摘要:
The voltage-gated calcium channel (VGCC) beta subunit (Ca-v beta) protein is a kind of cytosolic auxiliary subunit that plays an important role in regulating the surface expression and gating characteristics of high-voltage-activated (HVA) calcium channels. Ditylenchus destructor is an important plant-parasitic nematode. In the present study, the putative Ca-v beta subunit gene of D. destructor, namely, DdCa(v)beta, was subjected to molecular characterization. In situ hybridization assays showed that DdCa(v)beta was expressed in all nematode tissues. Transcriptional analyses showed that DdCa(v)beta was expressed during each developmental stage of D. destructor, and the highest expression level was recorded in the third-stage juveniles. The crucial role of DdCa(v)beta was verified by dsRNA soaking-mediated RNA interference (RNAi). Silencing of DdCa(v)beta or HVA Ca-v alpha(1) alone and co-silencing of the DdCa(v)beta and HVA Ca-v alpha(1) genes resulted in defective locomotion, stylet thrusting, chemotaxis, protein secretion and reproduction in D. destructor. Co-silencing of the HVA Ca-v alpha(1) and Ca-v beta subunits showed stronger interference effects than single-gene silencing. This study provides insights for further study of VGCCs in plant-parasitic nematodes.
作者:
Wang, Ya Rong*;Hu, Zhao*;Zhong, Jie*;Chen, Yi*;Zhu, Jun Zi*
期刊:
Plant Disease,2022年106(1):316 ISSN:0191-2917
通讯作者:
Wang, Ya Rong;Hu, Zhao;Zhong, Jie;Chen, Yi;Zhu, Jun Zi
作者机构:
[Wang, Ya Rong; Hu, Zhao] Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Changsha, China;[Wang, Ya Rong] 1357928347@qq.com;[Hu, Zhao] 1373961913@qq.com;[Zhong, Jie] plant pathology, bHunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Nongda Road 1, Furong District, Changsha City, Hunan Province, 410128, P.R. China, Changsha, China, 410128;[Zhong, Jie] 1601389330@qq.com
通讯机构:
Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Furong District, Hunan Province, Changsha, China
摘要:
Tobacco (Nicotiana tabacum L.) is an annual, leafy, herb of the genus Nicotiana in the family Solanaceae. It is an important commercial crop in China. In 2020, a leaf spot disease was observed on tobacco leaves in commercial fields in the Hunan Province of China. Symptoms appeared as water-soaked, yellow-green spots, then turned dark brown, and coalesced into larger necrotic lesions, often leading to leaf wilt. Approximately 20% of the plants in a 50-ha area were infected, exhibiting symptomatic spots on 60% of these leaves. Symptomatic leaf samples were collected and cut into small pieces, sterilized with 70% ethanol for 10 s, 0.1% HgCl2 for 40s, rinsed with sterile distilled water for three times, plated on potato dextrose agar (PDA) and incubated at 26°C in the dark. Isolates with similar morphology were developed from ten samples. Fungal isolates produced densely, white to dark green, aerial mycelium. Conidia were straight, hyaline, aseptate, cylindrical, contained oil globules, and 15 to 25 µm × 3.0 to 4.0 µm (n=50). Appressoria were dark brown, irregularly shaped, 5.5 to 10.0 μm × 4.5 to 6.5 μm (n=50). These morphological characteristics were typical of Colletotrichum cliviicola (Yang et al. 2009). For molecular identification, the internal transcribed spacer (ITS) region of rDNA, actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and chitin synthase (CHS-1) genes of a representative isolate CS16-2 were amplified and sequenced using the primer pairs as described previously (Weir et al. 2012). These sequences were deposited in GenBank (GenBank Accession Nos. MW649137 for ITS, MW656181 for ACT, MW656182 for GAPDH and MW656183 for CHS-1). BLAST analysis showed that they had 99.46% to 100% identity to the corresponding sequences of C. cliviicola strains. A concatenated phylogenetic tree was generated, using the ACT, GAPDH and CHS-1 sequences of the isolate CS16-2 and other closely matching Colletotrichum species obtained from the GenBank. We found that the CS16-2 was grouped with the C. cliviicola clade with 97% bootstrap support, including the C. cliviicola strain AH1B6 (Wang et al. 2016). Pathogenicity was tested spraying 2-month-old potted tobacco plants until runoff with a conidial suspension (105 spores/ml). Leaves were mock inoculated with sterilized water. The pathogenicity tests were performed twice, with three replicate plants each. Plants were kept in humid chambers at 26°C with a 12-h photoperiod. Five days post-inoculation, the inoculated plants developed symptoms of consisting of the yellow-brown necrotic lesion resembling the symptoms that were observed in fields, while the control plants remained symptomless. C. cliviicola was re-isolated and identified by morphological and molecular methods as described above. Currently, C. cliviicola has been reported to be the causal agent of anthracnose in some plants, such as soybean (Zhou et al. 2017) and Zamioculcas zamiifolia (Barbieri et al. 2017). However, to our knowledge, this is the first report of C. cliviicola causing leaf spot on tobacco in China and even in the word. Given that the may greatly affect the yield and quality of tobacco production, growers should be prepared to manage this new disease. This work might provide further insight for disease diagnosis on tobacco as some other Colletotrichum species, such as C. fructicola (Wang et al. 2016) and C. karsti (Zhao et al. 2020), have also been responsible for anthracnose.
