Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development
Pine wilt disease (PWD) caused by the pine wood nematode (PWN), <i>Bursaphelenchus xylophilus</i>, is a globally distributed destructive disease of pine forest. To study the PWD pathogenic mechanism, the cinnamyl alcohol dehydrogenase gene (<i>BxCAD-1</i>) from <i>B. xy...
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MDPI AG
2023-07-01
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| Series: | Forests |
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| Online Access: | https://www.mdpi.com/1999-4907/14/7/1379 |
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| author | Guosong Dong Qunqun Guo Wenjun Deng Yu Zhang Hongzheng Tai Chao Wang Guicai Du Ronggui Li |
| author_facet | Guosong Dong Qunqun Guo Wenjun Deng Yu Zhang Hongzheng Tai Chao Wang Guicai Du Ronggui Li |
| author_sort | Guosong Dong |
| collection | DOAJ |
| description | Pine wilt disease (PWD) caused by the pine wood nematode (PWN), <i>Bursaphelenchus xylophilus</i>, is a globally distributed destructive disease of pine forest. To study the PWD pathogenic mechanism, the cinnamyl alcohol dehydrogenase gene (<i>BxCAD-1</i>) from <i>B. xylophilus</i> was selected. The <i>BxCAD-1</i> gene was amplified by PCR from the cDNA library of <i>B. xylophilus</i> and cloned into the expression vector pET-15b to construct the recombinant vector pET-15b-<i>BxCAD-1</i>. The recombinant cinnamyl alcohol dehydrogenase protein was purified by Ni-NTA affinity chromatography from <i>Escherichia coli</i> BL21 (DE3) harboring pET-15b-<i>BxCAD-1</i> induced by IPTG. The effects of pH, temperature, metal ions and substrates on the activity of BxCAD-1 were determined, showing the highest catalytic activity at pH 8.0 and 40 °C with cinnamyl alcohol as substrate and Zn<sup>2+</sup> as an activator. To elucidate the functions of <i>BxCAD-1</i> in <i>B. xylophilus</i>, the expression of the gene was down-regulated by RNA interference. Results showed that the movement, feeding, reproduction, spawning rate, hatching rate, lifespan, infectivity and sensitivity to ethanol decreased compared with negative controls. RNA interference also affected the development of <i>B. xylophilus</i> from the larval stage to the adult stage. In situ hybridization showed that the gene was expressed in the digestive tract of male and female adults. This study revealed a promising target for PWD control. |
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| language | English |
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| publishDate | 2023-07-01 |
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| series | Forests |
| spelling | doaj.art-5e26809b3b1145e08310d06f7ad5102a2023-11-18T19:23:25ZengMDPI AGForests1999-49072023-07-01147137910.3390/f14071379Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and DevelopmentGuosong Dong0Qunqun Guo1Wenjun Deng2Yu Zhang3Hongzheng Tai4Chao Wang5Guicai Du6Ronggui Li7College of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaCollege of Life Science, Qingdao University, Qingdao 266071, ChinaPine wilt disease (PWD) caused by the pine wood nematode (PWN), <i>Bursaphelenchus xylophilus</i>, is a globally distributed destructive disease of pine forest. To study the PWD pathogenic mechanism, the cinnamyl alcohol dehydrogenase gene (<i>BxCAD-1</i>) from <i>B. xylophilus</i> was selected. The <i>BxCAD-1</i> gene was amplified by PCR from the cDNA library of <i>B. xylophilus</i> and cloned into the expression vector pET-15b to construct the recombinant vector pET-15b-<i>BxCAD-1</i>. The recombinant cinnamyl alcohol dehydrogenase protein was purified by Ni-NTA affinity chromatography from <i>Escherichia coli</i> BL21 (DE3) harboring pET-15b-<i>BxCAD-1</i> induced by IPTG. The effects of pH, temperature, metal ions and substrates on the activity of BxCAD-1 were determined, showing the highest catalytic activity at pH 8.0 and 40 °C with cinnamyl alcohol as substrate and Zn<sup>2+</sup> as an activator. To elucidate the functions of <i>BxCAD-1</i> in <i>B. xylophilus</i>, the expression of the gene was down-regulated by RNA interference. Results showed that the movement, feeding, reproduction, spawning rate, hatching rate, lifespan, infectivity and sensitivity to ethanol decreased compared with negative controls. RNA interference also affected the development of <i>B. xylophilus</i> from the larval stage to the adult stage. In situ hybridization showed that the gene was expressed in the digestive tract of male and female adults. This study revealed a promising target for PWD control.https://www.mdpi.com/1999-4907/14/7/1379<i>Bursaphelenchus xylophilus</i>RNAicinnamyl alcohol dehydrogenasefunctions |
| spellingShingle | Guosong Dong Qunqun Guo Wenjun Deng Yu Zhang Hongzheng Tai Chao Wang Guicai Du Ronggui Li Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development Forests <i>Bursaphelenchus xylophilus</i> RNAi cinnamyl alcohol dehydrogenase functions |
| title | Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development |
| title_full | Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development |
| title_fullStr | Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development |
| title_full_unstemmed | Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development |
| title_short | Cinnamyl Alcohol Dehydrogenase Gene Regulates <i>Bursaphelenchus xylophilus</i> Reproduction and Development |
| title_sort | cinnamyl alcohol dehydrogenase gene regulates i bursaphelenchus xylophilus i reproduction and development |
| topic | <i>Bursaphelenchus xylophilus</i> RNAi cinnamyl alcohol dehydrogenase functions |
| url | https://www.mdpi.com/1999-4907/14/7/1379 |
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