长牡蛎壳橙性状遗传参数评估及与生长性状的关联性

长牡蛎壳橙快速生长品系是结合群体选育与种内群体间杂交方法,以壳色性状和生长性状为选育指标,经过连续3代选育获得的新品系。本研究以第1代长牡蛎壳橙快速生长品系(G1)为亲本,采用巢式设计建立了48个混养家系,共获得863个个体。利用微卫星标记对混养家系进行了系谱分析,并基于REML法评估了长牡蛎壳橙快速生长品系壳色性状的遗传参数及与生长性状的相关性。结果显示,863个个体中,有851个个体被准确鉴定其所属家系;长牡蛎壳橙快速生长品系壳色性状颜色参数指标a*(红绿轴色品指数)、b*(黄蓝轴色品指数)和ΔE(色差)等均具中高等遗传力,大小依次为0.47±0.23、0.42±0.21和0.56±0.29。4个颜色参数指标间的遗传相关和表型相关范围分别为−0.79~0.86和−0.45~0.48。壳色性状与各生长性状的遗传相关和表型相关较低,分别为−0.33~0.17和−0.04~0.11。研究表明,在长牡蛎壳橙快速生长品系育种过程中,对壳色性状进行选育可以得到预期的改良效果。此外,壳橙性状与生长性状应分别作为目标性状进行协同选择,以实现同时改良两个性状的目的。本研究可为长牡蛎壳橙快速生长品系选育提供基础资料。

温度和盐度对长牡蛎“海大1号”二倍体和三倍体幼虫生长与存活的影响

为确定长牡蛎(Crassostrea gigas)“海大1号”苗种规模化繁育的最适条件,本研究设置4个温度梯度(20、25、30和35℃)和5个盐度梯度(20、25、30、35和40),分析了不同温度和盐度对长牡蛎“海大1号”二倍体和三倍体幼虫生长和存活的影响。研究表明,当温度25℃和盐度30时,长牡蛎“海大1号”二倍体和三倍体幼虫的受精率、孵化率和变态率最高。不同温度处理下,三倍体幼虫的生长速度和累计存活率始终高于二倍体幼虫。随着温度升高幼虫生长速度增加,但存活率下降,在25℃时三倍体幼虫存活率最高(52.00%±6.56%),温度为30℃时,三倍体幼虫壳高增长最快(14.82μm·d^(-1))。不同盐度下,三倍体幼虫的生长速度和累计存活率显著高于二倍体幼虫。幼虫的存活率随着盐度升高呈先升高再下降的趋势,三倍体幼虫累计存活率在盐度30时最高(52.00%±6.56%);幼虫的生长速度随着盐度升高呈下降的趋势,三倍体幼虫壳高在盐度20时增长最快(18.22μm·d^(-1))。研究结果表明,长牡蛎“海大1号”三倍体幼虫比二倍体幼虫表现出更强的温度、盐度的耐受性;长牡蛎“海大1号”三倍体苗种培育的适宜盐度为20~30,适宜温度为20~25℃。本文可为长牡蛎“海大1号”三倍体苗种的规模化繁育提供参考。

长牡蛎“海大1号”与壳橙品系杂交子代的杂种优势及配合力

为培育生长快速、存活率高的长牡蛎新品种,以第11代长牡蛎“海大1号”(H)和第8代壳橙品系(O)为亲本,采用完全双列杂交设计,建立自交组HH[H(♀)×H(♂)]和OO[O(♀)×O(♂)]与杂交组HO[H(♀)×O(♂)]和OH[O(♀)×H(♂)],分析了4个组合的生长与存活杂种优势以及一般配合力(GCA)和特殊配合力(SCA)效应值。结果显示,20日龄时,杂交组存活中亲优势MF1为58.03%,HO和OH的存活与HH相比分别提高31.48%和35.80%。360日龄时,4个组合壳高由高到低依次为OH>HO>HH>OO,杂交组的壳高和湿重中亲杂种优势MF1分别为24.65%和46.02%。与HH相比,OH的壳高和湿重分别提高了23.51%和39.60%。杂交组存活中亲优势MF1、高亲优势HHO和HOH分别为68.31%、40.29%和53.96%。除180日龄时存活率性状的父本GCA外,长牡蛎“海大1号”在其余时期的GCA均为正效应值,为优质高产杂交育种亲本;OH的生长和存活SCA方差均显著高于HO。研究表明,以壳橙选育品系为母本,“海大1号”选育品种为父本的杂交组合OH具有较高杂种优势与特殊配合力,可为培育生长快、存活率高的优质杂交牡蛎品种提供素材。

