Table 3.

Comparisons a–f of MS control and WPBS media, callus culture regimes and transformation treatments in Agrobacterium mediated transformations of various grass species using genes of interest or gus A

Species	Medium and treatment	No. of explants or embryos	Timing	Total weight of callus (g)	GUS or PCR pos plants or (calli)	Plants per g callus	Plants per explant	Significance between plant numbers per replicate dish
(a) Comparison of MS control and WPBS media and different callus culture regimes using gus A
L. perenne	MS	9	6, 3, 2, 1  weeks	13.23	30	2.27	3.33	NS
L. perenne	WPBS	10	6, 3, 2, 1 weeks	34.01	100	2.94	10
L. temulentum	MS	425	10–12 day	10.81	3 (27)	0.28	0.01	P ≤ 0.01
L. temulentum	WPBS	566	10–12 day	17.63	23 (70)	1.3	0.04
B. distachyon	MS	50	3, 1 weeks	4.34	3	0.69	0.06	P ≤ 0.01
B. distachyon	WPBS	55	3, 1 weeks	9.25	16	1.73	0.29
B. distachyon	MS	43	3, 2, 1 weeks	27.09	1	0.04	0.02	P ≤ 0.01
B. distachyon	WPBS	47	3, 2, 1 weeks	41.09	18	0.44	0.38
F. arundinacea	MS	n/r	6, 3, 2, 1 weeks	3.59	3	0.84	n/r	NS
F. arundinacea	WPBS	n/r	6, 3, 2, 1 weeks	6.68	9	1.35	n/r
M. sinensis	MS	9	6, 2, 2, 1 weeks	9.97	0	0	0	NS
M. sinensis	WPBS	15	6, 2, 2, 1 weeks	36.73	25	0.68	1.67
M. floridulus	MS	8	6, 2, 2, 1 weeks	12.66	0	0	0	NS
M. floridulus	WPBS	25	6, 2, 2, 1 weeks	28.07	1	0.04	0.04
(b) Comparison of adding or leaving out copper (Cu) in infection and co-cultivation media using gus A
L. perenne	MS	4	6, 3, 2, 1 weeks	6.4	14	2.19	3.5	MS and WPBS NS  + and − Cu NS
L. perenne	MS + Cu	5	6, 3, 2, 1 weeks	6.83	16	2.34	3.2
L. perenne	WPBS-Cu	5	6, 3, 2, 1 weeks	17.51	57	3.26	11.4
L. perenne	WPBS + Cu	5	6, 3, 2, 1 weeks	16.6	43	2.59	8.6
(c) Comparison of calli derived from shoot tips and OMM meristem tissue using genes of interest
F. arundinacea	WPBS	68	6, 2, 1 weeks	11.22	16	1.43	0.24	P ≤ 0.05
F. arundinacea	WPBS OMM	45	8, 1.5 weeks	15.03	13	0.86	0.29
L. perenne	WPBS	74	5, 2, 1 weeks	9.64	17	1.76	0.23	NS
L. perenne	WPBS OMM	19	5, 2, 1 weeks	20.05	11	0.55	0.58
L. temulentum	WPBS	1848	4–5, 1 weeks	311.02	4	0.01	0.002	P ≤ 0.01
L. temulentum	WPBS OMM	321	3–5, 1 weeks	70.55	12	0.17	0.037
(d) Comparison of callus culture regimes using genes of interest
B. distachyon	WPBS	124	3, 1 weeks	18.18	29	1.6	0.23	3,1 wk and 3,2,1wk P ≤ 0.05
B. distachyon	WPBS	88	4, 1 weeks	27.59	27	0.98	0.31
B. distachyon	WPBS	47	3, 2, 1 weeks	41.09	18	0.44	0.38
L. perenne	WPBS	45	4, 2, 1 weeks	10.51	20	1.9	0.44	NS
L. perenne	WPBS	39	5, 2, 1, 1 weeks	40.99	56	1.37	1.44
(e) Comparison of vacuum treatment (vac) and immersion (imm) of calli using genes of interest
L. temulentum	MS vac	110	10 day	2.63	1 (4)	0.38	0.01	MS and WPBS P ≤ 0.01 Vacuum and immersion NS
L. temulentum	MS imm	154	10 day	3.69	1 (5)	0.27	0.01
L. temulentum	WPBS vac	154	10 day	4.71	6 (19)	1.27	0.04
L. temulentum	WPBS imm	220	10 day	6.73	8 (25)	1.19	0.04
(f) Comparison of heat shock (HS) and vacuum treatment (vac) of calli using genes of interest
B. distachyon	WPBS HS	29	4, 1 weeks	8.25	6	0.73	0.21	P = 0.054
B. distachyon	WPBS vac	29	4, 1 weeks	8.25	15	1.82	0.52

NS not significant, n/r not recorded