Evaluation of Predation on Phytophagous Insects by a Phytozoophagous Mirid Bug, Apolygus lucorum
Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Insect Rearing
2.2. Laboratory Predation Assays
- (a)
- Predation on H. armigera eggs: Cotton gauze pieces (3 cm × 3 cm) containing H. armigera eggs (laid within 24 h) were prepared. Non-viable or misshapen eggs were removed under a stereomicroscope using an insect pin, and the number of eggs on each piece was recorded. Each gauze piece was placed in a glass tube (7.5 cm × 2 cm), and one A. lucorum individual (2nd instar nymph, 4th instar nymph, 5-day-old female, or 5-day-old male) was introduced using a soft brush. The tube opening was covered with an 80-mesh nylon net and secured with a rubber band to prevent escape. A moistened cotton ball was placed on the net to provide moisture. Prey densities tested were 5, 10, 20, 30, 40, and 50 eggs per tube, with five replicates per density. After 24 h, the number of consumed eggs was recorded under a stereomicroscope based on their shriveled, flattened appearance. Any eggs that had hatched within 24 h (which did not occur) or not shriveled were excluded from consumption counts.
- (b)
- Predation on A. gossypii nymphs: Cotton leaves infested with A. gossypii nymphs were collected from the field and brought back to the laboratory. Under a stereomicroscope, healthy, unparasitized, and uninfected aphid nymphs (without distinction of instar to better reflect natural field conditions) were carefully selected using a fine brush and transferred onto cotton gauze pieces (3 cm × 3 cm). Each gauze piece containing the designated number of aphids was placed in a glass tubes (7.5 cm × 2 cm), and one A. lucorum individual (2nd instar nymph, 4th instar nymph, 5-day-old female, or 5-day-old male) was introduced into each tube. The tube opening was covered with an 80-mesh nylon net and secured with a rubber band to prevent escape, while a moistened cotton ball was placed on the net to provide moisture. Prey densities tested were 2, 4, 8, 12, 16, and 20 nymphs per tube, with five replicates per density. After 24 h, the predator was removed, and the number of consumed aphids (identified by their darkened, shriveled appearance) was recorded.
- (c)
- Predation on B. tabaci nymphs: Cotton leaves infested with B. tabaci nymphs were collected from the field and brought back to the laboratory. The leaves were cut into leaf discs (3 cm × 3 cm). Under a stereomicroscope, excess nymphs, as well as unhealthy, parasitized, or infected individuals, were removed, leaving only healthy 4th instar nymphs to ensure uniformity of prey size. Each leaf disc was placed in a glass tube (7.5 cm × 2 cm), and one A. lucorum individual (2nd instar nymph, 4th instar nymph, 5-day-old female, or 5-day-old male) was introduced into each tube. The tube opening was covered with an 80-mesh nylon net and secured with a rubber band to prevent escape, while a moistened cotton ball was placed on the net to provide moisture. The prey densities tested were 2, 4, 8, 12, 16, and 20 nymphs per tube, with five replicates per density. After 24 h, the number of consumed nymphs (identified by their shriveled appearance) was recorded.
2.3. Field Detection of Predation by A. lucorum
2.3.1. COI-Based Molecular Detection Method
- (a)
- DNA template preparation: Genomic DNA was extracted from individual A. lucorum using a Blood/Cell/Tissue Genomic DNA Extraction Kit (Tiangen Biotech, Beijing, China) following the manufacturer’s instructions. The DNA extracts were stored at −20 °C until use. For PCR amplification, 1 µL of DNA solution was used as the template.
- (b)
- Species-specific primers: Species-specific primers were designed based on mitochondrial COI gene sequences from H. armigera (GenBank accession No. AY264944) and A. gossypii (GenBank accession No. AY842502). Primers were synthesized by Sangon Biotech (Shanghai, China) and purified by PAGE.Primers for H. armigera:Forward primer MLCF: 5′-GGTGATCCTATTTTATATCAC-3′Reverse primer MLCR: 5′-GAGTATCAATATCTATACCAG-3′Amplicon length: 239 bp.Primers for A. gossypii:Forward primer MYF: 5′-TTCACATCAGCAACTATAATC-3′Reverse primer MYR: 5′-ACTACATAATAAGTGTCATGC-3′Amplicon length: 208 bp.
