Comprehensive analysis of sesame LRR-RLKs: structure, evolution and dynamic expression profiles under Macrophomina phaseolina stress.

Leucine-rich repeat receptor-like kinases (LRR-RLKs) can participate in the regulation of plant growth and development, immunity and signal transduction. Sesamum indicum, one of the most important oil crops, has a significant role in promoting human health. In this study, 175 SiLRR-RLK genes were identified in S. indicum, and they were subdivided into 12 subfamilies by phylogenetic analysis. Gene duplication analysis showed that the expansion of the SiLRR-RLK family members in the sesame was mainly due to segmental duplication. Moreover, the gene expansion of subfamilies IV and III contributed to the perception of stimuli under M. phaseolina stress in the sesame. The collinearity analysis with other plant species revealed that the duplication of SiLRR-RLK genes occurred after the differentiation of dicotyledons and monocotyledons. The expression profile analysis and functional annotation of SiLRR-RLK genes indicated that they play a vital role in biotic stress. Furthermore, the protein-protein interaction and coexpression networks suggested that SiLRR-RLKs contributed to sesame resistance to Macrophomina phaseolina by acting alone or as a polymer with other SiLRR-RLKs. In conclusion, the comprehensive analysis of the SiLRR-RLK gene family provided a framework for further functional studies on SiLRR-RLK genes.

Full-length transcriptome and RNA-Seq analyses reveal the resistance mechanism of sesame in response to Corynespora cassiicola.

BACKGROUND: Corynespora leaf spot is a common leaf disease occurring in sesame, and the disease causes leaf yellowing and even shedding, which affects the growth quality of sesame. At present, the mechanism of sesame resistance to this disease is still unclear. Understanding the resistance mechanism of sesame to Corynespora leaf spot is highly important for the control of infection. In this study, the leaves of the sesame resistant variety (R) and the sesame susceptible variety (S) were collected at 0-48 hpi for transcriptome sequencing, and used a combined third-generation long-read and next-generation short-read technology approach to identify some key genes and main pathways related to resistance. RESULTS: The gene expression levels of the two sesame varieties were significantly different at 0, 6, 12, 24, 36 and 48 hpi, indicating that the up-regulation of differentially expressed genes in the R might enhanced the resistance. Moreover, combined with the phenotypic observations of sesame leaves inoculated at different time points, we found that 12 hpi was the key time point leading to the resistance difference between the two sesame varieties at the molecular level. The WGCNA identified two modules significantly associated with disease resistance, and screened out 10 key genes that were highly expressed in R but low expressed in S, which belonged to transcription factors (WRKY, AP2/ERF-ERF, and NAC types) and protein kinases (RLK-Pelle_DLSV, RLK-Pelle_SD-2b, and RLK-Pelle_WAK types). These genes could be the key response factors in the response of sesame to infection by Corynespora cassiicola. GO and KEGG enrichment analysis showed that specific modules could be enriched, which manifested as enrichment in biologically important pathways, such as plant signalling hormone transduction, plant-pathogen interaction, carbon metabolism, phenylpropanoid biosynthesis, glutathione metabolism, MAPK and other stress-related pathways. CONCLUSIONS: This study provides an important resource of genes contributing to disease resistance and will deepen our understanding of the regulation of disease resistance, paving the way for further molecular breeding of sesame.

First Report of Root Rot Caused by Pythium myriotylum on Sesame in China.

