The role of <i>mab‐3</i> in spermatogenesis and ontogenesis of pinewood nematode, <i>Bursaphelenchus xylophilus</i>

Zhou, Lifeng; Ma, Xinxin; Zhu, Najie; Zou, Qingchi; Guo, Kai; Bai, Liqun; Yu, Hongshi; Hu, Jiafu

doi:10.1002/ps.6001

Cited by 13 publications

(9 citation statements)

References 67 publications

(116 reference statements)

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“…In recent years, the sex-determining genes of B. xylophilus have been extensively studied. For example, mab-3 is essential for spermatogenesis in B. xylophilus ( Zhou et al, 2020 ). In addition, more and more evidence indicates that the sex determination mechanism of B. xylophilus is also affected by environmental factors, such as nutrient conditions ( Wei et al, 2021 ) and volatile substances concentration ( Cui et al, 2018 ).…”

Section: Discussionmentioning

confidence: 99%

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Regulatory roles of Bxy-laf-1 in reproductive behaviour of Bursaphelenchus xylophilus

Sun¹,

Wang

Liu³

et al. 2022

Front. Physiol.

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Bursaphelenchus xylophilu is a worldwide quarantine nematode, causing huge economic losses and ecological disasters in many countries. The sex ratio of B. xylophilus plays an important role in the nematode infestation. The laf-1-related genes are highly conserved in animals, playing crucial roles in sex determination. Therefore, we investigated the expression pattern and biological function of its orthologue, Bxy-laf-1 in B. xylophilus. Bxy-laf-1 has two typical conserved DNA-binding domains, DEAD and Helicase C. The real-time quantitative PCR data revealed that Bxy-laf-1 expression was required throughout the entire life of B. xylophilus, with the maximum expression in the J2 stage and the lowest expression in the adult stage. mRNA in situ hybridization showed that Bxy-laf-1 is mainly located in the cephalopharynx and reproductive organs of B. xylophilus. RNA interference (RNAi) indicated that the head swing frequency was dramatically decreased. The RNA interference results displayed that a significant reduction in motility was observed in the hatched larvae. The female to male sex ratio was also decreased in the F0 and F1 generations, but recovered in the F2 generation. The tail of female adults with eggs in the belly appeared deformities. This phenomenon appeared in the F0 and F1 generations, but recovered in the F2 generation. Bxy-laf-1 is a typical sex-determination gene with distinct expression patterns in males and females. As demonstrated in other species, the sex ratio was altered after knocking down Bxy-laf-1 expression. The results of this study contribute to our understanding of the molecular processes of Bxy-laf-1 in B. xylophilus, which may provide clues about how to control pine wilt disease by inhibiting ontogenic growth and reducing nematode fertility.

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Section: Discussionmentioning

confidence: 99%

“…Under a microscope, a virgin male and virgin female were placed in a drop of water on a concave glass slide. In a humidifier, the slides were stored in the dark at 25°C ( Zhou et al, 2020 ). A zoom stereomicroscope (Zeiss, Thornwood, United States) was used to investigate the mating behavior of male and female adults.…”

Section: Methodsmentioning

confidence: 99%

Regulatory roles of Bxy-laf-1 in reproductive behaviour of Bursaphelenchus xylophilus

Sun¹,

Wang

Liu³

et al. 2022

Front. Physiol.

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“…When a large number of females were in an egg-bearing state, nematodes were separated and added to sterilized double-distilled water (ddH 2 O), and eggs were laid in an incubator set at 25°C for 1 h. The nematode suspension was gently removed and washed once or twice, and synchronized nematode embryos were obtained by adding ddH 2 O to the Petri dishes. Synchronized second-stage juvenile (J2) larvae were obtained by adding ddH 2 O to the synchronized embryos and placing them in an incubator at 25°C for 24 h. Synchronized third and fourth parasitic stage (J3 and J4) larvae and adults were obtained by culturing synchronized J2 larvae on gray mold plates and incubating them at 25°C for 36, 48, and 60 h, respectively ( Zhou et al, 2021 ).…”

Section: Methodsmentioning

confidence: 99%

“…They discovered that the Bxy-fuca gene is primarily expressed in the body wall muscles and intestine. Subsequently, Zhou et al (2021) also successfully utilized whole-mount in situ hybridization for intact nematode using DIG-labeled single-stranded RNA (ss RNA) probes. Wen et al (2021) applied the DIG-labeled antisense cDNA probes and the whole-mount in situ hybridization to the intact pine wood nematode and demonstrated that BxSCD1 was specifically expressed in the dorsal glands and intestine of PWN.…”

Section: Introductionmentioning

confidence: 99%

Comparative effects of two in situ hybridization methods for the pinewood nematode (Bursaphelenchus xylophilus)

Wang,

Zhuge,

Tang

et al. 2023

Front. Microbiol.

