First Report of Bursaphelenchus pinophilus on Korean pine (Pinus koraiensis)

The genus Bursaphelenchus Fuchs, 1937 contains approximately 90 species (3) that are morphologically similar. Pine wood nematode, Bursaphelenchus xylophilus (4) Nickle, 1970, is the causal organism of pine wilt disease and accurate identification is essential for diagnosis of the disease. In Korea, pine wilt disease was first reported in 1988 and devastated 6,800 ha of pine forest through 2008. For a survey of trees with pine wilt disease, wood samples were taken randomly from dead Pinus koraiensis in Namyangju, Gyeonggi Province in Korea. The extracted nematodes from dead trees were maintained in culture on Botrytis cinerea and morphological characteristics were observed with an inverted light microscope (Leica DE/DMI 3000B). Identification of Bursaphelenchus spp. based on morphological characteristics is difficult, especially for identification of juveniles that carry few morphological features for species identification. The internal transcribed spacer (ITS) region in ribosomal DNA provides useful molecular diagnostic markers for this genus (1). The nematodes were provisionally identified as Bursaphelenchus pinophilus based on the characteristic long and arcuate body shape, male spicule with distinctive rostrum and small cucullus, female vulval flap, and mucronate conical tail. Other Bursaphelenchus spp. with vulval flaps and spicules with cucullus are B. xylophius, B. mucronatus, B. abruptus, and B. pinophilus. For molecular diagnosis, DNA was extracted from more than 30 individual nematodes with a DNeasy Kit (Qiagen, Valencia, CA) and ITS regions 1, 2, and 5.8S in rDNA were amplified by PCR (US/PTC-0220; Bio Rad, Hercules, CA). The ITS-restriction fragment length polymorphism pattern was consistent with that of B. pinophilus (2). The ITS rDNA sequence of B. pinophilus from Korean pines had a 98% sequence homology to that of B. pinophilus in GenBank (Accession No. AM160664). The pathogenicity of B. pinophilus has not been determined. To our knowledge, this is the first report of B. pinophilus on P. koraiensis, but it was previously reported from Poland, Germany, and Portugal on P. sylvestris and P. pinaster (1). References: (1) H. Braasch. EPPO Bull. 31:127, 2001. (2) W. Burgermeister et al. Russ. J. Nematol. 13:29, 2005. (3) R. Sriwati et al. Nematology 10:1, 2008. (4) G. Steiner and E. M. Buhrer. J. Agric. Res. 48:946, 1934.

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First Report of Pine Wilt Disease on Pinus koraiensis in Korea

Plant Disease ◽

10.1094/pdis-92-8-1251a ◽

2008 ◽

Vol 92 (8) ◽

pp. 1251-1251 ◽

Cited By ~ 12

Author(s):

H. Han ◽

Y.-J. Chung ◽

S.-C. Shin

Keyword(s):

