Socket Array Irregularities and Wing Membrane Distortions at the Eyespot Foci of Butterfly Wings Suggest Mechanical Signals for Color Pattern Determination.

Eyespot foci on butterfly wings function as organizers of eyespot color patterns during development. Despite their importance, focal structures have not been examined in detail. Here, we microscopically examined scales, sockets, and the wing membrane in the butterfly eyespot foci of both expanded and unexpanded wings using the Blue Pansy butterfly Junonia orithya. Images from a high-resolution light microscope revealed that, although not always, eyespot foci had scales with disordered planar polarity. Scanning electron microscopy (SEM) images after scale removal revealed that the sockets were irregularly positioned and that the wing membrane was physically distorted as if the focal site were mechanically squeezed from the surroundings. Focal areas without eyespots also had socket array irregularities, but less frequently and less severely. Physical damage in the background area induced ectopic patterns with socket array irregularities and wing membrane distortions, similar to natural eyespot foci. These results suggest that either the process of determining an eyespot focus or the function of an eyespot organizer may be associated with wing-wide mechanics that physically disrupt socket cells, scale cells, and the wing membrane, supporting the physical distortion hypothesis of the induction model for color pattern determination in butterfly wings.

Phenotypic plasticity in tropical butterflies is linked to climatic seasonality on a macroevolutionary scale.

Phenotypic plasticity can be adaptive in fluctuating environments by providing rapid environment-phenotype matching and this applies particularly in seasonal environments. African Bicyclus butterflies have repeatedly colonized seasonal savannahs from ancestral forests around the late Miocene, and many species now exhibit seasonal polyphenism. On a macroevolutionary scale, it can be expected that savannah species will exhibit higher plasticity because of experiencing stronger environmental seasonality than forest species. We quantified seasonality using environmental niche modeling and surveyed the degree of plasticity in a key wing pattern element (eyespot size) using museum specimens. We showed that species occurring in highly seasonal environments display strong plasticity, while species in less seasonal or aseasonal environments exhibit surprisingly variable degrees of plasticity, including strong to no plasticity. Furthermore, eyespot size plasticity has a moderate phylogenetic signal and the ancestral Bicyclus likely exhibited some degree of plasticity. We propose hypotheses to explain the range of plasticity patterns seen in less seasonal environments and generate testable predictions for the evolution of plasticity in Bicyclus. Our study provides one of the most compelling cases showing links between seasonality and phenotypic plasticity on a macroevolutionary scale and the potential role of plasticity in facilitating the colonization of novel environments.

Genome Assembly and Annotation of the Dark-Branded Bushbrown Butterfly Mycalesis mineus (Nymphalidae: Satyrinae).

We report a high-quality genome draft assembly of the dark-branded bushbrown, Mycalesis mineus, a member of the Satyrinae subfamily of nymphalid butterflies. This species is emerging as a promising model organism for investigating the evolution and development of phenotypic plasticity. Using 45.99 Gb of long-read data (N50 = 11.11 kb), we assembled a genome size of 497.4 Mb for M. mineus. The assembly is highly contiguous and nearly complete (96.8% of Benchmarking Universal Single-Copy Orthologs lepidopteran genes were complete and single copy). The genome comprises 38.71% of repetitive elements and includes 20,967 predicted protein-coding genes. The assembled genome was super-scaffolded into 28 pseudo-chromosomes using a closely related species, Bicyclus anynana, with a chromosomal-level genome as a template. This valuable genomic tool will advance both ongoing and future research focused on this model organism.

Eyespot variation and field temperature in the Meadow Brown butterfly.

Since the classic work of E.B. Ford, explanations for eyespot variation in the Meadow Brown butterfly have focused on the role of genetic polymorphism. The potential role of thermal plasticity in this classic example of natural selection has therefore been overlooked. Here, we use large daily field collections of butterflies from three sites, over multiple years, to examine whether field temperature is correlated with eyespot variation, using the same presence/absence scoring as Ford. We show that higher developmental temperature in the field leads to the disappearance of the spots visible while the butterfly is at rest, explaining the historical observation that hindwing spotting declines across the season. Strikingly, females developing at 11°C have a median of six spots and those developing at 15°C only have three. In contrast, the large forewing eyespot is always present and scales with forewing length. Furthermore, in contrast to the smaller spots, the size of the large forewing spot is best explained by calendar date (days since 1st March) rather than the temperature at pupation. As this large forewing spot is involved in startling predators and/or sexual selection, its constant presence is therefore likely required for defence, whereas the disappearance of the smaller spots over the season may help with female crypsis. We model annual total spot variation with phenological data from the UK and derive predictions as to how spot patterns will continue to change, predicting that female spotting will decrease year on year as our climate warms.

