Genomic Sequence Resource of Kabatiella zeae, the Causative Pathogen of Corn Eyespot Disease.

Kabatiella zeae is the causative pathogen of corn eyespot disease, which is an important leaf disease that damages corn (Zea mays L.) worldwide. In this study, we provided an annotated draft of the assembled genome of the K. zeae field strain KZ1 through PacBio and Illumina sequencing. The assembled KZ1 genome size is 23,602,820 bp, and its GC content is 50.71%. The completeness of the assembled genome is 97.6% in this study. The assembly obtained in this study has 94 contigs and the length of N50 is 720,243 bp. This study is the first report of the K. zeae genome, which contributes to further research on the genetic variation and pathogenic mechanism of this important fungal pathogen.[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.

Genome Sequence Resource for Nigrospora oryzae, an Important Pathogenic Fungus Threatening Crop Production.

Nigrospora oryzae is an important phytopathogenic fungus with a broad host range. Here, we report an annotated draft of the genome of N. oryzae field strain GZL1 collected from maize assembled from PacBio and Illumina sequencing reads. The assembly we obtained has 15 scaffolds with an N50 length of 4,037,616 bp. The resulting GZL1 draft genome is 43,214,190 bp, with GC content of 58.19%. The completeness of GZL1 genome assembly is 99.30%. This study is the first report of the genome sequence of N. oryzae, which can facilitate future study of the genetic variation and pathogenic mechanism of this important fungal pathogen.[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.

Immunocompetence of Gynaikothrips uzeli (Thysanoptera: Phlaeothripidae) populations from different latitudes against Beauveria bassiana (Hypocreales: Cordycipitaceae).

Gynaikothrips uzeli gall thrips are protected from insecticide exposure by their leaf gall habitat. A biocontrol strategy based on entomopathogenic fungi is an alternative approach for the control of G. uzeli. Higher temperatures can promote the reproduction and spread of pests; however, the impact of higher temperatures on biological control is unclear. We studied the immunocompetence of thrips from different latitudes and determined the effect of degree days on thrips immunity. We examined the potential impact of temperature on the biocontrol provided by entomopathogenic fungi. Beauveria bassiana pathogenicity against thrips increased with decreasing latitude, suggesting that immunity of thrips increased as latitude increased. The phenoloxidase activity of G. uzeli increased with increasing latitude but there was no significant change in hemocyte concentration. This indicated that the humoral immunity of thrips was significantly associated with degree days, and this was confirmed by transcriptome data. Transcriptome and RT-PCR results showed that the expression of key genes in eight toll pathways increased with increasing latitude. The relative expression of key genes in the Toll pathway of thrips and the activity of phenoloxidase decreased with increasing degree days that are characteristic of lower latitudes. These changes led to a decrease in humoral immunity. The immunity of G. uzeli against entomopathogenic fungi increased as degree days characteristic of lower latitudes decreased. Increased temperatures associated with lower latitude may therefore increase biocontrol efficacy. This study clarified immune level changes and molecular mechanisms of thrips under different degree days.

Specific and sensitive detection of the guava fruit anthracnose pathogen (Colletotrichum gloeosporioides) by loop-mediated isothermal amplification (LAMP) assay.

Anthracnose of guava, caused by the fungus Colletotrichum gloeosporioides, is a major factor limiting worldwide guava production. Timely and accurate detection of the pathogen is important in developing a disease management strategy. Herein, a loop-mediated isothermal amplification (LAMP) assay for the specific and sensitive detection of C. gloeosporioides was developed using primers targeting the ß-tubulin 2 (TUB2) gene. The optimal reaction conditions were 64 °C for 60 min. The specificity of the method was tested against C. gloeosporioides isolates, Colletotrichum spp. isolates, and isolates of other genera. Positive results were obtained only in the presence of C. gloeosporioides, whereas no cross-reaction was observed for other species. The detection limit of the LAMP assay was 10 fg of genomic DNA in a 25 µL reaction. The LAMP assay successfully detected C. gloeosporioides in guava fruit collected in the field. The results indicate that the developed LAMP assay is a simple, cost-effective, rapid, highly sensitive, and specific tool for the diagnosis of guava anthracnose caused by C. gloeosporioides and could be useful for disease management.

The Subcellular Localization and Functional Analysis of Fibrillarin2, a Nucleolar Protein in Nicotiana benthamiana.

Nucleolar proteins play important roles in plant cytology, growth, and development. Fibrillarin2 is a nucleolar protein of Nicotiana benthamiana (N. benthamiana). Its cDNA was amplified by RT-PCR and inserted into expression vector pEarley101 labeled with yellow fluorescent protein (YFP). The fusion protein was localized in the nucleolus and Cajal body of leaf epidermal cells of N. benthamiana. The N. benthamiana fibrillarin2 (NbFib2) protein has three functional domains (i.e., glycine and arginine rich domain, RNA-binding domain, and α-helical domain) and a nuclear localization signal (NLS) in C-terminal. The protein 3D structure analysis predicted that NbFib2 is an α/ß protein. In addition, the virus induced gene silencing (VIGS) approach was used to determine the function of NbFib2. Our results showed that symptoms including growth retardation, organ deformation, chlorosis, and necrosis appeared in NbFib2-silenced N. benthamiana.

A New Diagnostic System for Detection of 'Candidatus Liberibacter asiaticus' Infection in Citrus.

In this study, two polyclonal antibodies were produced against the Omp protein of 'Candidatus Liberibacter asiaticus'. First, omp genes were sequenced to exhibit 99.9% identity among 137 isolates collected from different geographical origins. Then, two peptides containing the hydrophobic polypeptide-transport-associated (POTRA) domain and ß-barrel domain, respectively, were identified on Omp protein. After that, these two peptides were overexpressed in Escherichia coli and purified by affinity chromatography to immunize the white rabbits. Finally, the antiserum was purified by affinity chromatography. The two Omp antibodies gave positive results (0.454 to 0.633, 1:1,600 dilution) in enzyme-linked immunosorbent assay against 'Ca. L. asiaticus'-infected samples collected from different geographical origins but revealed negative results against other pathogen-infected, nutrient-deficient and healthy samples. The antibody against the POTRA domain of Omp protein could detect 'Ca. L. asiaticus' in 45.7% of the symptomatic samples compared with a 56.2% detection rate with a polymerase chain reaction assay. These new antibodies will provide a very useful supplement to the current approaches to 'Ca. L. asiaticus' detection and also provide powerful research tools for tracking distribution of this pathogen in vivo.