Dataset: PDFR gene expression and phenotypic data in Hyphantria cunea

This dataset supports the findings presented in the manuscript:

The Pigment-Dispersing Factor Receptor (PDFR) gene is involved in circadian rhythm and molting in Hyphantria cunea

Authors

Xue Li¹, Ge Zhao¹, Rongxia Zhao, Zhengxuan Wang, Lili Sun*, Chuanwang Cao*

¹ These authors contributed equally to this work.

Corresponding authors

Affiliation

Key Laboratory of Sustainable Forest Ecosystem Management-Ministry of Education, School of Forestry, Northeast Forestry University, Harbin, China

---

Data collection methods

Organism: Hyphantria cunea (fall webworm)

Treatments: dsPDFR (RNA interference targeting the PDFR gene) and dsGFP (control)

Measurements:

PDFR expression: measured by qPCR, presented as 2⁻ΔΔCt

Mortality rate: percentage of dead individuals at indicated time points

Malformation rate: percentage of individuals with developmental abnormalities

Pupation rate: percentage of individuals that successfully pupated

Behavioral/activity data: recorded at hourly intervals under different light regimes

Files and variables

1.Raw_data_for-Figure1-Relative-expression-level-of-PDFR-development-stage

Data brief description: In Figure 1A, we sampled Hyphantria cunea individuals at different developmental stages: egg stage, 1st to 7th instar larvae, female pupae (♀P) on day 1, male pupae (♂P) on day 1, female adults (♀A) on day 1, and male adults(♂A) on day 1. Quantitative real-time PCR (qRT-PCR) was performed to assess changes in PDFR expression levels. Each treatment had three mechanical and three biological replicates. The ct value is the original data detected by the instrument. After calculation by the formula, 2^-∆∆Ct is obtained. We calculated the relative expression of the target gene according to the 2^-∆∆Ct. We selected EF-1a and RPL13 as internal reference genes. The expression levels are shown relative to those in the egg stage.

Ct: The number of threshold cycles, the number of cycles experienced when the fluorescence signal in each reaction tube reaches the set threshold.

2^-∆∆Ct: The gene expression level change of the treatment group relative to the control group.

2.Raw_data_for-Figure1-Relative-expression-level-of-PDFR-tissue

Data brief description: In Figure 1B，samples were collected from the head, epidermis, fatbody, foregut, midgut, hindgut, silk gland, malpighian tubules, testis and ovary of H. cunea larvae. Quantitative real-time PCR (qRT-PCR) was performed to assess changes in PDFR expression levels. Each treatment had three mechanical and three biological replicates. The ct value is the original data detected by the instrument. After calculation by the formula, 2^-∆∆Ct is obtained.  The bars represent 2^−ΔΔCT values (± SEs) normalized to the expression of two internal control genes (EF-1α and RPL13).

3.Raw_data_for-Figure2-Relative-expression-level-of-PDFR-tissue-heads-5th instar-16L8D

Data brief description: In Figure 2A，it shows the effects of photoperiod on the expression rhythm of PDFR gene in the heads of the 5th instar H. cunea larvae under 16L:8D. Under photoperiods of 16L:8D, nine head samples from the 1st-3rd day of the 5th H. cunea larvae were collected at each of the following time points: ZT0h, ZT4h, ZT8h, ZT12h, ZT16h, ZT20h, ZT24h，ZT28h, ZT32h, ZT36h, ZT40h, ZT44h, ZT48h, ZT52h，ZT56h, ZT60h, ZT64h, ZT68h and ZT72h. Different lowercase letters represent the significance of difference, which was calculated by one-way ANOVA followed by Duncan’s test (P<0.05). The cosine analysis online tool analyses cosine (https://cosinor.online/app/cosinor.php) was used to evaluate the circadian rhythm of gene expression.

