Volatile compounds in pine stands suffering from pine wilt disease: qualitative and quantitative evaluation

Nematology ◽  
2006 ◽  
Vol 8 (6) ◽  
pp. 869-879 ◽  
Author(s):  
Kazuyoshi Futai ◽  
Natsumi Kanzaki ◽  
Yuko Takeuchi
Keyword(s):  
Volatile Compounds ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Economic Damage ◽  
Qualitative And Quantitative ◽  
Pine Wilt ◽  
Pine Trees ◽  
Quantitative Properties ◽  
Pine Stands

AbstractPine wilt disease causes ecological and economic damage in Japanese pine forests in spite of intensive effort to protect them from the pine wood nematode, Bursaphelenchus xylophilus. Pine trees infected with B. xylophilus emit a characteristic bouquet of volatile compounds bioactive to the vector beetle of the nematode, Monochamus alternatus, and potentially affecting symptom development inside the trees. To investigate the qualitative and quantitative properties of volatile compounds in the field, we profiled the volatile emissions in two Japanese black pine stands, one naturally suffering from pine wilt disease and the other artificially inoculated with B. xylophilus. In both pine stands, the emission of some terpenoids from the infected trees such as (−)-α-pinene, began to increase in summer, overlapping the oviposition season of the vector beetle, but peaked in the summer and autumn. These data suggest that the beetles may not necessarily depend on the tremendous quantity of volatiles alone when they search for suitable trees on which to oviposit.

scholarly journals The specialization of Esteya vermicola hyphae in infection to Bursaphelenchus xylophilus and its colonization of pine tree

2020 ◽  
Author(s):  
Hai-Hua Wang ◽  
Can Yin ◽  
Jie Gao ◽  
Ran Tao ◽  
Piao-Piao Dai ◽  
...  
Keyword(s):  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Fungal Colonization ◽  
Pinewood Nematode ◽  
Pine Tree ◽  
Pine Wilt ◽  
Pine Trees ◽  
Colonization Patterns ◽  
Esteya Vermicola

AbstractPine wilt disease (PWD) caused by the nematode Bursaphelenchus xylophilus is a serious problem on pines, and there is currently no effective control strategy for this disease. Although the endoparasitic fungus Esteya vermicola showed great effectiveness in controlling pine wilt disease, the colonization patterns of the host pine tree xylem by this fungus are unknown. To investigate the colonization patterns of pine xylem by this fungus, the species Pinus koraiensis grown in a greenhouse was used as an experimental host tree. The fungal colonization of healthy and wilting pine trees by E. vermicola was quantified using PCR with a TaqMan probe, and a green fluorescence protein (GFP) transformant was used for visualization. The results reported a specific infection approach used by E. vermicola to infect B. xylophilus and specialized fungal parasitic cells in PWN infection. In addition, the inoculated blastospores of E. vermicola germinated and grew inside of healthy pine xylem, although the growth rate was slow. Moreover, E. vermicola extended into the pine xylem following spray inoculation of wounded pine seedling stems, and a significant increase in fungal quantity was observed in response to B. xylophilus invasion. An accelerated extension of E. vermicola colonization was shown in PWN-infected wilting pine trees, due to the immigration of fungal-infected PWNs. Our results provide helpful knowledge about the extension rate of this fungus in healthy and wilting PWN-susceptible pine trees in the biological control of PWD and will contribute to the development of a management method for PWD control in the field.Author summaryPine wilt disease, caused by Bursaphelenchus xylophilus, has infected most pine forests in Asian and European forests and led to enormous losses of forest ecosystem and economy. Esteya vermicola is a bio-control fungus against pinewood nematode, showed excellent control efficient to pine wilt disease in both of greenhouse experiments and field tests. Although this bio-control agent was well known for the management of pine wilt disease, the infection mechanism of fungal infection and colonization of host pine tree are less understand. Here, we use GFP-tagged mutant to investigate the fungal infection to pinewood nematode; additionally, the temporal and spatial dynamics of E. vermicola colonize to pine tree were determined by the TaqMan real-time PCR quantification, as well as the response to pinewood nematode invasion. We found a specific infection approach used by E. vermicola to infect B. xylophilus and specialized fungal parasitic cells in PWN infection. In addition, the fungal germination and extension inside of pine tree xylem after inoculation were revealed. In addition, the quantity of E. vermicola increased as response to pinewood nematode invasion was reported. Our study provides two novel technologies for the visualization and detection of E. vermicola for the future investigations of fungal colonization and its parasitism against pinewood nematode, and the mechanisms of the bio-control process.

