熊本牡蛎中国群体与美国群体杂交效应及杂交三倍体优势分析

为了评估熊本牡蛎(Crassostreasikamea)中国群体与美国群体的杂交效应与杂交三倍体优势,构建了杂交群体和自交群体,并使用细胞松弛素B诱导了杂交三倍体,比较分析了幼虫、稚贝与成贝的生长与存活优势。自交群体和杂交群体的幼虫、稚贝与成贝的培养条件均相同,室内培育幼虫,培育密度为4～5个/mL,自然海区养殖稚贝与成贝,养殖密度为40～45个/吊。结果表明,与自交组相比,杂交二倍体具有较高的卵裂率(13.61%)与D幼率(5.67%)(P0.05),但杂交二倍体幼虫的壳高生长表现杂种劣势(–0.43%),而稚贝、成贝的壳长与壳高的生长表现杂种优势,平均优势率分别为3.96%与6.65%。杂交三倍体诱导组幼虫的壳高生长优势率为3.69%,稚贝及成贝的壳长与壳高平均生长优势率分别为12.69%与13.64%,并且日龄越大生长优势越显著。总体上,杂交三倍体诱导组在3～180日龄具有存活劣势,并且15日龄存活劣势率最大(–48.72%),而在360日龄存活优势率为6.70%。杂交二倍体幼虫与成贝均具有存活优势,平均优势率分别为10.44%与4.59%。本研究表明熊本牡蛎中美地理群体杂交二倍体的生长和存活优于自交二倍体,而杂交三倍体诱导组的生长优势较显著,并且在成贝期具有显著的成活率优势,表明杂交三倍体诱导组的优势来源于三倍体优势和部分杂种优势。     

菲律宾蛤仔福建与广东群体的双列杂交

2015年9月–12月,以菲律宾蛤仔福建养殖群体和广东野生群体子一代为亲本,开展了双列杂交实验,建立了两个自交组和两个杂交组,研究了杂交子代幼虫和稚贝的生长与存活的杂种优势。结果显示,各实验组均有较高的受精率和孵化率,但无显著差异。总体上,幼虫期的生长受母体效应影响显著,稚贝期的生长杂交效应主要受交配方式影响。相反,幼虫期的存活受配对方式影响最显著,稚贝期的存活受卵源影响显著。在幼虫期,杂交组与自交组在生长和存活方面差异不显著。在稚贝期,生长表现出明显的杂种优势,壳长和壳高在30日龄以后,正反交组的平均生长速率显著快于自交组。在40日龄时,壳高和壳长总杂交优势值达到最大,分别为25.64和27.00。这可能是因为杂种优势的表达具有时期差异性。在幼虫期,福建自交组表现出最高的存活率,为36.45%±1.85%;稚贝期,广东自交组存活率最高,为52.27%±2.13%。在幼虫期和稚贝期的存活率方面,未观测到杂种优势,这可能是由于两个杂交亲本群体与存活相关的基因频率无差异或者检测次数较少所致。     

太平洋牡蛎与葡萄牙牡蛎两亚种间杂交及其早期阶段生长与存活的杂种优势

利用山东青岛养殖的太平洋牡蛎(N)与汕头本地养殖的葡萄牙牡蛎(S)两个近缘种为亲本,采用正交设计建立了杂种组NS(N♂×S♀)和SN(S♂×N♀)与纯种组NN(N♂×N♀)和SS(S♂×S♀)4个不同的遗传组合,通过比较不同阶段(幼虫期、稚贝期、养成阶段)的生长和存活数据,研究了牡蛎近缘种间的杂种优势,目的为改良牡蛎的生产性状。结果表明,这两个近缘种之间杂交能够产生显著的杂种优势,杂交后代的生长与存活两个表型性状都得到改良。杂交组比近交组生长得快,杂种优势在幼虫期为37.44%,稚贝期为42.47%。杂交组也比近交组存活率高,8日龄幼虫存活率的杂种优势为76.80%、14日龄幼虫存活率的杂种优势可达107.70%,60、90和105日龄稚贝存活率的杂种优势分别为17.30%、15.62%和9.08%。研究表明,通过太平洋牡蛎和葡萄牙牡蛎两个近缘种间的杂交有望解决牡蛎养殖产业存在的育苗难、存活率低、生长慢、个体小等问题。     