Dna Sequences ◽

Pinus Densiflora ◽

Pine Wilt Disease ◽

Wilt Disease ◽

Internal Transcribed Spacers ◽

Pinus Koraiensis ◽

Korean Pine ◽

Accurate Identification ◽

First Report ◽

Pine Wilt

Pine wilt disease is one of the most important forest tree diseases, especially in the East Asian countries of Japan, China, and Korea. The causal agent, Bursaphelenchus xylophilus (3), is transmitted by the insect vectors, pine sawyer beetles (Monochamus spp.). The vectors mainly attack coniferous trees and the infected trees die within several weeks. In Korea, pine wilt disease was first reported in Busan City in 1988, and now, the damaged area covers 7,820 ha and more than 60 cities in Korea. The main host trees are Pinus densiflora and P. thunbergii, which are the most common pines distributed in Korea. In 2006, however, we found pine wilt disease in a forest of P. koraiensis located in Gwangju City in Gyeonggi Province. Symptoms were systemic and leaves turned brown or yellowish. Wood samples were collected from the affected trees, and nematodes were extracted by a modified Baermann's funnel method. After 24 h, nematodes were collected from wood chips, and morphology was observed with an inverted light microscope (Leica DE/DMI 3000B, Wetzlar, Germany). Morphology was characterized by a typical Aphelenchoid-type esophagus, head constriction, female vulva flap, female tail, and a male spicule shape similar to B. xylophilus. For more accurate identification, DNA was extracted from individual nematodes with a DNeasy Kit (Qiagen, Valencia, CA). The internal transcribed spacers (ITS) 1 and 2 and 5.8S regions were amplified by PCR (1). Sequenced nucleotide information was compared with the sequences of B. xylophilus already reported in GenBank (Accession Nos. AB294736, AB277208, AM 157747, AY 347913, and BXU92464). ITS DNA sequences of the nematode from Korean pine was >99% identical to B. xylophilus in GenBank. The B. xylophilus from Korean pine tree was also successfully cultured in Botrytis cinerea medium and pathogenicity was tested from June to October 2007. More than 95% mortality was observed with the inoculation of 20 replicate 15-year-old trees of P. koraiensis and P. densiflora with 15,000 nematodes per tree. Therefore, we confirmed that pine wilt disease occurred in P. koraiensis in Korea. P. koraiensis is an endemic species in Korea and distribution is limited to the northern Korean Peninsula and some locations in Russia. It has been shown that P. koraiensis is susceptible to the pine wood nematode by an inoculation test in Japan (2); however, to our knowledge, this is the first report of pine wilt disease on P. koraiensis under field conditions in Korea. References: (1) W. K. Burgermeister et al. Russ. J. Nematol. 13:29, 2005. (2) K. Futai and T. Furuno. Bull. Kyoto Univ. For. 51:3, 1979. (3) G. Steiner and E. M. Buhrer. J. Agr. Res. 48:946, 1934.

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First Report of Bursaphelenchus mucronatus kolymensis Associated with Pinus sylvestris in Serbia

Plant Disease ◽

10.1094/pdis-06-14-0594-pdn ◽

2014 ◽

Vol 98 (12) ◽

pp. 1745-1745

Author(s):

J. Bačić ◽

B. G. Stare ◽

G. Urek ◽

S. Širca

Keyword(s):

Pinus Sylvestris ◽

Morphological Difference ◽

Pine Wilt Disease ◽

Its Region ◽

Wilt Disease ◽

Morphological Identification ◽

First Report ◽

Pine Wilt ◽

Bursaphelenchus Mucronatus ◽

Female Tail

Bursaphelenchus mucronatus kolymensis (Korentchenko) Braasch et al. (1), also referred to as the European type of B. mucronatus, is morphologically very similar to the pine wood nematode (PWN) B. xylophilus (Steiner & Buhrer) Nickle, the causal agent of pine wilt disease. The main morphological difference between the two species is the shape of the female tail mucro (3); however, some populations of PWN vary in mucro shape, which can lead to misidentification. Since PWN was found and identified for the first time in Portugal in 1999 (4), concern about the spread of PWN to pines and other conifers all over Europe has increased. Therefore, the PWN survey in every European country is essential. In 2011, the Serbian Ministry of Agriculture, Forestry and Water Management implemented phytosanitary measures for detection of PWN in the territory of the Republic of Serbia. In September 2013, wood samples were collected from a 40-year-old wilted Pinus sylvestris located on Divcibare Mountain, western Serbia. Nematodes were extracted by a modified Baermann funnel method. Nematode specimens (10 females and 10 males) were characterized by an Aphelenchoid-type esophagus, female vulva flap, mucro on the end of female tail, and males with paired arcuate spicules. Morphological identification matched that of B. mucronatus kolymensis (1). The specific morphometric evidence for this European type subspecies of B. mucronatus was sub-cylindrical female tail with mucro 3 to 4 μm long, digitate, well offset from tail (5). Species identification was confirmed by PCR-RFLP of the rDNA ITS region using one female and one male nematode separately (2). The sizes of restricted DNA fragments as determined with agarose gel and capillary electrophoresis were in compliance with reported data in the literature (2,4) and support the identification of the nematode as B. mucronatus kolymensis. To our knowledge, this is the first report of any species belonging to Bursaphelenchus genus in Serbia. The presence of B. mucronatus kolymensis in Serbia indicates the possibility of establishment and spread of PWN. Therefore, the measures taken in case of PWN occurrence should be prepared and training foresters to identify pine wilt disease symptoms should be encouraged in Serbia. References: (1) H. Braasch et al. J. Nematode Morphol. Syst. 14:77, 2011. (2) W. Burgermeister et al. Nematology 11:649, 2009. (3) Y. Mamiya and N. Enda. Nematologica 25:353, 1979. (4) M. M. Mota et al. Nematology 1:727, 1999. (5) M. M. Mota and P. Vieira. Page 146 in: Pine Wilt Disease: A Worldwide Threat to Forest Ecosystems. Springer Science and Business Media, 2008.

