Abstract
Artificial induction has been widely employed to generate genetic resources with increased mutation rates for subsequent selection. To assess the potential of physical mutagenesis in shrimp breeding, we utilized Atmospheric and Room Temperature Plasma (ARTP) to induce mutations in fertilized eggs from four different families of Litopenaeus vannamei. Treatment durations ranged from 2 to 12 min. The results revealed a significant decrease in hatching rates among the ARTP-treated groups, ranging from 3.22% to 67.81%, with more substantial reductions observed in longer treatment durations. Following a 90-day mixed rearing test, we found no statistically significant differences (P > 0.05) in the average body weight of 743 shrimps across the 13 mutagenesis groups compared to the control group. However, variations in weight were observed in the 22 largest or smallest sample groups, with 12 groups displaying an average increase of 8.69% and 10 groups showing an average decrease of 9.97% compared to their respective controls. Regarding fitness traits, seven treatment groups from two families exhibited higher median lethal times or mean survival times against WSSV compared to the control group, while two treatment groups demonstrated improved resistance to ammonium nitrogen. Genomic resequencing analysis was conducted on growth sampling groups, identifying a total of 2,718,961 SNPs and 331,172 Indels in the treatment group. The mutation rate in the treatment group was 2.148 × 10−3, significantly higher than the control group's rate of 5.275 × 10−5. Within the treatment group, 17,104 specific non-synonymous SNPs were identified in the exons of 7780 genes, markedly surpassing the control group's count of 121 SNPs and 110 genes. GO annotation analysis revealed an even distribution of mutations induced by ARTP across various biological processes, cellular components, and functions. Furthermore, KEGG enrichment analysis demonstrated a significant enrichment trend in metabolic pathways, both in terms of gene count and FDR value. A total of 42 KEGGs and several potential growth-related genes, including IGF1 and AMY2, previously confirmed to be highly correlated with growth in L. vannamei, were identified. In conclusion, a feasible method for whole-genome mutagenesis in shrimp zygotes was developed in this study. Tests on economic traits revealed that ARTP can generate mutant individuals with either a greater degree of variation or superior traits. Resequencing revealed the presence of numerous mutation SNPs involved in various biological processes and contribute to a higher level of genetic diversity. These results suggest that ARTP can be utilized to enhance the available genetic variation in shrimp breeding, thereby providing valuable materials and methods for genetic breeding in whiteleg shrimp.
Published Version
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