Abstract:During a 2019–2020 survey of plant pathogenic oomycetes in Nanjing, China, a cluster of five adjacent Rhododendron pulchrum plants in Xuanwuhu Park exhibited symptoms including crown and root rot and wilting. foliage blight caused due to collar and had rotting crown and root tissues resultingrot foliage blight. Diseased roots were rinsed in water, cut into 10 mm pieces, immersed in 70% ethanol for 60 sec, and plated onto clarified V8 juice agar (cV8A) containingamended with pimaricin (20 mg/liter), ampicillin … Show more
“…(2019) developed LAMP assays for detecting P. cinnamomi in soil and targeted a new target gene ( Pcinn100006 ) that had been identified from genomic sequencing data. Over recent years, there have been many reports of pathogens causing crown and root rot in Rhododendron pulchrum , including P. pini ( Xu et al., 2021 ), Phytopythium littorale ( Li et al., 2021 ), and Phytopythium helicoides ( Chen et al., 2021 ) except P. cinnamomi . Therefore, more isolates need to be used to verify the specificity of the RPA-LFD assay, especially those causing diseases in the same host.…”
Phytophthora cinnamomi causes crown and root wilting in more than 5,000 plant species and represents a significant threat to the health of natural ecosystems and horticultural crops. The early and accurate detection of P. cinnamomi is a fundamental step in disease prevention and appropriate management. In this study, based on public genomic sequence data and bioinformatic analysis of several Phytophthora, Phytopythium, and Pythium species, we have identified a new target gene, Pcinn13739; this allowed us to establish a recombinase polymerase amplification–lateral flow dipstick (RPA-LFD) assay for the detection of P. cinnamomi. Pcinn13739-RPA-LFD assay was highly specific to P. cinnamomi. Test results for 12 isolates of P. cinnamomi were positive, but negative for 50 isolates of 25 kinds of Phytophthora species, 13 isolates of 10 kinds of Phytopythium and Pythium species, 32 isolates of 26 kinds of fungi species, and 11 isolates of two kinds of Bursaphelenchus species. By detecting as little as 10 pg.µl−1 of genomic DNA from P. cinnamomi in a 50-µl reaction, the RPA-LFD assay was 100 times more sensitive than conventional PCR assays. By using RPA-LFD assay, P. cinnamomi was also detected on artificially inoculated fruit from Malus pumila, the leaves of Rhododendron pulchrum, the roots of sterile Lupinus polyphyllus, and the artificially inoculated soil. Results in this study indicated that this sensitive, specific, and rapid RPA-LFD assay has potentially significant applications to diagnosing P. cinnamomi, especially under time- and resource-limited conditions.
“…(2019) developed LAMP assays for detecting P. cinnamomi in soil and targeted a new target gene ( Pcinn100006 ) that had been identified from genomic sequencing data. Over recent years, there have been many reports of pathogens causing crown and root rot in Rhododendron pulchrum , including P. pini ( Xu et al., 2021 ), Phytopythium littorale ( Li et al., 2021 ), and Phytopythium helicoides ( Chen et al., 2021 ) except P. cinnamomi . Therefore, more isolates need to be used to verify the specificity of the RPA-LFD assay, especially those causing diseases in the same host.…”
Phytophthora cinnamomi causes crown and root wilting in more than 5,000 plant species and represents a significant threat to the health of natural ecosystems and horticultural crops. The early and accurate detection of P. cinnamomi is a fundamental step in disease prevention and appropriate management. In this study, based on public genomic sequence data and bioinformatic analysis of several Phytophthora, Phytopythium, and Pythium species, we have identified a new target gene, Pcinn13739; this allowed us to establish a recombinase polymerase amplification–lateral flow dipstick (RPA-LFD) assay for the detection of P. cinnamomi. Pcinn13739-RPA-LFD assay was highly specific to P. cinnamomi. Test results for 12 isolates of P. cinnamomi were positive, but negative for 50 isolates of 25 kinds of Phytophthora species, 13 isolates of 10 kinds of Phytopythium and Pythium species, 32 isolates of 26 kinds of fungi species, and 11 isolates of two kinds of Bursaphelenchus species. By detecting as little as 10 pg.µl−1 of genomic DNA from P. cinnamomi in a 50-µl reaction, the RPA-LFD assay was 100 times more sensitive than conventional PCR assays. By using RPA-LFD assay, P. cinnamomi was also detected on artificially inoculated fruit from Malus pumila, the leaves of Rhododendron pulchrum, the roots of sterile Lupinus polyphyllus, and the artificially inoculated soil. Results in this study indicated that this sensitive, specific, and rapid RPA-LFD assay has potentially significant applications to diagnosing P. cinnamomi, especially under time- and resource-limited conditions.
