| Plutella xylostella（Linnaeus）,belonging to the order Lepidoptera.Plutella xylostella is an important pest of cruciferous vegetables worldwide,and it is one of the most serious pests with resistance development.Plutella xylostella has strong reproductive ability,short generation,fast mutation and good genetic plasticity.Long-term use of chemical methods to correct Plutella xylostella not only caused damage to the environment,but also caused the resistance of Plutella xylostella to various pesticides to increase rapidly.It is imperative to develop the green prevention and control technolo of diamondback moth.At present,the research on Plutella xylostella parasitism is mostly at the physiological and biochemical level,but the genetic and molecular mechanisms of split-shot infertility are still unclear.In this study,using the TMT proteomics method,we studied the proteomic changes of the spermatozoa plutella after 0 Gy,200 Gy and 400 Gy dose60Co-γirradiation,and combined with the bioinformatics analysis method,preliminarily studied the molecular mechanism of irradiation affecting the male reproductive ability of the Plutella xylostella.The main research results are as follows:Quantitative proteomics based on TMT labeling technology identified a total of 5145proteins in the spermatozoa plutella tissues,and 606 differential proteins in the 200 Gy/CK comparison group,among which 296 proteins were significantly up-regulated and 310proteins were significantly down-regulated.A total of 1843 differential proteins were identified in the 400 Gy/CK comparison group,of which 970 were significantly up-regulated and 873 were significantly down-regulated.A total of 2057 differential proteins were identified in the 400 Gy/200 Gy comparison group,of which 1116 were significantly up-regulated and 941 were significantly down-regulated.According to the GO analysis results,the differential proteins are mainly involved in the metabolic process and the cellular process,and mainly perform the functions of binding and catalytic activity.The KEGG pathway enrichment analysis showed that the oxidative phosphorylation,TCA cycle,phagocytosis and various amino acid anabolism pathways of the moth were significantly up-regulated after 200 Gy irradiation.The spliceosome,RNA transport,m RNA monitoring pathway and other pathways were significantly enriched and down-regulated.After 400 Gy dose irradiation,the glycolysis pathway,ribosomal generating pathway and various carbohydrate metabolic pathways were significantly up-regulated in the spermatozoa plutellae,and the mitochondrial pathway,homologous recombination pathway,autophagy and apoptosis pathway were also activated.The ribosomal pathway and various amino acid metabolic pathways were inhibited.Compared with the group of 400 Gy and 200 Gy dose irradiation,400 Gy irradiation promoted the expression of signal pathways related to protein hydrolysis and gene expression in tissue cells,and inhibited the expression of signal pathways related to protein synthesis and cytoskeleton synthesis.At the same time,Westetn Blot showed that irradiation had an effect on the phosphorylation level of tyrosine.With the increase of irradiation dose,the phosphorylation level of tyrosine gradually decreased.When the male pupae of diamondback moth was treated with different radiation doses,different response mechanisms were activated in the spermatozoa.After 200 Gy irradiation dose stimulation,the cells in the testis of the moth can resist the damage caused by irradiation by accelerating the metabolism,increasing the energy supply and protein synthesis.However,the inhibition of transcription-related pathways may result in the increase of defective RNA,which will be passed on to the offspring and affect the growth and reproduction of the offspring.However,the stimulation of 400 Gy radiation dose caused autophagy and apoptosis of spermatozoa,which weakened the motility and fertilization ability of spermatozoa,resulting in reproductive toxicity. |
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