4.Raw_data_for-Figure2-Relative-expression-level-of-PDFR-tissue-heads-5th instar-12L12D

Data brief description: In Figure 2B，it shows the effects of photoperiod on the expression rhythm of PDFR gene in the heads of the 5th instar H. cunea larvae under 12L:12D. Under photoperiods of 12L:12D, nine head samples from the 1st-3rd day of the 5th H. cunea larvae were collected at each of the following time points: ZT0h, ZT4h, ZT8h, ZT12h, ZT16h, ZT20h, ZT24h，ZT28h, ZT32h, ZT36h, ZT40h, ZT44h, ZT48h, ZT52h，ZT56h, ZT60h, ZT64h, ZT68h and ZT72h. Different lowercase letters represent the significance of difference, which was calculated by one-way ANOVA followed by Duncan’s test (P<0.05). The cosine analysis online tool analyses cosine (https://cosinor.online/app/cosinor.php) was used to evaluate the circadian rhythm of gene expression.

5.Raw_data_for-Figure2-Relative-expression-level-of-PDFR-tissue-heads-5th instar-24L0D

Data brief description: In Figure 2C，it shows the effects of photoperiod on the expression rhythm of PDFR gene in the heads of the 5th instar H. cunea larvae under 24L:0D. Under photoperiods of 24L:0D, nine head samples from the 1st-3rd day of the 5th H. cunea larvae were collected at each of the following time points: ZT0h, ZT4h, ZT8h, ZT12h, ZT16h, ZT20h, ZT24h，ZT28h, ZT32h, ZT36h, ZT40h, ZT44h, ZT48h, ZT52h，ZT56h, ZT60h, ZT64h, ZT68h and ZT72h. Different lowercase letters represent the significance of difference, which was calculated by one-way ANOVA followed by Duncan’s test (P<0.05). The cosine analysis online tool analyses cosine (https://cosinor.online/app/cosinor.php) was used to evaluate the circadian rhythm of gene expression.

6.Raw_data_for-Figure3-Silencing-efficiency-of-PDFR-5th

Data brief description: In Figure 3，larvae were injected once with dsRNA on the first day of the 5th instar. Samples were collected at 24h, 48 h and 72 h post-injection to evaluate the RNAi silencing efficiency of PDFR. Larvae injected with dsGFP at corresponding time points served as the control. Data are presented as mean ± standard error (SE) (n = 3). Quantitative real-time PCR (qRT-PCR) was performed to assess changes in PDFR expression levels. Each treatment had three mechanical and three biological replicates. The ct value is the original data detected by the instrument. After calculation by the formula, 2-∆∆Ct is obtained. We calculated the relative expression of the target gene according to the 2-∆∆Ct. We selected EF-1a and RPL13 as internal reference genes. The expression levels are shown relative to those in dsGFP at the same point. Statistical significance of gene expression differences between the two groups was calculated using Student's t-test ( **P < 0.01; *** P < 0.001).

7.Raw_data_for-Figure4-Survival-rates-of-larvae-16L8D

Data brief description: In Figure 4A，using injection of 2 μg dsGFP as a negative control, 2 μg of dsPDFR were injected into first-day 4th instar larvae of H. cunea under 16L:8D. Survival rates of larvae in each treatment group and the control group were recorded at 24, 48 and 72 hours post-injection, and survival curves were plotted accordingly (0 represents surviving individuals, 1 represents dead individuals). Each treatment consisted of 30 larvae and was independently replicated three times. The log‑rank (Mantel‑Cox) test was used to analyze the significance of differences among the survival curves (ns P>0.05, *** P < 0.001).

8.Raw_data_for-Figure4-Survival-rates-of-larvae-12L12D

Data brief description: In Figure 4B ，using injection of 2 μg dsGFP as a negative control, 2 μg of dsPDFR were injected into first-day 4th instar larvae of H. cunea under 12L:12D. Survival rates of larvae in each treatment group and the control group were recorded at 24, 48 and 72 hours post-injection, and survival curves were plotted accordingly (0 represents surviving individuals, 1 represents dead individuals). Each treatment consisted of 30 larvae and was independently replicated three times. The log‑rank (Mantel‑Cox) test was used to analyze the significance of differences among the survival curves (ns P>0.05, *** P < 0.001).

