Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference
RNA interference is of value in determining gene function in many organisms. Plant parasitic nematodes are refractory to microinjection as a means of introducing RNA and do not show any oral uptake until they are within plants. We have used octopamine to stimulate uptake by prepara-sitic second stag...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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The American Phytopathological Society
2002-08-01
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| Series: | Molecular Plant-Microbe Interactions |
| Online Access: | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.8.747 |
| _version_ | 1811264532349190144 |
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| author | P. E. Urwin Catherine J. Lilley Howard J. Atkinson |
| author_facet | P. E. Urwin Catherine J. Lilley Howard J. Atkinson |
| author_sort | P. E. Urwin |
| collection | DOAJ |
| description | RNA interference is of value in determining gene function in many organisms. Plant parasitic nematodes are refractory to microinjection as a means of introducing RNA and do not show any oral uptake until they are within plants. We have used octopamine to stimulate uptake by prepara-sitic second stage juveniles of two cyst nematodes, Heterodera glycines and Globodera pallida. This new technique was used to facilitate uptake of double stranded RNA (dsRNA) together with fluoroscein isothiocyanate as a visual marker. Targeting cysteine proteinases did not reduce the number of parasites but caused a shift from the normal female/male ratio of 3:1 to 1:1 by 14 days postinfection (dpi). Exposure of H. glycines to dsRNA corresponding to a newly characterized protein with homology to C-type lectins did not affect sexual fate, but 41% fewer parasites were recovered from the plants. As expected, treatment with dsRNA corresponding to the major sperm protein (MSP) had no effect on either parasite development or sexual fate over 14 days. Northern analysis showed lower transcript abundance for the two targeted mRNAs that occur in J2, plus a later inhibition for MSP transcripts when males developed sperm at 15 dpi. These findings establish a procedure for RNAi of plant parasitic nematodes. |
| first_indexed | 2024-04-12T20:06:03Z |
| format | Article |
| id | doaj.art-af53c45bc85b477b831257c08050d37e |
| institution | Directory Open Access Journal |
| issn | 0894-0282 1943-7706 |
| language | English |
| last_indexed | 2024-04-12T20:06:03Z |
| publishDate | 2002-08-01 |
| publisher | The American Phytopathological Society |
| record_format | Article |
| series | Molecular Plant-Microbe Interactions |
| spelling | doaj.art-af53c45bc85b477b831257c08050d37e2022-12-22T03:18:24ZengThe American Phytopathological SocietyMolecular Plant-Microbe Interactions0894-02821943-77062002-08-0115874775210.1094/MPMI.2002.15.8.747Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA InterferenceP. E. UrwinCatherine J. LilleyHoward J. AtkinsonRNA interference is of value in determining gene function in many organisms. Plant parasitic nematodes are refractory to microinjection as a means of introducing RNA and do not show any oral uptake until they are within plants. We have used octopamine to stimulate uptake by prepara-sitic second stage juveniles of two cyst nematodes, Heterodera glycines and Globodera pallida. This new technique was used to facilitate uptake of double stranded RNA (dsRNA) together with fluoroscein isothiocyanate as a visual marker. Targeting cysteine proteinases did not reduce the number of parasites but caused a shift from the normal female/male ratio of 3:1 to 1:1 by 14 days postinfection (dpi). Exposure of H. glycines to dsRNA corresponding to a newly characterized protein with homology to C-type lectins did not affect sexual fate, but 41% fewer parasites were recovered from the plants. As expected, treatment with dsRNA corresponding to the major sperm protein (MSP) had no effect on either parasite development or sexual fate over 14 days. Northern analysis showed lower transcript abundance for the two targeted mRNAs that occur in J2, plus a later inhibition for MSP transcripts when males developed sperm at 15 dpi. These findings establish a procedure for RNAi of plant parasitic nematodes.https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.8.747 |
| spellingShingle | P. E. Urwin Catherine J. Lilley Howard J. Atkinson Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference Molecular Plant-Microbe Interactions |
| title | Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference |
| title_full | Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference |
| title_fullStr | Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference |
| title_full_unstemmed | Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference |
| title_short | Ingestion of Double-Stranded RNA by Preparasitic Juvenile Cyst Nematodes Leads to RNA Interference |
| title_sort | ingestion of double stranded rna by preparasitic juvenile cyst nematodes leads to rna interference |
| url | https://apsjournals.apsnet.org/doi/10.1094/MPMI.2002.15.8.747 |
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