Standardized Genetic Transformation Protocol for Chrysanthemum cv. ‘Jinba’ with TERMINAL FLOWER 1 Homolog CmTFL1a
| Autor: | Yaohui Gao, Yike Gao, Saba Haider |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
0301 basic medicine lcsh:QH426-470 Chrysanthemum Transgene Chrysanthemum morifolium Flowers Genetically modified crops Biology transgenic plants 01 natural sciences Article 03 medical and health sciences Transformation Genetic Plant Growth Regulators Gene Expression Regulation Plant meropenem transformation efficiency Genetics medicine Genetics (clinical) Plant Proteins CmTFL1a fungi food and beverages Kanamycin Agrobacterium tumefaciens Plants Genetically Modified biology.organism_classification humanities Plant Leaves Plant Breeding lcsh:Genetics Transformation (genetics) Horticulture 030104 developmental biology expression pattern Shoot 010606 plant biology & botany medicine.drug Transformation efficiency |
| Zdroj: | Genes Volume 11 Issue 8 Genes, Vol 11, Iss 860, p 860 (2020) |
| ISSN: | 2073-4425 |
| DOI: | 10.3390/genes11080860 |
| Popis: | Chrysanthemum (Chrysanthemum x morifolium Ramat.) cultivar Jinba is a distinctive short-day chrysanthemum that can be exploited as a model organism for studying the molecular mechanism of flowering. The commercial value of Jinba can be increased in global flower markets by developing its proper regeneration and genetic transformation system. By addressing typical problems associated with Agrobacterium-mediated transformation in chrysanthemum, that is, low transformation efficiency and high cultivar specificity, we designed an efficient, stable transformation system. Here, we identify the features that significantly affect the genetic transformation of Jinba and standardize its transformation protocol by using CmTFL1a as a transgene. The appropriate concentrations of various antibiotics (kanamycin, meropenem and carbenicillin) and growth regulators (6-BA, 2,4-D and NAA) for the genetic transformation were determined to check their effects on in vitro plant regeneration from leaf segments of Jinba thus, the transformation protocol was standardized through Agrobacterium tumefaciens (EHA105). In addition, the presence of the transgene and its stable expression in CmTFL1a transgenic plants were confirmed by polymerase chain reaction (PCR) analysis. The CmTFL1a transgene constitutively expressed in the transgenic plants was highly expressed in shoot apices as compared to stem and leaves. Overexpression of CmTFL1a led to a delay in transition to the reproductive phase and significantly affected plant morphology. This study will help to understand the biological phenomenon of TFL1 homolog in chrysanthemum. Moreover, our findings can explore innovative possibilities for genetic engineering and breeding of other chrysanthemum cultivars. |
| Databáze: | OpenAIRE |
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