Bacterial Communities Associated With the Pine Wilt Disease Vector Monochamus alternatus (Coleoptera: Cerambycidae) During Different Larval Instars

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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Pine wood nematode, pine wilt disease, vector beetle and pine tree: how a multiplayer system could reply to climate change.

Climate change and insect pests ◽

10.1079/9781780643786.0220 ◽

2015 ◽

pp. 220-234 ◽

Cited By ~ 7

Author(s):

A. Roques ◽

L Zhao ◽

J Sun ◽

C. Robinet

Keyword(s):

Climate Change ◽

Pine Wilt Disease ◽

Wilt Disease ◽

Pine Wood Nematode ◽

Pine Wood ◽

Disease Vector ◽

Pine Tree ◽

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/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 Communities and Virulence Associated with Pine Wood Nematode Bursaphelenchus xylophilus from Different Pinus spp.

International Journal of Molecular Sciences ◽

10.3390/ijms20133342 ◽

2019 ◽

Vol 20 (13) ◽

pp. 3342 ◽

Cited By ~ 6

Author(s):

Qi Xue ◽

Yang Xiang ◽

Xiao-Qin Wu ◽

Ming-Jie Li

Keyword(s):

Bacterial Communities ◽

Bacterial Species ◽

Pine Wilt Disease ◽

Bursaphelenchus Xylophilus ◽

Wilt Disease ◽

Sequencing Analysis ◽

Culturable Bacteria ◽

Pine Wilt ◽

Pinus Spp ◽

Pine Species

Bursaphelenchus xylophilus, the causal agent of pine wilt disease, is a destructive threat to pine forests. The role of bacteria associated with B. xylophilus in pine wilt disease has attracted widespread attention. This study investigated variation in bacterial communities and the virulence of surface-sterilized B. xylophilus from different Pinus spp. The predominant culturable bacteria of nematodes from different pines were Stenotrophomonas and Pseudomonas. Biolog EcoPlate analysis showed that metabolic diversity of bacteria in B. xylophilus from P. massoniana was the highest, followed by P. thunbergii and P. densiflora. High-throughput sequencing analysis indicated that bacterial diversity and community structure in nematodes from the different pine species varied, and the dominant bacteria were Stenotrophomonas and Elizabethkingia. The virulence determination of B. xylophilus showed that the nematodes from P. massoniana had the greatest virulence, followed by the nematodes from P. thunbergii and P. densiflora. After the nematodes were inoculated onto P. thunbergii, the relative abundance of the predominant bacteria changed greatly, and some new bacterial species emerged. Meanwhile, the virulence of all the nematode isolates increased after passage through P. thunbergii. These inferred that some bacteria associated with B. xylophilus isolated from different pine species might be helpful to adjust the PWN’s parasitic adaptability.

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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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