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Plant Disease Apr 2024Kiwifruit is widely cultivated for its high vitamin C content and nutritional value. In January 2022, root rot symptoms were found in about 30% of Actinidia chinensis...
Kiwifruit is widely cultivated for its high vitamin C content and nutritional value. In January 2022, root rot symptoms were found in about 30% of Actinidia chinensis cv. Jinyan plants grafted on A. deliciosa rootstocks in an orchard located in Sanming (26.32°N, 117.23°E), Fujian Province of China. The affected plants appeared stunted, with brown and decaying roots, some of which were covered with white hyphae. To isolate the pathogen, the surfaces of typical symptomatic roots were sterilized for 30 s using 75% ethanol, followed by four rinses in sterile water, placing on potato dextrose agar (PDA), and incubating away from light at 25°C for 7 days. 16 Globisporangium-like isolates were obtained through hyphal tip isolation, displaying a milky-white appearance with irregular protuberances on the surface, and yellow-white backs with radial fold lines. The isolates were then cultured on corn meal agar for 5 days at 25°C in dark for morphological characteristics. Under microscope, the hyphae appeared as long strips without septa and 4.1 to 8.2 µm wide (average 6.7 µm), containing irregularly sized spherical droplets. Both terminal and intercalary hyphae swellings were observed; these appeared either spherical or subspherical, with some having projections. Their dimensions were 12.3 to 27.6 µm (average 17.3 µm). The oospores were mostly spherical, either plerotic or aplerotic, 11.8 to 22.3 µm wide (average 18.9 µm), with occasional projections. The antheridia were rod-shaped and curved, with one end attached to the oogonia. Amplification of the sequences of internal transcribed spacer (ITS) regions and cytochrome c oxidase subunit I (COI) were conducted using the primers ITS1/ITS4 (White et al. 1990) and OomCoxI-Levlo/OomCoxI-Levup (Robideau et al. 2011), respectively. The sequencing results revealed identical ITS and COI sequences in all 16 isolates. BLASTn analysis of the 969-bp ITS sequence ON202808 showed 99.38-99.59% similarity (965/971bp, 967/971bp) with the KJ162353 and AY598701 sequences from Globisporangium spinosum isolates, while the 700-bp COI sequence ON075783 showed 100% and 99.41% identity (680/680bp, 676/680bp) with the GenBank sequences HQ708835 and HQ708832, respectively, from G. spinosum. Phylogenetic analysis also showed that the obtained isolate (termed MA16) clustered with isolates from G. spinosum on the same evolutionary branch. For pathogenicity testing, four-month-old healthy Jinyan (A. chinensis) plants grown in sterilized media were transferred to sterile petri dishes covered with wet filter paper, and their roots were inoculated with a 5-mm-wide disk of MA16 when cultivated on PDA medium for 5 days. Miliang-1 (A. deliciosa) and Hongyang (A. chinensis) plants were treated similarly. The control groups each included three plants that were inoculated with non-colonized PDA. The plants were kept at 25 °C with a 12-/12-h light/dark cycle for 10 days when the inoculated plants exhibited root rot symptoms similar to those seen in the field, together with rotting and browning of the leaves. The control plants appeared healthy with no symptoms. After re-isolated from infected tissues, the pathogen was verified to be G. spinosum according to its ITS sequence, thus fulfilling the Koch's postulates. Recently, Pythium spinosum has been classified as G. spinosum according to whole-genome sequencing and phylogenomic analysis (Nguyen et al. 2022). Based on the morphological features and pathogenicity results, MA16 was identified as G. spinosum (van der Plaats-Niterink 1981; Huo et al. 2023). This report appears to be the first description of kiwifruit root rots caused by G. spinosum in China, and its identification will assist the development of strategies to counteract the disease.
PubMed: 38687573
DOI: 10.1094/PDIS-12-23-2773-PDN -
Plant Disease Jul 2007During 2005 and 2006, eight declining, mature Fagus sylvatica trees with severe crown dieback were encountered at two stands near Ljubljana and Kamnik in central...
