Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe
To control Heterodera glycines Ichinohe (soybean cyst nematode) in Glycine max (L.) Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reprodu...
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| Veröffentlicht in: | Planta 2010-08, Vol.232 (3), p.775-785 |
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| creator | Li, Jiarui Todd, Timothy C Oakley, Tom R Lee, Junghoon Trick, Harold N |
| description | To control Heterodera glycines Ichinohe (soybean cyst nematode) in Glycine max (L.) Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reproduction or fitness (Cpn-1, Y25 and Prp-17) were PCR-amplified using specific primers and independently cloned into the pANDA35HK RNAi vector using a Gateway cloning strategy. Soybean roots were transformed with these constructions using a composite plant system. Confirmation of transformation was attained by PCR and Southern blot analysis. Transgene expression was detected using reverse transcription PCR (RT-PCR) and expression of siRNAs was confirmed in transgenic plants using northern blot analysis. Bioassays performed on transgenic composite plants expressing double-stranded RNA fragments of Cpn-1, Y25 and Prp-17 genes resulted in a 95, 81 and 79% reduction for eggs g⁻¹ root, respectively. Furthermore, we demonstrated a significant reduction in transcript levels of the Y25 and Prp-17 genes of the nematodes feeding on the transgenic roots via real-time RT-PCR whereas the expression of non-target genes were not affected. The results of this study demonstrate that over-expression of RNA interference constructs of nematode reproduction or fitness-related genes can effectively control H. glycines infection with levels of suppression comparable to conventional resistance. |
| doi_str_mv | 10.1007/s00425-010-1209-7 |
| format | Article |
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Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reproduction or fitness (Cpn-1, Y25 and Prp-17) were PCR-amplified using specific primers and independently cloned into the pANDA35HK RNAi vector using a Gateway cloning strategy. Soybean roots were transformed with these constructions using a composite plant system. Confirmation of transformation was attained by PCR and Southern blot analysis. Transgene expression was detected using reverse transcription PCR (RT-PCR) and expression of siRNAs was confirmed in transgenic plants using northern blot analysis. Bioassays performed on transgenic composite plants expressing double-stranded RNA fragments of Cpn-1, Y25 and Prp-17 genes resulted in a 95, 81 and 79% reduction for eggs g⁻¹ root, respectively. Furthermore, we demonstrated a significant reduction in transcript levels of the Y25 and Prp-17 genes of the nematodes feeding on the transgenic roots via real-time RT-PCR whereas the expression of non-target genes were not affected. The results of this study demonstrate that over-expression of RNA interference constructs of nematode reproduction or fitness-related genes can effectively control H. glycines infection with levels of suppression comparable to conventional resistance.</description><identifier>ISSN: 0032-0935</identifier><identifier>EISSN: 1432-2048</identifier><identifier>DOI: 10.1007/s00425-010-1209-7</identifier><identifier>PMID: 20582434</identifier><identifier>CODEN: PLANAB</identifier><language>eng</language><publisher>Berlin/Heidelberg: Berlin/Heidelberg : Springer-Verlag</publisher><subject>Agriculture ; Animals ; Base Sequence ; Bioassays ; Biological and medical sciences ; Biomedical and Life Sciences ; Blotting, Northern ; Blotting, Southern ; Cloning ; Composite plants ; DNA Primers ; Double stranded RNA ; Ecology ; Fecundity ; Fertility - genetics ; Forestry ; Fundamental and applied biological sciences. Psychology ; Genes ; Genetic vectors ; Glycine max ; Glycine max - parasitology ; Life Sciences ; Nematoda - genetics ; Nematoda - physiology ; Nematodes ; Original Article ; Phytopathology. Animal pests. Plant and forest protection ; Plant roots ; Plant Sciences ; Plants ; Plants, Genetically Modified ; Polymerase chain reaction ; Reverse Transcriptase Polymerase Chain Reaction ; Rhizobium rhizogenes ; RNA interference ; Roots ; Seedlings ; Soybeans ; Transgenic plants</subject><ispartof>Planta, 2010-08, Vol.232 (3), p.775-785</ispartof><rights>Springer-Verlag Berlin Heidelberg 2010</rights><rights>Springer-Verlag 