- (c)
- PCR amplification: PCR reactions were performed in a thermal cycler (Bioer, Hangzhou, China). Each 25 µL reaction mixture contained 0.25 µL of EX Taq polymerase (TaKaRa, Dalian, China), 2.5 µL of 10× PCR buffer (supplied with the Taq polymerase), 2 µL of dNTPs (Tiangen Biotech, Beijing, China), 1 µL of DNA template, 0.5 µL of each forward and reverse primer, and sterile distilled water to a final volume of 25 µL. Each sample was run in duplicate to ensure reliability.The PCR cycling program was as follows: initial denaturation at 94 °C for 5 min; 36 cycles of 94 °C for 30 s, 55 °C for 30 s, and 72 °C for 1 min; final extension at 72 °C for 10 min.Positive controls (DNA from H. armigera eggs or A. gossypii nymphs) and negative controls (sterile water instead of DNA template, and DNA from A. lucorum that had not fed on H. armigera eggs or A. gossypii) were included.
- (d)
- Amplicon detection: A 10 µL of each PCR product was electrophoresed on a 1.2% agarose gel containing 0.5 µg/mL GoldView nucleic acid stain (Solarbio, Beijing, China) at 180 V for 15 min. A DL2000 DNA molecular weight marker (TaKaRa, Dalian, China) was used as a reference. After electrophoresis, the gel was visualized and photographed under UV light using a gel imaging system (Tanon, Shanghai, China).
2.3.2. Field Investigation and Sampling
2.4. Statistical Analysis
3. Results
3.1. Functional Response Type
3.2. Predation Rates of A. lucorum on Three Prey Types
3.3. Detection of Prey Predation by A. lucorum in Cotton Fields
4. Discussion
5. Conclusions
Author Contributions
Funding
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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| Prey | Developmental Stage | Parameter | Estimate | Standard Error | p-Value | R2 |
|---|---|---|---|---|---|---|
| Helicoverpa armigera eggs | 2nd instar nymph | Linear (P1) | −0.0648 | 0.0940 | 0.4906 | 0.9199 |
| Quadratic (P2) | 0.0034 | 0.0035 | 0.3415 | |||
| 4th instar nymph | Linear (P1) | −0.6235 | 0.5008 | 0.2132 | 0.9736 | |
| Quadratic (P2) | 0.0160 | 0.0147 | 0.2743 | |||
| 5-day-old female adult | Linear (P1) | −0.6754 | 0.1924 | 0.0004 | 0.9784 | |
| Quadratic (P2) | 0.0198 | 0.0061 | 0.0012 | |||
| 5-day-old male adult | Linear (P1) | −0.1182 | 0.1104 | 0.2844 | 0.9965 | |
| Quadratic (P2) | 0.0029 | 0.0040 | 0.4744 | |||
| Aphis gossypii nymph | 2nd instar nymph | Linear (P1) | −0.3019 | 0.3630 | 0.4056 | 0.9655 |
| Quadratic (P2) | 0.0181 | 0.0336 | 0.5894 | |||
| 4th instar nymph | Linear (P1) | −0.3196 | 0.3735 | 0.3922 | 0.9802 | |
| Quadratic (P2) | 0.0356 | 0.0350 | 0.3086 | |||
| 5-day-old female adult | Linear (P1) | −0.1879 | 0.3728 | 0.6142 | 0.9637 | |
| Quadratic (P2) | 0.0350 | 0.0357 | 0.3246 | |||
| 5-day-old male adult | Linear (P1) | −0.3590 | 0.4513 | 0.4263 | 0.9573 | |
| Quadratic (P2) | 0.0451 | 0.0422 | 0.2858 | |||
| Besimia tabaci nymph | 2nd instar nymph | Linear (P1) | −0.1597 | 0.3687 | 0.6649 | 0.9141 |
| Quadratic (P2) | 0.0123 | 0.0359 | 0.7315 | |||
