scholarly journals A Virus Infecting Hibiscus rosa-sinensis Represents an Evolutionary Link Between Cileviruses and Higreviruses

2021 ◽  
Vol 12 ◽  
Author(s):  
Alejandro Olmedo-Velarde ◽  
John Hu ◽  
Michael J. Melzer

Hibiscus (Hibiscus spp.) are popular ornamental and landscape plants in Hawaii which are susceptible to foliar diseases caused by viruses belonging to the genera Cilevirus and Higrevirus (family Kitaviridae). In this study, a virus infecting H. rosa-sinensis plants displaying foliar symptoms consistent with infection by a kitavirus, including yellow chlorotic blotches with a green perimeter, was characterized. The genome consisted of two RNAs 8.4 and 4.4 kb in length, and was organized most similarly to cileviruses, but with important distinctions. These included the location of the p29 homolog as the 3′-terminal open reading frame (ORF) of RNA2 instead of its typical locus at the 3′-end of RNA1; the absence of a p15 homolog on RNA2 and the adjacent intergenic region which also harbors small putative ORFs of unknown function; and the presence of an ORF encoding a 10 kDa protein at the 3′-terminal end of RNA1 that was also found to be present in the hibiscus green spot virus 2 genome. Spherical particles approximately 55–65 nm in diameter were observed in infected leaf tissue, and viral RNA was detected by reverse-transcription PCR in individual mites collected from symptomatic plants tentatively identified as Brevipalpus yothersi. Although phylogenetic analyses placed this virus between the higrevirus and cilevirus clades, we propose the tentative taxonomic placement of this virus, designated hibiscus yellow blotch virus (HYBV), within the genus Cilevirus.

Forests ◽  
2021 ◽  
Vol 12 (11) ◽  
pp. 1574
Author(s):  
Thomas R. Gaskin ◽  
Max Tischendorf ◽  
Ines Günther ◽  
Marius Rehanek ◽  
Carmen Büttner ◽  
...  

We identified a novel virus in diseased European ash (Fraxinus excelsior) and manna ash (F. ornus) trees exhibiting chlorotic ringspots, mottle and leaf deformation such as curling and shoestring symptoms. High-throughput sequencing (HTS, Illumina RNASeq) of total RNA isolated from diseased leaf material in combination with RT-PCR-based amplification techniques and Sanger sequencing determined five complete genome segments, each encoding a single open reading frame. Sequence analyses of RNA1–RNA5 revealed a genome organization typical for emaraviruses, i.e., (i) conserved and complementary terminal 5′and 3′termini of each genome segment (ii) proteins showing significant homologies to the RNA-dependent RNA polymerase (RdRP) encoded by RNA1, the glycoprotein precursor (GPP) encoded by RNA2, the viral nucleocapsid protein (N, RNA3), the movement protein (MP, RNA4), and a protein of 26 kDA (P26, RNA5) highly similar to proteins of unknown function encoded by other emaraviruses. Furthermore, we identified spherical particles (double-membrane bodies, DMB) of different sizes (70–80 nm in diameter) which are typical for emaraviruses exclusively in virus-infected leaf tissue exhibiting mottle and leaf deformation. Sequence comparison and phylogenetic analyses confirmed the identified novel virus as a new member of the genus Emaravirus. We established a species-specific RT-PCR detection protocol and could associate the observed disease symptoms with the infection of the novel emaravirus in F. excelsior and F. ornus. Therefore, we propose the name ash shoestring-associated emaravirus (ASaV). Investigation of ASaV-infected sample trees originating from different locations in Switzerland, Germany, Italy and Sweden provided a wide geographical distribution of the virus in affected ash species. To our knowledge, this is the first confirmation of an emaravirus affecting ash tree species with shoestring symptoms of leaves in Europe.


2014 ◽  
Vol 15 (1) ◽  
pp. 36-40 ◽  
Author(s):  
Nina Shishkoff

Growth-inhibiting fungicides are used routinely to control common and regulated Oomycete pathogens. This study investigated whether such fungicides could affect detection of Phytophthora ramorum from plant tissue, both foliage and roots. Whole plants of Rhododendron × ‘Cunningham's White’ were inoculated with P. ramorum and treated 3 days later with fosetyl-Al, mefenoxam, or propamocarb. The foliage was sampled over time to see if fungicides prevented successful culturing of the pathogen from infected leaf tissue or interfered with detection using real-time PCR or ELISA. Mefenoxam significantly reduced the ability to culture the pathogen from leaves for the first 6 weeks while recovery from leaves treated with other fungicides did not differ from water-treated controls; detection using PCR or ELISA was not affected by fungicide application. The roots of Viburnum cuttings were inoculated with P. ramorum and then treated 4 days later with fosetyl-Al, mefenoxam, or propamocarb. The amount of inoculum in flow through water samples taken weekly for 5 weeks was quantified and percent root colonization determined at the end of the experiment. Propamocarb had no effect on inoculum production or root infection, while viable inoculum production was significantly decreased in fosetyl-Al- or mefenoxam-treated plants over 5 weeks, and root colonization was significantly decreased. Accepted for publication 23 January 2014. Published 18 March 2014.


