Fig 1.
Life cycle of Planococcus krauhniae.
E: Egg, N1: first-instar nymph (undifferentiated in males and females), N2: second-instar nymph (the first few days are undifferentiated), N2♂: male second-instar nymph (identifiable by filamentous secretions), N2♀: female second-instar nymph (filamentous secretions absent), N3: female third-instar nymph, Pre: male prepupa, Pu: male pupa, Ad: adult. Scale bars: 200 μm (E, N1 and N2), 500 μm (N2♀, N2♂, Pre, Pu, Ad). N1, N2, N3, Pre, Pu, and female adult specimens had their secretions removed in the photographs. Arrows: molting events. Red outline: non-feeding stages. Grey gradient: degree of sexual dimorphism.
Fig 2.
JH biosynthesis, JH signal transduction, and early responses.
JH biosynthesis, JH signal transduction and early response. Expression profiles of Pkjhamt (A), PkMet (B), Pktai (C), and PkKr-h1 (D) during male and female mealybug development after egg oviposition. E: Egg, N1: first-instar nymph, N2: early second-instar nymphs, N3: female third-instar nymph, f: female adult, pre: male prepupa, pu: male pupa, m: male adult. Samples were collected from E to N2D3, using the sex-bias strategy (see the Methods section).
Fig 3.
JHM pyriproxyfen effects on male development and JH signaling.
A. Treatment during prepupa. B. Treatment during pupa. (C-I) Effects of JHM on gene expression. PreD1: Treatment 24-48 hours after the prepupal molt, RNA extraction 5 days after the treatment, N = 6. PuD0: Treatment 0-24 hours after the pupal molt, RNA extraction 6 days after the treatment, N = 5. Boxplots were constructed with the ggplot2 R package (with upper and lower hinges: 1st and 3rd quartiles, middle line: median, dots: outlier values). Unpaired two-sample Student’s t-test with *: p-value < 0.05 and **: p-value < 0.01. C. PkMet; D. Pktai com; E. Pktai IN-1; F. Pktai DEL-1; G. PkKr-h1; H. PkKr-h1 A; I. PkKr-h1 B.
Table 1.
Effects of the pyriproxyfen (JHM) treatment on male metamorphosis in P. kraunhiae.
Fig 4.
br copies and zinc-finger isoforms in the Japanese mealybug.
A. Protein alignment of the zinc-finger motif for each identified isoform. B. Phylogeny of zinc-finger isoforms for different insect groups. See Supporting Information for the phylogenetic analysis method.
Fig 5.
Expression profiles and JHM effects on br.
A. Expression profiles of Pkbr copies identified in the present study. E: Egg, N1: first-instar nymph, N2: second-instar nymphs, N3: female third-instar nymph, Pre: male prepupa, Pu: male pupa. Samples were collected using the sex-bias strategy from E to N2D3. B-F. Effects of JHM on gene expression. PreD1: Treatment 24-48 hours after the prepupal molt, RNA extraction 5 days after the treatment, N = 6. PuD0: treatment 0-24 hours after the pupal molt, RNA extraction 6 days after the treatment, N = 5. Boxplots were constructed with the ggplot2 R package (with upper and lower hinges: 1st and 3rd quartiles, middle line: median, dots: outlier values). Unpaired two-sample Student’s t-test with *: p-value < 0.05 and **: p-value < 0.01. B. Pkbr1 com, C. Pkbr1 Z2, D. Pkbr1 Z4, E. Pkbr3 com, F. Pkbr3 Z2.
Fig 6.
Summary of differential JH titers and early response gene patterns when phenotypic differences arise during post-embryonic development in P. kraunhiae.
N2: Second-instar nymph, N3: female third-instar nymph, Pre: prepupa, Pu: pupa.