Management of pine wilt disease vectoring Monochamus alternatus adults using spray and soil application of Metarhizium anisopliae JEF isolates

Monochamus alternatus is an important insect pest in pine forests of southern China and the dispersing vector of the pine wood nematode, Bursaphelenchus xylophilus, which leads to pine wilt disease (PWD). Microbiome of M. alternatus may contribute to survival of larvae in the host pine trees. In order to investigate the intestinal bacterial structure of M. alternatus during the larvae and pupae stages in host trees, and infer the function of symbiotic bacteria, we used 16S rRNA gene Illumina sequencing to obtain and compare the bacterial community composition in the foregut, midgut, and hindgut of larvae, pupal intestines, larval galleries, and pupal chambers of M. alternatus. The diversity of the bacterial community in larval intestines and pupal intestines were similar, as well as was significantly greater in larval galleries and pupal chambers. Although there were differences in bacterial compositions in different samples, similar components were also found. Proteobacteria and Firmicutes were the two most dominant phyla in all samples, and genera Enterobacter, Raoultella, Serratia, Lactococcus, and Pseudomonas were dominant in both the intestinal samples and plant tissue samples. Enterobacter was the most abundant genus in larval intestines, and Serratia was dominant in pupal intestine. The functions of these dominant and specific bacteria were also predicted through metagenomic analyses. These bacteria may help M. alternatus degrade cellulose and pinene. The specific role of symbiotic bacteria in the infection cycle of PWD also warrants further study in the future.

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Characterization of bacterial communities associated with the pinewood nematode insect vector Monochamus alternatus Hope and the host tree Pinus massoniana

10.21203/rs.2.14055/v3 ◽

2020 ◽

Author(s):

Yajie Guo ◽

Qiannan Lin ◽

lvyi Chen ◽

Carballar-Lejarazú Rebeca ◽

Aishan Zhang ◽

...

Keyword(s):

Pine Wilt Disease ◽

Wilt Disease ◽

Insect Vector ◽

Insect Vectors ◽

Pinewood Nematode ◽

Monochamus Alternatus ◽

Form Complex ◽

Pine Wilt ◽

Bacterial Microbiota ◽

Habitat Niche

Abstract Background: Monochamus alternatus Hope is one of the insect vectors of pinewood nematode (Bursaphelenchus xylophilus), which causes the destructive pine wilt disease. The microorganisms within the ecosystem, comprising plants, their environment, and insect vectors, form complex networks. This study presents a systematic analysis of the bacterial microbiota in the M. alternatus midgut and its habitat niche.Methods: Total DNA was extracted from 20 types of samples (with three replicates each) from M. alternatus and various tissues of healthy and infected P. massoniana (pines). 16S rDNA amplicon sequencing was conducted to determine the composition and diversity of the bacterial microbiota in each sample. Moreover, the relative abundances of bacteria in the midgut of M. alternatus larvae were verified by counting the colony-forming units.Results: Pinewood nematode infection increased the microbial diversity in pines. Bradyrhizobium, Burkholderia, Dyella, Mycobacterium, and Mucilaginibacter were the dominant bacterial genera in the soil and infected pines. These results indicate that the bacterial community in infected pines may be associated with the soil microbiota. Interestingly, the abundance of the genus Gryllotalpicola was highest in the bark of infected pines. The genus Cellulomonas was not found in the midgut of M. alternatus, but it peaked in the phloem of infected pines, followed by the phloem of heathy pines. Moreover, the genus Serratia was not only present in the habitat niche, but it was also enriched in the M. alternatus midgut. The colony-forming unit assays showed that the relative abundance of Serratia sp. peaked in the midgut of instar II larvae (81%).Conclusions: Overall, the results indicate that the bacterial microbiota in the soil and in infected pines are correlated. The Gryllotalpicola sp. and Cellulomonas sp. are potential microbial markers of pine wilt disease. Additionally, Serratia sp. could be an ideal agent for expressing insecticidal protein in the insect midgut by genetic engineering, which represents a new use of microbes to control M. alternatus.

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Bacterial community associated to the pine wilt disease insect vectors Monochamus galloprovincialis and Monochamus alternatus

Scientific Reports ◽

10.1038/srep23908 ◽

2016 ◽

Vol 6 (1) ◽

Cited By ~ 15

Author(s):

Marta Alves ◽

Anabela Pereira ◽

Patrícia Matos ◽

Joana Henriques ◽

Cláudia Vicente ◽

...