2010</rights><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c446t-3e551040103558512f2e04f0ab9679e3895f0bfc509813f7418ceeb0adeb44d23</citedby><cites>FETCH-LOGICAL-c446t-3e551040103558512f2e04f0ab9679e3895f0bfc509813f7418ceeb0adeb44d23</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/23391740$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/23391740$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,776,780,799,27901,27902,41464,42533,51294,57992,58225</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23066939$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20582434$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, Jiarui</creatorcontrib><creatorcontrib>Todd, Timothy C</creatorcontrib><creatorcontrib>Oakley, Tom R</creatorcontrib><creatorcontrib>Lee, Junghoon</creatorcontrib><creatorcontrib>Trick, Harold N</creatorcontrib><title>Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe</title><title>Planta</title><addtitle>Planta</addtitle><addtitle>Planta</addtitle><description>To control Heterodera glycines Ichinohe (soybean cyst nematode) in Glycine max (L.) Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reproduction or fitness (Cpn-1, Y25 and Prp-17) were PCR-amplified using specific primers and independently cloned into the pANDA35HK RNAi vector using a Gateway cloning strategy. Soybean roots were transformed with these constructions using a composite plant system. Confirmation of transformation was attained by PCR and Southern blot analysis. Transgene expression was detected using reverse transcription PCR (RT-PCR) and expression of siRNAs was confirmed in transgenic plants using northern blot analysis. Bioassays performed on transgenic composite plants expressing double-stranded RNA fragments of Cpn-1, Y25 and Prp-17 genes resulted in a 95, 81 and 79% reduction for eggs g⁻¹ root, respectively. Furthermore, we demonstrated a significant reduction in transcript levels of the Y25 and Prp-17 genes of the nematodes feeding on the transgenic roots via real-time RT-PCR whereas the expression of non-target genes were not affected. The results of this study demonstrate that over-expression of RNA interference constructs of nematode reproduction or fitness-related genes can effectively control H. glycines infection with levels of suppression comparable to conventional resistance.</description><subject>Agriculture</subject><subject>Animals</subject><subject>Base Sequence</subject><subject>Bioassays</subject><subject>Biological and medical sciences</subject><subject>Biomedical and Life Sciences</subject><subject>Blotting, Northern</subject><subject>Blotting, Southern</subject><subject>Cloning</subject><subject>Composite plants</subject><subject>DNA Primers</subject><subject>Double stranded RNA</subject><subject>Ecology</subject><subject>Fecundity</subject><subject>Fertility - genetics</subject><subject>Forestry</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genes</subject><subject>Genetic vectors</subject><subject>Glycine max</subject><subject>Glycine max - parasitology</subject><subject>Life Sciences</subject><subject>Nematoda - genetics</subject><subject>Nematoda - physiology</subject><subject>Nematodes</subject><subject>Original Article</subject><subject>Phytopathology. Animal pests. Plant and forest protection</subject><subject>Plant roots</subject><subject>Plant Sciences</subject><subject>Plants</subject><subject>Plants, Genetically Modified</subject><subject>Polymerase chain reaction</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Rhizobium rhizogenes</subject><subject>RNA interference</subject><subject>Roots</subject><subject>Seedlings</subject><subject>Soybeans</subject><subject>Transgenic plants</subject><issn>0032-0935</issn><issn>1432-2048</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNp9kU2P0zAQhi0EYruFH8ABiJAQp8D4K4mPaMXSlVbiAHuOHGfcTZXaxU5A_fdMlcIiDpw81vu888nYCw7vOUD9IQMooUvgUHIBpqwfsRVXUpQCVPOYrQAoBiP1BbvMeQdAYl0_ZRcCdCOUVCv2cxPzVPaYhh_YF3k-HBLmPMRQRF8E3Nsp9lgkPKTYz24iqrChL_wwBeKKLdJT9OgS2kyRRzeHfpiOJ_sGJyQbJltsx6MbTuiNux9CvMdn7Im3Y8bn53fN7q4_fbvalLdfPt9cfbwtnVLVVErUmoOiCaXWjebCCwTlwXamqg3KxmgPnXcaTMOlrxVvHGIHtsdOqV7INXu35KUBvs-Yp3Y_ZIfjaAPGObe1lKZuaBtEvvmH3MU5BWqurbhsJGja3prxBXIp5pzQt4c07G06thza003a5SYtnP50EyqwZq_Oieduj_0fx-8jEPD2DNjs7OiTDW7ID5yEqjLSECcWLpMUtpgeOvxf9ZeLaZenmP5KKg2vFZD-etG9ja3dJip891UAl8CbqqbJ5S9EIrc9</recordid><startdate>20100801</startdate><enddate>20100801</enddate><creator>Li, Jiarui</creator><creator>Todd, Timothy C</creator><creator>Oakley, Tom R</creator><creator>Lee, Junghoon</creator><creator>Trick, Harold N</creator><general>Berlin/Heidelberg : Springer-Verlag</general><general>Springer-Verlag</general><general>Springer</general><general>Springer Nature B.V</general><scope>FBQ</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QP</scope><scope>7QR</scope><scope>7TM</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20100801</creationdate><title>Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe</title><author>Li, Jiarui ; Todd, Timothy C ; Oakley, Tom R ; Lee, Junghoon ; Trick, Harold