During 2005 and 2006, eight declining, mature Fagus sylvatica trees with severe crown dieback were encountered at two stands near Ljubljana and Kamnik in central Slovenia. Bleeding cankers were seen mostly in the lower parts of the stem. Fresh cankers showed orange brown necroses of the inner bark and cambium. Older cankers were dark brown and surrounded by cracks and shedding bark. Small pieces of marginal tissue were excised from fresh necrotic lesions and incubated in the dark at 20°C on PARP and cornmeal (CMA) agar plates. Soil samples collected from around the base of these declining trees were submerged in sterile distilled water. Leaves of Rhododendron catawbiense were used as bait to test for the presence of Phytophthora spp. Several similar isolates were obtained from trees and soil from both stands. Colonies were narrowly petaloid, slow growing on PARP, and fast growing on CMA. The colonies produced abundant sporangia when submerged in pond water. Semipapillate, noncaducous sporangia were ovoid to obpyriform, but were occasionally distorted, and 30 to 69 (49) μm long and 23 to 44 (34) μm wide. Neither hyphal swellings nor chlamydospores developed. Isolates were homothallic showing paragynous antheridia, spherical oogonia of 22 to 31 (25) μm diameter, and plerotic oospores of 20 to 27 (23) μm diameter. The morphological characters resembled those described for Phytophthora citricola (1). ITS rDNA, spanning ITS 1 and 2, plus the 5.8S rDNA were generated using primers ITS4 and ITS5. Isolates from symptomatic trees and the soil yielded identical sequences and were the same as various sequences deposited for P. citricola at GenBank. The ITS rDNA of one representative strain was deposited at GenBank (Accession No. EF423556). Mycelial plugs from one of the P. citricola strains grown on CMA were used to inoculate stem wounds of seven potted seedlings and wounds made on four freshly cut, healthy branches of F. sylvatica. Sterile agar plugs were used as controls. The test was carried out over 4 weeks at 20°C. Extensive necrotic lesions developed around inoculation points on seedlings and branches, whereas the controls showed no symptoms. P. citricola could also be reisolated from margins of these lesions. During the past decade, declining F. sylvatica trees were observed in an increasing number of stands in Germany (2). Several Phytophthora species appeared to be involved in this decline, but P. citricola was the most frequently recovered species (2,3). To our knowledge, this it the first report of P. citricola found associated with a decline of European beech in Slovenia. References: (1) D. C. Erwin and O. K. Ribeiro. Pages 282-287 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (2) T. Jung. Forst Holz. 60:131, 2005. (3) T. Jung et al. Mycologist 19:159, 2005.
PubMed: 30780412
DOI: 10.1094/PDIS-91-7-0907C -
Frontiers in Genetics 2022Ovarian development is a key physiological process that holds great significance in the reproduction of the Chinese mitten crab (), which is an economically important...
Ovarian development is a key physiological process that holds great significance in the reproduction of the Chinese mitten crab (), which is an economically important crab species for aquaculture. However, there is limited knowledge for the regulatory mechanisms of ovarian development. To study the molecular mechanisms of its ovarian development, transcriptome analysis was performed in the ovary and hepatopancreas of during ovarian stages I (oogonium proliferation), II (endogenous vitellogenesis), and III (exogenous vitellogenesis). The results showed that 5,520 and 226 genes were differentially expressed in the ovary and hepatopancreas, respectively. For KEGG enrichment analysis, the differentially expressed genes in the ovary were significantly clustered in phototransduction-fly, phagosome, and ECM-receptor interaction. Significantly enriched pathways in the hepatopancreas included fatty acid biosynthesis, fatty acid metabolism, and riboflavin metabolism. Further analysis showed that 25 genes and several pathways were mainly involved in oogenesis, including the ubiquitin-proteasome pathway, cyclic AMP-protein kinase A signaling pathway, and mitogen-activated protein kinase signaling pathway. Twenty-five candidate genes involved in vitellogenesis and endocrine regulation were identified, such as vitellogenin, vitellogenin receptor, estrogen sulfotransferase, ecdysone receptor, prostaglandin reductase 1, hematopoietic prostaglandin D synthase and juvenile hormone acid O-methyltransferase. Fifty-six genes related to nutritional metabolism were identified, such as fatty acid synthase, long-chain-fatty-acid-CoA ligase 4, 1-acyl-sn-glycerol-3-phosphate acyltransferase 4, fatty acid-binding protein, and glycerol-3-phosphate acyltransferase 1. These results highlight the genes involved in ovarian development and nutrition deposition, which enhance our understanding of the regulatory pathways and physiological processes of crustacean ovarian development.
