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Molecular Cloning and Sexually Dimorphic Expression of Two Dmrt Genes in the Giant Freshwater Prawn, Macrobrachium rosenbergii

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Abstract

The molecular mechanism of sex determination changed rapidly during evolution among phyla. One exception is the Dmrt (doublesex and mab-3 related transcription factor) family gene, which is widely conserved for its involvement in sex development from invertebrates to humans. In this study, we identified two DM-domain genes in the giant freshwater prawn, Macrobrachium rosenbergii, which we named MroDmrt11E and MroDmrt99B according to their high similarities and phylogenetically close relationships with arthropod Dmrt11E and Dmrt99B genes, respectively. Amino acid alignments and structural prediction uncovered conservation and putative active sites of their DM domains. Realtime PCR analysis exhibits a sexually dimorphic and embryonic development-dependent expression pattern of both genes. The transcription of both MroDmrt11E and MroDmrt99B was prominent in testis, and both mRNAs were localized in spermatogonia and spermatozoa during spermatogenesis. RNAi-targeting MroDmrt99B showed no effect on several male-reproduction related genes while the MroDmrt11E RNAi induced a significant decrease of the transcript of Iag, an important regulatory gene in the male determination mechanism of the prawn. The complicated expression profiles of Dmrt family genes and closely related subfamily members may provide clues of their rapid evolution and multi-functionality. Our study will contribute to understanding mechanistic and evolutionary dynamics of Dmrt family genes in metazoan bisexual development and embryogenesis.

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Acknowledgments

This work was supported by the 863 Program of China (2012AA10A401 and 2012AA092205), the National Natural Sciences Foundation of China (41206125), the Scientific Research Fund of Zhejiang Provincial Science and Technology Department (2012C12907-9) and the Scientific Research Fund of Zhejiang Provincial Education Department (Y200805042).

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Authors and Affiliations

  1. Key Laboratory of Conservation Biology for Endangered Wildlife of the Ministry of Education, College of Life Sciences, Zhejiang University, Zijingang Campus, 866 Yuhangtang Road, Hangzhou, 310058, Zhejiang, People’s Republic of China

    Yan-Qin Yu, Qing-Guo Zeng, Ye-Qing Qian, Jin-Shu Yang & Wei-Jun Yang

  2. Biology and Environment College, Zhejiang Wanli University, Ningbo, 315100, Zhejiang, People’s Republic of China

    Wen-Ming Ma

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  1. Yan-Qin Yu
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Correspondence to Wei-Jun Yang.

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40003_2014_106_MOESM1_ESM.tif

Schematic cloning strategies of the MroDmrt11E (a) and MroDmrt99B (b) cDNAs. Primer names and positions correspond to Table 1. 5′- and 3′-RACE primers are provided with FirstChoice™ RLM-RACE Kit (Ambion, Austin, TX, USA). Open reading frames (ORFs) are shaded, and DM domains are highlighted. Lengths are drawn to scale. (TIFF 1354 kb)

Protein sequences used in this study. (DOC 83 kb)

40003_2014_106_MOESM3_ESM.tif

Nucleotide and deduced amino acid sequences of MroDmrt11E (a) and MroDmrt99B (b) cDNAs. Nucleotides of open reading frames are shown in bold with the translated amino acids above. Asterisks indicate stop codons (TAA). Sequences of DM domains are shaded. The short 7aa short conserved motif of MroDMRT11E is boxed. (TIFF 1590 kb)

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Yu, YQ., Ma, WM., Zeng, QG. et al. Molecular Cloning and Sexually Dimorphic Expression of Two Dmrt Genes in the Giant Freshwater Prawn, Macrobrachium rosenbergii . Agric Res 3, 181–191 (2014). https://doi.org/10.1007/s40003-014-0106-x

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