Enhanced accumulation of pinosylvin stilbenes and related gene expression in Pinus strobus after infection of pine wood nematode

2021 ◽  
Author(s):  
Hwan-Su Hwang ◽  
Jung Yeon Han ◽  
Yong Eui Choi
Keyword(s):  
Transcription Factors ◽  
Defense Mechanisms ◽  
De Novo ◽  
Pinus Strobus ◽  
Pine Wilt Disease ◽  
Monomethyl Ether ◽  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Dependent Manner ◽  
Pine Wood

Abstract Pine wood nematodes (PWNs: Bursaphelenchus xylophilus) infect pine trees and cause serious pine wilt disease. Eastern white pine (Pinus strobus) has resistance to PWN. However, the detailed defense mechanisms of P. strobus against PWN are not well known. When P. strobus plants were infected with PWNs, the accumulation of stilbenoids, dihydropinosylvin monomethyl ether (DPME) and pinosylvin monomethyl ether (PME), were increased remarkably. DPME and PME had the high nematicidal activity. Interestingly, the nematicidal activity of the two compounds was resulted in a developmental stage-dependent manner. PME was more toxic to adult PWNs than juveniles, whereas DPME was found more toxic to juvenile PWNs than the adults. The genes involved in PME and DPME biosynthesis such as phenylalanine ammonia-lyase (PAL), 4-coumarate-CoA ligase (4CL), pinosylvin synthase (STS), and pinosylvin O-methyltransferase (PMT) were isolated using de novo sequencing of the transcriptome in P. strobus. In addition, transcription factors (bHLH, MYB and WRKY) related to stilbene biosynthesis were isolated. qPCR analyses of the selected genes (PAL, 4CL, STS, and PMT) including transcription factors (bHLH, MYB and WRKY) revealed that the expression level of the selected genes highly enhanced after PWN infection. Our results suggest that pinosylvin-type stilbenoid biosynthesis is highly responsive to PWN infection and plays an important role in PWN resistance of P. strobus trees.

scholarly journals Nematicidal Activity of Benzyloxyalkanols against Pine Wood Nematode

Biomolecules ◽  
2021 ◽  
Vol 11 (3) ◽  
pp. 384
Author(s):  
Junheon Kim ◽  
Su Jin Lee ◽  
Joon Oh Park ◽  
Kyungjae Andrew Yoon
Keyword(s):  
Forest Ecosystems ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Environmental Damage ◽  
Pine Wood Nematode ◽  
Glutathione S Transferase ◽  
Pine Wood ◽  
Chemical Treatments ◽  
Continuous Use

Pine wilt disease (PWD) is caused by the pine wood nematode (PWN; Bursaphelenchus xylophilus) and causes severe environmental damage to global pine forest ecosystems. The current strategies used to control PWN are mainly chemical treatments. However, the continuous use of these reagents could result in the development of pesticide-resistant nematodes. Therefore, the present study was undertaken to find potential alternatives to the currently used PWN control agents abamectin and emamectin. Benzyloxyalkanols (BzOROH; R = C2–C9) were synthesized and the nematicidal activity of the synthetic compounds was investigated. Enzymatic inhibitory assays (acetylcholinesterase (AChE) and glutathione S-transferase (GST)) were performed with BzOC8OH and BzOC9OH to understand their mode of action. The benzyloxyalkanols showed higher nematicidal activity than did benzyl alcohol. Among the tested BzOROHs, BzC8OH and BzC9OH showed the strongest nematicidal activity. The LD50 values of BzC8OH and BzC9OH were 246.1 and 158.0 ppm, respectively. No enzyme inhibitory activity was observed for BzC8OH and BzC9OH. The results suggested that benzyloxyalcohols could be an alternative nematicidal agent.

scholarly journals Enhanced production of pinosylvin stilbene with aging of Pinus strobus callus and nematicidal activity of callus extracts against pinewood nematodes