Genome-wide analysis and characterization of the TaTLP gene family in wheat and functional characterization of the TaTLP44 in response to Rhizoctonia cerealis.

Wheat sharp eyespot is a soil-borne disease caused by Rhizoctonia cerealis, which occurs in many countries worldwide and significantly reduces the yield. Thaumatin-like protein (TLP), also known as PR5, is a member of the pathogen response protein family and plays an essential role in plant resistance to pathogen infection. In this study, 131 TaTLP genes were identified from the wheat genome, of which 38 TaTLPs were newly discovered. The TaTLP gene family contains many tandem duplications and fragment duplications, which is a major pathway for gene amplification. Besides, we also analyzed the physicochemical properties, gene structure and promoter cis-acting regulatory elements of all the TaTLP genes. In addition, the expression patterns of nine TaTLPs in response to R. cerealis were analyzed by RT-qPCR. Six TaTLP proteins expressed in vitro had no significant inhibitory effect on R. cerealis, suggesting that these TaTLP proteins may function in other ways. Finally, we performed gene silencing of TaTLP44 in wheat, which increased the expression of some defense-associated genes and improved resistance to R. cerealis. In summary, we systematically analyzed TaTLP family members and demonstrated that TaTLP44 negatively regulates the resistance to R. cerealis by controlling expression of defense-associated genes. These results provide new insights into the functional mechanism of TaTLP proteins.

Illuminating the diversity of carotenoids in microalgal eyespots and phototaxis.

Photosynthetic organisms biosynthesize various carotenoids, a group of light-absorbing isoprenoid pigments that have key functions in photosynthesis, photoprotection, and phototaxis. Microalgae, in particular, contain diverse carotenoids and carotenoid biosynthetic pathways as a consequence of the various endosymbiotic events in their evolutionary history. Carotenoids such as astaxanthin, diadinoxanthin, and fucoxanthin are unique to algae. In microalgae, carotenoids are concentrated in the eyespot, a pigmented organelle that is important for phototaxis. A wide range of microalgae, including chlorophytes, euglenophytes, ochrophytes, and haptophytes, have an eyespot. In the chlorophyte Chlamydomonas reinhardtii, carotenoid layers in the eyespot reflect light to amplify the photosignal and shield photoreceptors from light, thereby enabling precise phototaxis. Our recent research revealed that, in contrast to the ß-carotene-rich eyespot of C. reinhardtii, the euglenophyte Euglena gracilis relies on zeaxanthin for stable eyespot formation and phototaxis. In this review, we highlight recent advancements in the study of eyespot carotenoids and phototaxis in these microalgae, placing special emphasis on the diversity of carotenoid-dependent visual systems among microalgae.

Trehalase Inhibitor Validamycin May Have Additional Mechanisms of Toxicology against Rhizoctonia cerealis.

