Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants
<i>Meloidogyne incognita</i> is a devastating plant parasitic nematode that causes root knot disease in a wide range of plants. In the present study, we investigated host-induced RNA interference (RNAi) gene silencing of chitin biosynthesis pathway genes (chitin synthase, glucose-6-phosp...
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MDPI AG
2020-09-01
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| Series: | International Journal of Molecular Sciences |
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| author | Vimalraj Mani Chinreddy Subramanyam Reddy Seon-Kyeong Lee Soyoung Park Hyoung-Rai Ko Dong-Gwan Kim Bum-Soo Hahn |
| author_facet | Vimalraj Mani Chinreddy Subramanyam Reddy Seon-Kyeong Lee Soyoung Park Hyoung-Rai Ko Dong-Gwan Kim Bum-Soo Hahn |
| author_sort | Vimalraj Mani |
| collection | DOAJ |
| description | <i>Meloidogyne incognita</i> is a devastating plant parasitic nematode that causes root knot disease in a wide range of plants. In the present study, we investigated host-induced RNA interference (RNAi) gene silencing of chitin biosynthesis pathway genes (chitin synthase, glucose-6-phosphate isomerase, and trehalase) in transgenic tobacco plants. To develop an RNAi vector, ubiquitin (UBQ1) promoter was directly cloned, and to generate an RNAi construct, expression of three genes was suppressed using the GATEWAY system. Further, transgenic <i>Nicotiana benthamiana</i> lines expressing dsRNA for chitin synthase (CS), glucose-6-phosphate isomerase (GPI), and trehalase 1 (TH1) were generated. Quantitative PCR analysis confirmed endogenous mRNA expression of root knot nematode (RKN) and revealed that all three genes were more highly expressed in the female stage than in eggs and in the parasitic stage. In vivo, transformed roots were challenged with <i>M</i>. <i>incognita</i>. The number of eggs and root knots were significantly decreased by 60–90% in RNAi transgenic lines. As evident, root galls obtained from transgenic RNAi lines exhibited 0.01- to 0.70-fold downregulation of transcript levels of targeted genes compared with galls isolated from control plants. Furthermore, phenotypic characteristics such as female size and width were also marginally altered, while effect of egg mass per egg number in RNAi transgenic lines was reduced. These results indicate the relevance and significance of targeting chitin biosynthesis genes during the nematode lifespan. Overall, our results suggest that further developments in RNAi efficiency in commercially valued crops can be applied to employ RNAi against other plant parasitic nematodes. |
| first_indexed | 2024-03-10T16:25:54Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T16:25:54Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-dc06b91acd5043db950bde386bd31ff02023-11-20T13:15:22ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-09-012118662610.3390/ijms21186626Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco PlantsVimalraj Mani0Chinreddy Subramanyam Reddy1Seon-Kyeong Lee2Soyoung Park3Hyoung-Rai Ko4Dong-Gwan Kim5Bum-Soo Hahn6Department of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, KoreaDepartment of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, KoreaDepartment of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, KoreaDepartment of Agricultural Biotechnology, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, KoreaCrop Protection Division, National Institute of Agricultural Sciences, Rural Development Administration, Wanju 55365, KoreaDepartment of Bio-Industry and Bio-Resource Engineering, Sejong University, Seoul 05006, KoreaNational Agrobiodiversity Center, National Institute of Agricultural Sciences, Rural Development Administration, Jeonju 54874, Korea<i>Meloidogyne incognita</i> is a devastating plant parasitic nematode that causes root knot disease in a wide range of plants. In the present study, we investigated host-induced RNA interference (RNAi) gene silencing of chitin biosynthesis pathway genes (chitin synthase, glucose-6-phosphate isomerase, and trehalase) in transgenic tobacco plants. To develop an RNAi vector, ubiquitin (UBQ1) promoter was directly cloned, and to generate an RNAi construct, expression of three genes was suppressed using the GATEWAY system. Further, transgenic <i>Nicotiana benthamiana</i> lines expressing dsRNA for chitin synthase (CS), glucose-6-phosphate isomerase (GPI), and trehalase 1 (TH1) were generated. Quantitative PCR analysis confirmed endogenous mRNA expression of root knot nematode (RKN) and revealed that all three genes were more highly expressed in the female stage than in eggs and in the parasitic stage. In vivo, transformed roots were challenged with <i>M</i>. <i>incognita</i>. The number of eggs and root knots were significantly decreased by 60–90% in RNAi transgenic lines. As evident, root galls obtained from transgenic RNAi lines exhibited 0.01- to 0.70-fold downregulation of transcript levels of targeted genes compared with galls isolated from control plants. Furthermore, phenotypic characteristics such as female size and width were also marginally altered, while effect of egg mass per egg number in RNAi transgenic lines was reduced. These results indicate the relevance and significance of targeting chitin biosynthesis genes during the nematode lifespan. Overall, our results suggest that further developments in RNAi efficiency in commercially valued crops can be applied to employ RNAi against other plant parasitic nematodes.https://www.mdpi.com/1422-0067/21/18/6626root knot nematode<i>Meloidogyne incognita</i>chitin biosynthesisRNA-interferencetransgenic plants |
| spellingShingle | Vimalraj Mani Chinreddy Subramanyam Reddy Seon-Kyeong Lee Soyoung Park Hyoung-Rai Ko Dong-Gwan Kim Bum-Soo Hahn Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants International Journal of Molecular Sciences root knot nematode <i>Meloidogyne incognita</i> chitin biosynthesis RNA-interference transgenic plants |
| title | Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants |
| title_full | Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants |
| title_fullStr | Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants |
| title_full_unstemmed | Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants |
| title_short | Chitin Biosynthesis Inhibition of <i>Meloidogyne incognita</i> by RNAi-Mediated Gene Silencing Increases Resistance to Transgenic Tobacco Plants |
| title_sort | chitin biosynthesis inhibition of i meloidogyne incognita i by rnai mediated gene silencing increases resistance to transgenic tobacco plants |
| topic | root knot nematode <i>Meloidogyne incognita</i> chitin biosynthesis RNA-interference transgenic plants |
| url | https://www.mdpi.com/1422-0067/21/18/6626 |
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