scholarly journals Assessment of the PCR technique for identification of Bursaphelenchus mucronatus Mamiya & Enda 1979, B. xylophilus Steiner & Buhrer 1934 (Nickle 1970) and B. fraudulentus Rühm 1956 (Nematoda, Aphelenchoididae) in crude extract of nematodes isolated from wood

2021 ◽  
Vol 61 (4) ◽  
pp. 346-351
Keyword(s):  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Coniferous Forests ◽  
Pine Wood Nematode ◽  
Direct Examination ◽  
Pine Wood ◽  
Pine Wilt ◽  
Detection And Identification ◽  
Pine Trees

The pine wood nematode, Bursaphelenchus xylophilus Steiner & Buhrer 1934 (Nickle 1970) is the major causative agent of the pine wilt disease which has become devastating to Asian and European coniferous forests. These regions are also naturally occupied by two other native but nonpathogenic species, i.e. B. mucronatus Mamiya & Enda 1979 and B. fraudulentus Rühm 1956 which are closely related to the invasive B. xylophilus. Moreover, all these three species can colonize pine trees, and potentially be extracted from the same wood samples. Due to the cosmopolitan character and wide genetic variation within- and between existing populations the taxonomic distinction of these species based exclusively on their morphology is difficult or, almost impossible. The present quarantine regulations related to B. xylophilus require the most credible and simple methods which could allow for a possibly earliest detection and precise identification of this species in wood shipments and conifer forests stands. The main objectives of the presently reported research were to simplify the presently available procedures for possibly fast and precise detection and identification of B. xylophilus examined in the background of the remaining Bursaphelenchus species of the xylophilus group and other bacterio- and mycetophagous nematodes naturally present in the pine wood samples. The developed method is based on a direct examination of the crude nematode extract from wood samples and subsequent use of PCR technique with earlier designed specific reaction starters amplifying ITS1–28S rDNA regions.

scholarly journals Comparative Secretome and Functional Analyses Reveal Glycoside Hydrolase Family 30 and Cysteine Peptidase as Virulence Determinants in the Pinewood Nematode Bursaphelenchus xylophilus

2021 ◽  
Vol 12 ◽  
Author(s):  
Ryoji Shinya ◽  
Haru Kirino ◽  
Hironobu Morisaka ◽  
Yuko Takeuchi-Kaneko ◽  
Kazuyoshi Futai ◽  
...  
Keyword(s):  
Glycoside Hydrolase ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Glycoside Hydrolase Family ◽  
Pinewood Nematode ◽  
Virulence Determinants ◽  
Pine Wilt ◽  
Pine Trees ◽  
Hydrolase Family

Pine wilt disease, caused by the pinewood nematode, Bursaphelenchus xylophilus, is one of the world’s most serious tree diseases. Although the B. xylophilus whole-genome sequence and comprehensive secretome profile have been determined over the past decade, it remains unclear what molecules are critical in pine wilt disease and govern B. xylophilus virulence in host pine trees. Here, a comparative secretome analysis among four isolates of B. xylophilus with distinct virulence levels was performed to identify virulence determinants. The four candidate virulence determinants of B. xylophilus highly secreted in virulent isolates included lipase (Bx-lip1), glycoside hydrolase family 30 (Bx-GH30), and two C1A family cysteine peptidases (Bx-CAT1 and Bx-CAT2). To validate the quantitative differences in the four potential virulence determinants among virulence groups at the protein level, we used real-time reverse-transcription polymerase chain reaction analysis to investigate these determinants at the transcript level at three time points: pre-inoculation, 3 days after inoculation (dai), and 7 dai into pine seedlings. The transcript levels of Bx-CAT1, Bx-CAT2, and Bx-GH30 were significantly higher in virulent isolates than in avirulent isolates at pre-inoculation and 3 dai. A subsequent leaf-disk assay based on transient overexpression in Nicotiana benthamiana revealed that the GH30 candidate virulent factor caused cell death in the plant. Furthermore, we demonstrated that Bx-CAT2 was involved in nutrient uptake for fungal feeding via soaking-mediated RNA interference. These findings indicate that the secreted proteins Bx-GH30 and Bx-CAT2 contribute to B. xylophilus virulence in host pine trees and may be involved in pine wilt disease.

How different is induced host resistance against the pine wood nematode, Bursaphelenchus xylophilus, by two avirulent microbes?