Sharp eyespot is a crucial disease affecting cereal plants, such as bread wheat (Triticum aestivum) and barley (Hordeum vulgare), and is primarily caused by the pathogenic fungus Rhizoctonia cerealis. As disease severity has increased, it has become imperative to find an effective and reasonable control strategy. One such strategy is the use of the trehalose analog, validamycin, which has been shown to have a potent inhibitory effect on several trehalases found in both insects and fungi, and is widely used as a fungicide in agriculture. In this study, we demonstrated that 0.5 µg/mL validamycin on PDA plates had an inhibitory effect on R. cerealis strain R0301, but had no significant impact on Fusarium graminearum strain PH-1. Except for its inhibiting the trehalase activity of pathogenic fungi, little is known about its mechanism of action. Six trehalase genes were identified in the genome of R. cerealis, including one neutral trehalase and five acidic trehalase genes. Enzyme activity assays indicated that treatment with 5 µg/mL validamycin significantly reduces trehalase activity, providing evidence that validamycin treatment does indeed affect trehalase, even though the expression levels of most trehalase genes, except Rc17406, were not obviously affected. Transcriptome analysis revealed that treatment with validamycin downregulated genes involved in metabolic processes, ribosome biogenesis, and pathogenicity in the R. cerealis. KEGG pathway analysis further showed that validamycin affected genes related to the MAPK signaling pathway, with a significant decrease in ribosome synthesis and assembly. In conclusion, our results indicated that validamycin not only inhibits trehalose activity, but also affects the ribosome synthesis and MAPK pathways of R. cerealis, leading to the suppression of fungal growth and pesticidal effects. This study provides novel insights into the mechanism of action of validamycin.

Extreme Diversity of Mycoviruses Present in Single Strains of Rhizoctonia cerealis, the Pathogen of Wheat Sharp Eyespot.

Rhizoctonia cerealis is the pathogen of wheat sharp eyespot, which occurs throughout temperate wheat-growing regions of the world. In this project, the genomes of viruses from four strains of R. cerealis were analyzed based on Illumina high-throughput transcriptome sequencing (RNA-Seq) data. After filtering out reads that mapped to the fungal genome, viral genomes were assembled. In total, 131 virus-like sequences containing complete open reading frames (ORFs), belonging to 117 viruses, were obtained. Based on phylogenetic analysis, some of them were identified as novel members of the families Curvulaviridae, Endornaviridae, Hypoviridae, Mitoviridae, Mymonaviridae, and Phenuiviridae, while others were unclassified viruses. Most of these viruses from R. cerealis were significantly different from the viruses already reported. We propose the establishment of a new family, Rhizoctobunyaviridae, and two new genera, Rhizoctobunyavirus and Iotahypovirus. We further clarified the distribution and coinfection of these viruses in the four strains. Surprisingly, 39 viral genomes of up to 12 genera were found in strain R1084. Strain R0942, containing the fewest viruses, also contained 21 viral genomes belonging to 10 genera. Based on the RNA-Seq data, we estimated the accumulation level of some viruses in host cells and found that the mitoviruses in R. cerealis generally have very high accumulation. In conclusion, in the culturable phytopathogenic fungus R. cerealis, we discovered a considerable diversity of mycoviruses and a series of novel viruses. This study expands our understanding of the mycoviral diversity in R. cerealis and provides a rich resource for the further use of mycoviruses to control wheat sharp eyespot. IMPORTANCE Rhizoctonia cerealis is a binucleate fungus that is widely distributed worldwide and can cause sharp eyespot disease in cereal crops. In this study, 131 virus-like sequences belonging to 117 viruses were obtained based on analysis of high-throughput RNA-Seq data from four strains of R. cerealis. Many of these viruses were novel members of various virus families, while others were unclassified viruses. As a result, a new family named Rhizoctobunyaviridae and two new genera, Rhizoctobunyavirus and Iotahypovirus, were proposed. Moreover, the discovery of multiple viruses coinfecting a single host and the high accumulation levels of mitoviruses have shed light on the complex interactions between different viruses in a single host. In conclusion, a significant diversity of mycoviruses was discovered in the culturable phytopathogenic fungus R. cerealis. This study expands our understanding of mycoviral diversity, and provides a valuable resource for the further utilization of mycoviruses to control wheat diseases.

Identification of Long Intergenic Noncoding RNAs in Rhizoctonia cerealis following Inoculation of Wheat.

