scholarly journals Molecular Cloning and Expression Analysis of the Endogenous Cellulase Gene MaCel1 in Monochamus alternatus

Forests ◽  
2020 ◽  
Vol 11 (12) ◽  
pp. 1372
Author(s):  
Yachao Li ◽  
Hao Chen ◽  
Xu Chu ◽  
Qiuyu Ma ◽  
Guanghong Liang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Developmental Stages ◽  
Sequence Similarity ◽  
Pupal Stage ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Cloning And Expression ◽  
Cellulase Gene ◽  
Monochamus Alternatus ◽  
Real Time Quantitative Pcr

The purpose of this study was to characterize the endogenous cellulase gene MaCel1 of Monochamus alternatus, which is an important vector of Bursaphelenchus xylophilus, a pine wood nematode, which causes pine wilt disease (PWD). In this study, MaCel1 was cloned by rapid amplification of cDNA end (RACE), and its expression analyzed by RT-qPCR (real-time quantitative PCR detecting). A total of 1778 bp of cDNA was obtained. The encoding region of this gene was 1509 bp in length, encoding a protein containing 502 amino acids with a molecular weight of 58.66 kDa, and the isoelectric point of 5.46. Sequence similarity analysis showed that the amino acids sequence of MaCel1 had high similarity with the β-Glucosinolate of Anoplophoraglabripennis and slightly lower similarity with other insect cellulase genes (GH1). The β-D-Glucosidase activity of MaCel1 was 256.02 ± 43.14 U/L with no β-Glucosinolate activity. MaCel1 gene was widely expressed in the intestine of M. alternatus. The expression level of MaCel1 gene in male (3.46) and female (3.51) adults was significantly higher than that in other developmental stages, and the lowest was in pupal stage (0.15). The results will help reveal the digestive mechanism of M. alternatus and lay the foundation for controlling PWD by controlling M. alternatus.

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 Mitogenome Analysis of Four Lamiinae Species (Coleoptera: Cerambycidae) and Gene Expression Responses by Monochamus alternatus When Infected with the Parasitic Nematode, Bursaphelenchus mucronatus

Insects ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 453
Author(s):  
Zi-Yi Zhang ◽  
Jia-Yin Guan ◽  
Yu-Rou Cao ◽  
Xin-Yi Dai ◽  
Kenneth B. Storey ◽  
...  
Keyword(s):  
Gene Expression ◽  
Phylogenetic Trees ◽  
Mitochondrial Gene ◽  
Mitochondrial Protein ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Monochamus Alternatus ◽  
Genome Database ◽  
Protein Coding ◽  
Nd5 Gene

We determined the mitochondrial gene sequence of Monochamus alternatus and three other mitogenomes of Lamiinae (Insect: Coleoptera: Cerambycidae) belonging to three genera (Aulaconotus, Apriona and Paraglenea) to enrich the mitochondrial genome database of Lamiinae and further explore the phylogenetic relationships within the subfamily. Phylogenetic trees of the Lamiinae were built using the Bayesian inference (BI) and maximum likelihood (ML) methods and the monophyly of Monochamus, Anoplophora, and Batocera genera was supported. Anoplophora chinensis, An. glabripennis and Aristobia reticulator were closely related, suggesting they may also be potential vectors for the transmission of the pine wood pathogenic nematode (Bursaphelenchus xylophilus) in addition to M. alternatus, a well-known vector of pine wilt disease. There is a special symbiotic relationship between M. alternatus and Bursaphelenchus xylophilus. As the native sympatric sibling species of B. xylophilus, B. mucronatus also has a specific relationship that is often overlooked. The analysis of mitochondrial gene expression aimed to explore the effect of B. mucronatus on the energy metabolism of the respiratory chain of M. alternatus adults. Using RT-qPCR, we determined and analyzed the expression of eight mitochondrial protein-coding genes (COI, COII, COIII, ND1, ND4, ND5, ATP6, and Cty b) between M. alternatus infected by B. mucronatus and M. alternatus without the nematode. Expression of all the eight mitochondrial genes were up-regulated, particularly the ND4 and ND5 gene, which were up-regulated by 4–5-fold (p < 0.01). Since longicorn beetles have immune responses to nematodes, we believe that their relationship should not be viewed as symbiotic, but classed as parasitic.

