Transcriptome profiling of banana shrimp (Fenneropenaeus merguiensis) ovaries and testes and its molecular markers 

Abstract

Banana shrimp (Fenneropenaeus merguiensis) is one of the popular shrimp consumed in Thailand. The development of the banana shrimp farming industry is therefore important. Since female shrimp grow faster and weigh more than males, female shrimp are therefore required for farming. A transcriptomic analysis of the testes and ovaries from the juvenile and adult shrimp was compared to search for the genes involved in sex differentiation and ovary development. The result showed a higher gene expression number in the testes of juvenile shrimp, while adult shrimp had a higher gene expression number in the ovaries. The differentially expressed genes (DEGs) between the testes and ovaries at each stage indicated that Sxl2, dsx, AGH, FEM-1, and Nrg-X2 are required for testes development during juvenile banana shrimp. In addition, SOP1, SOP2, Ptgr1, EST, Vgr, Vmol1, and TR-beta A are genes required for ovarian development during both juvenile and adult stages. The high expression of FoxL2 was found in the testis in this study, and FoxL2 was reported in other crustaceans involved in Vtg expression. Therefore, the interaction between FoxL2 of banana shrimp and the Vtg promoter silico modeling was identified with two binding sites and the highest relative binding score at 0.89. Knockdown FoxL2 gene with dsRNA in adult female banana shrimp stage can inhibit FoxL2 expression at 4 and 6 days and increase expression of Vtg as compared to the control group. This study indicates that FoxL2 plays a vital role in regulating Vtg expression in the ovary. In addition, randomly identifying postlarvae shrimp species for shrimp farming is vital as postlarvae shrimp, especially banana shrimp, are hard to locate using morphology. Identifying shrimp species in processed shrimp food is also essential in correctly labeling processed shrimp food products so as not to deceive consumers. Therefore, the markers VASA160, COI-RFLP, COI266, and Fm-15 were developed to identify F. merguiensis, Penaeus vannamei, Penaeus monodon, and Macrobrachium rosenbergii. VASA160, COI266, and Fm-15 were the specific marker to F. merguiensis. While COI-RFLP was used to identify each shrimp according to the restricted digestion of the COI PCR product. F. merguiensis and P. monodon were excluded from P. vannamei and M. rosenbergii using XbaI digestion. Then F. merguiensis was separated from P. monodon using SpeI, while P. vannamei was excluded from M. rosenbergii using HindIII. Furthermore, M. rosenbergii can be confirmed by digestion with ApaI. The higher expressed gene in the ovary in juvenile and adult shrimp will be useful in developing female banana shrimp and stimulating the ovary development of banana shrimp. Screening postlarvae-specific species such as F. merguiensis for farming using developed molecular markers can be utilized to identify shrimp species in processed shrimp foods for corrected labeling of the processed food.??????????(Fenneropenaeus merguiensis)???????????????????????????????? ??????????????????????????????????????????????????? ??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ???????????? ????????????? (transcriptome) ???????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ??????????????????????????????????????????????????????????????????????????? Sxl2 dsx AGH FEM-1 ??? Nrg-X2 ????????????????????????????????????????????????????????? ????????? SOP1 SOP2 Ptgr1 EST Vgr Vmol1 ??? TR-beta A ???????????????????????????????????????????????????????????????????????? ????????????? FoxL2 ????????????????????????????????????????????????? FoxL2 ???????????????????????? crustaceans ???????????????????????????????????????? Vtg ????????????????????????????????????? FoxL2 ???????????????? Vtg promoter ?????????????????????????????????????????????????????????????????????????????????????????????????????????? (relative binding score) ??????????????????????? 0.89 ??? knockdown ??? FoxL2 ???? dsRNA ??????????????????????????????????????????????????????????? FoxL2 ??? 4 ??? 6 ????????????????????????????? FoxL2 ???????????????????????? Vtg ????????????????????????????????????? ?????????????????????????? FoxL2 ?????????????????????????????????????????????? Vtg ???????????????????????????????????????????????????? (postlarvae) ?????????????????????????????????????? ??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ????????????????????????? VASA160 COI-RFLP COI266 ??? Fm-15 ???????????????????????????????? F. merguiensis Penaeus vannamei Penaeus monodon ??? Macrobrachium rosenbergii ?????????????????? VASA160 COI266 ??? Fm-15 ???????????????????????????????????????? F. merguiensis ?????? COI-RFLP ????????????????????????????????????????????????????????????? COI ????????? PCR ??????????????????? COI ???? XbaI ???????? F. merguiensis ??? P. monodon ?????? P. vannamei ??? M. rosenbergii ??????? F. merguiensis ???????????????? P. monodon ??????????????????? COI ???? SpeI ?????? P. vannamei ???????????????? M. rosenbergii ??????????????????? COI ???? HindIII. ????????? M. rosenbergii ???????????????????????????????????????????? COI ???? ApaI??????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????? ??????????????????????????????????????????? ???? F. merguiensis ?????????????????????????????????????????????????????????????????????????? ???????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????