Wheat sharp eyespot caused by Rhizoctonia cerealis is primarily a severe threat to worldwide wheat production. Currently, there are no resistant wheat cultivars, and the use of fungicides is the primary method for controlling this disease. Elucidating the mechanisms of R. cerealis pathogenicity can accelerate the pace of the control of this disease. Long intergenic noncoding RNAs (lincRNAs) that function in plant-pathogen interactions might provide a new perspective. We systematically analyzed lincRNAs and identified a total of 1,319 lincRNAs in R. cerealis. We found that lincRNAs are involved in various biological processes, as shown by differential expression analysis and weighted correlation network analysis (WGCNA). Next, one of nine hub lincRNAs in the blue module that was related to infection and growth processes, MSTRG.4380.1, was verified to reduce R. cerealis virulence on wheat by a host-induced gene silencing (HIGS) assay. Following that, RNA sequencing (RNA-Seq) analysis revealed that the significantly downregulated genes in the MSTRG.4380.1 knockdown lines were associated mainly with infection-related processes, including hydrolase, transmembrane transporter, and energy metabolism activities. Additionally, 23 novel microRNAs (miRNAs) were discovered during small RNA (sRNA) sequencing (sRNA-Seq) analysis of MSTRG.4380.1 knockdown, and target prediction of miRNAs suggested that MSTRG.4380.1 does not act as a competitive endogenous RNA (ceRNA). This study performed the first genome-wide identification of R. cerealis lincRNAs and miRNAs. It confirmed the involvement of a lincRNA in the infection process, providing new insights into the mechanism of R. cerealis infection and offering a new approach for protecting wheat from R. cerealis. IMPORTANCE Rhizoctonia cerealis, the primary causal agent of wheat sharp eyespot, has caused significant losses in worldwide wheat production. Since no resistant wheat cultivars exist, chemical control is the primary method. However, this approach is environmentally unfriendly and costly. RNA interference (RNAi)-mediated pathogenicity gene silencing has been proven to reduce the growth of Rhizoctonia and provides a new perspective for disease control. Recent studies have shown that lincRNAs are involved in various biological processes across species, such as biotic and abiotic stresses. Therefore, verifying the function of lincRNAs in R. cerealis is beneficial for understanding the infection mechanism. In this study, we reveal that lincRNAs could contribute to the virulence of R. cerealis, which provides new insights into controlling this pathogen.

First Report of Sharp Eyespot of Oat (Avena sativa) Caused by Ceratobasidium cereale in Korea.

Along with barley and wheat, oats (Avena sativa) are cultivated as winter crops in Korea, and the total area for oat cultivation is 103 ha in 2021. From late March to early April 2021, sharp eyespot symptoms on oat (cv. Choyang) leaf sheaths and straws were observed in two commercial fields located in Haenam (N34°38'35.04588/E126°38'31.00668) and Gangjin (N34°38'9.46788/E126°37'19.44984), Jeollanam-do, Korea. The incidence was 5% and 7%, respectively. Small brown spots were irregular circles that began to appear on the lower sheaths, and the spots gradually enlarged in the upper part of the sheaths. The center of each lesion turned whitish-brown with dark brown margins, resulting in a blight of the sheaths. Three plants displaying typical sharp eyespot lesions were collected from each of two individual regions, Haenam and Gangjin. To isolate the causal pathogen, two infected tissues (5  5 mm) from the collected plants were surface-sterilized by treating them with 70% ethanol for 1 min and 1% NaClO for 1 min immediately after being treated with 95% ethanol for 1 min. Subsequently, the samples were rinsed three times with distilled water, dried with sterile filter paper, transferred to 1.5% water agar supplemented with 100 ppm streptomycin, and then incubated in the dark at 25°C. Hyphae emerging from the randomly selected three independent tissues from each location were subcultured on potato dextrose agar (PDA, Sparks, MD 21152, USA), resulting in three independent isolates (HNO-1, HNO-2, HNO-3) from Haenam and three (KJO1-1, KJO1-2, KJO1-3) from Ganjin after single-hypha-tip purification. Colonies on the PDA were pigmented white at first and subsequently changed to light brown after 2 weeks. All collected isolates formed globose and irregular dark brown to black sclerotia on PDA after 2 weeks. Binuclear hyphae were white to dark brown in color, branched at right angles with a septum near the branch, and multinucleate cells, suggesting that these isolates belonged to Ceratobasidium cereale (Boerema et al., 1977; Burpee, 1980; Sharon et al.,2008). For molecular identification, the ITS (GenBank accession nos. MW691851-53 for HNO-1 to HNO-3; MW691857-59 for KJO1-1 to KJO1-3), LSU (OQ397530-35), rpb2 (OQ409878-83), tef1 (OQ409884-89), and atp6 (OQ409890-95) regions of six isolates were amplified using the primer pairs ITS4/5 (White et al., 1990), LROR/LR5 (Vilgalys and Hester, 1990), bRPB2-6F/bRPB2-7.1R (Matheny, 2005; Reeb et al., 2004), TEF1-F/TEF1-R (Litvintseva et al., 2006), and ATP61/ATP62 (Kretzer and Bruns, 1999), respectively. The sequences of ITS region showed 99.7% identity with C. cereale strain WK137-56 (KY379365) and 99.8% with Ceratobasidium sp. AG-D (KP171639). Using the MEGA X program (Kumar et al. 2018), a maximum likelihood phylogenetic analysis based on the concatenated ITS-LSU, rpb2, tef1 and atp6 sequences placed the six isolates within a clade comprising C. cereale (Gónzalez et al.,2016; Ji et al., 2017; Tomioka et al., 2021; Li et al., 2014). Two representative isolate, HNO-1 and KJO1-1, were deposited in the Korean Agriculture Culture Collection (Accession No. KACC 49887 and 410268, respectively). For pathogenicity, the six isolates were cultured on sterilized ray grains at 25°C in the dark for 3 weeks as the inoculum. Five oat (cv. Choyang) seeds were sown per pot containing 80 g of the infected ray grains mixed with 150 g of composite soil and 150 ml of water (Baroker Garden Soil, Seoul Bio Co., LTD). The control was treated with 80 g of the sterilized ray grains mixed with 150 g of composite soil and 150 ml of water. All inoculated and control pots were placed in a 20°C growth chamber with a 12-h photoperiod and 65% humidity. Typical sharp eyespot symptoms were observed on the oat sheath of seedlings three weeks post-inoculation. No symptoms were observed in the control seedlings. The infection assays were repeated thrice, with similar results. The pathogen was successfully re-isolated, and its identity was confirmed via morphological and molecular analyses. In Korea, few etiological studies have been conducted on oats because they are less economical than barley and wheat. Sharp eyespot disease caused by C. cereale has already been reported in barley and wheat (Kim et al., 1991); however, this is the first report of this disease in oats in Korea.

