PHOSPHOMANNOSE ISOMERASE/MANNOSE SELECTION SYSTEM TO SCREEN TRANSGENIC CHRYSANTHEMUM PLANTS (CHRYSANTHEMUM MORIFOLIUM)

The objective was to develop the selection system for transgenic chrysanthemum by Agrobacterium tumefaciens carrying genes related in saline alkaline tolerance without using antibiotic resistance genes.
The selection system was based on the use of Escherichia coli phosphomannose isomerase (PMI) gene as a selectable marker gene and mannose as a selective nutrient.
Leaf explants from in vitro germinated plants of chrysanthemum ‘Huoyan’ were infected with A. tumefaciens EHA105 pCAMBIA1301. The explants were selected on medium supplemented with a combination of mannose and sucrose as a carbon source.
The chrysanthemum plants were transformed with the PtDHAR gene, which was screened through cDNA micro-array analysis in Puccinellia tenuiflora as a highly expressed gene under saline alkaline stress.
Among the 847 leaf explants which were infected, there were 155 shoots surviving after selection by the medium containing 10 g L^-1 mannose.
PCR analysis confirmed that 5 independent transformed lines were positive for PMI and PtDHAR gene.
Four were confirmed by Southern blot analysis.
The pmi/mannose selection system was efficient for producing transgenic chrysanthemum plants without using antibiotics or herbicides.

XXVIII International Horticultural Congress on Science and Horticulture for People (IHC2010): International Symposium on Advances in Ornamentals, Landscape and Urban Horticulture

L.N. Xie, J. Wang, L.G. Yu, Y. Zhang, B. Zhou , Y.H. Li, S. Kawabata

Chrysanthemum, pmi/mannose selection system, saline, alkaline, tolerance

https://doi.org/10.17660/ActaHortic.2012.937.44