Keyword(s):

Bacterial Community ◽

Pine Wilt Disease ◽

Wilt Disease ◽

Insect Vectors ◽

Monochamus Alternatus ◽

Monochamus Galloprovincialis ◽

Pine Wilt

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Characterization of bacterial communities associated with the pinewood nematode insect vector Monochamus alternatus Hope and the host tree Pinus massoniana

10.21203/rs.2.14055/v2 ◽

2020 ◽

Author(s):

Yajie Guo ◽

Qiannan Lin ◽

lvyi Chen ◽

Carballar-Lejarazú Rebeca ◽

Aishan Zhang ◽

...

Keyword(s):

Pine Wilt Disease ◽

Wilt Disease ◽

Insect Vector ◽

Insect Vectors ◽

Pinewood Nematode ◽

Monochamus Alternatus ◽

Pine Wilt ◽

Bacterial Microbiota ◽

Habitat Niche ◽

Monochamus Alternatus Hope

Abstract Background Monochamus alternatus Hope is one of the insect vectors of pinewood nematode ( Bursaphelenchus xylophilus ), which causes the destructive pine wilt disease. The microorganisms within the ecosystem, comprising plants, their environment, and insect vectors, form complex networks. This study presents a systematic analysis of the bacterial microbiota in the M. alternatus midgut and its habitat niche. Methods Total DNA was extracted from 20 types of samples (with three replicates each) from M. alternatus and various tissues of healthy and infected P. massoniana (pines). 16S rDNA amplicon sequencing was conducted to determine the composition and diversity of the bacterial microbiota in each sample. Moreover, the relative abundances of bacteria in the midgut of M. alternatus larvae were verified by counting the colony-forming units. Results Pinewood nematode infection increased the microbial diversity in pines. Bradyrhizobium , Burkholderia , Dyella , Mycobacterium , and Mucilaginibacter were the dominant bacterial genera in the soil and infected pines. These results indicate that the bacterial community in infected pines may be associated with the soil microbiota. Interestingly, the abundance of the genus Gryllotalpicola was highest in the bark of infected pines. The genus Cellulomonas was not found in the midgut of M. alternatus , but it peaked in the phloem of infected pines, followed by the phloem of heathy pines. Moreover, the genus Serratia was not only present in the habitat niche, but it was also enriched in the M. alternatus midgut. The colony-forming unit assays showed that the relative abundance of Serratia sp. peaked in the midgut of instar II larvae (81%). Conclusions Overall, the results indicate that the bacterial microbiota in the soil and in infected pines are correlated. The Gryllotalpicola sp. and Cellulomonas sp. are potential microbial markers of pine wilt disease. Additionally, Serratia sp. could be an ideal agent for expressing insecticidal protein in the insect midgut by genetic engineering, which represents a new use of microbes to control M. alternatus .

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Relationship between pine wilt disease development in asymptomatic carrier trees of Bursaphelenchus xylophilus (Nematoda: Apelenchoididae) and their use by Monochamus alternatus (Coleoptera: Cerambycidae)

Applied Entomology and Zoology ◽

10.1007/s13355-014-0299-2 ◽

2014 ◽

Vol 50 (1) ◽

pp. 33-39 ◽

Cited By ~ 1

Author(s):

Noritoshi Maehara ◽

Takuya Aikawa ◽

Natsumi Kanzaki

Keyword(s):

Asymptomatic Carrier ◽

Pine Wilt Disease ◽

Bursaphelenchus Xylophilus ◽

Wilt Disease ◽

Disease Development ◽

Monochamus Alternatus ◽

Pine Wilt

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Possible altitude and temperature limits on pine wilt disease: the reproduction of vector sawyer beetles (Monochamus alternatus), survival of causal nematode (Bursaphelenchus xylophilus), and occurrence of damage caused by the disease

European Journal of Forest Research ◽

10.1007/s10342-013-0742-x ◽

2013 ◽

Vol 133 (2) ◽

pp. 225-233 ◽

Cited By ~ 5

Author(s):

Masashi Ohsawa ◽

Mitsuteru Akiba

Keyword(s):

