Growth inhibition of Spodoptera frugiperda larvae by camptothecin correlates with alteration of the structures and gene expression profiles of the midgut

- alteration of the structures and gene expression profiles of the midgut.
- frugiperda larvae..
- Histological and cytological changes were examined in the midgut of larvae fed on an artificial diet supplemented with 1.0 and 5.0 µg/g CPT.
- A total of 915 and 3560 differentially expressed genes (DEGs) were identified from samples treated with 1.0 and 5.0 µg/g CPT, respectively..
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- Plants are considered the most abundant natural re- source in the world for the identification of chemicals with insecticidal activity [7, 8].
- Field tests with 0.2 % camptothecin emulsifiable concentrate (EC) have shown high mortality on three important agricultural pests Nilaparvata lugens (Ståhl) Brevicoryne brassicae (L.
- Changes in the weight of S.
- Histopathological and ultrastructural changes in the midgut of larvae fed on diets.
- containing 1.0 and 5.0 µg/g CPT, respectively, were exam- ined.
- frugiperda larvae and has the potential as an insecticide for controlling this import- ant insect pest in the field..
- After 7 days of feeding, the larvae from the control group developed to sixth instar larvae, while the larvae treated with CPT grew slowly, and developed only into fourth or fifth instars.
- Histopathological changes were observed in the larval midgut fed on diets containing 1.0 and 5.0 µg/g CPT for 7 days based on hematoxylin-eosin (HE) staining.
- In comparison, many cells were disappeared and only a thin intestinal barrier was ob- served in the larval midgut fed on a 1.0 µg/g CPT diet (Fig.
- In larvae fed on a diet containing 5.0 µg/g CPT, only the basement membrane was left in the intes- tinal wall of the midgut, and nearly all functional cells disappeared (Fig.
- Similar phenomena were ob- served in the gut structure under TEM.
- In control lar- vae, chromatin was evenly distributed in the nucleus..
- Mitochondria and endoplasmic reticulum were abun- dant and distributed evenly in the cytoplasm.
- Microvilli were ordinally distributed in the gut (Fig.
- In con- trast, the number of mitochondria and endoplasmic reticulum decreased in midgut cells in larvae treated with 1.0 µg/g CPT.
- In larvae fed on the 5.0 µg/g CPT diet, chromatin con- densation occurred and chromatins were located close to the nuclear envelope.
- Midguts dissected from larvae treated with CPT (1.0 and 5.0 µg/g) for 7 days and control larvae were used for tran- scriptomic analyses.
- Q20 and Q30 refer to the percentage of bases with sequencing quality above 99 and 99.9 % to the total bases in the transcriptome.
- frugiperda larvae.
- A total of 915 unigenes were expressed differentially be- tween controls and samples treated with 1.0 µg/g CPT..
- Compared to the control group, 612 unigenes were up- regulated and 291 unigenes were down-regulated in the group treated with 1.0 µg/g CPT (Fig.
- The number of DEGs between control and 5.0 µg/g CPT-treated sam- ples increased to 3560.
- Compara- tive analyses revealed that 683 unigenes were differen- tially expressed in both 1.0 and 5.0 µg/g CPT-treated samples when compared to control.
- As shown in Table 1 and 39 detoxification genes were differentially expressed between controls and sam- ples treated with 1.0 µg/g CPT.
- The number of detoxification genes expressed differentially between controls and sam- ples treated with 5.0 µg/g CPT increased to 108, includ- ing genes encoding 57 P450s, 13 GSTs, 8 COEs, 11 UGTs, and 19 ABCs.
- Two and 18 genes en- coding mucins were differentially expressed between controls and samples treated with 1.0 and 5.0 µg/g CPT, respectively.
- 2 Histopathological and ultrastructural changes in the midgut of larvae fed on the diet supplemented with 1.0 and 5.0 µg/g CPT.