PubMed: 35685440
DOI: 10.3389/fgene.2022.910682 -
Plant Disease Aug 2023The eastern redbud ( L.) is an esthetically and economically important landscape tree with vibrant blossoms and attractive heart-shaped leaves. One-year-old eastern...
The eastern redbud ( L.) is an esthetically and economically important landscape tree with vibrant blossoms and attractive heart-shaped leaves. One-year-old eastern redbud seedlings grown in field condition in two commercial nurseries in Warren Co., Tennessee exhibited severe root rot in October 2021. Dark brown to black lesions and rot were observed in the affected roots (Fig. 1a). Disease severity was 50-75% of root area and disease incidence was approximately 30-40% of 10,000 plants. Surface sterilized (10% NaOCl; 1 min) symptomatic tissues were plated on V8-PARPH and incubated at 25°C. Whitish cottony mycelia with radiate and chrysanthemum flower-like growth patterns were observed within 4 days of incubation. Subglobose papillate sporangia (10.24 to 20.98 µm, =50), filamentous to globose smooth oogonia, bell-shaped antheridia and spherical zoospores that are characteristic of (de Cock et al. 2015) were observed in older cultures that were subjected to specific growth conditions as previously described by Ghimire & Baysal-Gurel (2023). Pathogen identification was confirmed by extracting total DNA using the DNeasy PowerLyzer Microbial Kit from 7-day-old cultures of isolates FBG0874, FBG1998, FBG2009 grown on V8-PARPH. specific LAMP assay was conducted for the rapid molecular screening and confirmation of the isolates (Ghimire et al. 2023). Primer pairs ITS1/ITS4 (White et al. 1990), NL1/NL4 (Baten et al. 2014), Levup and Fm85mod (Robideau et al. 2011) were used to amplify and sequence the internal transcribed spacer (ITS), 28S large subunit (LSU) of ribosomal RNA and the cytochrome c oxidase subunit I (CoxI) of mitochondrial DNA genetic markers, respectively. The sequences (GenBank accession nos. OR204701, OR205212, OR205213: ITS; OR205214, OR205215, OR205216: LSU; OR220805, OR220806, OR220807: CoxI) were 100% similar to ITS, LSU, and CoxI genetic markers of isolates in the NCBI database (MK011121: ITS, KX092469: LSU and KT692908: CoxI). Pathogenicity tests were performed on one-year-old eastern redbud seedlings grown in 1 gal containers to fulfill Koch's postulate. Eastern redbud seedlings were drench inoculated (150 ml/plant) with pathogen slurry (two plates of 7-day-old culture/liter) (Panth et al. 2021) of isolates FBG0874, FBG1998, and FBG2009 (five plants/isolate). Control plants were drenched with agar slurry without pathogen. The study was conducted in a greenhouse maintained at 21 to 23°C, 70%RH, with 16-h photoperiod and irrigated twice a day for 2 min using an overhead irrigation system. Fourteen days after inoculation dark brown to black lesions developed in the root of all inoculated plants that were identical to the symptoms observed in the original samples (Fig. 1b), while the roots of non-inoculated plants remained asymptomatic (Fig. 1c). Isolates resembling morphological characteristics were recovered from inoculated plants, and their identity was confirmed as using LAMP assay. has been previously reported to cause root and crown rot in flowering cherry, ginkgo, and red maple in Tennessee (Baysal-Gurel et al. 2021, Panth et al. 2021). To our knowledge, this is the first report of causing root rot of eastern redbud in Tennessee and the United States. Identification of this pathogen as the causal agent is important in designing and implementing effective management practices to mitigate this threat to redbud production.
PubMed: 37622272
DOI: 10.1094/PDIS-07-23-1276-PDN -
Cells Nov 2023(1) Fshβ and Lhβ showed stronger signals and higher transcript levels from 590 to 1050 dph than at earlier stages, implying their active involvement during primary...
Chronological Changes in Gonadotropin-Releasing Hormone 1, Gonadotropins, and Sex Steroid Hormones along the Brain-Pituitary-Gonadal Axis during Gonadal Sex Differentiation and Development in the Longtooth Grouper, .