Plant Disease ◽  
2010 ◽  
Vol 94 (12) ◽  
pp. 1508-1508 ◽  
Author(s):  
D. M. Sether ◽  
W. B. Borth ◽  
R. S. Shimabuku ◽  
H. R. Pappu ◽  
M. J. Melzer ◽  
...  

Onion (Allium spp.) production in Hawaii is mostly comprised of green onion and the locally prized sweet bulb onions (Allium cepa L.) that include short- and medium-day cultivars. Iris yellow spot virus (IYSV; family Bunyaviridae, genus Tospovirus) is an important constraint to bulb and seed onion production in many onion-growing regions of the continental United States and the world (3). In June 2010, straw-colored, diamond-shaped lesions with occasional green islands were observed on leaves of sweet onion ‘Linda Vista’ in an insecticide trial on Maui for onion thrips (Thrips tabaci) control. Collapse and lodging occurred when lesions on leaves were severe. Seven bulbs with green leaves exhibiting lesions were collected from this onion field in the Pulehu Region of the lower Kula District on Maui. Leaf samples that included a lesion or were within 1 cm of a lesion were found to be positive in indirect ELISA with IYSV-specific polyclonal antisera (2). A405nm readings after 1 h ranged from 0.263 to 2.067 for positive samples and 0.055 to 0.073 for healthy onion controls. Four samples that were prepared from leaf tissue several centimeters away from a lesion tested negative in ELISA. Such uneven virus distribution in the plants has been previously reported (4). In July 2010, symptomatic sweet onion from a commercial farm in upper Kula, Maui at the 1,060 to 1,220 m (3,500 to 4,000 foot) elevation tested positive for IYSV by ELISA. Green onion samples collected from a commercial farm in Omaopio, Maui, located approximately 0.8 km (0.5 mile) north of Pulehu, have tested negative, suggesting distribution may be limited at this time. RNA was isolated from leaf tissue from the seven ‘Linda Vista’ sweet onions collected from the Maui insecticide trial. Reverse transcription (RT)-PCR with forward and complementary primers 5′-CTCTTAAACACATTTAACAAGCAC-3′ and 5′-TAAAACAAACATTCAAACAA-3′ flanking the nucleocapsid (N) gene encoded by the small RNA of IYSV was conducted as previously described (1). Amplicons approximately 1.1 kb long were obtained from all seven symptomatic onion samples but not from healthy samples or water controls. Sequencing of selected amplicons confirmed IYSV infection. Three sequence variants (GenBank Accession Nos. HM776014–HM776016) were identified from two RT-PCR reactions. Phylogenetic analyses of the three sequence variants with the neighbor-joining procedure available through NCBI-BLASTn Tree View showed that the highest nucleotide identities of 97 to 98% were shared with IYSV isolates from New Zealand (EU477515), Nevada (FJ713699), and northern California (FJ713700). Phylogenetic analyses with the N-gene showed the sequences from Hawaii are most closely related to isolates from the western United States, Texas, and New Zealand. To date, to our knowledge, IYSV has not been detected on the islands of Kauai, Oahu, Molokai, or Hawaii. The distribution and economic consequences of this disease to Hawaii's onion production are under investigation. References: (1) H. R. Pappu et al. Arch Virol. 151:1015, 2006. (2) H. R. Pappu et al. Plant Dis. 92:588, 2008. (3) H. R. Pappu et al. Virus Res. 141:219, 2009. (4) T. N. Smith et al. Plant Dis. 90:729, 2006.


1977 ◽  
Vol 25 (6) ◽  
pp. 585 ◽  
Author(s):  
RD Pares ◽  
AD Greenwood

Infected leaf tissue was examined at 3, 4, 5 and 6 days after inoculation, after different fixing and staining techniques. One example of stomata1 penetration was seen. Examples of cell penetration and haustorium development were examined in detail. Haustoria penetrate host cells by altering host cell wall structure, and lomasomes are frequently present in the haustorium neck. Haustoria do not have nuclei and in early stages have abundant mitochondria that gradually decrease in number as infection advances.