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Hyo Bin Koo ◽  
Hwan-Su Hwang ◽  
Jung Yeon Han ◽  
Eun Ju Cheong ◽  
Yong-Soo Kwon ◽  
...  
Keyword(s):  
Pinus Strobus ◽  
Monomethyl Ether ◽  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Zygotic Embryos ◽  
Enhanced Production ◽  
Pinewood Nematodes ◽  
Ethanolic Extraction ◽  
Ethanol Extracts

AbstractPinosylvin stilbenes are phenolic compounds mainly occurring in the Pinaceae family. We previously reported that the accumulation of two pinosylvin stilbene compounds, dihydropinosylvin methyl ether (DPME) and pinosylvin monomethyl ether (PME), in Pinus strobus trees was highly enhanced by infection with pine wood nematodes (PWNs: Bursaphelenchus xylophilus), and these two compounds showed strong nematicidal activity against PWNs. In this work, we established a system of pinosylvin stilbene (DPME and PME) production via the in vitro culture of P. strobus calli, and we examined the nematicidal activity of callus extracts. Calli were induced from the culture of mature zygotic embryos of P. strobus. Optimized growth of calli was obtained in 1/2 Litvay medium with 1.0 mg/L 2,4-D and 0.5 mg/L BA. DPME and PME accumulation did not occur in nonaged (one-month-old) calli but increased greatly with prolonged callus culture. The concentrations of DPME and PME in three-month-old dark-brown calli were 6.4 mg/g DW and 0.28 mg/g DW, respectively. The effect of methyl jasmonate treatment on the accumulation of DPME and PME was evaluated in cell suspension culture of P. strobus. However, the treatment appeared to show slight increase of DPME accumulation compared to callus browning. A test solution prepared from crude ethanol extracts from aged calli (three months old) containing 120 µg/ml DPME and 5.16 µg/ml PME treated with PWNs resulted in 100% immobilization of the adult PWNs and 66.7% immobilization of the juvenile PWNs within 24 h. However, nonaged callus extracts did not show any nematicidal activity against juvenile PWNs and showed less than 20% nematicidal activity against adult PWNs. These results indicate that pinosylvin stilbenes can be effectively produced by prolonged culture of P. strobus calli, can be isolated using simple ethanolic extraction, and are applicable as beneficial eco-friendly compounds with nematicidal activity against PWNs.

scholarly journals Enhanced Production of Pinosylvin Stilbene with Aging of Pinus strobus Callus and Nematicidal Activity of Callus Extracts against Pinewood Nematodes

2021 ◽  
Author(s):  
Hyo Bin Koo ◽  
Hwan-Su Hwang ◽  
Jung Yeon Han ◽  
Eun Ju Cheong ◽  
Yong-Soo Kwon ◽  
...  
Keyword(s):  
Pinus Strobus ◽  
Monomethyl Ether ◽  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Zygotic Embryos ◽  
Enhanced Production ◽  
Pinewood Nematodes ◽  
Ethanolic Extraction ◽  
Ethanol Extracts

Abstract Pinosylvin stilbenes are phenolic compounds mainly occurring in the Pinaceae family. We previously reported that the accumulation of two pinosylvin stilbene compounds, dihydropinosylvin methyl ether (DPME) and pinosylvin monomethyl ether (PME), in Pinus strobus trees was highly enhanced by infection with pine wood nematodes (PWNs: Bursaphelenchus xylophilus), and these two compounds showed strong nematicidal activity against PWNs. In this work, we established a system of pinosylvin stilbene (DPME and PME) production via the in vitro culture of P. strobus calli, and we examined the nematicidal activity of callus extracts. Calli were induced from the culture of mature zygotic embryos of P. strobus. Optimized growth of calli was obtained in 1/2 Litvay medium with 1.0 mg/L 2,4-D and 0.5 mg/L BA. DPME and PME accumulation did not occur in nonaged (one-month-old) calli but increased greatly with prolonged callus culture. The concentrations of DPME and PME in three-month-old dark-brown calli were 6.4 mg/g DW and 0.28 mg/g DW, respectively. A test solution prepared from crude ethanol extracts from aged calli (three months old) containing 120 µg/ml DPME and 5.16 µg/ml PME treated with PWNs resulted in 100% immobilization of the adult PWNs and 66.7% immobilization of the juvenile PWNs within 24 hr. However, nonaged callus extracts did not show any nematicidal activity against juvenile PWNs and showed less than 20% nematicidal activity against adult PWNs. These results indicate that pinosylvin stilbenes can be effectively produced by prolonged culture of P. strobus calli, can be isolated using simple ethanolic extraction, and are applicable as beneficial eco-friendly compounds with nematicidal activity against PWNs.