魁蚶(Scapharca broughtonii)中国群体与韩国群体杂交子一代的生长和存活比较

以魁蚶(Scapharca broughtonii)中国群体(C)和韩国群体(K)为材料,进行了4个组合的杂交和自交实验,分析了各实验组的受精率、孵化率、幼虫和稚贝的生长、存活以及杂交子一代的杂种优势。结果显示,4个组的受精率和孵化率均较高(80%以上);自交组的受精率略高于杂交组,但孵化率无显著差异。在幼虫期,KK组和KC组的壳长平均值大于CK组和CC组,但存活率和变态率无显著差异;在稚贝期,KK组和KC组的壳长平均值也均大于CK组和CC组,但CC组和CK组的保苗率比KC组和KK组高。综合分析比较表明,杂交KC组的杂种优势率在幼虫期和稚贝期的壳长生长上均表现为正值,在稚贝期的存活率方面也表现为正值,杂交KC组结合了两群体的部分优良性状,是理想的育种材料。     

魁蚶(Scapharca broughtonii)中国群体与韩国群体杂交子一代的生长和存活比较

以魁蚶(Scapharca broughtonii)中国群体(C)和韩国群体(K)为材料,进行了4个组合的杂交和自交实验,分析了各实验组的受精率、孵化率、幼虫和稚贝的生长、存活以及杂交子一代的杂种优势.结果显示,4个组的受精率和孵化率均较高(80％以上);自交组的受精率略高于杂交组,但孵化率无显著差异.在幼虫期,KK组和KC组的壳长平均值大于CK组和CC组,但存活率和变态率无显著差异;在稚贝期,KK组和KC组的壳长平均值也均大于CK组和CC组,但CC组和CK组的保苗率比KC组和KK组高.综合分析比较表明,杂交KC组的杂种优势率在幼虫期和稚贝期的壳长生长上均表现为正值,在稚贝期的存活率方面也表现为正值,杂交KC组结合了两群体的部分优良性状,是理想的育种材料.     

壳金长牡蛎自交和杂交家系生长与存活比较

选择具有生长优势的长牡蛎(Crassostrea gigas)G2(A)家系及具有明显生长和存活优势的G19(B)和G28(C)家系进行完全双列杂交,得到包括3个自交组(AA、BB、CC)和6个杂交组(AB、AC、BA、BC、CA、CB)共9个壳金长牡蛎实验组,分析各实验组在幼虫期及稚贝期不同日龄的各生长指标和存活性能,并评估了杂交组的杂种优势。结果表明,在整个培育阶段,大多数杂交组在不同生长时期均表现出较高的生长和存活性能,其中,浮游幼虫期5日龄,所有杂交组的壳高和壳长均显著大于自交组(P0.05);10日龄后,杂交组CB、BC和AC的壳高显著大于相应的自交组(P0.05),杂种优势明显;稚贝期85和130日龄,杂交组CB的壳高性能大于其他实验组。浮游幼虫期10日龄,CB组存活率显著大于AA、CA组(P0.05);稚贝期85日龄后,杂交组AC、CA和CB的存活率杂种优势逐渐增大;190日龄,杂交组AC、BC与自交组AA、CC之间存活率差异显著(P0.05)。研究结果为壳金长牡蛎新品种的培育及杂种优势的充分利用奠定了重要基础。     

壳紫长牡蛎家系生长存活比较及生长性状的遗传参数评估

为了培育壳色和生长性状优良的壳紫长牡蛎(Crassostreagigas)新品系,2021年通过家系选育的方式,以壳色性状为主要选育指标,建立了第1代壳紫长牡蛎选育群体。本研究以壳紫F1为亲本,采用平衡巢式设计建立了36个全同胞家系,并以野生长牡蛎为亲本建立对照组,比较分析了各家系在生长和存活方面的性能表现;同时利用多性状动物模型对壳紫家系幼虫和稚贝期的生长性状遗传参数进行了评估。结果显示,不同生长阶段壳紫长牡蛎家系的存活率和壳高均值均高于对照组,幼虫期分别提高了1.49%～10.18%和3.75%～15.94%,稚贝期分别提高了4.05%～16.94%和15.95%～18.25%。不同壳紫家系的生长和存活情况也存在差异,没有发现在存活率和生长性状方面同时具有优势的家系。壳紫长牡蛎幼虫期壳高和壳长的狭义遗传力范围分别为0.43～0.84和0.49～0.80,稚贝期壳高和壳长的遗传力分别为0.15～0.33和0.18～0.37,均属于中高等遗传力。不同生长阶段壳紫长牡蛎家系壳高和壳长的遗传相关与表型相关均为正相关,幼虫期的相关系数分别为0.67～0.97和0.17～0.51,稚贝期为0....     