Boarding abilities of Bursaphelenchus mucronatus and B. xylophilus (Nematoda: Aphelenchoididae) on Monochamus alternatus (Coleoptera: Cerambycidae)

Nematology ◽  
2003 ◽  
Vol 5 (6) ◽  
pp. 843-849 ◽  
Author(s):  
Shota Jikumaru ◽  
Katsumi Togashi
Keyword(s):  
Pine Wilt Disease ◽  
Nematode Species ◽  
Bursaphelenchus Xylophilus ◽  
Insect Vector ◽  
Species Replacement ◽  
Monochamus Alternatus ◽  
Bursaphelenchus Mucronatus ◽  
Initial Load ◽  
Pine Trees ◽  
The Difference

Abstract Bursaphelenchus mucronatus is closely related to Bursaphelenchus xylophilus, the causative agent of pine wilt disease. Both nematodes are transmitted between host pine trees as the fourth-stage dispersal juveniles (JIV) by insect vectors. After the invasion of Japan by B. xylophilus, B. mucronatus, native to Japan, appears to have been replaced in the pine forests during the spread of the disease. To help understand this species replacement, the number of JIV carried by an insect vector (the initial nematode load) was compared between the two nematode species by using the beetle, Monochamus alternatus, in the laboratory. The initial load of B. mucronatus was significantly smaller than that of B. xylophilus although the number of third-stage dispersal juveniles (JIII) concentrated at the pupal chambers did not differ. Statistical analysis showed that the proportion of JIII moulting to JIV was the most important among three components explaining the difference in the initial load of B. mucronatus while the number of JIII concentrated at the pupal chamber was the most important for B. xylophilus. The phoretic affinity between the nematode and its vector is discussed in relation to its role in the species replacement.

Seasonal change in susceptibility of Pinus densiflora to Bursaphelenchus xylophilus infection, determined from the number of nematodes passing through branch sections

Nematology ◽  
2009 ◽  
Vol 11 (3) ◽  
pp. 409-418 ◽  
Author(s):  
Koji Matsunaga ◽  
Katsumi Togashi
Keyword(s):  
Seasonal Change ◽  
Developmental Stages ◽  
Pinus Densiflora ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Control Measures ◽  
Effective Control ◽  
Dispersal Ability ◽  
Second Stage ◽  
Individual Branch

AbstractPine wilt disease by Bursaphelenchus xylophilus has been causing devastating damage to Pinus densiflora and P. thunbergii forests in Japan. Reforestation using resistant trees is one of the most effective control measures. In this study we examined the seasonal change in the number of nematodes passing through 5 cm long P. densiflora branch sections during 24 h, to determine the season of greatest susceptibility when resistant candidates can be identified easily. We inoculated 200 nematodes of each of four B. xylophilus and one B. mucronatus isolate on the cut end of individual branch sections. The overall seasonal change in the number of nematodes passing was similar among the five isolates. Peaks occurred in August and between December and February for four isolates. A virulent isolate, T-4, exhibited remarkable fluctuations, with the highest peak in August, suggesting that a test in August with T-4 is most informative. Another study showed that the passage ability, which is a measure of the dispersal ability in branches, was low for eggs and second-stage propagative juveniles and high for other developmental stages. This indicates that the composition of developmental stages in the inoculum is important when comparing the numbers of passing nematodes obtained on different occasions and in different places.

Characteristics of the emergence of Monochamus alternatus, the vector of Bursaphelenchus xylophilus (Nematoda: Aphelenchoididae), from Pinus thunbergii logs in Nanjing, China, and of the transmission of the nematodes through feeding wounds

Nematology ◽  
2007 ◽  
Vol 9 (6) ◽  
pp. 807-816 ◽  
Author(s):  
Peiyin Shen ◽  
Hongmei Li ◽  
Maurice Moens ◽  
Maosong Lin ◽  
Peng Fu
Keyword(s):  
Adult Emergence ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Pinus Thunbergii ◽  
Transmission Curve ◽  
Visual Estimation ◽  
Monochamus Alternatus ◽  
Peak Period ◽  
Nematode Transmission ◽  
Unimodal Pattern