Nematicidal activity against Bursaphelenchus xylophilus of isoobtusilactone A isolated from Persea americana

Nematology ◽  
2010 ◽  
Vol 12 (2) ◽  
pp. 247-253 ◽  
Author(s):  
Quang Le Dang ◽  
Hyeok Ran Kwon ◽  
Yong Ho Choi ◽  
Gyung Ja Choi ◽  
Kyoung Soo Jang ◽  
...  
Keyword(s):  
Methanol Extract ◽  
Pine Wilt Disease ◽  
Stem Bark ◽  
Bursaphelenchus Xylophilus ◽  
Persea Americana ◽  
Nematicidal Activity ◽  
Nerium Oleander ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Thespesia Populnea

Abstract Methanol extracts of 50 plant species were assessed for nematicidal activity against the pine wood nematode (PWN), Bursaphelenchus xylophilus. Out of these, five extracts: Justicia diclipteroides subsp. usambarica; Nerium oleander; Persea americana; Scadoxus multiflorus subsp. multiflorus; and Thespesia populnea, caused 96-100% mortalities of pine wood nematode at 1000 μg ml–1 for 48 h. Methanol extract of the stem bark of P. americana (avocado) and the n-hexane layer obtained from the methanol extract by solvent partitioning caused 100% mortality of PWN at low concentrations of 125 and 63 μg ml–1, respectively. One potent nematicidal compound was isolated from the stem bark of P. americana and its chemical structure was determined to be isoobtusilactone A (1) by mass and NMR spectral analyses. The chemical showed a very strong nematicidal activity against PWN; it caused mortalities of over 97% at concentrations higher than 50 μg ml–1. Compared with a commercial synthetic nematicide, morantel tartrate, compound 1 showed a little more potent activity. The results suggested that compound 1 may have the potential to be explored as a natural nematicide or be useful as a lead molecule for development of new nematicidal agents for controlling the pine wilt disease caused by B. xylophilus.

Nematicidal Activity of Teleocidin B4 Isolated from Streptomyces sp. against Pine Wood Nematode, Bursaphelenchus xylophilus

2020 ◽  
Author(s):  
Min‐Kyoung Kang ◽  
Min‐Hee Kim ◽  
Hae‐Ryong Park ◽  
Min‐Jiao Liu ◽  
Chun Zhi Jin ◽  
...  
Keyword(s):  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Streptomyces Sp

scholarly journals Cathepsin L-like Cysteine Proteinase Genes Are Associated with the Development and Pathogenicity of Pine Wood Nematode, Bursaphelenchus xylophilus

2019 ◽  
Vol 20 (1) ◽  
pp. 215 ◽  
Author(s):  
Qi Xue ◽  
Xiao-Qin Wu ◽  
Wan-Jun Zhang ◽  
Li-Na Deng ◽  
Miao-Miao Wu
Keyword(s):  
Developmental Stages ◽  
Cysteine Proteinase ◽  
Cathepsin L ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Parasitic Nematodes ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Real Time Quantitative Pcr ◽  
Rapid Amplification

