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Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

Asadi-Aghbolaghi, Masoumeh and Dedicova, Beata and Ranade, Sonali Sachin and Le, Kim-Cuong and Sharifzadeh, Farzad and Omidi, Mansoor and Egertsdotter, Ulrika (2021). Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter. Plant Methods. 17 , 70
[Research article]

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Abstract

BackgroundStipagrostis 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.ResultsA 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.ConclusionTo 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.

Authors/Creators:Asadi-Aghbolaghi, Masoumeh and Dedicova, Beata and Ranade, Sonali Sachin and Le, Kim-Cuong and Sharifzadeh, Farzad and Omidi, Mansoor and Egertsdotter, Ulrika
Title:Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter
Series Name/Journal:Plant Methods
Year of publishing :2021
Volume:17
Article number:70
Number of Pages:14
Publisher:BMC
ISSN:1746-4811
Language:English
Publication Type:Research article
Article category:Scientific peer reviewed
Version:Published version
Copyright:Creative Commons: Attribution 4.0
Full Text Status:Public
Subjects:(A) Swedish standard research categories 2011 > 1 Natural sciences > 106 Biological Sciences (Medical to be 3 and Agricultural to be 4) > Genetics (medical genetics to be 30107 and agricultural genetics to be 40402)
Keywords:Grass, Stipagrostis pennata (Trin, ) De Winter, Somatic embryogenesis, Plant regeneration, SSR markers, Agrobacterium
URN:NBN:urn:nbn:se:slu:epsilon-p-113101
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-p-113101
Additional ID:
Type of IDID
DOI10.1186/s13007-021-00768-9
Web of Science (WoS)000671269900002
ID Code:25063
Faculty:S - Faculty of Forest Sciences
Department:(S) > Dept. of Forest Genetics and Plant Physiology
Deposited By: SLUpub Connector
Deposited On:26 Aug 2021 14:13
Metadata Last Modified:26 Aug 2021 14:21

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