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First Report of Postharvest Blue Mold decay caused by Penicillium expansum on Lemon (Citrus limon) fruit in China

Plant Disease ◽

10.1094/pdis-10-20-2239-pdn ◽

2021 ◽

Author(s):

Ibatsam Khokhar ◽

Jianming Chen ◽

Junhuan Wang ◽

Yang Jia ◽

Yanchun Yan ◽

...

Keyword(s):

Morphological Characteristics ◽

Its Region ◽

Penicillium Expansum ◽

Citrus Limon ◽

Conidial Suspension ◽

Blue Mold ◽

Causal Organism ◽

Lemon Fruit ◽

First Report ◽

Fungal Isolates

Lemon (Citrus limon) is one of the most important commercial (both dried and fresh) citrus fruits in China. In the spring of 2019, postharvest blue mold decay was observed at an incidence of 3-5% on lemon fruit at the local markets in Beijing, China. Fruit lesions were circular, brown, soft, and watery, and rapidly expanded at 25°C. To isolate the causal organism, small pieces (2 mm3) were cut from the lesions, surface-sterilized for 1 min in 1.5% NaOCl, rinsed three times with sterilized water, dried with sterile filter paper, placed onto potato dextrose agar (PDA) medium, and incubated at 25°C for 6 days. Eight morphologically similar single-colony fungal isolates were recovered from six lemon fruit. Colony surfaces were bluish-green on the upper surface and cream to yellow-brown one the reverse. Hyphae on colony margins were entirely subsurface and cream in color. Mycelium was highly branched, septate, and colorless, and conidiophores were 250 to 450 × 3.0 to 4.0 µm in size. Stipe of conidiophores were smooth-walled, bearing terminal penicilli, typically terverticillate or less commonly birverticillate, rami occurring singly, 16 to 23 × 3.0 to 4.0 µm, metulae in 3 to 6, measuring 12 to 15 × 3.0 to 4.0 µm. Phialides were ampulliform to almost cylindrical, in verticils of 5 to 8, measuring 8 to 11 × 2.5 to 3.2 µm with collula. Conidia were smooth-walled, ellipsoidal, measuring 3.0 to 3.5 × 2.5 to 3.0 µm. According to morphological characteristics, the fungus was identified as Penicillium expansum (Visagie et al. 2014). For molecular identification, genomic DNA of eight fungal isolates was extracted, regions of the beta-tubulin (TUB), and calmodulin (CAL) genes and ITS region, were amplified using Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 primers respectively. Obtained sequences of all isolates were identical to sequences of the representative isolate YC-IK12, which was submitted in the GenBank. BLAST results of YC-IK12 sequences (ITS; MT856700: TUB; MT856958: CAL; MT856959) showed 98 to 100% similarity with P. expansum accessions (NR-077154, LN896428, JX141581). For pathogenicity tests, 10 μl of conidial suspension (10 × 105 conidia/ml) from seven-day-old YC-IK12 culture was inoculated using a sterilized needle into the surface of each five asymptomatic disinfected lemons. As a control, three lemons were inoculated using sterile distilled water. All inoculated lemons were placed in plastic containers and incubated at 25°C for 7 days. Decay lesions, identical to the original observations, developed on all inoculated lemons, while control lemons remained asymptomatic. Fungus re-isolated from the inoculated lemon was identified as P. expansum on the basis morphology and Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 sequences. Previously, Penicillium spp. including P. expansum have been reported as post-harvest pathogens on various Citrus spp. (Louw & Korsten 2015). However, P. digitatum has been reported on lemons and P. expansum has been reported on stored Kiwifruit (Actinidia arguta), Malus, and Pyrus species in China (Tai, 1979; Wang et al. 2015). To our knowledge, this is the first report of blue mold caused by P. expansum on lemons in China. References Louw, J. P., Korsten, L. 2015. Plant Dis. 99:21-30. Tai, F.L. 1979. Sylloge Fungorum Sinicorum. Sci. Press, Acad. Sin., Peking, 1527 pages. 8097 Visagie, C.M. et al. 2014. Studies. Mycol.78: 343. Wang, C. W. et al. 2015. Plant Dis. 99:1037.

