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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography <t>(HPLC)</t> analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).
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Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography (HPLC) analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).

Journal: bioRxiv

Article Title: A single PLAT domain protein couples reproductive arrest and carotenoid pigmentation during diapause in the two-spotted spider mite, Tetranychus urticae Koch

doi: 10.64898/2026.05.13.724795

Figure Lengend Snippet: Body coloration phenotypes and carotenoid contents following RNAi-mediated gene silencing in Tetranychus urticae females reared under diapause-inducing conditions. (A) Representative images of the three body color phenotypes scored after dsRNA treatment: orange (typical diapause phenotype with high carotenoid accumulation), yellow (intermediate, altered carotenoid composition), and green (reduced diapause-associated carotenoid accumulation). Scale bars, 0.1 mm. The stacked bar chart represents the proportion of individuals classified as orange, yellow, or green within each treatment group following dsRNA treatments targeting the TuPDS ( tetur01g11270 ), TuLCPS ( tetur01g11260 ), TuCYP384A1 ( tetur38g00650 ), or TuPLAT10 ( tetur11g05720 ) genes (ds TuPDS , ds TuLCPS , ds TuCYP384A1 , or ds TuPLAT10 , respectively), compared with the dsRNA treatment targeting an intergenic region used as a negative control (dsNC). Different letters above the bars indicate significant differences among treatments (Fisher’s exact test, p < 0.001); n , total number of mites scored. Of females classified by body coloration phenotype, the body-color subgroups containing sufficient individuals were subjected to high-performance liquid chromatography (HPLC) analysis: yellow only for ds TuLCPS ; yellow and green for ds TuPDS , ds TuCYP384A1 , and ds TuPLAT10 . dsNC females (all orange) and non-diapausing females (all green) were each analyzed as a single group. HPLC analysis for quantifying the contents of (B) β-carotene and (C, D) astaxanthin in dsRNA-treated mites and non-diapausing mites. (C) Cholesterol esterase treatment (+CE) hydrolyses esterified astaxanthin into its free forms. In (D), −CE indicates samples without esterase treatment (free astaxanthin only). Data are mean ± standard error of the mean (SEM), with individual dots indicating biological replicates. Statistical significance relative to dsNC was determined using Dunnett’s test ( p < 0.001).

Article Snippet: For HPLC analysis, standard β-carotene (Nacalai Tesque, Inc., Kyoto, Japan) and astaxanthin (Dr. Ehrenstorfer GmbH, Augsburg, Germany) were dissolved in chloroform to prepare stock solutions of each standard at 10 mg mL −1 .

Techniques: Negative Control, High Performance Liquid Chromatography