Pine Wilt Disease ◽

Bursaphelenchus Xylophilus ◽

Wilt Disease ◽

Monochamus Alternatus ◽

Pine Wilt

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Genetic differentiation of the pine wilt disease vector Monochamus alternatus (Coleoptera: Cerambycidae) over a mountain range – revealed from microsatellite DNA markers

Bulletin of Entomological Research ◽

10.1017/s000748530700483x ◽

2007 ◽

Vol 97 (2) ◽

pp. 167-174 ◽

Cited By ~ 13

Author(s):

E. Shoda-Kagaya

Keyword(s):

Isolation By Distance ◽

Spatial Scales ◽

Pine Wilt Disease ◽

Wilt Disease ◽

Secondary Contact ◽

Mountain Range ◽

Monochamus Alternatus ◽

Damage Area ◽

Pine Wilt ◽

Geographical Barrier

AbstractTo study the dispersal process of the pine sawyer Monochamus alternatus (Hope) in frontier populations, a microsatellite marker-based genetic analysis was performed on expanding populations at the northern limit of its range in Japan. In Asian countries, M. alternatus is the main vector of pine wilt disease, the most serious forest disease in Japan. Sawyers were collected from nine sites near the frontier of the pine wilt disease damage area. A mountain range divides the population into western and eastern sides. Five microsatellite loci were examined and a total of 188 individuals was genotyped from each locus with the number of alleles ranged from two to nine. The mean observed heterozygosity for all loci varied from 0.282 to 0.480 in the nine sites, with an overall mean of 0.364. None of the populations have experienced a significant bottleneck. Significant differentiation was found across the mountain range, but the genetic composition was similar amongst populations of each side. It is believed that the mountain range acts as a geographical barrier to dispersal and that gene flow without a geographical barrier is high. On the west side of the mountain range, a pattern of isolation by distance was detected. This was likely to be caused by secondary contact of different colonizing routes on a small spatial scale. Based on these data, a process linking genetic structure at local (kilometres) and regional spatial scales (hundreds of kilometres) was proposed.

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Characterization of bacterial communities associated with the pine wilt disease insect vectors Monochamus alternatus Hope and host trees Pinus massoniana

10.21203/rs.2.14055/v1 ◽

2019 ◽

Author(s):

Yajie Guo ◽

Qiannan Lin ◽

lvyi Chen ◽

Carballar-Lejarazú Rebeca ◽

Ensi Shao ◽

...

Keyword(s):

Microbial Community ◽

Pine Wilt Disease ◽

Pinus Massoniana ◽

Wilt Disease ◽

Microbial Flora ◽

Pinewood Nematode ◽

Monochamus Alternatus ◽

Transmission Cycle ◽

Pine Wilt ◽

Monochamus Alternatus Hope

Abstract Background Pine wilt disease (PWD) is a destructive disease caused by the pinewood nematode Bursaphelenchus xylophilus . Monochamus alternatus Hope is the main vector of this disease. The symbiotic microorganisms can play an important role in the transmission cycle mechanism. However, the role of bacterial microorganisms in the transmission of pine wood nematode by M. alternatus is not clear currently. The main purpose of this study is to reveal the composition and diversity of microbial flora in the gut of M. alternatus , as well as healthy and infected Pinus massoniana and its peripheral environment to discover the important microbial flora contributing to the transmission cycle.Methods In this study, total DNA was extracted from 60 samples, including 20 samples of M. alternatus gut from different larval instars, healthy P. massoniana , nematode-infected P. massoniana and their peripheral environment (needles, bark, phloem, xylem, root, surface soil and rhizosphere soil), by triplicate. Samples were used for 16S rDNA Amplicon sequencing to determine the composition and diversity of microbial flora in each sample.Results Infection of pinewood nematode resulted in an increase of the microbial community in the nematode-infected P. massoniana and its peripheral environment when compared with healthy P. massoniana , the microbial community in different tissues changed. Among them, Gryllotalpicola and Cellulomonas showed to be endemic microorganisms in nematode-infected P. massoniana , which can be used as indicators to detect the disease. Serratia was shown as an opportunistic pathogen, and was found to be enriched in M. alternatus gut and was also detected in the host plant tissues.Conclusions This study clarified the change of microbial community in the transmission of pine wilt disease by M. alternatus . An important theoretical basis for the prevention of pine wilt disease was structured by our research.

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