(1) Fshβ and Lhβ showed stronger signals and higher transcript levels from 590 to 1050 dph than at earlier stages, implying their active involvement during primary oocyte development. (2) Fshβ and Lhβ at lower levels were detected during the phases of ovarian differentiation and oogonial proliferation. (3) E concentrations increased significantly at 174, 333, and 1435 dph, while T concentrations exhibited significant increases at 174 and 333 dph. These findings suggest potential correlations between serum E concentrations and the phases of oogonial proliferation and pre-vitellogenesis.
Topics: Female; Animals; Bass; Sex Differentiation; Gonadotropin-Releasing Hormone; Gonadal Steroid Hormones; Follicle Stimulating Hormone, beta Subunit; Luteinizing Hormone, beta Subunit; Brain
PubMed: 37998369
DOI: 10.3390/cells12222634 -
Insects Jul 2020The silkworm, Linnaeus, is an important economic insect and a representative model organism of Lepidoptera, which has been widely used in the study of reproduction and...
The silkworm, Linnaeus, is an important economic insect and a representative model organism of Lepidoptera, which has been widely used in the study of reproduction and development. The development of the silkworm's reproductive gland is easily affected by many external factors, such as chemical insecticides. After the silkworm larvae were treated with different concentrations of pyriproxyfen, the results showed that the number of eggs and hatching rate of eggs in the silkworm can be reduced by pyriproxyfen, and the concentration effects were displayed. Pyriproxyfen exposure could affect the normal development of the ovary tissue by reducing the number of oocytes and oogonia in the ovaries of silkworm fed with pyriproxyfen. We employed qRT-PCR, to detect the expressions of genes related to ovary development (, , and ) and hormone regulation ( and ) in silkworm. Our study showed that the transcription levels of , , and in the treatment group were lower than those in the control group (6.08%, 61.99%, 83.51%, 99.31% and 71.95%, respectively). The transcription level of was 70.22% for the control group, while that of was upregulated by 3.92-fold. Changes of transcription levels of these genes caused by pyriproxyfen exposure ultimately affect the absorption of nutrients, energy metabolism, ovary development and egg formation of the silkworm, thus leading to reproductive disorders of the silkworm. In general, our study revealed the response of silkworm reproduction to pyriproxyfen exposure and provided a certain reference value for the metabolism of the silkworm to pyriproxyfen.
PubMed: 32722009
DOI: 10.3390/insects11080467 -
International Journal of Molecular... Aug 2020TSP1 was reported to be involved in multiple biological processes including the activation of TGF-β signaling pathways and the regulation of angiogenesis during wound...
TSP1 was reported to be involved in multiple biological processes including the activation of TGF-β signaling pathways and the regulation of angiogenesis during wound repair and tumor growth, while its role in ovarian folliculogenesis remains to be elucidated. In the present study, Tsp1a was found to be expressed in the oogonia and granulosa cells of phase I to phase IV follicles in the ovaries of Nile tilapia by immunofluorescence. homozygous mutants were generated by CRISPR/Cas9. Mutation of resulted in increased oogonia, reduced secondary growth follicles and delayed ovary development. Expression of the cell proliferation marker PCNA was significantly up-regulated in the oogonia of the mutant ovaries. Furthermore, transcriptomic analysis revealed that expressions of DNA replication related genes were significantly up-regulated, while cAMP and MAPK signaling pathway genes which inhibit cell proliferation and promote cell differentiation were significantly down-regulated. In addition, aromatase (Cyp19a1a) expression and serum 17β-estradiol (E2) concentration were significantly decreased in the mutants. These results indicated that lacking resulted in increased proliferation and inhibited differentiation of oogonia, which in turn, resulted in increased oogonia, reduced secondary growth follicles and decreased E2. Taken together, our results indicated that was essential for ovarian folliculogenesis in Nile tilapia.
Topics: Animals; Aromatase; Female; Fish Proteins; MAP Kinase Signaling System; Ovarian Follicle; Proliferating Cell Nuclear Antigen; Thrombospondin 1; Tilapia
PubMed: 32824362
DOI: 10.3390/ijms21165893 -
Pathogens (Basel, Switzerland) Aug 2021Alder dieback remains a major problem in European alder stands and its spread continues to threaten their existence. The causal agent of this disease is the so-called...