通讯机构:
[Zhou, Xinyu] H;Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Nongda Road 1, Furong District, 410128, Changsha City, Hunan Province, P.R. China.
摘要:
A novel polymycovirus isolated from the plant-pathogenic fungus Colletotrichum gloeosporioides was identified. The viral genome is composed of nine double-stranded RNA segments, ranging in size from 699 bp to 2,444 bp. With the exception of dsRNA5, which contains two open reading frames (ORF5-1 and ORF5-2), the other dsRNA segments each contain one ORF. The proteins encoded by ORFs 1-8 are homologous to the proteins encoded by ORFs 1-8 of Colletotrichum camelliae filamentous virus 1 (CcFV-1). The amino acid sequences of the RNA-dependent RNA polymerase (RdRp) encoded by ORF1 and the viral methyltransferase encoded by ORF3 share 87.6% and 83.3% identity with CcFV-1. The proline-alanine-serine-rich protein (PASrp) encoded by ORF4 shares 86.6% sequence identity with that of CcFV-1. The proteins encoded by ORFs 2, 5 - 1, 6, 7, and 8 share 86.6%, 82.5%, 89.0%, 45.7%, and 95.5% sequence identity, respectively, with the corresponding proteins of CcFV-1. dsRNA9 is a defective copy of dsRNA2 that lacks a stretch of 1556 bp (nt 519 to nt 2074). Phylogenetic analysis based on the RdRp protein indicated that the novel virus clustered with members of the family Polymycoviridae, and based on the above results, we have tentatively named it "Colletotrichum gloeosporioides polymycovirus virus 1" (CgPmV1). To our knowledge, this is the first report of a polymycovirus with a defective dsRNA genome in C. gloeosporioides.
关键词:
CDK1;cell cycle;cyclin B;Heliothis virescens ascovirus 3h;SeFB cell line
摘要:
Ascoviruses are double-stranded DNA viruses that are pathogenic to noctuid larvae. In vitro infection causes the cells to fail to replicate and proliferate normally. However, the molecular mechanisms are unclear. In this study, the transmission electron microscopydata of infected-Spodoptera exigua (Hübner) fat body cells (SeFB, IOZCAS-SpexII-A cells) showed that virions were internalized in phagocytic vesicles, but not in the nucleus. FACS of cell-cycle progression was performed in SeFB cells infected with Heliothis virescens ascovirus 3h (HvAV-3h). The cell cycle phase distributions of the SeFB cells were G(1) = 29.52 ± 1.10%, S = 30.33 ± 1.19%, and G(2) /M = 40.06 ± 0.75%. The cell culture doubling time was approximately 24 h. The G(1) , S, and G(2) /M phases were each approximately 8 h. The unsynchronized or synchronized cells were arrested at G(2) /M phase after infection with HvAV-3h. Our data also showed that cells with more than 4N DNA content appeared in the HvAV-3h-treated group. While the mRNA levels of cyclin B(1) , cyclin H, and cyclin-dependent kinase 1 (CDK1) were downregulated after HvAV-3h infection, the mRNA expression levels of cyclin A, cyclin D, and cyclin B(2) were not significantly changed. Western blotting results showed that the expression of cyclin B(1) and CDK1 in infected SeFB cells within 24 h postinfection(hpi), and HvAV-3h infection inhibited the expression of cyclin B(1) and CDK1 at 12-24 hpi. Overall, these data implied that HvAV-3h infection leads to an accumulation of cells in the G(2) /M phases by downregulating the expression of cyclin B(1) and CDK1.