| 4th instar nymph | Linear (P1) | −0.1005 | 0.2703 | 0.7100 | 0.9540 | |
| Quadratic (P2) | 0.0108 | 0.0257 | 0.6750 | |||
| 5-day-old female adult | Linear (P1) | −0.2890 | 0.2994 | 0.3344 | 0.9029 | |
| Quadratic (P2) | 0.0331 | 0.0282 | 0.2404 | |||
| 5-day-old male adult | Linear (P1) | −0.0530 | 0.2984 | 0.8593 | 0.9556 | |
| Quadratic (P2) | 0.0025 | 0.0288 | 0.9320 |
| Prey | Developmental Stage | Functional Response | p | a | Th (d) | Maximum Consumption | a/Th |
|---|---|---|---|---|---|---|---|
| Helicoverpa armigera eggs | 2nd instar nymph | Na = N/(3.1989 + 0.0239 N) | 0.0010 | 0.3126 | 0.0239 | 41.84 | 13.08 |
| 4th instar nymph | Na = N/(0.9109 + 0.0145 N) | 0.0002 | 1.0978 | 0.0145 | 68.97 | 75.71 | |
| 5-day-old female adult | Na = N/(0.9113 + 0.0176 N) | <0.0001 | 1.0973 | 0.0176 | 56.82 | 62.35 | |
| 5-day-old male adult | Na = N/(1.0055 + 0.0136 N) | <0.0001 | 0.9945 | 0.0136 | 73.53 | 73.13 | |
| Aphis gossypii nymph | 2nd instar nymph | Na = N/(0.9358 + 0.1254 N) | 0.0048 | 1.0686 | 0.1254 | 7.97 | 8.52 |
| 4th instar nymph | Na = N/(1.4069 + 0.0037 N) | <0.0001 | 0.7108 | 0.0037 | 270.27 | 192.11 | |
| 5-day-old female adult | Na = N/(1.4640 + 0.0025 N) | <0.0001 | 0.6831 | 0.0025 | 400.00 | 273.24 | |
| 5-day-old male adult | Na = N/(1.2265 + 0.0012 N) | <0.0001 | 0.8153 | 0.0012 | 833.33 | 679.42 | |
| Besimia tabaci nymph | 2nd instar nymph | Na = N/(5.9340 + 0.2886 N) | <0.0001 | 0.1685 | 0.2886 | 3.47 | 0.58 |
| 4th instar nymph | Na = N/(3.7406 + 0.0597 N) | 0.0003 | 0.2673 | 0.0597 | 16.75 | 4.48 | |
| 5-day-old female adult | Na = N/(5.2068 + 0.2926 N) | 0.0200 | 0.1921 | 0.2926 | 3.42 | 0.66 | |
| 5-day-old male adult | Na = N/(3.6654 + 0.1742 N) | <0.0001 | 0.2728 | 0.1742 | 5.74 | 1.57 |
| Year | Area | Total No. of Detections | Positive Detection Rate of Helicoverpa armigera Eggs | Positive Detection Rate of Aphis gossypii |
|---|---|---|---|---|
| 2009 | Langfang | 126 | 0.00 | 7.04 ± 3.73 a |
| Xiajin | 141 | 0.00 | 1.22 ± 1.26 a | |
| Xinxiang | 42 | 0.00 | 0.00 ± 2.47 a | |
| 2010 | Langfang | 135 | 0.00 | 3.70 ± 2.47 a |
| Xiajin | 114 | 0.00 | 0.00 ± 1.23 a | |
| Xinxiang | 41 | 0.00 | 0.00 ± 1.23 a |
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Wang, L.; Liu, B.; Wu, K. Evaluation of Predation on Phytophagous Insects by a Phytozoophagous Mirid Bug, Apolygus lucorum. Insects 2026, 17, 397. https://doi.org/10.3390/insects17040397
Wang L, Liu B, Wu K. Evaluation of Predation on Phytophagous Insects by a Phytozoophagous Mirid Bug, Apolygus lucorum. Insects. 2026; 17(4):397. https://doi.org/10.3390/insects17040397
Chicago/Turabian StyleWang, Lili, Baoyou Liu, and Kongming Wu. 2026. "Evaluation of Predation on Phytophagous Insects by a Phytozoophagous Mirid Bug, Apolygus lucorum" Insects 17, no. 4: 397. https://doi.org/10.3390/insects17040397
APA StyleWang, L., Liu, B., & Wu, K. (2026). Evaluation of Predation on Phytophagous Insects by a Phytozoophagous Mirid Bug, Apolygus lucorum. Insects, 17(4), 397. https://doi.org/10.3390/insects17040397