Slit-Robo expression in the leech nervous system: insights into eyespot evolution.

BACKGROUND: Slit and Robo are evolutionarily conserved ligand and receptor proteins, respectively, but the number of slit and robo gene paralogs varies across recent bilaterian genomes. Previous studies indicate that this ligand-receptor complex is involved in axon guidance. Given the lack of data regarding Slit/Robo in the Lophotrochozoa compared to Ecdysozoa and Deuterostomia, the present study aims to identify and characterize the expression of Slit/Robo orthologs in leech development. RESULTS: We identified one slit (Hau-slit), and two robo genes (Hau-robo1 and Hau-robo2), and characterized their expression spatiotemporally during the development of the glossiphoniid leech Helobdella austinensis. Throughout segmentation and organogenesis, Hau-slit and Hau-robo1 are broadly expressed in complex and roughly complementary patterns in the ventral and dorsal midline, nerve ganglia, foregut, visceral mesoderm and/or endoderm of the crop, rectum and reproductive organs. Before yolk exhaustion, Hau-robo1 is also expressed where the pigmented eye spots will later develop, and Hau-slit is expressed in the area between these future eye spots. In contrast, Hau-robo2 expression is extremely limited, appearing first in the developing pigmented eye spots, and later in the three additional pairs of cryptic eye spots in head region that never develop pigment. Comparing the expression of robo orthologs between H. austinensis and another glossiphoniid leech, Alboglossiphonia lata allows to that robo1 and robo2 operate combinatorially to differentially specify pigmented and cryptic eyespots within the glossiphoniid leeches. CONCLUSIONS: Our results support a conserved role in neurogenesis, midline formation and eye spot development for Slit/Robo in the Lophotrochozoa, and provide relevant data for evo-devo studies related to nervous system evolution.

A cell wall invertase modulates resistance to fusarium crown rot and sharp eyespot in common wheat.

