A systemic gene silencing method suitable for high throughput, reverse genetic analyses of gene function in fern gametophytes

Table 1 Frequencies of gametophyte phenotypes after bombardment with various plasmids.

Plasmids introduced	% colorless & GUS positive^a,b	% colorless & GUS negative^a	% green & GUS positive^a	n^c
35S:irintCrChl+pFF19G	88.8 (+/-1.36)^d	6.8 (+/-0.46)^d	4 (+/- 1.41)^d	1263
irintCrChl+pFF19G	81.7 (+/-0.87)	8.3 (+/-1.20)	10 (+/-1.53)	775
35S:intantiCrChl+pFF19G	61.0 (+/-1.73)	11.0 (+/-1.73)	28.0 (+/-3.21)	564
35S:antiCrChl+pFF19G	12.0 (+/-1.53)	0	88.0 (+/-1.53)	724
antiCrChl+pFF19G	0.7 (+/-0.33)	0	99.3 (+/-0.33)	479
35S:senseCrChlint+pFF19G	37.7 (+/-1.12)	6.3 (+/-0.19)	56 (+/-1.00)	559
35S:senseCrChl+pFF19G	2.3 (+/-0.88)	0	97.7 (+/-0.88)	761
senseCrChl+pFF19G	1.7 (+/-0.33)	0	98.7 (+/-0.42)	677
CrChl cDNA+pFF19G	13.7 (+/-0.88)	0	86.0 (+/-1.16)	563
Δ35SpFF19G+	48.5 (+/-6.36)	51.0 (+/-5.66)	0.5 (+/-0.71)	470
35SinintCrChl
pFF19G	0	0	100	600
	% with lesions & GUS positive^a,b	% with lesions & GUS negative^a	% without lesions & GUS positive	n^c
CrUrod cDNA+pFF19G	61.0 (+/-0.71)	20.2 (+/-0.69)	18.9 (+/-0.43)	774
35S:intantiCrUrod+pFF19G	55.7 (+/-1.40)	13.5 (+/-1.61)	31.1 (+/- 2.25)	472

^aAll gametophytes were stained 7 days after shooting; each percent represents the average of three replicates; standard error is given in parentheses. ^bthe differences between treatments for all pairwise comparisons are statistically significant (z>3.4, P < 0.0003) with the exception of 35S:senseCrChl+pFF19G and senseCrChl+pFF19G. ^ctotal number of gametophytes displaying a phenotype and scored. ^dthe average of 5 replicates.

ISSN: 1471-2229