Reduced ALP activity correlates with reduced levels of HaALP transcripts in Cry1Ac-resistant <i>H. armigera</i> larvae.

<p>BBMV proteins from susceptible (96S) and Cry1Ac-resistant (BtR) <i>H. armigera</i> strains were used in specific ALP (A) or APN (B) activity assays. Different letters indicate statistically significant differences (p<0.05; Holm-Sidak method) among the samples. C) Mean relative transcript quantity of HaALP1 and HaALP2 isoforms. Data shown are the mean transcript quantity relative to the 96S sample with the highest transcript amounts from three independent biological replicates for each HaALP isoform and strain. All reactions were performed with triplicate technical replicates. Bars denote standard error of the mean. Different letters indicate significant differences (P<0.05; Holm-Sidak method).</p

BBMV from Cry1Ac-resistant <i>H. virescens</i> larvae display reduced ALP levels.

<p>(A) BBMV proteins from <i>H. virescens</i> strains as indicated were used in specific ALP or APN activity assays. Different letters indicate statistically significant differences (p<0.05; Holm-Sidak method) among the samples. (B) Western blot analysis of HvmALP, APN, or HevCaLP in BBMV proteins from <i>H. virescens</i> strains: Lane 1, YDK; lane 2, YHD2-B; lane 3, CXC; lane 4, KCBhyb.</p

Reduced ALP activity correlates with reduced levels of mALP in BBMV from Cry1Fa-resistant <i>S. frugiperda</i> larvae.

<p>BBMV proteins from susceptible (Mon and Ben) and Cry1Fa-resistant (456 and 512) strains of <i>S. frugiperda</i> were used in specific ALP (A) or APN (B) activity assays. Different letters indicate statistically significant differences (p<0.05; Holm-Sidak method) among the samples. C) Detection of mALP in BBMV from susceptible (Mon and Ben) and resistant (456 and 512) strains of <i>S. frugiperda</i> using Western blotting. Lane 1, Mon, lane 2, 456, lane 3, 512, lane 4, Ben. Arrow points to ALP protein band.</p

Reduced levels of HvmALP1 and HvmALP2 transcripts in Cry1Ac-resistant <i>H. virescens</i> larvae relative to YDK as detected by qRT-PCR.

<p>Data shown are the mean transcript quantity relative to the YDK sample with the highest transcript amounts from three independent biological replicates for each HvmALP isoform and strain. All reactions were performed with triplicate technical replicates. Bars denote standard error of the mean; different letters on each bar indicate significant differences (P<0.05; Holm-Sidak method).</p

Detection of reduced HvmALP expression in Cry-resistant strains of <i>H. virescens</i> using quantitative differential proteomic analysis (2D-DIGE).

<p>BBMV proteins from susceptible (YDK) and resistant (YHD2-B, CXC, and KCBhyb) larvae were fluorescently labeled and the corresponding sub-proteome analyzed using 2D-DIGE. A representative gel image is presented with arrows pointing to the four HvmALP spots detected with lower expression levels. The identity of these spots as HvmALP was obtained by peptide mass fingerprinting, de novo sequencing, and Western blotting with specific antisera (data not shown). The standardized log abundance for each spot in all three BBMV samples (labeled with Cy3 or Cy5) from each strain used is shown. Differences in protein levels between HvmALP spots in BBMV from susceptible and resistant larvae were statistically significant (p<0.001; Student T-test). HvmALP expression levels among BBMV samples from resistant larvae were not significantly different.</p