深凹壳型香港牡蛎家系生长与存活性状比较

为了培育壳型和生长性状优良的深凹壳型香港牡蛎(Crassostrea hongkongensis)新品系,以广东台山镇海湾香港牡蛎野生群体为基础群体,以壳型和生长为主要选育目标,依据壳型指数D和巢式设计建立了30个全同胞家系和1个对照组,评估各家系和对照组在幼虫期和稚贝期的生长和存活性能。结果表明,整个生长阶段,各家系生长性状和存活率的平均值均高于对照组。幼虫期,所有家系壳高、生长速度以及存活率的平均值均高于对照组,所有家系壳高平均值比对照组提高3.78%～7.73%,生长速度提高7.86%,存活率提高2.85%～7.32%。稚贝期,所有家系壳高、壳长以及生长速度、存活率的平均值均高于对照组平均值,壳高提高5.23%～16.32%,壳长提高7.94%～10.69%,生长速度提高9.55%～20.16%,存活率提高3.45%～12.25%。不同家系在不同时期的生长性状和存活性能也表现出较大的差异,G1在整个稚贝期的生长性状、生长速度以及存活率的平均值均显著高于所有家系和对照组的平均值, 12月龄G1家系的壳高、壳长、壳宽、壳型指数D、总重、壳重、软体重、存活率比所有家系平均值提高9.83%～54.75%,相比对照组平均值提高23.34%～80.77%,同时G3、G10、G12、G15、G19、G23、G26在稚贝期也表现出较大的生长优势和存活优势。研究表明,深凹壳型香港牡蛎家系选育群体在生长性状和存活性能上均具有较大的优势, G1、G3、G10、G12、G15、G19、G23、G26可作为后期培育生长性能优良的深凹壳型香港牡蛎新品系的育种材料。本研究为中国华南地区培育出深凹壳型、生长性能良好的香港牡蛎优良品种提供了理论基础和数据材料。     

太平洋牡蛎与近江牡蛎的种间杂交

为了评估太平洋牡蛎与近江牡蛎能否产生远缘杂种优势,于2010年5月,以成熟的两种牡蛎亲本为材料开展了2×2远缘杂交研究,由长牡蛎自繁组GG(Crassostrea gigas♀×C.gigas♂)、近江牡蛎自繁组AA(C.ariakensis♀×C.ariakensis♂)、正交组GA(Crassostreagigas♀×C.ariakensis♂)、反交组AG(C.ariakensis♀×C.gigas♂)4个实验组组成。分析了子代早期表型性状和杂种优势,并对杂交子代进行了遗传鉴定。结果表明:GA杂交组与AG组的受精强度具有不对称性。幼虫浮游期间,表型性状的中亲杂种优势几乎为0,GA组生长与存活性状表现出积极的单亲杂种优势,而AG组则具有明显的远交衰退现象;幼虫早期表型性状受到母本效应影响,而后减弱。变态期间,GA组变态率较高,得到了大量杂交稚贝;而AG组变态率极低,仅获得了72个杂交稚贝。稚贝培育期间,稚贝表现出中亲生长劣势与存活优势;GA组具有明显的单亲生长与存活优势,而AG组则表现出生长劣势并具有一定程度的存活优势。利用复合COI及ITS2鉴定了种间杂交子结果表明:正反交组杂交子均为真正意义上的两性融合杂交子。实验获得了具有显著杂种优势的GA组杂交子,为现有牡蛎的遗传改良提供了新的方向。     