Sesame (Sesamum indicum L.) is a very important oilseed crop and cultivated on 11.7 million hectares, producing 6.02 million tons of seeds with an average seed yield of 512 kg ha-1 in the world (Yadav et al. 2022). In June of 2021, diseased roots were observed on sesame in the villages of Mada and Hanba, Xiangcheng city (114.88°N, 33.13°E), Henan province, China. The diseased plants appeared stunted and wilted at the seedling stage. Approximately 7.1% to 17.7% of plants were affected in two fields, 0.6 ha in total, and disease severity in each affected plant ranged from 50% to 80%. Twenty-four disease plants were collected to confirm the pathogen. The diseased roots were cut into small fragments (2 to 5 mm long), surface sterilized with 75% ethanol (for 1 min), and 10% sodium hypochlorite (for 1 min) and then rinsed in sterilized water three times (1 min each rinse). The fragments were blotted dry and transferred to a potato dextrose agar (PDA) medium (potato 200 g/L, glucose 20 g/L, agar 18 g/L) amended with streptomycin (50 µg/mL). After incubation at 28°C for 24 h, white mycelium grew out from plant fragments. Then, a total of seven morphologically similar strains were transferred onto fresh V8 agar by hyphal tip transfer (Rollins 2003). By light microscope observations, the sporangia were filamentous or digitated, and undifferentiated or inflated lobulate. The oospores were mostly aplerotic, globose or subglobose in shape, and 20.4 to 42.6 µm in diameter (n = 90, n: Total number of oospores measured). Furthermore, antheridia were bulbous-like or clavate-like and were observed attached to the surface of the oospores. The zoospores were abundant and ranged from 8.5 to 14.2 µm in diameter. The morphology characteristics of all strains were consistent with those of Pythium myriotylum (Watanabe et al. 2007). Genomic DNA was extracted from the representative strain 20210628 using the CTAB method (Wangsomboondee et al. 2002). The complete internal transcribed spacer (ITS) and cytochrome oxidase subunit I gene (COI, COX1) can be valid and useful barcodes for accurate identification of many oomycetes (Robideau et al. 2011). The ITS and COI were amplified with the primers ITS1/ITS4 (Riit et al. 2016) and primers OomCox-Levup/OomCox-Levlo (Robideau et al. 2011), respectively. The nucleotide sequences obtained were deposited in the GenBank database under the accession numbers OM230138.2 (ITS) and ON500503.1 (COI). GenBank BLAST search identified the sequences as P. myriotylum ITS and COI sequences (e.g., HQ237488.1 and MK510848.1, respectively) with 100% coverage and 100% identity. To evaluate the pathogenicity, sesame seeds (cultivar: Jinzhi No.3) were planted in 12-cm-diameter plastic pots containing a mixture of sterilized soil, vermiculite and peat mossat a ratio of 3:1:1. Oospores were collected following the procedure of Raftoyannis et al. (2006) with minor modifications. Three-leaf stage sesame roots were soaked with 5 mL of oospore suspension at 1 × 106/mL of the 20210628 strain, and the control plants were inoculated with sterilized water. All plants were maintained in a greenhouse (28±2°C, > 80% R. H.). The experiment was repeated twice with three replications. The plants inoculated with P. myriotylum showed the water soak symptom on the stem base 7 days after inoculation, while control plants were symptomless. Three weeks after inoculation, the plants showed root tissue necrosis, root rot, and dwarfing symptoms that were similar to those observed on sesame plants in the field, while control plants remained healthy. P. myriotylum was re-isolated from the inoculated plants and the morphology was the same as the original strain 20210628. These results suggest that P. myriotylum is the causal agent of sesame root rot. Previous studies have revealed that P. myriotylum can cause root rot in peanuts (Yu et al. 2019), chili pepper (Hyder et al. 2018), green bean (Serrano et al. 2008) and aerial blight of tomato (Roberts et al. 1999). To the best of our knowledge, this is the first report of P. myriotylum causing root rot on sesame. This pathogen can infect plant roots and develop rapidly if no effective control measures are implemented. Once the disease breaks out in a large area, the yield of sesame will be seriously threatened. The results provide important implications for the prevention and management of this disease.

Wheat Thioredoxin (TaTrxh1) Associates With RD19-Like Cysteine Protease TaCP1 to Defend Against Stripe Rust Fungus Through Modulation of Programmed Cell Death.