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The gene localization technique of Bursaphelenchus xylophilus (pinewood nematode, PWN) is used for study gene expression in PWNs. Two in situ hybridization methods, namely, whole-mount in situ hybridization and the cut-off method are used widely. To compare the effects of these two in situ hybridization methods, the present study investigated the patterns of two functional genes expression in PWNs. The Bx-vap-2 gene (GenBank accession number: OR228482), related to pathogenicity, and the fem-2 gene (GenBank accession number: OR228481), related to sex determination, were selected to map related genes in the whole-mount and amputated PWNs at different ages using these in situ hybridization methods. Based on the overall statistical comparison, we found that compared to the cut-off method, the whole-mount method exhibited higher staining rates and correct staining rates for the fem-2 gene and Bx-vap-2 gene. However, considering the correct staining aspect, the cut-off method yielded better staining effects on pinewood nematode sections than the whole-mount method, with clearer hybridization signal locations and less non-specific staining. In other words, the cut-off method demonstrated more precise gene localization. Both methods are applicable for gene localization, but considering the overall staining pattern, analysis of experimental results, and comprehensive experimental operations, we believe that the whole-mount method is more suitable for gene localization and expression analysis of development-related genes in pinewood nematodes. This is because intact pinewood nematodes are better suited for showcasing the continuous developmental process of development-related genes. On the other hand, considering the experimental time, accuracy of staining site, and the amount of non-specific staining, the cut-off method is more suitable for disease-related genes. Additionally, to achieve better performance, the cut-off method can be selectively applied to samples during the experimental process.

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“…B. xylophilus strain used in the current study was initially isolated from the wilted Pinus massoniana in China, and then raised on the mycelia of Botrytis cinerea (non-sporulating strain) grown on potato dextrose agar (PDA) Petri dishes at 25 °C. C. elegans N2 strain was raised on nematode growth medium Petri dishes inoculated with Escherichia coli OP50 strain at 20 °C 36 . Nematodes were extracted from the Petri dishes via Baermann funnel technique 37 .…”

Section: Methodsmentioning

confidence: 99%

madd-4 plays a critical role in light against Bursaphelenchus xylophilus

Zhou

Sheng

Zhang

et al. 2022

Sci Rep

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Bursaphelenchus xylophilus is a notorious invasive species, causing extensive losses to pine ecosystems globally. Previous studies had shown that the development of B. xylophilus was seriously suppressed by light. However, the mechanism involved in the inhibition is unknown. Here, it is the first report that Bxy-madd-4 is a light-regulated gene, plays a potential role in B. xylophilus in responding to the blue light. Transcriptome sequencing revealed that the expression level of Bxy-madd-4 declined by 86.39% under blue light. The reverse transcription quantitative real-time PCR results were in accord with the transcriptome sequencing, confirming the expression level of Bxy-madd-4 was suppressed by blue light. Bxy-madd-4 promoter::mCherry reporter constructed in Caenorhabditis elegans were utilized to mimic the spatiotemporal expression patterns of Bxy-madd-4. Bxy-madd-4A promoter activity had a strong continuity throughout all development stages in C. elegans. Further RNA interference indicated that only 36.8% of the Bxy-madd-4 dsRNA treated embryos were hatched. Moreover, 71.6% of the hatched nematodes were abnormal, such as particles on the body surface and concave tissues. Our findings contribute towards a better understanding of the mechanism of light against the destructive invasive nematode, providing a promising hint for control of the destructive invasive nematode.

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The role of mab‐3 in spermatogenesis and ontogenesis of pinewood nematode, Bursaphelenchus xylophilus

Cited by 13 publications

References 67 publications

Regulatory roles of Bxy-laf-1 in reproductive behaviour of Bursaphelenchus xylophilus

Regulatory roles of Bxy-laf-1 in reproductive behaviour of Bursaphelenchus xylophilus

Comparative effects of two in situ hybridization methods for the pinewood nematode (Bursaphelenchus xylophilus)

madd-4 plays a critical role in light against Bursaphelenchus xylophilus

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