-induced gene silenced wheat plants, and pathology test on these silenced plants revealed that Lr35-mediated resistance phenotype was obviously altered, indicating that Lr35-mediated resistance was compromised. Conclusions All these findings strongly suggest that TaLr35PR5 is involved in Lr35-mediated adult wheat defense in response to leaf rust attack.

Common wheat (Triticum aestivum L.) is the major crop cultivated in Xinjiang and Anhui provinces of China. The climate in these two provinces is favourable for wheat leaf rust (Puccinia triticina) (Pt) infection. Here, we demonstrate a detailed investigation on the leaf rust resistance of 60 major wheat cultivars cultivated in these two regions. A mixture of high virulent Pt races (THTT, THTS, THTQ and PHPS) were used to phenotype all the collected wheat cultivars at an adult plant stage. Phenotypic disease severity (FDS) and the area under the disease progress curve (AUDPC) for each of these wheat cultivars were calculated. Among all the tested wheat cultivars, three cultivars (Xindong20, Xindong 29 and 99AR142-1) with the lowest FDS and AUDPC may carry major resistance genes. Twenty-seven cultivars (45% of the total tested ones) showed a relatively lower resistance with an average of 12.52% FDS and 126.3 AUDPC. Minor resistance or slow rusting genes may be present in this group of cultivars. Molecular markers for leaf rust resistance genes Lr1, Lr9, Lr19, Lr24, Lr26 and Lr34 were further used for the genotypic screening. Lr1, Lr19, Lr26 and Lr34 were detected in 19 (31.7%), 1 (1.7%), 12 (20%) and 6 (10%) wheat cultivars, respectively. Neither Lr9 nor Lr24 could be detected in any of the tested cultivars. These results will greatly improve wheat molecular breeding for leaf rust resistance in these areas.

Pathogenesis-related (PR) protein-5, is involved in host defense system against both biotic and abiotic stresses as well as the regulation of physiological processes in numerous plant species. Our earlier studies have reported the isolation of a full-length TaLr19TLP1 gene (516 bp, GenBank accession No. KJ764822) from wheat infected by leaf rust. Quantitative real-time polymerase chain reaction analyses revealed that TaLr19TLP1 transcript was significantly induced and upregulated during incompatible interaction, while a relatively low level of the transcript was detected during compatible interaction. In the current study, we demonstrate that the accumulation of TaLr19TLP1 transcript is significantly different in tested wheat organs. TaLr19TLP1 was induced by salicylic acid (SA), methyl jasmonic (MeJA), ethephon (ETH) and abscisic acid (ABA). The transcripts of TaLr19TLP1 accumulated at higher levels following pretreatment with SA, MeJA and ABA prior to infection with P. triticina. A slight induction was observed in ETH pretreated seedlings compared with the treatment without inoculation. In addition, TaLr19TLP1 was found to be predominately localized to extracellular spaces of onion epidermal cell. Knocking down the expression of TaLr19TLP1 through virus-induced gene silencing reduced wheat resistance against leaf rust pathogen. These results suggested that TaLr19TLP1 mediated disease resistance in wheat exposed to leaf rust pathogen.

The wheat (Triticum aestivum L.) line 19HRWSN-129 from CIMMYT showed resistance to the major Puccinia triticina (Pt) races collected in China. A 144 F2:3 population was generated by crossing the resistant line 19HRWSN-129 with the susceptible line Zhengzhou5389. All the F2:3 plants were phenotyped by inoculating with the Pt race PHGN in the greenhouse. Segregation analysis indicated the presence of a single dominant gene conferring resistance to leaf rust in the wheat line 19HRWSN-129, temporarily designated as Lrshi. Using molecular markers, we mapped this resistance locus on chromosome 7BL. It was closely linked to the SSR markers Xgwm344 and Xcfa2040 with genetic distances of 2.8 cM and 1.4 cM, respectively. Compared with other chromosome 7BL-located leaf rust resistance genes (Lr14a, Lr14b, Lr68), we demonstrate, that Lrshi is a novel wheat leaf rust resistance gene.