Thioredoxins (Trxs) function within the antioxidant network through modulation of one or more redox reactions involved in oxidative-stress signaling. Given their function in regulating cellular redox, Trx proteins also fulfill key roles in plant immune signaling. Here, TaTrxh1, encoding a subgroup h member of the Trx family, was identified and cloned in wheat (Triticum aestivum), which was rapidly induced by Puccinia striiformis f. sp. tritici invasion and salicylic acid (SA) treatment. Overexpression of TaTrxh1 in tobacco (Nicotiana benthamiana) induced programmed cell death. Silencing of TaTrxh1 in wheat enhanced susceptibility to P. striiformis f. sp. tritici in different aspects, including reactive oxygen species accumulation and pathogen-responsive or -related gene expression. Herein, we observed that the cellular concentration of SA was significantly reduced in TaTrxh1-silenced plants, indicating that TaTrxh1 possibly regulates wheat resistance to stripe rust through a SA-associated defense signaling pathway. Using a yeast two-hybrid screen to identify TaTrxh1-interacting partners, we further show that interaction with TaCP1 (a RD19-like cysteine protease) and subsequent silencing of TaCP1 reduced wheat resistance to P. striiformis f. sp. tritici. In total, the data presented herein demonstrate that TaTrxh1 enhances wheat resistance against P. striiformis f. sp. tritici via SA-dependent resistance signaling and that TaTrxh1 interaction with TaCP1 is required for wheat resistance to stripe rust.[Formula: see text] Copyright © 2021 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

ShNPSN11, a vesicle-transport-related gene, confers disease resistance in tomato to Oidium neolycopersici.

Tomato powdery mildew, caused by Oidium neolycopersici, is a fungal disease that results in severe yield loss in infected plants. Herein, we describe the function of a class of proteins, soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs), which play a role in vesicle transport during defense signaling. To date, there have been no reports describing the function of tomato SNAREs during resistance signaling to powdery mildew. Using a combination of classical plant pathology-, genetics-, and cell biology-based approaches, we evaluate the role of ShNPSN11 in resistance to the powdery mildew pathogen O. neolycopersici. Quantitative RT-PCR analysis of tomato SNAREs revealed that ShNPSN11 mRNA accumulation in disease-resistant varieties was significantly increased following pathogen, compared with susceptible varieties, suggesting a role during induced defense signaling. Using in planta subcellular localization, we demonstrate that ShNPSN11 was primarily localized at the plasma membrane, consistent with the localization of SNARE proteins and their role in defense signaling and trafficking. Silencing of ShNPSN11 resulted in increased susceptibility to O. neolycopersici, with pathogen-induced levels of H2O2 and cell death elicitation in ShNPSN11-silenced lines showing a marked reduction. Transient expression of ShNPSN11 did not result in the induction of a hypersensitive cell death response or suppress cell death induced by BAX. Taken together, these data demonstrate that ShNPSNl11 plays an important role in defense activation and host resistance to O. neolycopersici in tomato LA1777.

A Simple and Effective Technique for Production of Pycnidia and Pycnidiospores by Macrophomina phaseolina.

Production of pycnidia and pycnidiospores by Macrophomina phaseolina is not often seen in vitro. The objective of this study is to develop a simple and effective technique to obtain pycnidiospores of M. phaseolina isolates in vitro and to evaluate germination rates and pathogenicity of pycnidiospores. We found M. phaseolina isolates can produce pycnidia on oatmeal agar (OMA) under ultraviolet light with 365 nm wavelength (UV). For evaluating the effect of OMA and UV on growth of M. phaseolina, combinations of two agar media and three lighting conditions were tested. The results confirm that all six M. phaseolina isolates produced pycnidia only on OMA under UV. The pycnidiospores produced on OMA under UV had germination rates higher than 90%. In pathogenicity tests, inoculation with the pycnidiospores showed symptoms later than inoculation with hypha-colonized toothpicks. Significant differences in the pathogenicity is detected between isolates Mp2014003 and Mp2014024 when inoculation is done with the pycnidiospores (P < 0.001), but not when hypha-colonized toothpicks are used as inoculum (P = 0.091). This study provides a new method for obtaining pycnidiospores of M. phaseolina for future investigations.

Complete genome sequence of a novel victorivirus isolated from the sesame charcoal rot fungus Macrophomina phaseolina.