Fusarium crown rot (FCR) and sharp eyespot (SE) are serious soil-borne diseases in wheat and its relatives that have been reported to cause wheat yield losses in many areas. In this study, the expression of a cell wall invertase gene, TaCWI-B1, was identified to be associated with FCR resistance through a combination of bulk segregant RNA sequencing and genome resequencing in a recombinant inbred line population. Two bi-parental populations were developed to further verify TaCWI-B1 association with FCR resistance. Overexpression lines and ethyl methanesulfonate (EMS) mutants revealed TaCWI-B1 positively regulating FCR resistance. Determination of cell wall thickness and components showed that the TaCWI-B1-overexpression lines exhibited considerably increased thickness and pectin and cellulose contents. Furthermore, we found that TaCWI-B1 directly interacted with an alpha-galactosidase (TaGAL). EMS mutants showed that TaGAL negatively modulated FCR resistance. The expression of TaGAL is negatively correlated with TaCWI-B1 levels, thus may reduce mannan degradation in the cell wall, consequently leading to thickening of the cell wall. Additionally, TaCWI-B1-overexpression lines and TaGAL mutants showed higher resistance to SE; however, TaCWI-B1 mutants were more susceptible to SE than controls. This study provides insights into a FCR and SE resistance gene to combat soil-borne diseases in common wheat.

A Novel Wall-Associated Kinase TaWAK-5D600 Positively Participates in Defense against Sharp Eyespot and Fusarium Crown Rot in Wheat.

Sharp eyespot and Fusarium crown rot, mainly caused by soil-borne fungi Rhizoctonia cerealis and Fusarium pseudograminearum, are destructive diseases of major cereal crops including wheat (Triticum aestivum). However, the mechanisms underlying wheat-resistant responses to the two pathogens are largely elusive. In this study, we performed a genome-wide analysis of wall-associated kinase (WAK) family in wheat. As a result, a total of 140 TaWAK (not TaWAKL) candidate genes were identified from the wheat genome, each of which contains an N-terminal signal peptide, a galacturonan binding domain, an EGF-like domain, a calcium binding EGF domain (EGF-Ca), a transmembrane domain, and an intracellular Serine/Threonine protein kinase domain. By analyzing the RNA-sequencing data of wheat inoculated with R. cerealis and F. pseudograminearum, we found that transcript abundance of TaWAK-5D600 (TraesCS5D02G268600) on chromosome 5D was significantly upregulated, and that its upregulated transcript levels in response to both pathogens were higher compared with other TaWAK genes. Importantly, knock-down of TaWAK-5D600 transcript impaired wheat resistance against the fungal pathogens R. cerealis and F. pseudograminearum, and significantly repressed expression of defense-related genes in wheat, TaSERK1, TaMPK3, TaPR1, TaChitinase3, and TaChitinase4. Thus, this study proposes TaWAK-5D600 as a promising gene for improving wheat broad resistance to sharp eyespot and Fusarium crown rot (FCR) in wheat.

Mapping of Major Resistance Gene Qse.xn-2BL for Sharp Eyespot in the Wheat-Psathyrostachys huashanica Introgression Line H83.

Sharp eyespot, a soil-borne disease of wheat (Triticum aestivum L.), is one of the most devastating diseases and severely affects grain production. The most efficient and economical method of controlling the disease is the utilization of genetic resistance. In this study, the wheat-Psathyrostachys huashanica introgression line H83 processed the enhanced resistance to Rhizoctonia cerealis isolate R0301 than its wheat parent 7182. A resistance locus in the 600 to 800 Mb interval of chromosome 2BL was screened using 244 segregation population F2 plants of H83×Huixianhong with bulked segregant analysis and wheat axiom 660K genotyping array. Furthermore, by using 12 kompetitive allele-specific PCR markers, a major resistance gene, designated as Qse.xn-2BL, was identified in a secondary segregating population with 138 F3:4 lines and initially mapped to a 765.6 to 775.5 Mb interval on chromosome 2BL. Molecular cytology analysis revealed that H83 probably has an alien introgression at the distal of chromosome 2BL, where it overlapped with the mapping target gene. Above all, H83 showed great potential to improve wheat resistance to sharp eyespot and can be expected to improve resistance in wheat breeding.