- Hematoxylin – eosin staining of the midgut obtained from larvae fed on a normal diet.
- B: Histopathological changes in the midgut dissected from larvae fed on the diet supplemented with 1.0 µg/g CPT.
- C: Histopathological changes of the midgut dissected from larvae fed on the diet supplemented with 5.0 µg/g CPT.
- D: The ultrastructure of the midgut obtained from larvae fed on normal diet.
- E: The ultrastructure of the midgut dissected from larvae fed on the diet supplemented with 1.0 µg/g CPT.
- F: The ultrastructure of the midgut dissected from larvae fed on the diet supplemented with 5.0 µg/g CPT.
- (DN34507_c0_g1) and mucin-17 (DN3811_c0_g1) were up-regulated in samples treated with 1.0 µg/g CPT when compared to control.
- Most (16) DEGs encoding mucin proteins were up-regulated in samples treated with 5.0 µg/g CPT, and only two mucin genes were down- regulated (Fig.
- Specifically, Four genes encoding larval cuticle protein LCP-17 (DN2495_c0_g1), cuticle protein 6.4-like (DN2113_c0_g1), cuticle protein CP14.6-like (DN38621_c0_g1) and cuticular protein RR- 2 (DN1220_c0_g1) were down-regulated in samples treated with 1.0 µg/g CPT.
- Interestingly, 26 unigenes en- coding cuticle proteins were up-regulated in samples treated with 5.0 µg/g CPT, whereas there were only two cuticle protein-encoding genes that were down- regulated (Fig.
- A total of 553 DEGs between controls and samples treated with 1.0 µg/g CPT were assigned to 175 GO terms.
- A total of 2000 DEGs between controls and samples treated with 5.0 µg/g CPT were assigned to 215 GO terms.
- KEGG analysis revealed that 369 DEGs between con- trols and samples treated with 1.0 µg/g CPT were assigned to 178 pathways.
- There were 683 unigenes that exhibited differential expression between samples treated with 1.0 and 5.0 µg/g CPT.
- There were 464 unigenes that exhibited up-regulated expressions in samples treated with 1.0 and 5.0 µg/g CPT when compared to control.
- There were 217 unigenes that exhibited down-regulated expressions in samples treated with 1.0 and 5.0 µg/g CPT when compared to control.
- Purple ring represents DEGs identified from the comparison between controls and samples treated with 1.0 µg/g CPT.
- Green ring represents DEGs identified from the comparison between controls and samples treated with 5.0 µg/g CPT.
- metabolism” (15 DEGs), and “Ribosome biogenesis in eu- karyotes” (14 DEGs).
- KEGG analysis assigned 1401 DEGs between controls and samples treated with 5.0 µg/g CPT to 232 pathways.
- “DNA replication” was the most significantly enriched KEGG pathway in both sam- ples treated with either 1.0 or 5.0 µg/g CPT..
- frugiperda has become a serious insect pest in China in the past couple of years [26].
- Various chemicals such as chlorantraniliprole, spinetoram, emamectin benzoate, spinetoram, acephate, and pyraquinil have been evalu- ated to control this pest in the field [27 – 29].
- frugiperda larvae treated with CPT, but no mortality was observed.
- More efficient derivatives with improved solubility and hydrophobicity may be devel- oped in the future for pest control..
- CPT has been reported to induce alterations in the midguts of Trichoplusia ni and S.
- In this study, we ob- served the loss of epithelial cells, abnormal cell structure, and intestinal wall degradation in the midgut of S.
- For ex- ample, transcriptomic analyses have been carried out to identify DEGs in the Chinese populations of S.
- frugiperda larvae treated with azadirachtin were also ini- tially analyzed [36].
- frugiperda larvae treated with CPT were analyzed for the first time..
- For example, the tran- scription levels of detoxifying-related genes including P450s and GSTs were up-regulated by low-dose of acet- amiprid in the midgut of B.