Macrophomina phaseolina is an important phytopathogenic fungus with a broad host range. Here, the complete genome sequence of a novel victorivirus, tentatively named Macrophomina phaseolina victorivirus 1 (MpV1), was identified from strain 2012-019 of M. phaseolina. The MpV1 genome is 5,128 nucleotides long with a predicted GC content of 62%. Sequence analysis indicated that two open reading frames (ORF 1 and 2) overlap at a tetranucleotide AUGA sequence. Proteins encoded by ORF1 and ORF2 showed significant sequence similarity to coat proteins and the RNA-dependent RNA polymerases, respectively, of members of the family Totiviridae. Analysis of the genomic structure of MpV1, homolog searches of the deduced amino acid sequences, and phylogenetic analysis indicated that MpV1 is a new member of the genus Victorivirus. As far as we know, this is the first report of the full-length nucleotide sequence of the genome of a novel victorivirus that infects M. phaseolina.

A novel double-stranded RNA mycovirus that infects Macrophomina phaseolina.

Macrophomina phaseolina is a pathogenic fungus of the family Botryosphaeriaceae that causes stem rot or leaf blight in many economically important plants. Mycoviruses exist widely in fungi, but there are only a limited number of reports on mycovirus infection in M. phaseolina. A novel dsRNA virus, tentatively named "Macrophomina phaseolina fusagravirus 1" (MpFV1), was isolated from strain 2012-19 of M. phaseolina, and its molecular features were examined. The full-length cDNA of MpFV1 comprises 9,289 nucleotides with a predicted GC content of 48.1% and two discontinuous open reading frames (ORF 1 and 2). A-1 frameshift region with two typical factors, including a shifty heptamer (GGAAAAC) and an H-type pseudoknot, was predicted in the junction region of ORF1 and ORF2. The protein encoded by ORF1 shows significant similarity to a hypothetical protein, whereas ORF2 encodes an RNA-dependent RNA polymerase (RdRp) via a ribosomal frameshifting mechanism. Homology searches and phylogenetic analysis based on the RdRp sequence suggested that MpFV1 is a new member of the proposed family "Fusagraviridae".

Health Services Use during Transition from Pediatric to Adult Care for Inflammatory Bowel Disease: A Population-Based Study Using Health Administrative Data.

OBJECTIVES: To evaluate the impact of the transfer from pediatric to adult care on health services use for adolescents with inflammatory bowel disease (IBD). STUDY DESIGN: A population-based retrospective cohort study identified all children diagnosed with IBD from 1994 to 2008 and treated by pediatric gastroenterologists in Ontario, Canada, using health administrative data. Self-controlled case series analyses compared health service use in the 2 years before and 2 years after transfer with adult gastroenterologists, with a 6-month washout period at transfer. Outcomes evaluated included IBD-specific and IBD-related hospitalizations, emergency department use, outpatient visits, and laboratory use. The relative incidence (RI) in the post-transfer period was compared with pretransfer periods using Poisson regression analysis controlling for transfer starting age. Analyses were stratified by IBD type: Crohn's disease (CD) and ulcerative colitis (UC). RESULTS: There were 536 patients included in the study (388 CD, 148 UC). Emergency department use rate was higher after transfer for both CD (RI, 2.12; 95% CI, 1.53-2.93) and UC (RI, 2.34; 95% CI, 1.09-5.03), as were outpatient visits (CD: RI, 1.56; 95% CI, 1.42-1.72; UC: RI, 1.48; 95% CI, 1.24-1.76), and laboratory investigations (CD: RI, 1.43; 95% CI, 1.26-1.63; UC: 1.38; 95% CI, 1.13-1.68). There was no change in the hospitalization rate (CD: RI, 0.70; 95% CI, 0.42-1.18; UC: RI, 2.41; 95% CI, 0.62-9.40). CONCLUSIONS: Health services use in Canada increases after transfer from pediatric to adult care for outpatient visits, emergency department use, and laboratory tests, but not hospitalizations. This study has implications for the planning and budgeting of care for adolescents transitioning to adult care.

