Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter
Abstract Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation....
Main Authors: | , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2021-06-01
|
Series: | Plant Methods |
Subjects: | |
Online Access: | https://doi.org/10.1186/s13007-021-00768-9 |
id |
doaj-928ae706514547d8b91a453832e1ccec |
---|---|
record_format |
Article |
spelling |
doaj-928ae706514547d8b91a453832e1ccec2021-07-04T11:12:48ZengBMCPlant Methods1746-48112021-06-0117111410.1186/s13007-021-00768-9Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De WinterMasoumeh Asadi-Aghbolaghi0Beata Dedicova1Sonali Sachi Ranade2Kim-Cuong Le3Farzad Sharifzadeh4Mansoor Omidi5Ulrika Egertsdotter6Department of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of TehranDepartment of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural SciencesDepartment of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural SciencesDepartment of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural SciencesDepartment of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of TehranDepartment of Agronomy and Plant Breeding, College of Agriculture and Natural Resources, University of TehranDepartment of Forest Genetics and Plant Physiology, Umeå Plant Science Centre, Swedish University of Agricultural SciencesAbstract Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. Results A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L–1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L–1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. Conclusion To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes.https://doi.org/10.1186/s13007-021-00768-9GrassStipagrostis pennata (Trin.) De WinterSomatic embryogenesisPlant regenerationSSR markersAgrobacterium |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Masoumeh Asadi-Aghbolaghi Beata Dedicova Sonali Sachi Ranade Kim-Cuong Le Farzad Sharifzadeh Mansoor Omidi Ulrika Egertsdotter |
spellingShingle |
Masoumeh Asadi-Aghbolaghi Beata Dedicova Sonali Sachi Ranade Kim-Cuong Le Farzad Sharifzadeh Mansoor Omidi Ulrika Egertsdotter Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter Plant Methods Grass Stipagrostis pennata (Trin.) De Winter Somatic embryogenesis Plant regeneration SSR markers Agrobacterium |
author_facet |
Masoumeh Asadi-Aghbolaghi Beata Dedicova Sonali Sachi Ranade Kim-Cuong Le Farzad Sharifzadeh Mansoor Omidi Ulrika Egertsdotter |
author_sort |
Masoumeh Asadi-Aghbolaghi |
title |
Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter |
title_short |
Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter |
title_full |
Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter |
title_fullStr |
Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter |
title_full_unstemmed |
Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter |
title_sort |
protocol development for somatic embryogenesis, ssr markers and genetic modification of stipagrostis pennata (trin.) de winter |
publisher |
BMC |
series |
Plant Methods |
issn |
1746-4811 |
publishDate |
2021-06-01 |
description |
Abstract Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. Results A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L–1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L–1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. Conclusion To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes. |
topic |
Grass Stipagrostis pennata (Trin.) De Winter Somatic embryogenesis Plant regeneration SSR markers Agrobacterium |
url |
https://doi.org/10.1186/s13007-021-00768-9 |
work_keys_str_mv |
AT masoumehasadiaghbolaghi protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT beatadedicova protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT sonalisachiranade protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT kimcuongle protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT farzadsharifzadeh protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT mansooromidi protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter AT ulrikaegertsdotter protocoldevelopmentforsomaticembryogenesisssrmarkersandgeneticmodificationofstipagrostispennatatrindewinter |
_version_ |
1721320545121730560 |