Identification of a Novel Streptomyces sp. Strain HU2014 Showing Growth Promotion and Biocontrol Effect Against Rhizoctonia spp. in Wheat.

Wheat sharp eyespot is a serious disease caused by the phytopathogens Rhizoctonia cerealis and R. solani. Some species in the genus Streptomyces have been identified as potential biocontrol agents against phytopathogens. In this investigation, the physiological, biochemical, phylogenetic, and genomic characteristics of strain HU2014 indicate that it is a novel Streptomyces sp. most closely related to Streptomyces albireticuli. Strain HU2014 exhibited strong antifungal activity against R. cerealis G11 and R. solani YL-3. Ultraperformance liquid chromatography-mass spectrometry on the four extracts from the extracellular filtrate of strain HU2014 identified 10 chemical constituents in the Natural Products Atlas with high match levels (more than 90%). In an antifungal efficiency test on wheat sharp eyespot, two extracts significantly reduced the lesion areas on bean leaves infected by R. solani YL-3. The drenching of wheat in pots with spore suspension of strain HU2014 demonstrated a control efficiency of 65.1% against R. cerealis G11 (compared with 66.9% when treated by a 30% hymexazol aqueous solution). Additionally, in vitro and pot experiments demonstrated that strain HU2014 can produce indoleacetic acid, siderophores, extracellular enzymes, and solubilized phosphate, and it can promote plant growth. We conclude that strain HU2014 could be a valuable microbial resource for growth promotion of wheat and biological control of wheat sharp eyespot.

The somatic embryogenesis receptor kinase TaSERK1 participates in the immune response to Rhizoctonia cerealis infection by interacting and phosphorylating the receptor-like cytoplasmic kinase TaRLCK1B in wheat.

The sharp eyespot, caused by necrotrophic pathogen Rhizoctonia cerealis, often causes serious yield loss in wheat (Triticum aestivum). However, the mechanisms underlying wheat resistant responses to the pathogen are still limited. In this study, we performed a genome-wide analysis of somatic embryogenesis receptor kinase (SERK) family in wheat. As a result, a total of 26 TaSERK candidate genes were identified from the wheat genome. Only 6 TaSERK genes on the chromosomes 2A, 2B, 2D, 3A, 3B, and 3D showed obvious heightening expression patterns in resistant wheat infected with R. cerealis compared than those un-infected wheat. Of them, the transcripts of 3 TaSERK1 homoeologs on the chromosomes 2A, 2B, and 2D were significantly up-regulated in the highest level compared to other TaSERKs. Importantly, silencing of TaSERK1 significantly impaired wheat resistance to sharp eyespot. Further bio-molecular assays showed that TaSERK1 could interact with the defence-associated receptor-like cytoplasmic kinase TaRLCK1B, and phosphorylated TaRLCK1B. Together, the results suggest that TaSERK1 mediated resistance responses to R. cerealis infection by interacting and phosphorylating TaRLCK1B in wheat. This study sheds light on the understanding of the wheat SERKs in the innate immunity against R. cerealis, and provided a theoretical fulcrum to identify candidate resistant genes for improving wheat resistance against sharp eyespot in wheat.

Identification of Quantitative Trait Loci for Resistance to Wheat Sharp Eyespot in the Einstein × Tubbs Recombinant Inbred Line Population.

Wheat sharp eyespot (SES), caused by the soilborne pathogen Rhizoctonia cerealis Van der Hoeven (teleomorph: Ceratobasidium cereale), is a common stem disease of wheat globally. The disease caused a severe and extensive epidemic throughout the Willamette Valley of Oregon in 2014 and has remained one of the most important wheat diseases in this region. However, little was known about the genetics of host resistance to this disease. A recombinant inbred line (RIL) population with 257 lines developed from a cross of Einstein × Tubbs was used to study SES resistance of wheat. The phenotyping was conducted at two locations and in 3 years. Genotyping by sequencing was done by using Illumina HiSeq 3000. Low broad-sense heritability across four environments was obtained. The results of analysis of variance demonstrated that disease severity was significantly different among RILs for the data combined over environments and for one of the individual environments. Four SES resistance quantitative trait loci (QTL) were detected, including QSES-1A, QSES-2B, QSES-6A, and QSES-7A, and explained 5.9, 5.9, 8.8, and 8.3%, respectively, of the phenotypic variance. All four QTL overlapped or are in close proximity with one or more plant defense genes, and could lay the foundation for marker-assisted breeding.