The pine wood nematode (PWN), Bursaphelenchus xylophilus, is the pathogen of pine wilt disease (PWD), resulting in huge losses in pine forests. However, its pathogenic mechanism remains unclear. The cathepsin L-like cysteine proteinase (CPL) genes are multifunctional genes related to the parasitic abilities of plant-parasitic nematodes, but their functions in PWN remain unclear. We cloned three cpl genes of PWN (Bx-cpls) by rapid amplification of cDNA ends (RACE) and analyzed their characteristics using bioinformatic methods. The tissue specificity of cpl gene of PWN (Bx-cpl) was studied using in situ mRNA hybridization (ISH). The functions of Bx-cpls in development and pathogenicity were investigated using real-time quantitative PCR (qPCR) and RNA interference (RNAi). The results showed that the full-length cDNAs of Bx-cpl-1, Bx-cpl-2, and Bx-cpl-3 were 1163 bp, 1305 bp, and 1302 bp, respectively. Bx-cpls could accumulate specifically in the egg, intestine, and genital system of PWN. During different developmental stages of PWN, the expression of Bx-cpls in the egg stage was highest. After infection, the expression levels of Bx-cpls increased and reached their highest at the initial stage of PWD, then declined gradually. The silencing of Bx-cpl could reduce the feeding, reproduction, and pathogenicity of PWN. These results revealed that Bx-cpls play multiple roles in the development and pathogenic processes of PWN.

scholarly journals Autophagy contributes to the feeding, reproduction, and mobility of Bursaphelenchus xylophilus at low temperatures

2019 ◽  
Vol 51 (8) ◽  
pp. 864-872 ◽  
Author(s):  
Hongbin Liu ◽  
Xiaoqin Wu ◽  
Yaqi Feng ◽  
Lin Rui
Keyword(s):  
Defense Mechanisms ◽  
Feeding Rate ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Reproductive Rate ◽  
Reproduction Rate ◽  
Marker Proteins ◽  
Physiological Homeostasis ◽  
Autophagic Activity ◽  
Growth And Reproduction

Abstract The pine wood nematode (PWN), Bursaphelenchus xylophilus, is the pathogen causing pine wilt disease (PWD), which is a devastating forest disease. At present, little is known about the defense mechanisms of the PWN, which limits PWD control. Although autophagy plays an important role in the physiological and pathological processes of eukaryotes, its significance in the PWN remains unknown. In this study, we prepared an anti-BxATG8 polyclonal antibody and identified two PWN autophagy marker proteins: BxATG8-I and BxATG8-II. By western blot analysis, we found that the ratio of BxATG8-II to BxATG8-I, which represents autophagic activity, was decreased significantly when samples were treated with the autophagy inhibitor 3-methyladenine. As such, we were able to successfully detect and quantify autophagic activity in the PWN. Thereafter, we investigated the effects of low and high temperatures on PWN growth and reproduction. The results revealed that feeding rate, reproduction rate, and mobility decreased at 15°C and increased at 35°C. By contrast, autophagic activity was high at 15°C and low at 35°C, suggesting that the PWN regulates autophagic activity in response to changes in temperature to maintain physiological homeostasis. When autophagy was inhibited at 15°C, feeding rate, reproductive rate, and mobility declined further, indicating that autophagy is crucial for PWN growth and reproduction at low temperature. These results indicate that autophagy in the PWN is an important response mechanism to temperature changes.

Identification of a novel effector BxSapB3 that enhances the virulence of pine wood nematode Bursaphelenchus xylophilus

2019 ◽  
Vol 51 (10) ◽  
pp. 1071-1078
Author(s):  
Xin Huang ◽  
Longjiao Hu ◽  
Xiaoqin Wu
Keyword(s):  
Virulent Strain ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Wilt Disease ◽  
Pcr Analysis ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Early Stages ◽  
Pine Wilt ◽  
Weakly Virulent

