Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)
In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for imp...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
University of Debrecen
2014-04-01
|
| Series: | International Journal of Horticultural Science |
| Subjects: | |
| Online Access: | https://ojs.lib.unideb.hu/IJHS/article/view/1111 |
| _version_ | 1828230719343689728 |
|---|---|
| author | N. M. Ali Bacha M. Batha A. M. Abdul-Kader |
| author_facet | N. M. Ali Bacha M. Batha A. M. Abdul-Kader |
| author_sort | N. M. Ali Bacha |
| collection | DOAJ |
| description | In the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants.
Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of 0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance . |
| first_indexed | 2024-04-12T18:52:51Z |
| format | Article |
| id | doaj.art-3974a06232f04a28b5689351de63cc87 |
| institution | Directory Open Access Journal |
| issn | 1585-0404 2676-931X |
| language | English |
| last_indexed | 2024-04-12T18:52:51Z |
| publishDate | 2014-04-01 |
| publisher | University of Debrecen |
| record_format | Article |
| series | International Journal of Horticultural Science |
| spelling | doaj.art-3974a06232f04a28b5689351de63cc872022-12-22T03:20:25ZengUniversity of DebrecenInternational Journal of Horticultural Science1585-04042676-931X2014-04-01201-210.31421/IJHS/20/1-2/1111Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae)N. M. Ali Bacha0M. Batha1A. M. Abdul-Kader2General Commission for Scientific Agricultural Research (GCSAR), Biotechnology Department, Douma, P.O. Box 113, Damascus, SyriaDamascus University, Faculty of Agriculture, Department of Horticulture, Damascus, P.O. Box 30621, SyriaGeneral Commission for Scientific Agricultural Research (GCSAR), Biotechnology Department, Douma, P.O. Box 113, Damascus, SyriaIn the present study, g2ps1 gene from Gerbera hybrida coding for 2-pyrone synthase which contribute for fungal and insect resistance was used. The aim was to work out an efficient approach of genetic transformation for apple cvs. ‘Golden Delicious’, ‘Royal Gala’ and ‘MM111’, ‘M26’ rootstocks for improving their fungal resistance using genetic engineering techniques. Adventitious shoot formation from leaf pieces of apples studied was achieved using middle leaf segments taken from the youngest leaves from in vitro-grown plants. Optimum conditions for ‚direct’ shoot organogenesis resulted in high regeneration efficiency of 0%, 95%, 92%, 94% in the studied apples respectively. Putative transgenic shoots could be obtained on MS media with B5 Vitamins, 5.0 mg l-1 BAP, or 2.0 mg l-1 TDZ with 0.2 mg l-1 NAA in the presence of the selection agent “PPT” at 3.0-5.0 mgl-1. Shoot multiplication of transgenic shoots was achieved on: MS + B5 vitamins + 1.0 mg l-1 BAP + 0.3 mg l-1 IBA, 0.2 mg l-1 GA3+1.0 g/l MES+ 30 g/l sucrose + 7.0 g/l Agar, with the selection agent PPT at 5.0 mg l-1 and were subcultured every 4 weeks in order to get sufficient material to confirm transformation of the putative shoots obtained. Six, seven, one and six transgenic clones of the apples studied respectively have been obtained and confirmed by selection on the media containing the selection agent “PPT” and by PCR analysis using the suitable primers in all clones obtained for the presence of the selection” bar gene (447 bp) and the gene-of- interest “g2PS1” (1244 bp), with transformation efficiency of 0.4%, 0.6%, 0.1% and 0.3% respectively. These transgenic clones were multiplied further in vitro in the presence of the selection agent ‘PPT’ and rooted in vitro. Rooted transgenic plantlets were successfully acclimatized and are being kept under-containment conditions according to the biosafety by-law in Syria to evaluate their performance for fungal resistance .https://ojs.lib.unideb.hu/IJHS/article/view/1111Agrobacterium tumefaciensGenetic transformationg2ps1 geneGerbera hybridIn vitro cultureOrganogenesis |
| spellingShingle | N. M. Ali Bacha M. Batha A. M. Abdul-Kader Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) International Journal of Horticultural Science Agrobacterium tumefaciens Genetic transformation g2ps1 gene Gerbera hybrid In vitro culture Organogenesis |
| title | Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) |
| title_full | Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) |
| title_fullStr | Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) |
| title_full_unstemmed | Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) |
| title_short | Genetic engineering of apple (Malus domestica Borkh.) for resistance to fungal diseases using g2ps1 gene from Gerbera hybrida (Asteraceae) |
| title_sort | genetic engineering of apple malus domestica borkh for resistance to fungal diseases using g2ps1 gene from gerbera hybrida asteraceae |
| topic | Agrobacterium tumefaciens Genetic transformation g2ps1 gene Gerbera hybrid In vitro culture Organogenesis |
| url | https://ojs.lib.unideb.hu/IJHS/article/view/1111 |
| work_keys_str_mv | AT nmalibacha geneticengineeringofapplemalusdomesticaborkhforresistancetofungaldiseasesusingg2ps1genefromgerberahybridaasteraceae AT mbatha geneticengineeringofapplemalusdomesticaborkhforresistancetofungaldiseasesusingg2ps1genefromgerberahybridaasteraceae AT amabdulkader geneticengineeringofapplemalusdomesticaborkhforresistancetofungaldiseasesusingg2ps1genefromgerberahybridaasteraceae |