Abstract
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.
Highlights
IntroductionChrysanthemum (Chrysanthemum x morifolium Ramat) has the largest floriculture commodity in
Chrysanthemum (Chrysanthemum x morifolium Ramat) has the largest floriculture commodity inChina and is a popular traditional flower of esthetic and cultural importance
The results show that CmTFL1a was absent in control WT plants, comprehensively
Summary
Chrysanthemum (Chrysanthemum x morifolium Ramat) has the largest floriculture commodity in. China and is a popular traditional flower of esthetic and cultural importance. There is a growing demand for high quality chrysanthemum flowers and cut flowers in the local market. The plants can be regenerated from adventitious shoots from various chrysanthemum tissues or calli using in vitro culture methods [1]. There are various cultivars of chrysanthemum being commercially propagated, but Jinba holds significance due to its fast-growing habit. Jinba is a short-day chrysanthemum cultivar with relatively low transformation efficiency compared with other cultivars, making it difficult to obtain transgenic plants.
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