- litura lar- val midguts after being treated with tomatine [42].
- A: The top 14 pathways enriched with DEGs obtained from midgut samples from larvae treated with 1.0 µg/g CPT with FDR values.
- B: The 14 pathways significantly enriched with DEGs obtained from midgut samples from larvae treated with 5.0 µg/g CPT (corrected P-values <.
- Here we found that a large number of detoxification-related genes were coordinately up-regulated in the midgut of S.
- The dose-dependent upregulation of these genes further indicates their roles in the detoxification of CPTs in S..
- armigera after being treated with aba- mectin, indoxacarb, and lambda-cyhalothrin [47].
- mori treated with NaF [46].
- In the present study, eleven ABC transporters were up-regulated in S.
- frugiperda larvae treated with 5.0 µg/g CPT.
- frugiperda larvae treated with 1.0 and 5.0 µg/g CPT, respectively.
- Two UGT genes were previously found involved in the detoxification of benzoxazinoids from maize in S.
- frugiperda larvae [54].
- frugiperda, mucins are located in the peritrophic membrane and thus may protect epithelial cells and immobilize proteolytic en- zymes [57].
- Multiple genes encoding different mucins were mostly up-regulated in samples treated with CPT, suggesting that mucins may have participated in repairing damages caused by CPT in the peritrophic membrane as reported previously [58, 59].
- The up-regulation of genes en- coding both mucins and cuticle proteins suggested that the peritrophic membrane in the midgut of S..
- frugiperda larvae treated with different concentra- tions of CPT.
- Damage in the midgut was found after CPT treatments based on HE staining and TEM observation.
- Genes involved in detoxification, DNA replication, and structural components of the peritrophic membrane were up-regulated signifi- cantly.
- frugiperda larvae treated with 1.0 and 5.0 µg/g CPT was dissected and washed with cold phosphate-buffered saline (PBS)..
- Histopathological changes in the midgut of larvae fed on CPT were visual- ized on a microscope (Nikon, Japan)..
- Larval midguts exposed to 1.0 and 5.0 µg/g CPT for 7 days were dissected, washed with cold PBS, and then fixed in glutaraldehyde solution at 4 °C for 4 h.
- Ten to fifteen larval midgut samples exposed to 1.0 and 5.0 µg/g CPT for 7 days were collected, Total RNA was extracted using TRIzol® Reagent (Invitrogen, USA).
- The transcripts that shared sequence content were clustered and the longest transcript in the cluster was selected as the unigene.
- The raw reads of transcriptomes in this study have been deposited in the NCBI SRA database with the accession number of SRP242660..
- Estimation of the potential infestation area of newly-invaded fall armyworm Spodoptera frugiperda in the yangtze river valley of China.
- Effect of the flavonoid rutin on the biology of Spodoptera frugiperda (Lepidoptera:.
- The roles of mucus-forming mucins, peritrophins and peritrophins with mucin domains in the insect midgut.
- Stability of selected reference genes in Sf9 cells treated with extrinsic apoptotic agents.
- Azadirachtin A inhibits the growth and development of Bactrocera dorsalis larvae by releasing cathepsin in the midgut.
- Responses of detoxification enzymes in the midgut of Bombyx mori after exposure to low-dose of acetamiprid.
- detoxification in the diamondback moth, Plutella xylostella.
- Analysis of digital gene expression profiling in the gonad of male silkworms (Bombyx mori) under fluoride stress.
- Transcriptional response of ATP-binding cassette (ABC) transporters to insecticides in the cotton bollworm, Helicoverpa armigera.
- Identification and transcriptional response of ATP-binding cassette transporters to chlorantraniliprole in the rice striped stem borer, Chilo suppressalis.
- Chitin is a necessary component to maintain the barrier function of the peritrophic matrix in the insect midgut.
- Identification of cuticular protein genes in the Colorado potato beetle Leptinotarsa decemlineata (Coleoptera:

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