Simultaneous gene expression and multi-gene silencing in Zea mays using maize dwarf mosaic virus

BMC Plant Biology

Table 3 GFP and VIGS insert stability in constructs pWX27, pWX68, and pWX56

Construct	Insert	Rep.^a	Time (dpi)	Total plants	a^b	b	c	d	e
pWX27	GFP	1	7	20	9	10	0	0	1
		2	7	20	6	13	1	0	0
		3	7	20	11	8	0	1	0
		1	14	20	0	18	2	0	0
		2	14	20	0	13	4	2	1
		3	14	20	0	16	4	0	0
		1	21	20	0	5	12	2	1
		2	21	20	0	3	9	8	0
		3	21	20	0	3	12	5	0
pWX68	GFP	1	7	20	14	4	2	0	0
		2	7	20	2	11	4	3	0
		3	7	20	3	10	5	2	0
		1	14	20	10	6	2	2	0
		2	14	20	3	6	5	5	1
		3	14	20	2	4	6	8	0
		1	21	20	3	9	4	3	1
		2	21	20	0	0	12	7	1
		3	21	20	0	2	7	9	2
pWX56	GFP	1	7	20	7	6	1	4	2
		2	7	20	11	4	0	3	2
		3	7	20	8	5	1	1	5
		1	14	20	9	7	2	2	0
		2	14	20	3	14	1	2	0
		3	14	20	8	7	3	0	2
		1	21	20	2	8	2	4	4
		2	21	20	5	10	0	4	1
		3	21	20	0	12	2	5	1
pWX56	VIGS	1	7	20	6	9	5	0	0
		2	7	20	17	0	0	3	0
		3	7	20	0	16	0	0	4
		1	14	20	13	2	3	0	2
		2	14	20	6	12	0	1	1
		3	14	20	11	4	0	5	0
		1	21	20	12	3	0	1	4
		2	21	20	12	3	0	1	4
		3	21	20	4	4	1	1	10^c

^aThree experimental replicates (20 plants per construct in each), were tested by RT-PCR. Wild type pWX6 and healthy plants were used as positive and negative controls
^bRT-PCR results per sample were scored into 5 different categories (a,b,c,d and e), based on band(s) observed: a full length insertion single band observed, b multiple bands observed; including expected full-length insertion and smaller bands indicating insert loss, c multiple bands indicating insert loss, with none representing full length insertion; d single band detected at less than full-length insertion size, usually close to size of wild type virus amplicon; e no bands detected with primer pair
^cThis replicate was excluded from statistical analyses since no amplification was observed in half of the samples

ISSN: 1471-2229