Functional genomics of the horn fly, Haematobia irritans (Linnaeus, 1758)

BMC Genomics

Table 5 Results of RNAi experiments in female horn flies.

Group No.^a	Cumulative percent mortality			Oviposition (eggs per survived fly)^b	Expression silencing (average% ± SD) with respect to group 10 control^c				dsRNA injected^d
	12 hpi	24 hpi	36 hpi		6 hpi	12 hpi	24 hpi	36 hpi
1 (Serine protease)	57 ± 6	69 ± 11	78 ± 2	0.88 ± 0.23	ND	ND	57 ± 50 0 ± 0	ND	90 230
2* (Protease inhibitor)	73 ± 11	84 ± 7	94 ± 1	2.56 ± 1.80	ND	ND	83 ± 3**	ND	2_B12
3 (Vitellogenin)	55 ± 11	67 ± 8	74 ± 8	0.31 ± 0.9**	ND	ND	0 ± 0 0 ± 0 100 ± 2*	ND	7 20 76
4 (Ubiquitination)	42 ± 12	67 ± 11	83 ± 16	0.64 ± 0.63	ND	ND	92 ± 6* 70 ± 24 46 ± 43	ND	84 146 4_E04
5* (Ferritin)	24 ± 23	39 ± 37	47 ± 41	0.22 ± 0.17**	ND	ND	68 ± 13* 86 ± 86	ND	26 154
6* (vATPase)	17 ± 13	24 ± 11	34 ± 3	0.08 ± 0.05**	ND	ND	78 ± 6* 100 ± 1* 99 ± 8*	ND	7_F08 9_A08 17_H03
7 (Proteasome component)	64 ± 3	76 ± 8	84 ± 11	0.38 ± 0.03**	98 ± 2* 100 ± 6*	100 ± 2* 67 ± 9*	ND ND	0 ± 0 ND	7_A04 12_H09
8* (Immune response)	66 ± 7	ND	99 ± 8	0.23 ± 0.09**	100 ± 8* 0 ± 0	92 ± 5 0 ± 0	ND ND	96 ± 9* 98 ± 8*	6_F11 10_G05
9* (5'-nucleotidase)	50 ± 11	ND	91 ± 22	0.12 ± 0.09**	0 ± 0	98 ± 9*	ND	70 ± 7*	13_D07
10 (negative control)	45 ± 24	57 ± 30	64 ± 27	1.26 ± 0.90	---	---	---	---	191 2_E12
Injection buffer	46 ± 28	57 ± 30	65 ± 29	1.50 ± 1.01	0 ± 0	0 ± 0	0 ± 0	0 ± 0	None

^aThe average ± S.D. of two independent RNAi experiments for each of the test groups 1-9 and 14 experiments for the negative control and injection buffer groups is shown. One hundred flies were used on each RNAi experiment. Cumulative percent mortality was evaluated in female horn flies at 12, 24 and 36 hpi. Survival curves (temporal rates of mortality) were compared between different treatments and the control group 10 using Cox Proportional Hazards Survival Regression analysis (*P < 0.05). Abbreviation: ND, not determined.
^bOviposition data from test groups 1-9 and the injection buffer control were compared with the unrelated dsRNA-injected control group 10 by Student's t-test (**P < 0.005).
^cFor analysis of expression silencing, the mRNA levels of each knockdown gene were determined by real-time RT-PCR in 4 individual flies each at 6, 12 and 24 or 36 hpi. Percent expression silencing shown at different hpi corresponds to each individual gene targeted by dsRNA injection. The mRNA levels were normalized against horn fly 16S rRNA and the mean of the duplicate normalyzed Ct values from test dsRNA-injected groups 1-9 and in the injection buffer control were compared with the unrelated dsRNA-injected control group 10 by Student's t-test (*P < 0.05, **P < 0.005).
^dUnigenes GenBank accession numbers are shown in Table 6.

ISSN: 1471-2164