Alder dieback remains a major problem in European alder stands and its spread continues to threaten their existence. The causal agent of this disease is the so-called alder species complex, which includes the hybrid × and its parental species and . Little is known about the survival of these species in alder. The aim of our investigations was to find out whether, and if so where, the pathogen survives. The subject of these studies was alder roots. Therefore, artificial infection studies and histological studies with . × and were carried out on seedlings of black alder (). These histological studies revealed oogonia and oospores of . × and in different parts of the root tissue.
PubMed: 34451441
DOI: 10.3390/pathogens10080977 -
Animal Reproduction 2024Steroids and gonadotrophins are essential for the regulation of late stages of preantral development and antral follicular development. Although the luteinizing hormone...
Steroids and gonadotrophins are essential for the regulation of late stages of preantral development and antral follicular development. Although the luteinizing hormone receptor (LHCGR) has been detected in the preantral follicles of rats, rabbits, and pigs, its expression, in bovine fetal ovary, has not been demonstrated. Based on this, we aimed to investigate the expression of the and mRNA binding protein (), as well as, to quantify bta-miR-222 (a regulatory microRNA of the LHCGR gene) during the development of bovine fetal ovary. In summary, expression was observed in the preantral follicle in bovine fetal ovary, from oogonias to primordial, primary and secondary stages, and the mRNA abundance was lower on day 150 than day 60. However, the mRNA abundance of followed the opposite pattern. Similar to LRBP, the abundance of bta-miR-222 was higher on day 150 than day 60 or 90 of gestation. The LHCGR protein was detected in oogonia, primordial, primary, and secondary follicles. Moreover, both oocytes and granulosa cells showed positive immunostaining for LHCGR. In conclusion, we suggest the involvement of LHCGR/LRBP/bta-mir222 with mechanisms related to the development of preantral follicles in cattle.
PubMed: 38628494
DOI: 10.1590/1984-3143-AR2023-0112 -
Cells & Development Sep 2021P-Element-induced wimpy testis (Piwi) subfamily proteins form complexes that bind to Piwi-interacting RNA. This interaction is crucial for stem cell regulation and...
P-Element-induced wimpy testis (Piwi) subfamily proteins form complexes that bind to Piwi-interacting RNA. This interaction is crucial for stem cell regulation and formation, maintenance of germline stem cells, and gametogenesis in several metazoans. Planarians are effective models for studying stem cells. In the planarian Dugesia ryukyuensis, DrPiwi-1 is essential for the development of germ cells, but not somatic cells and sexual organs. DrPiwi-2 is indispensable for regeneration. In this study, we aimed to investigate the effects of Piwi on the differentiation of germ cells using monoclonal antibodies against DrPiwi-1 and DrPiwi-2. DrPiwi-1 and DrPiwi-2 co-localized more in immature germ cells than in mature germ cells in the ovary. DrPiwi-1 was found in the cytoplasm of early oogonia as undifferentiated germ cells, whereas DrPiwi-2 was found to localize not only in the nuclei but also in the cytoplasm of early oogonia. In descendant germ cells (oocytes), DrPiwi-2 was not present in the cytoplasm, but was strongly detected in the nucleolus. Moreover, we found that DrPiwi-1 forms a complex with DrPiwi-2. The cause of DrPiwi-1 depletion may be the severe reduction in the DrPiwi-2 level in the cytoplasm of oogonia. These results suggest that the formation of the DrPiwi-1 and DrPiwi-2 complex in the cytoplasm of oogonia is essential for oocyte differentiation. Our findings support the conclusion that DrPiwi-1 forms a complex with DrPiwi-2 in the cytoplasm of undifferentiated germ cells, and it signifies the start of gametogenesis. In contrast, in the testes, Drpiwi-1 was found in undifferentiated germ cells (spermatogonia), whereas DrPiwi-2 was found in descendant germ cells (spermatocytes). The process of germ cell differentiation from adult stem cells in planarians may be regulated in different ways in female and male germ lines by the Piwi family.
Topics: Animals; Cell Differentiation; Cytoplasm; Female; Male; Oocytes; Oogonia; Ovary; Planarians; Proteins; Testis
PubMed: 34171535
DOI: 10.1016/j.cdev.2021.203710