A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil
Rice water weevil, Lissorhoptrus brevirostris Suffrian (Coleoptera: Curculionidae), is one of the most important rice pests in Cuba. Here, we describe the production of transgenic rice plants expressing barley cystatin HvCPI-1 (Icy1 gene) to explore the potential of this protein for the control of r...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elfos Scientiae
|
| Series: | Biotecnología Aplicada |
| Subjects: | |
| Online Access: | http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S1027-28522009000400004&lng=en&tlng=en |
| _version_ | 1818083144157364224 |
|---|---|
| author | Raúl Armas Carlos Hernández Daymi Abreu Maylin Pérez Yeosvany Cabrera Merardo Pujol Julio Alfonso-Rubi |
| author_facet | Raúl Armas Carlos Hernández Daymi Abreu Maylin Pérez Yeosvany Cabrera Merardo Pujol Julio Alfonso-Rubi |
| author_sort | Raúl Armas |
| collection | DOAJ |
| description | Rice water weevil, Lissorhoptrus brevirostris Suffrian (Coleoptera: Curculionidae), is one of the most important rice pests in Cuba. Here, we describe the production of transgenic rice plants expressing barley cystatin HvCPI-1 (Icy1 gene) to explore the potential of this protein for the control of rice water weevil. Rice plants (Oryza sativa L. cv IACuba-28) were transformed via Agrobacterium tumefaciens with a plasmid carrying the Icy1 gene fused to the 35S promoter and the first exon/intron/exon from rice actin-1 gene. From 65 independent transgenic lines, 62 were positive in the PCR-Southern blot analyses. The transgene was correctly translated as indicated by western- and dot-blot assays with level of expression in T¹ plants of up to 2% of the total extracted protein. The functional integrity of the protein was confirmed in vitro by a reduction of up to 90% of the cysteine-proteinase activity in the gut of rice water weevils exposed to rice leaf extracts. Moreover, proteins extracted from T² transgenic rice roots showed a significant inhibition of up to 70% at pH 4.5 and 45% at pH 6.0 of the cathepsin B-like activity in the L. brevirostris larvae gut. These results demonstrate the potential of barley cystatin as an effective compound that may be combined with other pest control methods as an alternative in the struggle against insect resistance. |
| first_indexed | 2024-12-10T19:33:19Z |
| format | Article |
| id | doaj.art-f45a5461787848e6ad589be39f32cc4c |
| institution | Directory Open Access Journal |
| issn | 1027-2852 |
| language | English |
| last_indexed | 2024-12-10T19:33:19Z |
| publisher | Elfos Scientiae |
| record_format | Article |
| series | Biotecnología Aplicada |
| spelling | doaj.art-f45a5461787848e6ad589be39f32cc4c2022-12-22T01:36:12ZengElfos ScientiaeBiotecnología Aplicada1027-2852264321327S1027-28522009000400004A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevilRaúl Armas0Carlos Hernández1Daymi Abreu2Maylin Pérez3Yeosvany Cabrera4Merardo Pujol5Julio Alfonso-Rubi6Center for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyCenter for Genetic Engineering and BiotechnologyRice water weevil, Lissorhoptrus brevirostris Suffrian (Coleoptera: Curculionidae), is one of the most important rice pests in Cuba. Here, we describe the production of transgenic rice plants expressing barley cystatin HvCPI-1 (Icy1 gene) to explore the potential of this protein for the control of rice water weevil. Rice plants (Oryza sativa L. cv IACuba-28) were transformed via Agrobacterium tumefaciens with a plasmid carrying the Icy1 gene fused to the 35S promoter and the first exon/intron/exon from rice actin-1 gene. From 65 independent transgenic lines, 62 were positive in the PCR-Southern blot analyses. The transgene was correctly translated as indicated by western- and dot-blot assays with level of expression in T¹ plants of up to 2% of the total extracted protein. The functional integrity of the protein was confirmed in vitro by a reduction of up to 90% of the cysteine-proteinase activity in the gut of rice water weevils exposed to rice leaf extracts. Moreover, proteins extracted from T² transgenic rice roots showed a significant inhibition of up to 70% at pH 4.5 and 45% at pH 6.0 of the cathepsin B-like activity in the L. brevirostris larvae gut. These results demonstrate the potential of barley cystatin as an effective compound that may be combined with other pest control methods as an alternative in the struggle against insect resistance.http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S1027-28522009000400004&lng=en&tlng=enlissorhoptrus brevirostriscystatintransgenic ricegut proteinase activitycurculionids |
| spellingShingle | Raúl Armas Carlos Hernández Daymi Abreu Maylin Pérez Yeosvany Cabrera Merardo Pujol Julio Alfonso-Rubi A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil Biotecnología Aplicada lissorhoptrus brevirostris cystatin transgenic rice gut proteinase activity curculionids |
| title | A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| title_full | A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| title_fullStr | A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| title_full_unstemmed | A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| title_short | A barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| title_sort | barley cystatin stably expressed in rice exhibits strong in vitro inhibitory activity against gut proteinases of rice water weevil |
| topic | lissorhoptrus brevirostris cystatin transgenic rice gut proteinase activity curculionids |
| url | http://scielo.sld.cu/scielo.php?script=sci_arttext&pid=S1027-28522009000400004&lng=en&tlng=en |
| work_keys_str_mv | AT raularmas abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT carloshernandez abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT daymiabreu abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT maylinperez abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT yeosvanycabrera abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT merardopujol abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT julioalfonsorubi abarleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT raularmas barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT carloshernandez barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT daymiabreu barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT maylinperez barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT yeosvanycabrera barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT merardopujol barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil AT julioalfonsorubi barleycystatinstablyexpressedinriceexhibitsstronginvitroinhibitoryactivityagainstgutproteinasesofricewaterweevil |