AbstractThe pine wood nematode (PWN), Bursaphelenchus xylophilus, which in China and East Asia is vectored by Monochamus alternatus, is the causal agent of pine wilt disease of Japanese black pine, Pinus thunbergii. The vector emergence and the transmission of nematodes through vector feeding on pine twigs were monitored during 2004 and 2005 in Nanjing, China. The emergence started from late April to the end of June and peaked from late May to early June. There were 438 and 927 adults collected in 2004 and 2005, respectively, and approximately 70% of the beetles emerged during the peak period. Visual estimation of the nematode burden on vectors by observation of the atrium of the first abdominal spiracle gave unreliable information. The percentage of beetles carrying PWN of the total number of emerged beetles was between 20 and 30%. Seventy percent of the nematodes were released from beetle cadavers after 3 days of extraction. The sex and longevity of the beetle had no significant relationship with the number of nematodes that remained in cadavers. Transmission of nematodes into pine twigs through beetle feeding started 10 days after emergence of the vector. The period of nematode transmission could last for up to 79 days after beetle emergence. Two types of nematode-transmission curve were found by measuring the number of nematodes transmitted into pine twigs per day. The unimodal pattern peaked between 3 and 6 weeks after adult emergence; the bimodal pattern had two transmission peaks: one between 2 and 3 weeks, and another between 5 to 7 weeks after beetle emergence.

Is Bursaphelenchus mucronatus a weak pathogen to the Japanese red pine?

Nematology ◽  
2006 ◽  
Vol 8 (4) ◽  
pp. 485-489 ◽  
Author(s):  
Kazuyoshi Futai ◽  
Natsumi Kanzaki
Keyword(s):  
Pinus Densiflora ◽  
Pine Wilt Disease ◽  
Bursaphelenchus Xylophilus ◽  
Transmission Efficiency ◽  
Monochamus Alternatus ◽  
Disease Symptoms ◽  
Host Trees ◽  
Bursaphelenchus Mucronatus ◽  
Pine Trees ◽  
Mutualistic Relationship

AbstractThe pine wood nematode, Bursaphelenchus xylophilus, is the causal agent of pine wilt disease. It can invade healthy pine trees through the feeding wound of its vector beetle, Monochamus alternatus, and can enter a dead or dying tree through the vector's oviposition mark. By contrast, B. mucronatus, which is closely related to B. xylophilus but has been considered to have no pathogenicity to pine species, also invades living host trees through the same process as B. xylophilus, although the transmission efficiency is lower than that of B. xylophilus. In order to clarify the reason why Bursaphelenchus mucronatus invades a living host, we inoculated shaded and unshaded host seedlings of Pinus densiflora with B. mucronatus. Bursaphelenchus mucronatus displayed pathogenicity in the host seedlings but only with those that were shaded. In addition, B. mucronatus was recovered from the seedlings that had survived until the end of the experiment. Bursaphelenchus mucronatus seems to survive in the living healthy host tree without causing disease symptoms. These results suggest a mutualistic relationship between B. mucronatus and its vector beetle.

scholarly journals Cloning and expression of diadenosine 5′,5‴-P1,P4-tetraphosphate hydrolase from Lupinus angustifolius L

1998 ◽  
Vol 329 (2) ◽  
pp. 313-319 ◽  
Author(s):  
Danuta MAKSEL ◽  
Andrzej GURANOWSKI ◽  
C. Steven ILGOUTZ ◽  
Arthur MOIR ◽  
G. Michael BLACKBURN ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Sequence Similarity ◽  
Lupinus Angustifolius ◽  
Cloning And Expression ◽  
Lupinus Luteus ◽  
Higher Plant ◽  
Polyadenylated Rna

The first isolation, cloning and expression of cDNA encoding an asymmetric diadenosine 5ʹ,5‴P1,P4-tetraphosphate pyrophosphohydrolase (Ap4A hydrolase) from a higher plant is described. Ap4A hydrolase protein was purified from seeds of both Lupinus luteus and Lupinus angustifolius and partially sequenced. The Ap4A hydrolase cDNA was cloned from L. angustifolius cotyledonary polyadenylated RNA using reverse transcription and PCR with primers based on the amino acid sequence. The cDNA encoded a protein of 199 amino acids, molecular mass 22982 Da. When expressed in Escherichia coli fused to a maltose-binding protein, the enzyme catalysed asymmetric cleavage of Ap4A to AMP and ATP which was inhibited at concentrations of F- as low as 3 μM. These are properties characteristic of Ap4A hydrolase (asymmetrical) (EC 3.6.1.17). Comparison of the Ap4A hydrolase sequences derived from the four known cDNAs from pig, human, lupin and fission yeast showed that, like the mammalian hydrolase, the lupin enzyme possesses a Mut T motif but no other significant similarities. No sequence similarity to the human fragile histidine triad protein, as found in the Ap4A hydrolase from Schizosaccharomyces pombe, was detected in the Ap4A hydrolase from lupin.

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
Sign in / Sign up

Export Citation Format

Share Document