At present, De Man, Rogosa and Sharpe (MRS) broth is the medium of choice for promoting bacteriocin production. However, this medium is expensive and not applicable for large-scale production. Therefore, a low-cost and high-efficiency culture medium for bacteriocin Lac-B23 production by Lactobacillus plantarum J23 was developed. First, the effects of the composition of MRS broth on bacteriocin Lac-B23 production and bacterial growth were researched by a one variable at a time approach. Then, a Plackett-Burman design was used to screen significant components for production. Finally, the steepest ascent and central composite designs were used to obtain an optimum medium. The final composition of the modified MRS was much simpler than MRS broth, and the modified MRS contained only glucose, yeast extract, dipotassium phosphate, manganese sulfate monohydrate, Tween 80 and sodium acetate anhydrous. The highest bacteriocin Lac-B23 production reached 2560 activity units (AU)/mL in the modified MRS, which is nine times higher than that in MRS broth (280 AU/mL). Meanwhile, the cost per liter of the modified MRS (8.56 Ren Min Bi (RMB)/L) is 34.70% the cost of MRS broth (13.11 RMB/L), and the cost per arbitrary units of bacteriocin Lac-B23 in the modified MRS is approximately fourteen times more convenient (3.34 RMB/106 AU) than in the MRS broth (46.82 RMB/106 AU).

Subcritical water can effectively hydrolyze pectin into smaller molecules while still maintaining its functional regions. Pectic heteropolysaccharide can mediate immune regulation; however, the possible effects of subcritical water-hydrolyzed citrus pectin (SCP) on the immune response remain unclear. Therefore, the effects of SCP on immunomodulatory functions and intestinal microbial dysbiosis were investigated using a cyclophosphamide-induced immunosuppressed mouse model. In this research, immunosuppressed ICR mice were administrated with SCP at dosages of 300/600/1200 mg/kg.bw by oral gavage, and body weight, immune organ indexes, cytokines, and gut microbiota were determined. The results showed that subcritical water treatment decreased the molecular mass and increased the content of galacturonic acid in citrus pectin hydrolysates. Meanwhile, the treatment with SCP improved immunoregulatory functional properties and bioactivities over the original citrus pectin. For example, SCP protected immune organs (accelerated recovery of immune organ indexes) and significantly enhanced the expression of immune-related cytokines (IL-2, IL-6, IFN-γ, and TNF-α). The results of the 16S rDNA sequencing analysis on an IlluminaMiSeq platform showed that SCP normalized Cy-induced gut dysbiosis. SCP ameliorated Cy-dependent changes in the relative abundance of several taxa, shifting the balance back to normal status (e.g., SCP increased beneficial Muribaculaceae, Ruminococcaceae, Bacteroidaceae, and Prevotellaceae while decreasing pathogenic Brevundimonas and Streptococcus). The results of this study suggest an innovative application of citrus pectin as an immunomodulator.

Systemic acquired resistance (SAR) in Arabidopsis is established beyond the initial pathogenic infection or is directly induced by treatment with salicylic acid or its functional analogs (SA/INA/BTH). NPR1 protein and WRKY transcription factors are considered the master regulators of SAR. Our previous study showed that NPR1 homologs in wheat (Triticum aestivum L.) and barley (Hordeum vulgare L.) regulated the expression of genes encoding pathogenesis-related (PR) proteins during acquired resistance (AR) triggered by Pseudomonas syringae pv. tomato DC3000. In the present examination, AR induced by P. syringae DC3000 was also found to effectively improve wheat resistance to Puccinia triticina (Pt). However, with more complex genomes, genes associated with this SAR-like response in wheat and barley are largely unknown and no specific WRKYs has been reported to be involved in this biological process. In our subsequent analysis, barley transgenic line overexpressing wheat wNPR1 (wNPR1-OE) showed enhanced resistance to Magnaporthe oryzae isolate Guy11, whereas AR to Guy11 was suppressed in a barley transgenic line with knocked-down barley HvNPR1 (HvNPR1-Kd). We performed RNA-seq to reveal the genes that were differentially expressed among these transgenic lines and the wild-type barley plants during the AR. Several PR and BTH-induced (BCI) genes were designated as downstream genes of NPR1. The expression of few WRKYs was significantly associated with NPR1 expression during the AR events. The transient expression of three WRKY genes, including HvWRKY6, HvWRKY40, and HvWRKY70, in wheat leaves by Agrobacterium-mediated infiltration enhanced the resistance to Pt. In conclusion, a profile of genes associated with NPR1-mediated AR in barley was drafted and WRKYs discovered in the current study showed a substantial potential for improving wheat resistance to Pt.