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First Report of Podosphaera xanthii Causing Powdery Mildew on Mungbean (Vigna radiata) in Taiwan

Plant Disease ◽

10.1094/pdis-09-20-2092-pdn ◽

2021 ◽

Author(s):

Zong-ming Sheu ◽

Ming-hsueh Chiu ◽

Lawrence Kenyon

Keyword(s):

Powdery Mildew ◽

Vigna Radiata ◽

Morphological Characteristics ◽

Resistance Breeding ◽

Its Region ◽

Infected Leaf ◽

Pathogenicity Test ◽

Causal Organism ◽

First Report ◽

Powdery Mildew Pathogen

Mungbean (Vigna radiata L.) is routinely grown in the experimental fields at the headquarters of the World Vegetable Center (23°6'30.88"N, 120°17'51.31"E) for breeding, research and germplasm multiplication. In a spring 2016 mungbean trial, about 50% of the plants were affected with powdery mildew. The white, powdery-like patches first appeared on the upper leaf surfaces, and soon developed to grey patches on both sides of the leaves. Purple to brown discoloration appeared on the underside of the infected leaf. Microscopy examination revealed that the causal organism was not Erysiphe polygoni, which had previously been documented as the powdery mildew pathogen on mungbean in Taiwan (Hartman et al. 1993). The fungus produced typical structures of the powdery mildew Euoidium, anamorph of the genus Podosphaera. The mycelium consisted of septate, flexuous hyphae with indistinct appressoria. The erect conidiophores arising from superficial hyphae varied from straight or slightly curved to curled. Three to ten conidia were borne in long chains with crenate edges. Foot-cells were straight, cylindrical and measured 30 to 52 µm long. Conidia were hyaline, ellipsoid-ovoid to barrel-shaped, with fibrosin bodies, and measured 27 to 33 (mean = 30.4) × 15 to 20 (mean = 16.6) µm. Germ tubes were clavate and occasionally forked, and were produced from the lateral sites of the conidia. No chasmothecia were found in the samples. The morphological characteristics were consistent with P. xanthii (Castagne) U. Braun & Shishkoff (Braun & Cook 2012). To confirm the identity, the internal transcribed spacer (ITS) region of rDNA and partialβ-tubulin gene (TUB2) for the isolate MG3 were amplified with the primers ITS4/ITS5 (White et al. 1990) and BtuF5/BtuR7a (Ellingham et al. 2019), respectively. BLASTn analysis revealed the ITS sequence (MN833717) was 100% identical to many records of P. xanthii whereas the TUB2 sequence (MW363957) was 100% identical to a record of P. fusca (syn. P. xanthii; KC333362) in NCBI GenBank. A pathogenicity test was conducted by dusting conidia from an infected leaf onto six healthy four-week-old mungbean plants (cv ‘Tainan No. 3’). Another three plants were not inoculated and were used as control. All the plants were maintained in a greenhouse at 25 to 28°C. All inoculated plants developed powdery mildew symptoms after 10 days, whereas the control plants remained symptomless. To our knowledge, this is the first report of P. xanthii causing disease on mungbean in Taiwan. P. xanthii also has been reported on mungbean in Thailand (Meeboon et al. 2016), while other records referring to E. polygoni infecting Vigna spp. are from Brazil and Fiji (Farr & Rossman 2020). Although both P. xanthii and E. polygoni have now been reported as causing powdery mildew on mungbean in Taiwan, which species predominates or is more important remains unclear. A comprehensive survey with accurate species identification is required to develop effective management of the disease, particularly for resistance breeding.