Nematology ◽  
2006 ◽  
Vol 8 (3) ◽  
pp. 435-442 ◽  
Author(s):  
Yuko Takeuchi ◽  
Natsumi Kanzaki ◽  
Kazuyoshi Futai
Keyword(s):  
Induced Resistance ◽  
Host Resistance ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Pine Wilt ◽  
Pine Trees ◽  
Induce Resistance

AbstractPine wilt disease, caused by the pine wood nematode (PWN), Bursaphelenchus xylophilus, has been devastating pine forests in Japan for a century but an effective method for controlling the disease is still needed. Pre-inoculation of the host plant with an avirulent pathogen can induce resistance against subsequent infection with virulent pathogens; this is called 'induced resistance'. Host pine trees have this type of resistance against pine wilt disease but the detailed mechanism is unknown. In this study, 1-year-old potted seedlings of the susceptible Japanese black pine, Pinus thunbergii, were pre-inoculated with an avirulent isolate of PWN (C14-5), as well as an avirulent fungus, Botrytis cinerea, to induce host resistance against PWN. One, 2, 3, 4 and 5 weeks after inoculation, the seedlings were challenged with a virulent isolate of PWN (S10), and the survival of these seedlings was estimated by their external symptoms. Pre-inoculation with either C14-5 or Botrytis delayed the symptom development caused by subsequent inoculation with S10, suggesting that pre-inoculation induced host resistance against PWN. The resistance induced by pre-inoculation with C14-5 lasted for at least 5 weeks, while that induced by Botrytis weakened with time. It has been reported that the PWN survives inside the host seedlings for a long period without causing any symptoms under certain conditions, but Botrytis survives only for a while in the seedlings. Thus, the duration of host resistance induced by pre-inoculation may vary with the viability of the inoculum used for pre-inoculation. In order to obtain durable induced resistance, it may be necessary to repeat inoculations of avirulent microbes to be able to control pine wilt disease.

Suppressive effects of Bursaphelenchus mucronatus on pine wilt disease development and mortality of B. xylophilus-inoculated pine seedlings

Nematology ◽  
2014 ◽  
Vol 16 (2) ◽  
pp. 219-227 ◽  
Author(s):  
Si-Mi Liao ◽  
Satomi Kasuga ◽  
Katsumi Togashi
Keyword(s):  
Pine Wilt Disease ◽  
Nematode Species ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Reproductive Interference ◽  
Dependent Manner ◽  
Nematode Density ◽  
Pine Wilt ◽  
Bursaphelenchus Mucronatus ◽  
Pine Trees

Bursaphelenchus xylophilus causes pine wilt disease in Pinus trees whereas B. mucronatus has no or little virulence to the trees. Interspecific crossing experiments conducted so far suggest reproductive interference between the two nematode species. Theory predicts that one of the two competing species populations quickly displaces the other through reproductive interference in a frequency-dependent manner. Thus, it is anticipated that B. mucronatus suppresses the virulence of B. xylophilus against pine trees when B. mucronatus heavily outnumber B. xylophilus. To determine the suppressive effects of B. mucronatus, the two nematode species were inoculated simultaneously on 30 3-year-old Pinus thunbergii seedlings at three combinations of different numbers, and B. xylophilus alone was inoculated on 30 other seedlings at the corresponding numbers in early August. Seedlings were observed at intervals of 4 or 6 weeks and two stem sections were sampled from each seedling to determine the density and species composition of nematode populations after death or in December. Inoculation of B. mucronatus significantly retarded the speed of foliage discolouration from 0.170 ± 0.024 week−1 to 0.061 ± 0.017 week−1 and significantly prolonged the survival time of seedlings. Bursaphelenchus mucronatus significantly reduced the nematode density from 1799.7 ± 305.0 to 521.0 ± 148.4 (g dried seedling stem)−1. Analysis of rDNA genotype showed 1846 B xylophilus, no B. mucronatus and one hybrid.

scholarly journals Effects of cavitation on the development of pine wilt disease caused by Bursaphelenchus xylophilus.