N</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c446t-3e551040103558512f2e04f0ab9679e3895f0bfc509813f7418ceeb0adeb44d23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Agriculture</topic><topic>Animals</topic><topic>Base Sequence</topic><topic>Bioassays</topic><topic>Biological and medical sciences</topic><topic>Biomedical and Life Sciences</topic><topic>Blotting, Northern</topic><topic>Blotting, Southern</topic><topic>Cloning</topic><topic>Composite plants</topic><topic>DNA Primers</topic><topic>Double stranded RNA</topic><topic>Ecology</topic><topic>Fecundity</topic><topic>Fertility - genetics</topic><topic>Forestry</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genes</topic><topic>Genetic vectors</topic><topic>Glycine max</topic><topic>Glycine max - parasitology</topic><topic>Life Sciences</topic><topic>Nematoda - genetics</topic><topic>Nematoda - physiology</topic><topic>Nematodes</topic><topic>Original Article</topic><topic>Phytopathology. Animal pests. Plant and forest protection</topic><topic>Plant roots</topic><topic>Plant Sciences</topic><topic>Plants</topic><topic>Plants, Genetically Modified</topic><topic>Polymerase chain reaction</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Rhizobium rhizogenes</topic><topic>RNA interference</topic><topic>Roots</topic><topic>Seedlings</topic><topic>Soybeans</topic><topic>Transgenic plants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Jiarui</creatorcontrib><creatorcontrib>Todd, Timothy C</creatorcontrib><creatorcontrib>Oakley, Tom R</creatorcontrib><creatorcontrib>Lee, Junghoon</creatorcontrib><creatorcontrib>Trick, Harold N</creatorcontrib><collection>AGRIS</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Planta</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Jiarui</au><au>Todd, Timothy C</au><au>Oakley, Tom R</au><au>Lee, Junghoon</au><au>Trick, Harold N</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe</atitle><jtitle>Planta</jtitle><stitle>Planta</stitle><addtitle>Planta</addtitle><date>2010-08-01</date><risdate>2010</risdate><volume>232</volume><issue>3</issue><spage>775</spage><epage>785</epage><pages>775-785</pages><issn>0032-0935</issn><eissn>1432-2048</eissn><coden>PLANAB</coden><abstract>To control Heterodera glycines Ichinohe (soybean cyst nematode) in Glycine max (L.) Merr. (soybean), we evaluated the use of producing transgenic soybean seedlings expressing small interfering RNAs (siRNAs) against specific H. glycines genes. Gene fragments of three genes related to nematode reproduction or fitness (Cpn-1, Y25 and Prp-17) were PCR-amplified using specific primers and independently cloned into the pANDA35HK RNAi vector using a Gateway cloning strategy. Soybean roots were transformed with these constructions using a composite plant system. Confirmation of transformation was attained by PCR and Southern blot analysis. Transgene expression was detected using reverse transcription PCR (RT-PCR) and expression of siRNAs was confirmed in transgenic plants using northern blot analysis. Bioassays performed on transgenic composite plants expressing double-stranded RNA fragments of Cpn-1, Y25 and Prp-17 genes resulted in a 95, 81 and 79% reduction for eggs g⁻¹ root, respectively. Furthermore, we demonstrated a significant reduction in transcript levels of the Y25 and Prp-17 genes of the nematodes feeding on the transgenic roots via real-time RT-PCR whereas the expression of non-target genes were not affected. The results of this study demonstrate that over-expression of RNA interference constructs of nematode reproduction or fitness-related genes can effectively control H. glycines infection with levels of suppression comparable to conventional resistance.</abstract><cop>Berlin/Heidelberg</cop><pub>Berlin/Heidelberg : Springer-Verlag</pub><pmid>20582434</pmid><doi>10.1007/s00425-010-1209-7</doi><tpages>11</tpages></addata></record> |
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| subjects | Agriculture Animals Base Sequence Bioassays Biological and medical sciences Biomedical and Life Sciences Blotting, Northern Blotting, Southern Cloning Composite plants DNA Primers Double stranded RNA Ecology Fecundity Fertility - genetics Forestry Fundamental and applied biological sciences. Psychology Genes Genetic vectors Glycine max Glycine max - parasitology Life Sciences Nematoda - genetics Nematoda - physiology Nematodes Original Article Phytopathology. Animal pests. Plant and forest protection Plant roots Plant Sciences Plants Plants, Genetically Modified Polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction Rhizobium rhizogenes RNA interference Roots Seedlings Soybeans Transgenic plants |
| title | Host-derived suppression of nematode reproductive and fitness genes decreases fecundity of Heterodera glycines Ichinohe |
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