作者机构:
[Li, Xiao-Gang; Zhong, Jie; Li, Ping; Zhu, Jun-Zi] Hunan Agr Univ, Hunan Prov Key Lab Biol & Control Plant Dis & Ins, Nongda Rd 1, Changsha 410128, Peoples R China.;[Li, Xiao-Gang; Zhu, Jun-Zi] Hunan Agr Univ, Hunan Engn Res Ctr Agr Pest Early Warning & Contr, Nongda Rd 1, Changsha 410128, Peoples R China.
通讯机构:
[Jie Zhong; Xiao-Gang Li] H;Hunan Provincial Key Laboratory for Biology and Control of Plant Diseases and Insect Pests, Hunan Agricultural University, Nongda Road 1, Furong District, Changsha 410128, China<&wdkj&>Hunan Engineering Research Center of Agricultural Pest Early Warning and Control, Hunan Agricultural University, Nongda Road 1, Furong District, Changsha 410128, China
摘要:
Abstract: Luffa sponge gourd (Luffa cylindrica) is an important cucurbitaceous vegetable and is known as the source of loofah. From 2020 to 2021, a leaf disease occurred on luffa leaves in the Hunan Province of China. Symptoms were displayed as oval to irregular chlorotic lesions surrounded by yellow halos. The pathogens were isolated from the affected leaves. According to morphological characterization and molecular identification using multi-locus phylogenetic analysis of the internal transcribed spacer (ITS), actin (ACT), chitin synthase (CHS-1), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin (TUB2), and partial mating type (Mat1-2) gene (ApMAT) regions, the pathogens were identified as two Colletotrichum species: Colletotrichum fructicola and C. siamense. Koch’s postulates were identified by a pathogenicity test and re-confirmation. To the best of our knowledge, C. fructicola and C. siamense are two new species associated with luffa sponge gourd anthracnose. Keywords: morphological characterization; multi-locus phylogenetic analysis; Luffa cylindrica; Colletotrichum fructicola; Colletotrichum siamense
作者:
Ya Wang;Jing Tian;Zihao Wang;Chaonan Li;Xiaogang Li
期刊:
ACS Agricultural Science and Technology,2022年2(3):534-545 ISSN:2692-1952
通讯作者:
Xiaogang Li
作者机构:
[Li C.; Tian J.; Wang Z.; Wang Y.] College of Plant Protection, Hunan Agricultural University, Changsha, 410128, China;Hunan Prov. Engineering Technology Research Center for Bio Pesticide and Formulating Processing, Changsha, 410128, China;[Li X.] College of Plant Protection, Hunan Agricultural University, Changsha, 410128, China, Hunan Prov. Engineering Technology Research Center for Bio Pesticide and Formulating Processing, Changsha, 410128, China
通讯机构:
[Li, X.] C;College of Plant Protection, China
通讯机构:
[Hou, DH ] W;[Huang, GH ] H;Weifang Med Univ, Sch Life Sci & Technol, Weifang 261053, Peoples R China.;Hunan Agr Univ, Hunan Prov Key Lab Biol & Control Plant Dis & Ins, Changsha 410128, Hunan, Peoples R China.
摘要:
Analysis of orthology is important for understanding protein conservation, function, and phylogenomics. In this study, we performed a comprehensive analysis of gene orthology in the family Ascoviridae based on identification of 366 protein homologue groups and phylogenetic analysis of 34 non-single-copy proteins. Our findings revealed 90 newly annotated proteins, five newly identified core proteins for the family Ascoviridae, and 14 core proteins for the genus Ascovirus. A phylogenomic tree of 11 Ascoviridae members was constructed based on a concatenation of 35 of the 45 ortholog groups. In combination with phosphoproteomic results and conservation estimations, 30 conserved phosphorylation sites on 17 phosphoproteins were identified from a total of 176 phosphosites on 57 phosphoproteins from Heliothis virescens ascovirus 3h (HvAV-3h), providing potential research targets for investigating the role of these protein in the regulation of viral infection. This study will facilitate genome annotation and comparison of further Ascoviridae members as well as functional genomic investigations.