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水产养殖育种改良:使用重组类松弛素促性腺刺激肽促进海参成熟和产卵

Aquaculture Breeding Enhancement: Maturation and Spawning in Sea Cucumbers Using a Recombinant Relaxin-Like Gonad-Stimulating Peptide.

作者信息

Chieu Hoang Dinh, Turner Luke, Smith Meaghan K, Wang Tianfang, Nocillado Josephine, Palma Peter, Suwansa-Ard Saowaros, Elizur Abigail, Cummins Scott F

机构信息

Genecology Research Centre, University of the Sunshine Coast, Sippy Downs, QLD, Australia.

Research Institute for Marine Fisheries, HaiPhong, Vietnam.

出版信息

Front Genet. 2019 Feb 19;10:77. doi: 10.3389/fgene.2019.00077. eCollection 2019.

Abstract

Wild sea cucumber resources have been rapidly exhausted and therefore there is an urgent need to develop approaches that will help restocking. Currently, there is a lack of information regarding the genes involved in sea cucumber reproductive processes. The neurohormone relaxin-like gonad-stimulating peptide (RGP) has been identified as the active gonad-stimulating peptide in sea stars (Asteroidea), which could also be present in other echinoderm groups. In this study, a sea cucumber RGP was identified and confirmed by phylogenetic analysis. A recombinant RGP was produced in the yeast and confirmed by mass spectrometry. To assess bioactivity, four levels of purification were tested in an germinal vesicle breakdown (GVBD) bioassay. The most pure form induced 98.56 ± 1.19% GVBD in and 89.57 ± 1.19% GVBD in . Cruder levels of purification still resulted in some GVBD. Upon single injection into female , the recombinant RGP induced head waving behavior followed by spawning within 90-170 min. Spawned oocytes were fertilized successfully, larvae settled and developed into juveniles. Our results provide a key finding for the development of a break-through new artificial breeding approach in sea cucumber aquaculture.

摘要

野生海参资源已迅速枯竭,因此迫切需要开发有助于增殖放流的方法。目前,关于参与海参生殖过程的基因的信息匮乏。神经激素松弛素样促性腺刺激肽(RGP)已被确定为海星(Asteroidea)中的活性促性腺刺激肽,它也可能存在于其他棘皮动物类群中。在本研究中,通过系统发育分析鉴定并确认了一种海参RGP。在酵母中产生了重组RGP,并通过质谱法进行了确认。为了评估生物活性,在生发泡破裂(GVBD)生物测定中测试了四个纯化水平。最纯的形式在[具体物种1]中诱导了98.56±1.19%的GVBD,在[具体物种2]中诱导了89.57±1.19%的GVBD。较低纯化水平仍导致了一些GVBD。将重组RGP单次注射到雌性[具体物种]体内后,诱导了头部摆动行为,随后在90 - 170分钟内产卵。产出的卵母细胞成功受精,幼虫附着并发育成幼体。我们的研究结果为海参养殖中突破性的新型人工繁殖方法的开发提供了关键发现。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5b77/6389678/0a68f84507c2/fgene-10-00077-g0001.jpg

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