6-DMAP和低渗诱导长牡蛎“海大2号”三倍体的研究

为探索低渗和6-二甲基氨基嘌呤(6-DMAP)诱导长牡蛎(Crassostrea gigas)三倍体最佳条件以及6-DMAP和低渗配合使用诱导三倍体的可行性,本研究通过低渗、6-DMAP以及6-DMAP与低渗配合三种方法诱导长牡蛎“海大2号”三倍体。研究显示,低渗处理组中,当第一极体(PB1)出现40%时,在盐度为8的低渗海水中持续处理15 min,综合评价指数最高,为34.33%,此时2日龄长牡蛎的三倍体率和存活率分别为73.77%±1.89%和45.78%±6.23%,8日龄长牡蛎的三倍体率和存活率分别为50.65%±4.89%和21.17%±5.54%。6-DMAP处理组中,当PB1出现40%时,75 mg/L的6-DMAP持续处理15 min,综合评价指数最高(36.52%),此时2日龄长牡蛎的三倍体率和存活率分别为74.81%±0.68%和46.30%±1.54%,8日龄长牡蛎的三倍体率和存活率分别为60.37%±0.61%和30.25%±2.85%。6-DMAP和低渗配合组中,当PB1出现40%时,在盐度为8的低渗海水中,以50 mg/L的6-DMAP持续处理15 min,综合评价指数最高(36.18%),此时2日龄长牡蛎的三倍体率和存活率分别为74.34%±3.49%和45.07%±4.41%,8日龄长牡蛎的三倍体率和存活率分别为68.23%±2.52%和40.42%±4.30%。综合比较以上三种方法,通过6-DMAP和低渗配合诱导获得三倍体的倍率更稳定,幼虫存活率更高。研究结果可为长牡蛎“海大2号”三倍体培育提供参考资料。

Productive Traits and Triploid Rate Stability in Triploidy-Induced ‘Haida No. 2’ Strain of the Pacific Oyster, Crassostrea gigas

In order to evaluate the effects of triploidy induction on a selected strain‘Haida No.2’of the Pacific oyster Crassostrea gigas,which is characterized with golden shell color and high growth rate,the growth,survival rate and stability of triploid rate were analyzed at different development stages in the present study.Three different conditions inhibiting the release of polar body Ⅱ or polar body Ⅰ were tested:(A)Cytochalasin-B(CB),0.5mg L^(−1) at 10min post-insemination for 15 min;(B)CB,0.5mg L^(−1)at 15 min postinsemination for 20 min;and(C)CB,0.7mg L^(−1),at 15 min post-insemination for 20 min.The triploidy induction treatments significantly reduced the D-larvae and survival rates at the larvae stage but not at the juvenile and adult stages.Triploid rate dramatically decreased at the larval stage and did not significantly change at the juvenile and adult stages.Regarding the stability of the triploid rate,there was a significant difference between the three treatment groups.Larvae from the treatment A and control groups exhibited higher growth rates in shell height than those from the other two treatment groups at day 27.Triploid juveniles and adults from the treatment A group exhibited a higher wet weight than diploids from the control group and triploids from the other treatment groups.Triploidy induction did not affect the shell color of the progeny.The results obtained in the study demonstrate that triploidy induction has the potential to be used to increase the production of C.gigas variety‘Haida No.2’without modifying its golden shell color.