Identification of new resistance loci against wheat sharp eyespot through genome-wide association study.

Introduction: Wheat sharp eyespot caused by Rhizoctonia cerealis is a serious pathogenic disease affecting plants. The effective strategy for controlling this disease is breeding resistant cultivar. However, to date, no wheat varieties are fully resistant to sharp eyespot, and only a few quantitative trait loci (QTLs) have been shown to be associated with sharp eyespot resistance. Methods: To understand the genetic basis of this disease, a genome-wide association study (GWAS) of sharp eyespot resistance in 262 varieties from all China wheat regions was conducted. Results: After cultivation for three years, only 6.5% of the varieties were resistant to sharp eyespot. Notably, the varieties from the middle and lower Yangtze River displayed higher sharp eyespot resistance than those from Huanghuai wheat zone. Only two varieties had the same resistance level to the control Shanhongmai. The results of GWAS showed that 5 single nucleotide polymorphism (SNP) loci were markedly related to sharp eyespot resistance in the three years repeatedly, and two QTLs, qSE-6A and qSE-7B, on chromosome 6A and 7B were identified. Based on the 'CG' haplotypes of significant SNPs, we found that the two QTLs exhibited additive effects on attenuating sharp eyespot resistance. Discussion: These results provide novel insights into the genetic basis of sharp eyespot resistance in China wheat varieties. The SNPs related to sharp eyespot resistance can be applied for marker-assisted selection in plant breeding.

Ultrabithorax modifies a regulatory network of genes essential for butterfly eyespot development in a wing sector-specific manner.

Nymphalid butterfly species often have a different number of eyespots in forewings and hindwings, but how the hindwing identity gene Ultrabithorax (Ubx) drives this asymmetry is not fully understood. We examined a three-gene regulatory network for eyespot development in the hindwings of Bicyclus anynana butterflies and compared it with the same network previously described for forewings. We also examined how Ubx interacts with each of these three eyespot-essential genes. We found similar genetic interactions between the three genes in fore- and hindwings, but we discovered three regulatory differences: Antennapedia (Antp) merely enhances spalt (sal) expression in the eyespot foci in hindwings, but is not essential for sal activation, as in forewings; Ubx upregulates Antp in all hindwing eyespot foci but represses Antp outside these wing regions; and Ubx regulates sal in a wing sector-specific manner, i.e. it activates sal expression only in the sectors that have hindwing-specific eyespots. We propose a model for how the regulatory connections between these four genes evolved to produce wing- and sector-specific variation in eyespot number.

Butterfly Wing Color Pattern Modification Inducers May Act on Chitin in the Apical Extracellular Site: Implications in Morphogenic Signals for Color Pattern Determination.

Butterfly wing color patterns are modified by various treatments, such as temperature shock, injection of chemical inducers, and covering materials on pupal wing tissue. Their mechanisms of action have been enigmatic. Here, we investigated the mechanisms of color pattern modifications usingthe blue pansy butterfly Junoniaorithya. We hypothesized that these modification-inducing treatments act on the pupal cuticle or extracellular matrix (ECM). Mechanical load tests revealed that pupae treated with cold shock or chemical inducers were significantly less rigid, suggesting that these treatments made cuticle formation less efficient. A known chitin inhibitor, FB28 (fluorescent brightener 28), was discovered to efficiently induce modifications. Taking advantage of its fluorescent character, fluorescent signals from FB28 were observed in live pupae in vivo from the apical extracellular side and were concentrated at the pupal cuticle focal spots immediately above the eyespot organizing centers. It was shown that chemical modification inducers and covering materials worked additively. Taken together, various modification-inducing treatments likely act extracellularly on chitin or other polysaccharides to inhibit pupal cuticle formation or ECM function, which probably causes retardation of morphogenic signals. It is likely that an interactive ECM is required for morphogenic signals for color pattern determination to travel long distances.