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First Report of Bursaphelenchus xylophilus Causing Pine Wilt Disease on Pinus radiata in Spain

Plant Disease ◽

10.1094/pdis-03-15-0252-pdn ◽

2015 ◽

Vol 99 (10) ◽

pp. 1449-1449 ◽

Cited By ~ 19

Author(s):

P. Zamora ◽

V. Rodríguez ◽

F. Renedo ◽

A. V. Sanz ◽

J. C. Domínguez ◽

...

Keyword(s):

Pinus Radiata ◽

Pine Wilt Disease ◽

Bursaphelenchus Xylophilus ◽

Wilt Disease ◽

First Report ◽

Pine Wilt

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

Japanese Journal of Phytopathology ◽

10.3186/jjphytopath.54.606 ◽

1988 ◽

Vol 54 (5) ◽

pp. 606-615 ◽

Cited By ~ 34

Author(s):

Keiko KURODA ◽

Toshihiro YAMADA ◽

Kazuhiko MINEO ◽

Hirotada TAMURA

Keyword(s):

Pine Wilt Disease ◽

Bursaphelenchus Xylophilus ◽

Wilt Disease ◽

Pine Wilt

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

Forest Ecosystems ◽

10.1186/s40663-021-00328-6 ◽

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.

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Dynamical aspects of pine wilt disease and control measures

Chaos Solitons & Fractals ◽

10.1016/j.chaos.2021.110764 ◽

2021 ◽

Vol 145 ◽

pp. 110764

Author(s):

Takasar Hussain ◽

Adnan Aslam ◽

Muhammad Ozair ◽

Fatima Tasneem ◽

J.F. Gómez-Aguilar

Keyword(s):

Pine Wilt Disease ◽

Wilt Disease ◽

Control Measures ◽

Pine Wilt ◽

And Control

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Spatiotemporal Pattern of Pine Wilt Disease in the Yangtze River Basin

Forests ◽

10.3390/f12060731 ◽

2021 ◽

Vol 12 (6) ◽

pp. 731

Author(s):

Zhuoqing Hao ◽

Jixia Huang ◽

Yantao Zhou ◽

Guofei Fang

Keyword(s):

River Basin ◽

Yangtze River ◽

Yangtze River Basin ◽

Pine Wilt Disease ◽

Wilt Disease ◽

Spatiotemporal Pattern ◽

Pinewood Nematode ◽

The Yangtze River ◽

Pine Wilt ◽

The Yangtze River Basin

The Yangtze River Basin is among the river basins with the strongest strategic support and developmental power in China. As an invasive species, the pinewood nematode (PWN) Bursaphelenchus xylophilus has introduced a serious obstacle to the high-quality development of the economic and ecological synchronization of the Yangtze River Basin. This study analyses the occurrence and spread of pine wilt disease (PWD) with the aim of effectively managing and controlling the spread of PWD in the Yangtze River Basin. In this study, statistical data of PWD-affected areas in the Yangtze River Basin are used to analyse the occurrence and spread of PWD in the study area using spatiotemporal visualization analysis and spatiotemporal scanning statistics technology. From 2000 to 2018, PWD in the study area showed an “increasing-decreasing-increasing” trend, and PWD increased explosively in 2018. The spatial spread of PWD showed a “jumping propagation-multi-point outbreak-point to surface spread” pattern, moving west along the river. Important clusters were concentrated in the Jiangsu-Zhejiang area from 2000 to 2015, forming a cluster including Jiangsu and Zhejiang. Then, from 2015–2018, important clusters were concentrated in Chongqing. According to the spatiotemporal scanning results, PWD showed high aggregation in the four regions of Zhejiang, Chongqing, Hubei, and Jiangxi from 2000 to 2018. In the future, management systems for the prevention and treatment of PWD, including ecological restoration programs, will require more attention.

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Can insecticide applications used to kill vector insects prevent pine wilt disease?

Pest Management Science ◽

10.1002/ps.6532 ◽

2021 ◽

Author(s):

Jong‐Kook Jung ◽

Ung Gyu Lee ◽

Deokjea Cha ◽

Dong Soo Kim ◽

Chansik Jung

Keyword(s):

Pine Wilt Disease ◽

Wilt Disease ◽

Pine Wilt

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