Plant Disease ◽  
2002 ◽  
Vol 86 (10) ◽  
pp. 1115-1122 ◽  
Author(s):  
B. M. Pryor ◽  
J. O. Strandberg ◽  
R. M. Davis ◽  
J. J. Nunez ◽  
R. L. Gilbertson

Alternaria dauci was recovered in California from carrot crop residue and from volunteer carrot plants in fallow carrot fields. The fungus was not recovered from common weeds surrounding fallow fields. To evaluate further the survival of A. dauci on carrot crop residue, infected carrot leaf tissue was placed in fields or in soil in greenhouse pots, and recovered over time. In California, A. dauci was recovered from infected leaf tissue in both fallow and irrigated fields for as long as 1 year. In Florida, A. dauci was recovered from infected leaf tissue in fallow fields for up to 30 weeks. In greenhouse experiments, A. dauci was recovered from infected leaf tissue for as long as 1 year in dry soil, but only up to 30 weeks in soil that was watered weekly. To determine the infectivity of A. dauci borne on carrot crop residue, infected carrot crops were incorporated into organic and mineral field soils, and soil samples were collected over time. Carrot seed were planted in collected soil, and seedling infection by A. dauci was recorded. Seedling infection was detected up to 13 and 14 weeks after crop incorporation in organic and mineral soil, respectively. Seedling infection was detected for up to 5 weeks in soil that remained dry compared with 3 weeks in flooded soil.


1984 ◽  
Vol 62 (11) ◽  
pp. 2283-2289 ◽  
Author(s):  
M. E. C. Rey ◽  
H. M. Garnett

Detailed morphological studies of the leaf pathogen Phyllachora paspalicola (tarspot) on its host Panicum maximum Jacq. have been previously described. This research reports physiological changes in leaf tissue infected by P. paspalicola. Aspects of photosynthetic and nitrogen metabolism were studied in healthy and tarspot-infected leaf tissue. Concentrations of glucose, fructose, and sucrose in infected and noninfected leaves were also measured. Gross and net photosynthetic rates declined significantly in tarspot-infected leaves at high disease intensities (>25%). Chlorophyll content declined in leaves exhibiting >25% infection. Transmission electron microscopy revealed disruption of lamellae and formation of large lipid bodies in infected mesophyll and bundle sheath cells. Significant changes in some C4 enzymes, such as NADP malate dehydrogenase, aspartate aminotransferase, ribulose-1,5-bisphosphate carboxylase, phosphoenolpyruvate carboxylase, were noted in tarspot-infected leaves. Small differences in the activities of NADP–malate enzyme and alanine aminotransferase were also observed. Activity of the respiratory enzyme succinate dehydrogenase was also stimulated in intensely infected leaves. An increase in nitrate levels was measured as ascopore sporogenesis occurred. At sporulation and during postsporulation, total nitrogen declined in leaves exhibiting intense infection (>25%). A small increase in glucose and fructose content occurred in leaves exhibiting mature fruiting bodies, whereas a significant (p = 0.05) increase in sucrose content was observed in infected leaves.


2003 ◽  
Vol 30 (7) ◽  
pp. 821 ◽  
Author(s):  
J. Doug Dean ◽  
Paul H. Goodwin ◽  
Tom Hsiang

Among a collection of 840 expressed sequence tags of Malva pusilla leaves infected with Colletotrichum gloeosporioides f. sp. malvae (Cgm), a total of four different glutathione S-transferase (GST) (EC 2.5.1.18) genes were identified, each showing a different pattern of expression following infection. MpGSTU1 and MpGSTU2 were members of the class tau GSTs, MpGSTF1 was a member of the class phi GSTs, and MpGSTZ1 belonged to the class zeta GSTs. Infection by Cgm occurs by a hemibiotrophic process with an initial biotrophic phase preceding the necrotrophic phase and the appearance of symptoms. Expression of MpGSTZ1 progressively increased during infection, corresponding directly with the growth of the pathogen. Expression of MpGSTU2 was similar to that of MpGSTZ1, except for a greater increase during the late necrotrophic phase. MpGSTU1 expression remained relatively constant throughout the infection, whereas MpGSTF1 expression was induced primarily during the conversion from the biotrophic to necrotrophic phases of infection. Incubation of healthy mallow leaves in the dark resulted in decreased expression of MpGSTF1 and MpGSTU1, but not MpGSTZ1 and MpGSTU2. The differential expression patterns indicate that these mallow GST genes play a variety of roles in healthy and fungal-infected leaf tissue. The nucleotide sequences reported in this paper have been submitted to GenBank under the accession numbers AY206003, AY206001, AY206002, AY206000 and AY205999.


1989 ◽  
Vol 23 (2) ◽  
pp. 111-126 ◽  
Author(s):  
Martin Tabler ◽  
Isolde Günther ◽  
Rainer Kern ◽  
Heinz L. Sänger

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