高温胁迫对长牡蛎‘海大3号’品系存活及5项免疫指标的影响

夏季海水温度升高会对长牡蛎(Crassostrea gigas)体内生化反应产生重要影响,进而影响养殖牡蛎的存活率。本研究探讨了长牡蛎‘海大 3 号’在受到温度突升和温度渐升胁迫时的高温耐受性,以及在温度突升胁迫条件下过氧化氢酶(CAT)、超氧化物歧化酶(SOD)、溶菌酶(LSZ)、总抗氧化能力(T-AOC)和丙二醛(MDA)5 种相关免疫指标在 72 h 内的变化。温度渐升实验中,实验海水的温度从 16℃逐渐升温(2℃/d)。温度突升实验中,将长牡蛎从暂养水温(16℃)分别直接转移至 18℃、22℃、26℃、30℃和 34℃。实验结果显示,温度渐升时长牡蛎‘海大 3 号’的最高存活温度(survival temperature maximum, STMax)为 33.63℃,最高临界温度(critical temperature maximum, CTMax)为 40.13℃,存活率为 50%时的温度(50% critical temperature maximum, 50%CTMax)为 36.67℃。在温度突升胁迫实验中, 72 h 的半数致死高温(median lethal temperature)72-h LT50为 30.13℃。各实验组内脏团中 5 项相关免疫指标随时间增加变化显著。在前 12 h, CAT、SOD 及 LSZ 活性和 T-AOC 显著升高,之后逐渐下降恢复到初始水平。MDA含量在 6 9 h 达到最大值,之后逐渐下降。这些结果表明高温胁迫会引起长牡蛎‘海大 3 号’体内的抗氧化免疫和溶菌酶活性发生显著变化,进而对牡蛎的存活产生重要影响。本研究检测的高温环境的耐受能力,将为长牡蛎新品系‘海大 3 号’的应用推广提供参考。

Mass Selection for Growth Improvement in Black Shell Line of Pacific Oyster Crassostrea gigas

The Pacific oyster,Crassostrea gigas,naturally distributing along the coast of northwest Pacific,is one of the most important bivalve species due to its high economic value and fecundity.In China,we have initiated a selective breeding program on both shell color and growth rate of C.gigas since 2010.A black shell line was obtained through four-generation family selection.In this study,mass selection for growth improvement was conducted in the sixth generation and seventh generation of black shell lines.To assess the progress of potential genetic improvement,the progeny of two generations of black shell lines were selected to evaluate their shell heights via a 450-day farming experiment.As the results,after growing for 450 days,the sixth generation and seventh generation of selected lines were 9.03% and 11.42% larger than the control lines,respectively.During the grow-out stage,the genetic gain of two generations was 8.82%±0.18% and 11.54%±0.43%,respectively;and the corresponding realized heritability was 0.45±0.04 and 0.41±0.04,respectively.These results indicated that the mass selection for shell height achieved steady progress in the two generations of C.gigas.

Seasonal Changes of Reproductive Activity and Biochemical Composition of Pen Shell Atrina pectinata Linnaeus,1767 in Bohai Sea,China

Seasonal variations in the biochemical composition and the reproductive cycle of pen shell Atrina pectinata in Bohai Sea were investigated from May 2013 to April 2014. Histological analysis indicated that the reproductive cycle of A. pectinata can be divided into two phases, inactive stage and gametogenesis, which were equally and significantly influenced by seawater temperature and food availability. Gametogenesis began in late autumn(October), and completed in June and July. Spawning took place in August, coinciding with the highest water temperature and the richest phytoplankton. The significantly high glycogen content in adductor muscles sustained throughout the late active and ripeness stages, but plummeted during spawning and inactive stages. The protein content in female gonads exhibited a synchronous increase along with oocyte diameter and lipid content, suggesting that the female gonads could accumulate protein and lipid for vitelline in A. pectinata. Furthermore, the RNA/DNA ratio was found to be a useful index to indicate the level of gonad maturation in both males and females. The findings of the present study provided a foundation for the fishery resource administration and the aquaculture development of this species.

Effects of Temperature,Salinity and Stocking Density on Larval Survival and Growth of Reciprocal Crosses Between Two Strains of Pacific Oysters,Crassostrea gigas

The Pacific oyster Crassostrea gigas is one of the most widely cultivated aquaculture species and contributes signifi-cantly to total seafood production for human beings.However,mass mortality occurred frequently,and in some regions almost all oysters died during seed production and grow-out stage.In order to explore whether hybridization breeding can improve its growth and survival,a complete diallel cross between a selected strain‘Haida No.1’(S)and an orange shell variant(O)of C.gigas was carried out.The larval growth and survival were compared among hybrids and purebred strains at temperatures of 16,20,24,28 and 32℃;salinities of 15,20,25,30 and 35;and stocking densities of 0.5,1,2,4 and 8 larvae mL−1.Under different environments,the hybridization between two strains of C.gigas showed the heterosis of growth and survival.The mean shell height and survival rate of the two reciprocal crosses(OS,SO)were significantly higher than those of the two purebred strains(SS,OO)under all environ-mental conditions.In particular,OS showed greater heterosis than the purebred strains and SO progeny.The results showed that the productive traits of the‘Haida No.1’could be improved by crossing with the orange shell line.Meanwhile,the results from this study also indicated that hybridization between the two strains of C.gigas may be a promising way for breeding new variety with high survival rate.