The wheat cultivar Weimai 8 is a good source of adult-plant resistance to leaf rust in China. In order to understand the genetic background of the resistance genes, 179 F2:3 families derived from the cross Weimai 8 and Zhengzhou 5389 (susceptible to leaf rust) were used to construct a linkage map. Results showed that there was a major QTL on chromosome 2AS, temporarily named QLr.hbau-2AS, responsible for the resistance. QLr.hbau-2AS from the resistant parent Weimai 8 was between the SSR markers Xcfd36 and Xbarc1138, with an interval length of 2.58 cM. In the 2010-2011, 2011-2012 and 2012-2013 crop seasons, it explained 25.79, 71.55 and 60.72% of the phenotypic variation, respectively. The location of QLr.hbau-2AS was close to the leaf rust resistance gene Lr37. However, they may not be the same since QLr.hbau-2AS has a different virulence test response from Lr37. This study identified two closely linked flanking molecular markers QLr.hbau-2AS and these molecular markers will help marker-assisted selection in breeding resistant cultivars in the future.

Leaf rust (LR), caused by Puccinia triticina (Pt), is worldwide one of the most spread diseases in common wheat (Triticum aestivum L.). With the objective of identifying and mapping new genes for resistance to leaf rust, F1 and F2 plants from a cross between the resistant cultivar LB0288 and the susceptible cultivar Thatcher were inoculated with the Chinese P. triticina pathotype THTT in the greenhouse. A set of 1273 simple sequence repeat (SSR) primer pairs were used to test the parents and the resistant and susceptible bulks. The results indicated, that LB0288 carried a single dominant resistance gene LrLB88, closely linked to the CAPS marker of Lr1 (WR003) and the SSR marker Xbarc144, with genetic distances of 0 cM and 5.3 cM, respectively. Based on the chromosomal location and seedling reactions, it can be concluded that LrLB88 might be a new leaf rust resistance gene. These markers can be useful for marker assisted selection in breeding of leaf rust resistant wheat cultivars.

Leaf rust (LR), caused by Puccinia triticina, is one of the most widespread diseases of common wheat (Triticum aestivum L.) worldwide. The LR resistance gene LrBi16 has been mapped on chromosome arm 7BL in Chinese wheat cultivar Bimai 16 and was closely linked to SSR loci Xcfa2257 and Xgwm344 with genetic distances of 2.8 cM and 2.9 cM, respectively. In the present study, a total of 304 AFLP primer pairs were used to screen Bimai 16 and Thatcher and resistant and susceptible DNA bulks. The polymorphic AFLP marker P-ATT/M-CGC173 bp was used to genotype F2 and F3 populations to identify markers more closely linked to LrBi16. Marker P-ATT/M-CGC173 bp was tightly linked to LrBi16 with a genetic distance of 0.5 cM. As LrBi16 was mapped near the Lr14a locus, 809 F2 plants from the Bimai 16/RL6013 (Lr14a) cross were inoculated with the Pt pathotype FHNQ to test the allelism of Lr14a and LrBi16. All of the F2 plants were resistant to FHNQ (IT between; and 2), suggesting that Lr14a and LrBi16 are allelic.