长牡蛎3种壳色家系间杂交子代生长和存活比较

为了利用杂种优势培育长牡蛎优良新品种,实验以3种不同壳色长牡蛎家系(白色/W、黑色/B、紫色/P)为材料,采用3×3完全双列杂交法,建立了3个自交组合和6个正反杂交组合,分析了各实验组幼虫期和养成期的生长、存活以及杂交子代的杂种优势。结果表明,浮游幼虫期,杂交组PB表现出显著的生长优势;与自交组相比,各杂交组均有着较高的幼虫存活率;在幼虫存活率方面,所有杂交组均有较高的杂种优势率。在养成期,紫壳色自交组的壳高显著大于白壳色和黑壳色自交组;6个杂交组中,PB的壳高生长最快,BP次之,PW、WP的生长最慢;各杂交组与自交组的成活率差异均不显著;杂交组BP及其反交组PB的壳高、壳长、总重和存活率的杂种优势率分别介于3.71%~15.27%、-2.00%~13.10%、11.23%~41.56%、-2.77%~9.83%,其他4个杂交组在整个养成阶段没有表现出杂种优势。     

Farming triploid oysters

Although the commercial benefits of triploidy have been evaluated in the Pacific oyster Crassostrea gigas (Thunberg, 1793), eastern oyster C. virginica (Gmelin, 1791), Sydney rock oyster Saccostrea glomerata (Gould, 1850) and European flat oyster Ostrea edulis (Linnaeus, 1750), so far this technique has only been commercialised for Pacific oysters.

Commercial production of triploids on the West Coast of North America began in 1985. Since then production of triploids has greatly increased and the use of tetraploid males to fertilise eggs from diploids to produce batches of 100% triploids has been developed. In 1999/2000, triploid Pacific oysters made up 30% of all Pacific oysters farmed on the West Coast of North America. Some hatcheries now use tetraploid males instead of chemical or physical stress to produce triploids. The rapid uptake of triploid and tetraploidy techniques has been facilitated by the almost total dependence that these oyster industries have on hatcheries for the supply of seed. This industry in the Pacific Northwest of the US and in British Columbia, Canada, would not have developed to its current size without hatchery seed supplies. Triploids are preferred over diploids in summer because diploids are less marketable when in spawning condition.

There was only limited interest in triploidy production in France until 1999, when IFREMER began to make sperm from tetraploids available to commercial hatcheries. In 1999/2000, only 10% to 20% of all the hatchery-supplied Pacific oyster spat were triploids, but with the use of sperm from tetraploid oysters, this could increase sharply.

Elsewhere around the world, the commercial uptake of triploid oysters has been slow to develop. However, in countries where the production of Pacific oysters is based on hatchery supply of seed, it is likely that with the use of tetraploid oysters, the farming of triploid oysters will increase in the near future.     

A comparison of the gametogenic cycle between the Pacific oyster Crassostrea gigas and the Suminoe oyster Crassostrea rivularis in Washington State

The gamotogenic cycle of the Suminoe oyster, Crassostrea rivularis, was compared with that of two groups of Pacific oysters (Crassostrea gigas) on an intertidal oyster bed in Oakland Bay, Washington. The Pacific oysters exhibited an earlier and more extensive development of gonad relative to C. rivularis. Resulting carbohydrate depletion was much greater in C. gigas relative to C. rivularis. Both groups of Pacific oysters spawned completely in mid-August. C. rivularis did not spawn but, rather, initiated phagocytosis of gametes by leucocytes. The implications of the results on the development of a marketable “summer” oyster are discussed.     

Development and distribution of the non-indigenous Pacific oyster (<Emphasis Type="Italic">Crassostrea gigas</Emphasis>) in the Dutch Wadden Sea

Pacific oysters (Crassostrea gigas) were first observed in the Dutch Wadden Sea near Texel in 1983. The population increased slowly in the beginning but grew exponentially from the mid-1990s onwards, although now some stabilisation seems to be occurring. They occur on a variety of substrates such as mussel beds (Mytilus edulis), shell banks, dikes and poles. After initial settlement spat may fall on older individuals and congregate to dense clumps and subsequently form reefs. Individual Pacific oysters grow 3–4 cm long in their first year and 2–3 cm in their second year. Many mussel beds (Mytilus edulis) are slowly taken over by Pacific oysters, but there are also several reports of mussel spat settling on Pacific oyster reefs. This might in the end result in combined reefs. Successful Pacific oyster spat fall seems to be related to high summer temperatures, but also after mild summers much spat can be found on old (Pacific oyster) shells. Predation is of limited importance. Mortality factors are unknown, but every now and then unexplained mass mortality occurs. The gradual spread of the Pacific oyster in the Dutch Wadden Sea is documented in the first instance based on historical and anecdotal information. At the start of the more in-depth investigation in 2002, Pacific oysters of all size classes were already present near Texel. Near Ameland the development could be followed from the first observed settlement. On dense reefs each square metre may contain more than 500 adult Pacific oysters, weighing more than 100 kg per m2 fresh weight.     