“…To observe the morphology of various spores, several colony agar blocks taken off with a sterile puncher were placed in a liquid V8 at 26 • C for 3 days with a 12/12 h light-dark cycle, then liquid V8 was replaced with sterile water and 3-5 drops of soil extract (100 g (3~10 cm deep) of surface soil from a fertile vegetable garden was collected, and 100 mL of tap water was added, fully stirred and precipitated for several hours; the supernatant was filtered with ordinary filter paper to remove the coarse particles of the soil and was repeatedly filtered twice with a 0.22 µm microporous membrane) to stimulate sporangial production [49]. Species were identified based on morphological characteristics (colony morphology, color and texture, sporangia, chlamydospores and zoospores) of the three isolates on V8.…”
As an ornamental plant, Fatsia japonica has been widely used in gardens. From April 2021 to 2022, a disease that caused the wilting and root rot of F. japonica in a large area was observed, which eventually led to the plants wilting and dying, while the leaves did not fall off. This disease greatly reduced the landscape effect of plants. An oomycete species was isolated from the roots of the infected plants. This colony morphology was slightly radial to stellate, and the aerial mycelium was flocculent. Oval sporangia with papillae, apical chlamydospores and zoospores formed in sporangia were observed. The morphological characteristics were consistent with Phytophthora. For accurate identification, the internal transcribed spacer (ITS), cytochrome oxidase subunit II (COXII) and large ribosomal subunit (LSU) genes were amplified and sequenced. The species was identified as Phytophthora nicotianae using phylogenetic analysis. Finally, the disease was reproduced by inoculating healthy F. japonica with a zoospore suspension; the symptoms were consistent with those of natural infections, and the isolate obtained from artificially infected plants had the same morphological characteristics as the inoculated isolate. The results demonstrated that P. nicotianae is the pathogenic factor of root rot. of F. japonica. This is the first report of root rot on F. japonica caused by P. nicotianae in China.
“…As with food crops, ornamental cultivars with improved heat resistance are needed to resist the consequences of global climate change (Wang et al, 2022). To date, little work has been conducted on the evolution of heat resistance in R. × pulchrum, with most research focused on topics such as plastomics (Shen et al, 2019(Shen et al, , 2020, propagation (Si et al, 2012), pathology, and photosynthetic metabolism (Chen et al, 2021;Xu et al, 2021). Under heat stress, heat shock factors (HSFs) bind to heat shock elements (HSEs) in a heat shock response (HSE) gene promoter to initiate the transcription of the HSE genes (Lohmann et al, 2004).…”
Rhododendron is the largest genus in Ericaceae and is well known for its diversity and beauty of flowers present in different species, making it a much‐revered lineage of ornamental plants. Many species of Rhododendron are intolerant of high temperatures, which are becoming more common and intense in urban areas under global climate change. Therefore, the discovery and description of genes from heat‐tolerant Rhododendron lineages are essential in the development of new climate‐resilient cultivars. One such species known to be heat tolerant is Rhododendron × pulchrum Sweet. To better understand the genomics of heat tolerance in this species, we assembled a haplotype‐resolved and chromosome‐scale genome for R. × pulchrum, which had a genome size of 509 Mb; a scaffold N50 of 37 251 370 bp; and contained 35 610 genes. In addition, based on the same reannotation pipeline, we conducted pan‐genomic analyses for all seven available chromosome‐scale Rhododendron genomes and found 14 415 gene groups shared across all species and 18 018 gene groups distributed in the other species, including 1879 gene groups found in only a single species. Finally, we analyzed the transcriptomic data from heat‐treated and non‐heat‐treated R. × pulchrum plants to quantify the genes that are most important during heat stress in an effort to inform the development of climate‐resilient cultivars. This study provides insight into the genome diversity in Rhododendron and targets several genes related to agronomic traits that may help in further analysis.
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