Characterizing the Posttransfer Period Among Patients with Pediatric Onset IBD: The Impact of Academic Versus Community Adult Care on Emergent Health Resource Utilization.

BACKGROUND: Patients diagnosed with inflammatory bowel disease (IBD) during childhood require transfer to an adult gastroenterologist, in Ontario usually just before their 18th birthday. Pediatric onset IBD is a complex phenotype with demonstrated noncompliance risk that may require targeted measures to optimize health care outcomes in the adult care setting. PURPOSE: The purpose of this study was to determine the impact of posttransfer health care setting (academic versus community gastroenterologist) on emergent health resource utilization. METHODS: This was a population-based retrospective cohort study using health care administrative data from Ontario, Canada. A cohort of patients with Pediatric onset IBD was identified and health resource utilization during a 2-year pretransfer period, transfer of care period and 2-year posttransfer period was analyzed. Posttransfer health care setting was defined as academic (i.e., gastroenterologists providing care in a university affiliated tertiary care center) versus community. A third comparator group, loss to follow-up, was also identified. The primary outcome of this study comprised emergency department utilization. Secondary outcomes included hospitalizations, surgeries, ambulatory visits, endoscopic investigations, and radiological investigations. RESULTS: Overall, there were no significant differences found in emergency department use, ambulatory care visits (aside from the expected drop in the lost to follow-up group), hospitalizations, endoscopic procedures, or radiological procedures between exposure groups. CONCLUSIONS: Posttransfer health care setting does not seem to significantly impact emergent health resource utilization in the posttransfer period.

The roles of inoculants' carbon source use in the biocontrol of potato scab disease.

Despite the application of multiple strains in the biocontrol of plant diseases, multistrain inoculation is still constrained by its inconsistency in the field. Nutrients, especially carbons, play an important role in the biocontrol processes. However, little work has been done on the systematic estimation of inoculants' carbon source use on biocontrol efficacies in vivo. In the present study, 7 nonpathogenic Streptomyces strains alone and in different combinations were inoculated as biocontrol agents against the potato scab disease, under field conditions and greenhouse treatments. The influence of the inoculants' carbon source use properties on biocontrol efficacies was investigated. The results showed that increasing the number of inoculated strains did not necessarily result in greater biocontrol efficacy in vivo. However, single strains with higher growth rates or multiple strains with less carbon source competition had positive effects on the biocontrol efficacies. These findings may shed light on optimizing the consistent biocontrol of plant disease with the consideration of inoculants' carbon source use properties.

Mutations in Drosophila Greatwall/Scant reveal its roles in mitosis and meiosis and interdependence with Polo kinase.

Polo is a conserved kinase that coordinates many events of mitosis and meiosis, but how it is regulated remains unclear. Drosophila females having only one wild-type allele of the polo kinase gene and the dominant Scant mutation produce embryos in which one of the centrosomes detaches from the nuclear envelope in late prophase. We show that Scant creates a hyperactive form of Greatwall (Gwl) with altered specificity in vitro, another protein kinase recently implicated in mitotic entry in Drosophila and Xenopus. Excess Gwl activity in embryos causes developmental failure that can be rescued by increasing maternal Polo dosage, indicating that coordination between the two mitotic kinases is crucial for mitotic progression. Revertant alleles of Scant that restore fertility to polo-Scant heterozygous females are recessive alleles or deficiencies of gwl; they show chromatin condensation defects and anaphase bridges in larval neuroblasts. One recessive mutant allele specifically disrupts a Gwl isoform strongly expressed during vitellogenesis. Females hemizygous for this allele are sterile, and their oocytes fail to arrest in metaphase I of meiosis; both homologues and sister chromatids separate on elongated meiotic spindles with little or no segregation. This allelic series of gwl mutants highlights the multiple roles of Gwl in both mitotic and meiotic progression. Our results indicate that Gwl activity antagonizes Polo and thus identify an important regulatory interaction of the cell cycle.