Puccinia triticina (Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effector-mediated wheat susceptibility remains largely unexplored. In this study, an effector Pt_21 was identified to interact with the apoplast-localized wheat thaumatin-like protein TaTLP1 using a yeast two-hybrid assay and the Pt_21-TaTLP1 interaction was characterized. The interaction between Pt_21 and TaTLP1 was validated by in vivo co-immunoprecipitation assay. A TaTLP1 variant, TaTLP1C71A, that was identified by the site-directed mutagenesis failed to interact with Pt_21. Pt_21 was able to suppress Bax-mediated cell death in leaves of Nicotiana benthamiana and inhibit TaTLP1-mediated antifungal activity. Furthermore, infiltration of recombinant protein Pt_21 into leaves of transgenic wheat line overexpressing TaTLP1 enhanced the disease development of leaf rust compared to that in wild-type leaves. These findings demonstrate that Pt_21 suppresses host defense response by directly targeting wheat TaTLP1 and inhibiting its antifungal activity, which broadens our understanding of the molecular mechanisms underlying Pt effector-mediated susceptibility in wheat. [ABSTRACT FROM AUTHOR]

Wheat leaf rust is an important disease worldwide. Growing resistant cultivars is an effective means to control the disease. In the present study, 244 recombinant inbred lines from Zhou 8425B/Chinese Spring cross were phenotyped for leaf rust severities during the 2011–2012, 2012–2013, 2013–2014, and 2014–2015 cropping seasons at Baoding, Hebei province, and 2012–2013 and 2013–2014 cropping seasons in Zhoukou, Henan province. The population was genotyped using the high-density Illumina iSelect 90K SNP assay and SSR markers. Inclusive composite interval mapping identified eight QTL, designated as QLr.hebau-2AL, QLr.hebau-2BS, QLr.hebau-3A, QLr.hebau-3BS, QLr.hebau-4AL, QLr.hebau-4B, QLr.hebau-5BL, and QLr.hebau-7DS, respectively. QLr.hebau-2BS, QLr.hebau-3A, QLr.hebau-3BS, and QLr.hebau-5BL were derived from Zhou 8425B, whereas the other four were from Chinese Spring. Three stable QTL on chromosomes 2BS, 4B and 7DS explained 7.5–10.6%, 5.5–24.4%, and 11.2–20.9% of the phenotypic variance, respectively. QLr.hebau-2BS in Zhou 8425B might be the same as LrZH22 in Zhoumai 22; QLr.hebau-4B might be the residual resistance of Lr12, and QLr.hebau-7DS is Lr34. QLr.hebau-2AL, QLr.hebau-3BS, QLr.hebau-4AL, and QLr.hebau-5BL are likely to be novel QTL for leaf rust. These QTL and their closely linked SNP and SSR markers can be used for fine mapping, candidate gene discovery, and marker-assisted selection in wheat breeding.

The wheat (Triticum aestivum L.) line 5R618, bred at the China Agricultural University, is resistant in the seedling stage to the majority of the current Chinese pathotypes of wheat leaf rust (Puccinia triticina). To identify and map the leaf rust resistance gene in the 5R618 line, F2 plants and F2:3 families from a cross between 5R618 and Zhengzhou5389 (susceptible) were inoculated in the greenhouse with the Chinese P. triticina pathotype THJP. Results from the F2 and F2:3 populations indicate that a single dominant gene, temporarily designated Lr5R, conferred resistance. Using the molecular marker method, Lr5R was located on the 3DL chromosome. It was closely linked to the markers Xbarc71 and OPJ-09 with genetic distances of 0.9 cM and 1.0 cM, respectively. At present only one designated gene (Lr24) is located on the 3DL chromosome. The genetic distance between Lr5R and Lr24 confirms that Lr5R is a new leaf rust resistance gene.