Effective Agrobacterium-mediated transformation protocols for callus and roots of halophyte ice plant (Mesembryanthemum crystallinum)
| العنوان: | Effective Agrobacterium-mediated transformation protocols for callus and roots of halophyte ice plant (Mesembryanthemum crystallinum) |
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| المؤلفون: | Hau-Hsuan Hwang, Hsiao-Huei Chen, Fan-Chen Huang, Jia-Fang Ho, Chih-Hao Wang, Shin-Fei Chi, Hungchen Emilie Yen |
| المصدر: | Botanical Studies Botanical Studies, Vol 60, Iss 1, Pp 1-15 (2019) |
| سنة النشر: | 2018 |
| مصطلحات موضوعية: | 0106 biological sciences, 0301 basic medicine, Agrobacterium, Plant Science, Biology, 01 natural sciences, 03 medical and health sciences, Tissue culture, Halophyte, lcsh:Botany, Ice plant, Mesembryanthemum crystallinum, fungi, food and beverages, Agrobacterium tumefaciens, biology.organism_classification, lcsh:QK1-989, Horticulture, Transformation (genetics), 030104 developmental biology, Callus, Original Article, Transformation protocol, 010606 plant biology & botany, Transformation efficiency |
| الوصف: | Background Ice plant (Mesembryanthemum crystallinum L.) is a model plant for studying salt-tolerant mechanisms in higher plants. Many salt stress-responsive ice plant genes have been identified with molecular and biochemical approaches. However, no further functional characterization of these genes in host plant due to lack of easy and effective transformation protocols. Results To establish efficient transformation system of ice plants, three types of ice plant materials, hypocotyl-derived callus, aseptically-grown seedlings and pot-grown juvenile plants, were used to develop Agrobacterium-mediated transformation protocols. The highest transient transformation efficiency was with 5-day-old ice plant callus co-incubated with an Agrobacterium tumefaciens at 2.5 × 109 cells mL−1 for 48 h. The 3-day-old ice plant seedlings with root tip removed were successfully infected with A. tumefaciens or A. rhizogenes, and obtained 85% and 33–100% transient transformation rates, respectively. The transient transformation assays in ice plant callus and seedlings demonstrated that the concentrations of Agrobacteria, the durations of co-incubation time, and the plant growth stages were three important factors affecting the transient transformation efficiencies. Additionally, pot-grown juvenile plants were syringe-injected with two A. rhizogenes strains A8196 and NCPPB 1855, to establish transformed roots. After infections, ice plants were grown hydroponically and showed GUS expressions in transformed roots for 8 consecutive weeks. Conclusions Our Agrobacterium-mediated transformation protocols utilized hypocotyl-derived callus and seedlings as plant materials, which can be easily obtained in large quantity. The average successful transient transformation rates were about 2.4–3.0% with callus and 33.3–100.0% with seedlings. We also developed a rapid and efficient protocol to generate transgenic roots by A. rhizogenes infections without laborious and challenging tissue culture techniques. This protocol to establish composite ice plant system demonstrates excellent improvements in efficiency, efficacy, and ease of use over previous ice plant transformation protocols. These Agrobacterium-mediated transformation protocols can be versatile and efficient tools for exploring gene functions at cellular and organ levels of ice plants. Electronic supplementary material The online version of this article (10.1186/s40529-018-0249-3) contains supplementary material, which is available to authorized users. |
| تدمد: | 1817-406X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::384921384eaceb7579c7b8416801d769 https://pubmed.ncbi.nlm.nih.gov/30617933 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....384921384eaceb7579c7b8416801d769 |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 1817406X |
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