Effective Agrobacterium-mediated transformation protocols for callus and roots of halophyte ice plant (Mesembryanthemum crystallinum)

Botanical Studies

Table 1 Transient transformation rates of A. tumefaciens EHA105 strain harboring the pBISN1 plasmid in ice plant seedlings and distribution (%) of GUS staining in ice plant seedlings of different ages

Ages	Plant types	Transient transformation rates^a	% of tissues showing GUS activities^b
3-day-old	Intact seedling	66.7 ± 2.8*	Cotyledon	100.0 ± 2.8^#
	Intact seedling	66.7 ± 2.8*	Hypocotyl	14.3 ± 0.4
	Root tip removed	85.0 ± 1.1*	Cotyledon	88.9 ± 0.9^#
	Root tip removed	85.0 ± 1.1*	Hypocotyl	22.2 ± 0.2
5-day-old	Intact seedling	48.3 ± 3.8*	Cotyledon	63.4 ± 1.3^#
	Intact seedling	48.3 ± 3.8*	Hypocotyl	52.1 ± 2.5
	Root tip removed	80.0 ± 2.0*	Cotyledon	98.2 ± 2.0^#
	Root tip removed	80.0 ± 2.0*	Hypocotyl	25.7 ± 0.3
7-day-old	Intact seedling	0.0 ± 0.0	0.0 ± 0.0
	Root tip removed	13.3 ± 2.8	Cotyledon	0.0 ± 0.0^#
	Root tip removed	13.3 ± 2.8	Hypocotyl	100.0 ± 2.8

^aNumber of transiently transformed seedlings/total number of seedlings × 100%. The 60–80 ice plant seedlings were infected with A. tumefaciens strain for each independent transformation assay. Data are mean ± SD (standard deviation) from at least three independent experiments
^bNumber of GUS stained tissues/number of transiently transformed seedlings × 100%
* P < 0.05 comparing 3-, or 5-day-old infected seedlings and 7-day-old infected seedlings of the same ice plant type based on pairwise Student t-test
^#P < 0.05 comparing cotyledon and hypocotyl tissues of the same age and type ice plants based on pairwise Student t-test