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徐红琴, 马慧妹, 曾起, 胡蓓娟, 盛军庆, 洪一江. 温度对池蝶蚌雌雄同体和性逆转的影响[J]. 水生生物学报, 2022, 46(5): 741-753. DOI: 10.7541/2021.2021.058
引用本文: 徐红琴, 马慧妹, 曾起, 胡蓓娟, 盛军庆, 洪一江. 温度对池蝶蚌雌雄同体和性逆转的影响[J]. 水生生物学报, 2022, 46(5): 741-753. DOI: 10.7541/2021.2021.058
XU Hong-Qin, MA Hui-Mei, ZENG Qi, HU Bei-Juan, SHENG Jun-Qing, HONG Yi-Jiang. TEMPERATURE ON HERMAPHRODITISM AND SEX REVERSAL IN FRESHWATER PEAL MUSSEL, HYRIOPSIS SCHLEGELII[J]. ACTA HYDROBIOLOGICA SINICA, 2022, 46(5): 741-753. DOI: 10.7541/2021.2021.058
Citation: XU Hong-Qin, MA Hui-Mei, ZENG Qi, HU Bei-Juan, SHENG Jun-Qing, HONG Yi-Jiang. TEMPERATURE ON HERMAPHRODITISM AND SEX REVERSAL IN FRESHWATER PEAL MUSSEL, HYRIOPSIS SCHLEGELII[J]. ACTA HYDROBIOLOGICA SINICA, 2022, 46(5): 741-753. DOI: 10.7541/2021.2021.058

温度对池蝶蚌雌雄同体和性逆转的影响

TEMPERATURE ON HERMAPHRODITISM AND SEX REVERSAL IN FRESHWATER PEAL MUSSEL, HYRIOPSIS SCHLEGELII

  • 摘要: 为探究温度对池蝶蚌(Hyriopsis schlegelii)性腺发育的影响, 研究连续两年对池蝶蚌性腺进行雌雄同体现象筛选及跟踪观察, 并采用5个温度诱导池蝶蚌生殖滤泡分化。通过对不同时期的池蝶蚌的性腺进行qRT-PCR检测, 同时利用原位杂交技术对性别决定关键基因Dmrt1进行细胞定位。结果显示: 在自然条件下, 6月龄池蝶蚌出现了雌雄同体现象, 28月龄的雌雄同体蚌到30月龄左右会出现不同性别的分化(77.8%分化为雄蚌, 16.7%维持雌雄同体现象, 5.5%分化为雌蚌)。在不同温度刺激下, 14月龄池蝶蚌出现了雌雄同体和性逆转现象。32℃刺激, 14月龄雄蚌出现12.5%雌雄同体; 27℃刺激, 14月龄雄蚌和27月龄雄蚌分别出现37.5%和12.5%性逆转; 23℃刺激, 14月龄雌蚌出现14.29%性逆转; 19℃刺激, 14月龄雌蚌出现25%性逆转。qRT-PCR结果显示: Dmrt1在胚胎及6、27月龄表达显著高于其他时期, 32℃刺激, 在27月龄雄蚌中的表达显著高于其他温度, 19℃刺激, 在14、27月龄雌蚌中的表达显著高于其他温度; Fem1b、Fem1c的时空表达趋势基本一致, 与Tra2a大体一致, 在5、32月龄与Sox9相互抑制; 19℃、23℃和27℃刺激, Fem1b、Tra2aSox9三个基因在14月龄雄蚌中表达显著高于其他温度, 且23℃刺激, Sox9在27月龄雄蚌中表达也显著高于其他温度; Foxl2在27℃下的27月龄雌蚌中表达显著高于其他温度。Tra2a、Fem1b、Fem1c、Sox9Dmrt1五个基因在不同月龄的雌雄同体蚌内表达有显著差异, Foxl2在雌蚌和雌雄同体蚌内表达显著高于同期雄蚌。原位杂交技术结果显示, Dmrt1 mRNA阳性信号主要定位在精原细胞、初级精母细胞和成熟卵母细胞中。综上所述, 温度影响池蝶蚌生殖滤泡的分化, 27℃及以上的高温容易诱导雄蚌向雌蚌逆转或产生雌雄同体现象, 23℃及以下的低温容易诱导雌蚌向雄蚌逆转; 并且温度很可能是通过调控Dmrt1等性别相关基因的表达量, 从而调节生殖滤泡分化的。

     

    Abstract: In order to investigate the effects of temperature on gonadal development of Hyriopsis schlegelii, we screened and observed hermaphrodite phenomena in gonadal glands of H. schlegelii for two consecutive years, and induced reproductive follicular differentiation of H. schlegelii by five temperatures. In this study, qRT-PCR was used to detect the gonads of H. schlegelii in different periods, and in situ hybridization was used to locate the key gene Dmrt1. The results showed that the pool of natural conditions, 6 months H. schlegelii appeared the phenomenon of hermaphrodite and 28 months of hermaphrodite H. schlegelii had different gender differentiation about 30 months (77.8% differentiated into males, 16.7% remained hermaphroditism, and 5.5% differentiated into females). At 32℃, 12.5% of male clam were hermaphroditic at 14 months of age. When stimulated at 27℃, sex reversal of 14-month-old males and 27-month-old males was 37.5% and 12.5%, respectively. When stimulated at 23℃ and 19℃, sex reversal of 14 months old female H. schlegelii were 14.29% and 25%, respectively. The mRNA level of Dmrt1 was significantly higher in embryo of 6 and 27 months than that in other stages. The mRNA level of Dmrt1 of male H. schlegelii of 27 months at 32℃ was significantly higher than that at other temperatures, and its expression of female H. schlegelii of 14 and 27 months at 19℃ was significantly higher than that at other temperatures. The temporal and spatial expression trend of Fem1b and Fem1c was basically the same as that of Tra2a, which inhibited each other with Sox9 at 5 and 32 months of age. The expression of Fem1b, Tra2a and Sox9 in 14-month-old males at 19, 23 and 27℃ was significantly higher than those at other temperatures and the expression of Sox9 in 27-month-old males at 23℃ was also significantly higher than that at other temperatures. The expression of Foxl2 in 27-month-old female H. schlegelii at 27℃ was significantly higher than that at other temperatures. The expression of Tra2a, Fem1b, Fem1c, Sox9 and Dmrt1 were significantly different in the hermaphroditic mussels at different months of age, and the expression of Foxl2 in female hermaphroditic mussels was significantly higher than that in male mussels at the same period. The results of in situ hybridization showed that Dmrt1 mRNA positive signal was mainly located in spermatogonia, primary spermatocytes and mature oocytes. In conclusion, temperature affects the differentiation of reproductive follicles of H. schlegelii. High temperature above 27℃ can easily induce male to female reversal or hermaphrodite phenomenon, and low temperature below 23℃ can easily induce female to male reversal. Temperature probably regulates reproductive follicular differentiation by regulating the expression of Dmrt1 and other sex-related genes.

     

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