Table 1.
Genes from Acyrthosiphon pisum chosen for RNA interference in Myzus persicae.
Figure 1.
Transgenic tobacco lines expressing artificial hairpin RNAs or microRNAs have effects on aphid fecundity.
A–F and I–K represent transgenic plants expressing hpRNA corresponding to genes V-ATPaseE, CPRR1, Rps5, SMARCD1, TBCD, and delta-COP, and AChE2, Rps14 and Med31, respectively, of Myzus persicae, while G and H represent transgenic plants expressing amiRNA targeting AChE2. The control (con) transgenic plants contained an empty vector. Bars represent the means (±SE) of three lines of each genotype. The experiment was repeated at least twice with similar results. Asterisks represent significant differences between the transgenic and the wild type (wt) as determined by Student’s t-test (An asterisk denotes p<0.05, and a double asterisk indicates p<0.01). In each test, the fecundity of each construct was the average value from three independent lines, and the fecundity of each line was collected from three plants.
Figure 2.
Expression profiles of the acetylcholinesterase 2 coding gene (MpAChE2).
(A) qRT-PCR analysis of the MpAChE2 expression at different developmental stages of Myzus persicae. (B) Relative expression level of MpAChE2 in adult aphid organs with the expression level in the whole body of L1 instar nymph as one (L1). The housekeeping gene, α-actin, was used as an internal control. The data represent the means (±SE) of three replicates.
Figure 3.
Expression levels of artificial microRNAs and small interfering RNAs, targeting the MpAChE2 gene of Myzus persicae.
Relative expression levels of amiR-AChE2-1 (A) or amiR-AChE2-2 (B) in tobacco plants analyzed by qRT-PCR. The expression levels of amiRNAs were relative to the expression of tobacco 5.8S rRNA and normalized. The data represent the means (±SE) of three biological replicates. Student’s t-test was used to determine significant differences in transgenic plants compared to wild type plants (An asterisk denotes p<0.01, and a double asterisk indicates p<0.001). (C) Northern blot detection of AChE2 siRNAs in plants. rRNA/tRNA stained with ethidium bromide was used as a loading control.
Figure 4.
Knock-down of MpAChE2 expression by plant-mediated RNAi.
Transgenic tobacco lines expressing amiR-AChE2 or hpAChE2 down-regulated the target gene expression compared to empty vector-transgenic plants (con) and wild type (wt). The data represent the means (±SE) of three biological replicates. Student’s t-test was used to determine significant differences in aphid fed on transgenic plants compared to aphid fed on wild type plants (An asterisk denotes p<0.05).