1988 ◽  
Vol 54 (5) ◽  
pp. 606-615 ◽  
Author(s):  
Keiko KURODA ◽  
Toshihiro YAMADA ◽  
Kazuhiko MINEO ◽  
Hirotada TAMURA
Keyword(s):  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Pine Wilt

scholarly journals Early detection of pine wilt disease in Pinus tabuliformis in North China using a field portable spectrometer and UAV-based hyperspectral imagery

2021 ◽  
Vol 8 (1) ◽  
Author(s):  
Run Yu ◽  
Lili Ren ◽  
Youqing Luo
Keyword(s):  
Early Stage ◽  
Vegetation Indices ◽  
Pine Wilt Disease ◽  
Wilt Disease ◽  
Pinus Tabulaeformis ◽  
Pine Tree ◽  
Preventative Measures ◽  
Pine Wilt ◽  
Pine Trees ◽  
Airborne Vehicle

Abstract Background Pine wilt disease (PWD) is a major ecological concern in China that has caused severe damage to millions of Chinese pines (Pinus tabulaeformis). To control the spread of PWD, it is necessary to develop an effective approach to detect its presence in the early stage of infection. One potential solution is the use of Unmanned Airborne Vehicle (UAV) based hyperspectral images (HIs). UAV-based HIs have high spatial and spectral resolution and can gather data rapidly, potentially enabling the effective monitoring of large forests. Despite this, few studies examine the feasibility of HI data use in assessing the stage and severity of PWD infection in Chinese pine. Method To fill this gap, we used a Random Forest (RF) algorithm to estimate the stage of PWD infection of trees sampled using UAV-based HI data and ground-based data (data directly collected from trees in the field). We compared relative accuracy of each of these data collection methods. We built our RF model using vegetation indices (VIs), red edge parameters (REPs), moisture indices (MIs), and their combination. Results We report several key results. For ground data, the model that combined all parameters (OA: 80.17%, Kappa: 0.73) performed better than VIs (OA: 75.21%, Kappa: 0.66), REPs (OA: 79.34%, Kappa: 0.67), and MIs (OA: 74.38%, Kappa: 0.65) in predicting the PWD stage of individual pine tree infection. REPs had the highest accuracy (OA: 80.33%, Kappa: 0.58) in distinguishing trees at the early stage of PWD from healthy trees. UAV-based HI data yielded similar results: the model combined VIs, REPs and MIs (OA: 74.38%, Kappa: 0.66) exhibited the highest accuracy in estimating the PWD stage of sampled trees, and REPs performed best in distinguishing healthy trees from trees at early stage of PWD (OA: 71.67%, Kappa: 0.40). Conclusion Overall, our results confirm the validity of using HI data to identify pine trees infected with PWD in its early stage, although its accuracy must be improved before widespread use is practical. We also show UAV-based data PWD classifications are less accurate but comparable to those of ground-based data. We believe that these results can be used to improve preventative measures in the control of PWD.

scholarly journals The Microbiome of Endophytic, Wood Colonizing Bacteria from Pine Trees as Affected by Pine Wilt Disease

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Diogo Neves Proença ◽  
Romeu Francisco ◽  
Susanne Kublik ◽  
Anne Schöler ◽  
Gisle Vestergaard ◽  
...  
Keyword(s):  
Pine Wilt Disease ◽  
Wilt Disease ◽  
Pine Wilt ◽  
Pine Trees

scholarly journals Study of symptoms and gene expression in four Pinus species after pinewood nematode infection

2011 ◽  
Vol 9 (2) ◽  
pp. 272-275 ◽  
Author(s):  
Albina R. Franco ◽  
Carla Santos ◽  
Mariana Roriz ◽  
Rui Rodrigues ◽  
Marta R. M. Lima ◽  
...  
Keyword(s):  
Virulent Strain ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Pinewood Nematode ◽  
Pine Trees ◽  
Severe Infections ◽  
Genomic Results ◽  
Avirulent Isolate

Pine wilt disease, caused by the pinewood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle, is originating severe infections in pine trees. The disease is detected when external symptoms appear (e.g. needle chlorosis), but trees could remain asymptomatic for long periods and serve as a long-term host. The primary goal of this study was to assess the effect of inoculation with an avirulent isolate of B. xylophilus (C14-5) on different Pinus spp. seedlings (P. sylvestris, P. nigra, P. pinea and P. pinaster). At the same time, seedlings were also inoculated with a virulent strain, HF, in order to compare the phenotypic and genomic results of the two types of inoculations. The effect of inoculation was determined in terms of expression of various Pinus genes potentially involved in the response to the disease.The results suggest that P. pinea and P. nigra are more resistant to infection by the nematode than P. sylvestris and P. pinaster. The phenotypic and genetic differences were more marked among P. pinea and P. pinaster.

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