Abstract Pine wilt disease, caused by the pine wood nematode Bursaphelenchus xylophilus, leads to severe damage to pine forests in China. In our previous study, effectors secreted by this pathogen were shown to play roles in the different infection stages of pine wilt disease, and a series of candidate effectors were predicted by transcriptome sequencing. This study identified and characterized a novel effector, BxSapB3, which was among these candidate effectors. Agrobacterium-mediated transient expression was used to identify BxSapB3. BxSapB3 was secreted by B. xylophilus and found to be capable of inducing cell death in Nicotiana benthamiana. Quantitative real-time PCR (qRT-PCR) analysis revealed that BxSapB3 was upregulated in a highly virulent strain of B. xylophilus and expressed at lower levels in a weakly virulent strain at the early stages of infection. When BxSapB3 was silenced in B. xylophilus, the process of infection was delayed. These results indicate that BxSapB3 acts as an effector and contributes to virulence at the early stages of B. xylophilus infection.

Nematicidal activity of onion (Allium cepa) oil and its components against the pine wood nematode (Bursaphelenchus xylophilus)

Nematology ◽  
2007 ◽  
Vol 9 (2) ◽  
pp. 231-235 ◽  
Author(s):  
Sang-Chul Shin ◽  
Il-Kwon Park ◽  
In-Ho Choi
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Allium Cepa ◽  
Bursaphelenchus Xylophilus ◽  
Nematicidal Activity ◽  
Gas Chromatography Mass Spectrometry ◽  
Pine Wood Nematode ◽  
Sulphur Compounds ◽  
Pine Wood ◽  
Weak Activity

AbstractOnion oil and its components were tested for their nematicidal activity against the pine wood nematode, Bursaphelenchus xylophilus, with 96-well plate bioassays. Responses varied with test materials and concentration. Good nematicidal activity against male, female and juvenile nematodes of B. xylophilus was achieved with onion oil. The LC50 values of onion oil against male, female and juvenile pine wood nematodes were 17.6, 13.8 and 12.1 μg ml−1, respectively. Analysis by gas chromatography-mass spectrometry led to the identification of four major sulphur compounds: propyl disulphide, propyl trisulphide, methyl propyl disulphide and methyl propyl trisulphide. These four compounds and propyl sulphide and methyl propyl sulphide were tested individually for their nematicidal activities against pine wood nematode. LC50 values of propyl trisulphide and methyl propyl trisulphide against juvenile nematode were 5.0 and 22.9 μg ml−1, respectively. Other sulphur compounds showed weak activity.

scholarly journals Sampling design for efficient detection of pine wood nematode, Bursaphelenchus xylophilus, in diseased trees using a DNA detection kit: variation across branch, trunk and tree

Nematology ◽  
2018 ◽  
Vol 20 (7) ◽  
pp. 641-652 ◽  
Author(s):  
Yuuki Nakabayashi ◽  
Takuya Aikawa ◽  
Michinari Matsushita ◽  
Kazuhiko Hoshizaki
Keyword(s):  
Sampling Design ◽  
Dna Detection ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Molecular Techniques ◽  
Effective Control ◽  
Pine Wood Nematode ◽  
Pine Wood ◽  
Efficient Detection ◽  
Minimum Number

Summary Detection of pine wood nematode, Bursaphelenchus xylophilus, is fundamental for effective control of pine wilt disease. Recent molecular techniques, such as DNA detection, have enhanced detectability of the nematodes whereas appropriate field sampling has received less attention. In order to elucidate a sampling design that most efficiently detects B. xylophilus using a commercially-distributed DNA detection kit, we compared detection levels of B. xylophilus using wood chips taken from various positions on dead trees. Results showed that the DNA kit had a higher detection level than the conventional method, and that trunk samples had higher levels than branch samples. Statistical model revealed that among-tree variation influenced the detectability more strongly than within-tree variation. Our results suggest that, in practice, with limited resources for control, it is more efficient to take samples from many trees with a minimum number from each tree, rather than taking many samples from a small number of trees.

Sign in / Sign up

Export Citation Format

Share Document