长牡蛎MITF基因表达及与壳色的关联

为了解长牡蛎MITF基因的表达及其与壳色的关联,验证了长牡蛎中的4个MITF基因,对长牡蛎MITF氨基酸序列进行了序列分析和多序列比对,分析了长牡蛎幼体发育各时期的转录组,采用荧光定量PCR的方法研究长牡蛎各组织及黑壳、白壳牡蛎特定部位mRNA表达情况。4个MITF基因中有3个基因可能为假基因,有表达的长牡蛎MITF基因共编码448个氨基酸,为亲水性不稳定蛋白,含有N端结构域(MITFTFEBC3N superfamily)和高度保守的功能性结构域HLH结构域。转录组分析发现MITF在长牡蛎个体发育的各个时期均有表达,在稚贝期达到最高。组织表达结果显示MITF在外套膜中的表达水平显著高于其他组织。MITF在黑壳牡蛎闭壳肌中的表达量显著低于白壳牡蛎,而在黑壳牡蛎外套膜中的表达量高于白壳牡蛎,但不显著。在黑壳、白壳牡蛎外套膜边缘的表达量都显著高于内侧。研究表明,长牡蛎MITF基因可能在牡蛎壳形成早期就参与了黑色素的生成调控和个体的生长发育,可调控牡蛎酪氨酸酶Tyr2基因,参与外套膜和贝壳中黑色素的形成。本研究为进一步研究牡蛎壳色形成机制奠定了基础。     

Production of Pacific oysters, Crassostrea gigas Thunberg, from wild-caught and hatchery-produced seed grown at several densities on oyster shells

Abstract. Production of Pacific oysters was studied under pilot-scale conditions in Baynes Sound, British Columbia, using common bottom culture strategies. Four seeding treatments, each with a different seed per cultch density were cultured: wild-caught seed 5mm in shell height at 10 seed per cultch piece and hatchery produced seed 1-2 mm in shell height at densities of 11, 40 and 105 per cultch piece. The cultch material for all treatments was Pacific oyster shell. All seed was reared for approximately 1 year in a seed nursery located at the 2.2-m tidal level then transferred to a 1-m tidal level grow-out plot until harvest 4 years later in May 1988, Clusters of large numbers of oysters were separated and evenly distributed within the plots when the oysters attained a shell height of 60-100 mm. During the first year, growth was slow and mortalities were relatively high. All treatments produced oysters of similar size at harvest. The proportion surviving at harvest was substantially higher for the wild oysters which were initially larger at time of planting. Within the hatchery treatments proportional survival per cultch piece was inversely related to initial density on the shell; however, total production per cultch piece was positively related to initial density. Most efficient use of seed is attained at lower densities per shell; however, most efficient use of cultch and effort to handle cultch is attained at high densities.     

The role of Uronema marinum (Protozoa) in oyster hatchery production

In Maine's hatchery production of oysters, significant mortalities at the early juvenile stage have been associated with ciliate infestations. The predominant ciliate isolated from live infested oysters was identified as Uronema marinum. Feeding experiments, designed to identify the food source of the ciliate in the oyster tank environment, determined that U. marinum is a bacteriophage and not a histophage, thereby clarifying its role in the oyster mortalities.     