9.Raw_data_for-Figure4-Survival-rates-of-larvae-24L0D

Data brief description: In Figure 4C，using injection of 2 μg dsGFP as a negative control, 2 μg of dsPDFR were injected into first-day 4th instar larvae of H. cunea under 24L:0D. Survival rates of larvae in each treatment group and the control group were recorded at 24, 48 and 72 hours post-injection, and survival curves were plotted accordingly (0 represents surviving individuals, 1 represents dead individuals). Each treatment consisted of 30 larvae and was independently replicated three times. The log‑rank (Mantel‑Cox) test was used to analyze the significance of differences among the survival curves (ns P>0.05, *** P < 0.001).

10.Raw_data_for-Figure5-ecdysis-rates-of-larvae-16L8D

Data brief description: In Figure 5A，shows the timing of molting of the 5th instar larvae under 16L:8D. The 1st day of 4th instar H. cunea larvae were microinjected per treatment (dsGFP/dsPDFR), the 100 living individuals were randomly selected for the hourly monitoring of timing of molting that the larvae reached the 5th instar larvae. The Zeitgeber Time is in 24h.

11.Raw_data_for-Figure5-ecdysis-rates-of-larvae-12L12D

Data brief description: In Figure 5B，shows the timing of molting of the 5th instar larvae under 12L:12D. The 1st day of 4th instar H. cunea larvae were microinjected per treatment (dsGFP/dsPDFR), the 100 living individuals were randomly selected for the hourly monitoring of timing of molting that the larvae reached the 5th instar larvae. The Zeitgeber Time is in 24h.

12.Raw_data_for-Figure5-ecdysis-rates-of-larvae-24L0D

Data brief description: In Figure 5C，shows the timing of molting of the 5th instar larvae under 24L:0D. The 1st day of 4th instar H. cunea larvae were microinjected per treatment (dsGFP/dsPDFR), the 100 living individuals were randomly selected for the hourly monitoring of timing of molting that the larvae reached the 5th instar larvae. The Zeitgeber Time is in 24h.

13.Raw_data_for-Figure6-mortality-malformation rate-pupation rate-mature larvae-16L8D

Data brief description: In Figure 6A，shows the mortality, malformation rate, and pupation rate in the mature larvae of the H. cunea under 16L:8D. The 4th day of 6th instar H. cunea larvae were microinjected with 2 μg dsRNA (dsGFP/dsPDFR). Each treatment group was consisted of 25 mature larvae, with four replicates for each treatment. Every 24 hours following injection, the mortality, malformation, and pupation rates of the 6th instar H. cunea larvae were determined for both the dsGFP group and the dsPDFR group. Statistical significance between the two groups was calculated using Student's t-test (ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P< 0.001; **** P < 0.0001).

14.Raw_data_for-Figure6-mortality-malformation rate-pupation rate-mature larvae-12L12D

Data brief description: In Figure 6B，shows the mortality, malformation rate, and pupation rate in the mature larvae of the H. cunea under 12L:12D. The 4th day of 6th instar H. cunea larvae were microinjected with 2 μg dsRNA (dsGFP/dsPDFR). Each treatment group was consisted of 25 mature larvae, with four replicates for each treatment. Every 24 hours following injection, the mortality, malformation, and pupation rates of the 6th instar H. cunea larvae were determined for both the dsGFP group and the dsPDFR group. Statistical significance between the two groups was calculated using Student's t-test (ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P< 0.001; **** P < 0.0001).

15.Raw_data_for-Figure6-mortality-malformation rate-pupation rate-mature larvae-24L0D

Data brief description: In Figure 6C，shows the mortality, malformation rate, and pupation rate in the mature larvae of the H. cunea under 24L:0D. The 4th day of 6th instar H. cunea larvae were microinjected with 2 μg dsRNA (dsGFP/dsPDFR). Each treatment group was consisted of 25 mature larvae, with four replicates for each treatment. Every 24 hours following injection, the mortality, malformation, and pupation rates of the 6th instar H. cunea larvae were determined for both the dsGFP group and the dsPDFR group. Statistical significance between the two groups was calculated using Student's t-test (ns, P > 0.05; * P < 0.05; ** P < 0.01; *** P< 0.001; **** P < 0.0001).

Contact

For questions about the dataset, please contact:

Lili Sun: sunlilinefu@126.com

Chuanwang Cao: chuanwangcao@nefu.edu.cn