Ostreid herpesvirus OsHV‐1 μVar in Pacific oysters Crassostrea gigas (Thunberg 1793) of the Wadden Sea,a UNESCO world heritage site

The Wadden Sea is an extensive wetland area, recognized as UNESCO world heritage site of international importance. Since the mid‐1990s, the invasive Pacific oyster Crassostrea gigas (Thunberg 1793) population in the area has grown exponentially, having a distinct impact on the ecosystem. The recent spread of the emerging oyster pathogen Ostreid herpesvirus OsHV‐1 μVar worldwide and specifically in the oyster culture areas in the south of the Netherlands raised the question whether the virus may also be present in the Wadden Sea. In the summer of 2012 juvenile Pacific oysters were collected from five locations in the Dutch Wadden Sea. The virus was shown to be present in three of the five locations by real‐time PCR and sequencing. It was concluded that OsHV‐1 μVar has settled itself in Pacific oyster reefs in the Wadden Sea. These results and the recent discoveries of OsHV‐1 microvariants in Australia and Korea indicate that OsHV‐1 μVar and related variants might be more widespread than can be deduced from current literature. In particular in regions with no commercial oyster culture, similar to the Wadden Sea, the virus may go undetected as wild beds with mixed age classes hamper the detection of mortality among juvenile oysters.     

美国熊本牡蛎基础群体构建及生产性能评估

为了获得优良熊本牡蛎(Crassosrea sikamea)种质资源,于2015年从美国Taylor贝类养殖公司引入熊本牡蛎,采用2种配对方式构建了核心基础群体;以中国熊本牡蛎选育群体为对照,评估了美国熊本牡蛎引种的生产性能及可行性。结果表明,美国熊本牡蛎相对于中国群体而言,表现出生长较快,商品规格比率较高,但存活力较低的特点。经过1周年养成,美国熊本牡蛎较中国群体子代壳高提高了11.48%,总重提高了48.12%。评估表明,群体来源对子代生产性能造成了显著的影响,而配对策略尚未产生显著影响。本研究发现美国熊本牡蛎在中国华南沿海具有较好的生产性能,这为熊本牡蛎种质改良提供了宝贵的繁育材料,也为华南牡蛎产业提供了一个可选择的优良品种。     

A herpes-like virus infecting Crassostrea gigas and Ruditapes philippinarum larvae in France

Concomitant sporadic high mortalities were reported in June 1997 among batches of larval Pacific oyster, Crassostrea gigas, and Manila clam, Ruditapes philippinarum , in a French commercial hatchery. Histological observation showed the presence of cellular abnormalities in affected animals. Electron transmission microscopy revealed the presence of herpes-like virus particles in infected larvae of both bivalve species. Viruses observed in C. gigas and R. philippinarum are closely related with respect to ultrastructure and morphogenesis. They were detected simultaneously in both bivalve species larvae indicating possible interspecific transmission. Moreover, PCR analysis using oyster herpes-like virus specific primers allowed amplification of fragments of expected sizes for both bivalve species and demonstrated the presence of viral DNA. The PCR products obtained for both bivalve species and digested by restriction enzymes displayed the same patterns. These data suggest that the same herpes-like virus may infect larval oysters and clams.     

Growth trials of Crassostrea gigas and Ostrea edulis in inshore waters of Malta (Central Mediterranean)

To investigate the possibility of establishing oyster culture in Malta, growth trials with C. gigas and O. edulis were undertaken. The oysters were placed in floating cages at depths of 3 and 8 m at Marsaxlokk bay, Mistra bay and Rinella creek. These oysters were sampled at two-month intervals for their growth, meat-shell index and survival. The temperature, salinity, oxygen tension, chlorophyll-a levels and the levels of particulate matter in suspension in the water at the test sites were monitored concurrently. A programme of qualitative and quantitative analyses of the natural phytoplankton population was also carried out.O. edulis suffered catastrophic summer mortalities at all locations when the ambient temperature of the water reached about 25°C; this virtually rules out any possibility of starting any flat oyster industry using this system. As for C. gigas, Marsaxlokk bay 3 m appears to hold some promise for its culture. At this location, this species showed reasonable growth, relatively low mortality and normal shell growth. Although at Rinella overall growth of C. gigas was comparable to that at Marsaxlokk at 3 m, the Rinella oysters accumulated copper and exhibited abnormal and thickened shell growth with the result that the oysters were of a much inferior condition. Growth at Mistra (both depths) and Marsaxlokk 8 m was stunted.No clear correlation could be established between the performance of the oysters and the environmental parameters monitored. Several factors that may have been responsible for the observed differences in oyster performance are discussed. In particular, the nature of the water movements is thought to have played a key role in dictating the growth of the oysters.