CN107873592A - 一种虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法 - Google Patents
一种虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法 Download PDFInfo
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Abstract
本发明公开了一种虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,包括以下步骤:家系建立、幼贝育成、获得高出柱率高品质脂肪酸闭壳肌家系,前述的高出柱率高品质脂肪酸闭壳肌家系,是通过生长性状及脂肪酸组分来评定,生长性状包括壳长、壳宽、壳高、扇贝湿重、闭壳肌与软体部干重并计算出柱率,脂肪酸组分测定包括油脂提取、转酯化及气相色谱检测;出柱率高且EPA与DHA含量高及ω‑3/ω‑6脂肪酸比值高的家系即可筛选出高出柱率高品质脂肪酸闭壳肌家系,高出柱率高品质脂肪酸闭壳肌家系经2~3年培育成可繁育成体,作为培育F2代的亲本,此家系具有抗逆性强、营养价值高、维护成本低、管理相对容易等优势。
Description
技术领域
本发明涉及一种虾夷扇贝高品质脂肪酸闭壳肌家系选育方法,属于农业海水养殖技术领域。
背景技术
虾夷扇贝(Mizuhopecten yessoensis)是全世界扇贝科中最适于增养殖的优良品种之一。自20世纪80年代初从日本成功引入该品种以来,历经30多年工厂化育苗及人工增养殖技术的不断完善与推广,虾夷扇贝已成为我国贝类养殖的主导品种。近年来,虾夷扇贝在北黄海主要养殖区的死亡率居高不下,制约了产业的健康稳定发展。研究表明种质衰退、环境变化、过度养殖、病原侵袭等可能与虾夷扇贝的大规模死亡有关培育优质抗逆新品种,是保证虾夷扇贝增养殖产业可持续发展的有效途径之一。当前虾夷扇贝养殖所用苗种的生长、抗逆性乃至品质质量等性状大多未经过系统的定向选育,养殖良种覆盖率低。我国重要经济扇贝的选择育种现阶段已取得一定进展,成功选育出了“中科红”海湾扇贝,“蓬莱红”栉孔扇贝、“海大金贝”虾夷扇贝等新品种,有力促进了扇贝养殖业的健康发展。
虾夷扇贝闭壳肌的高度不饱和脂肪酸以Omega-3(ω-3)系列为主,主要包括二十二碳六烯酸(docosahexaenoic acid,DHA)和二十碳五烯酸(eicosapentaenoic acid,EPA)。ω-3脂肪酸因其在生命体中发挥的重要作用而备受青睐,其适量摄入能够调节机体免疫和基因表达,增强机体大脑的前扣带皮层功能进而改善认知的灵活性。鱼类和海产品是自然状态下人类蛋白质和必需微量营养素的重要来源,其长链ω-3脂肪酸DHA和EPA是保障育龄妇女及婴幼儿生长发育的必需脂肪酸。目前我国虾夷扇贝的人工选育仍主要围绕在生长与壳色方面,还没有开展基于脂肪酸的抗逆优质品种的选育,因此虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育工作亟待开展以满足水产养殖的需要。
发明内容
为解决现有技术的不足,本发明的目的在于提供一种虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,采用该方法可以提高虾夷扇贝闭壳肌的出柱率及营养价值,从而促进扇贝种业的发展及高附加值海洋保健产品的开发。
为了实现上述目的,本发明采用如下的技术方案来实现:
一种虾夷扇贝高品质脂肪酸闭壳肌家系选育方法,包括以下步骤:
(1)、家系建立:在虾夷扇贝的苗种繁育季节,建立若干同胞或者半同胞家系;
(2)、幼贝育成:将步骤(1)所得的家系经常规的幼虫培育、中间暂养和海上育成环节后,获取壳长在5~6cm的扇贝,待用;
(3)、获得高出柱率高品质脂肪酸闭壳肌家系:将步骤(2)所得的扇贝中进行生长性状和脂肪酸组分测定,生长性状包括壳长、壳宽、壳高、扇贝湿重、闭壳肌与软体部干重,并利用闭壳肌与软体部干重比率计算出柱率,即出柱率=闭壳肌干重/软体部干重,脂肪酸组分测定包括油脂提取、转酯化及气相色谱检测,筛选闭壳肌出柱率高的家系,进一步筛选闭壳肌脂肪酸的EPA与DHA含量高并且ω-3/ω-6脂肪酸比值高的家系;其中闭壳肌脂肪酸的EPA 与DHA含量高并且ω-3/ω-6脂肪酸比值高的家系为高品质脂肪酸闭壳肌家系;
(4)、将步骤(3)所得的高出柱率高品质脂肪酸闭壳肌家系经2-3年培育成可繁育成体,作为培育F2代的亲本。
对于上文所述的技术方案,优选的情况下,所述步骤(3)中闭壳肌出柱率的值高于家系平均值10%以上。
对于上文所述的技术方案,优选的情况下,所述步骤(3)中闭壳肌脂肪酸的EPA与DHA 含量高于家系平均值5%以上,ω-3/ω-6脂肪酸比值高于家系平均值5%以上。
对于上文所述技术方案,优选的情况下,在筛选虾夷扇贝闭壳肌高出柱率的同时,进一步筛选出高品质脂肪酸闭壳肌家系。
对于上文所述技术方案,优选的情况下,每个家系所用于分析的幼贝数不少于30枚。
本发明的有益效果:通过比较各家系扇贝的出柱率及脂肪酸组成便可筛选出高品质脂肪酸闭壳肌家系,具有出柱率高和营养价值高等优势,即闭壳肌出柱率高于家系平均值10%以上,闭壳肌脂肪酸的EPA与DHA含量高于家系平均值5%以上,ω-3/ω-6脂肪酸比值高于家系平均值5%以上;采用本发明的方法筛选出的优良家系,既可以作为高品质脂肪酸虾夷扇贝育种材料加以直接利用,也可与其它优良品系进行杂交育种,选育出具有其它优良性状的扇贝新品种;另外,本发明的方法可广泛应用于扇贝优质种质资源的筛选与评价,特别是虾夷扇贝高品质脂肪酸闭壳肌新品种培育以及高脂肪酸QTL遗传解析、高脂肪酸功能基因定位。
附图说明
图1是虾夷扇贝各家系的生长性状与出柱率;
图2是虾夷扇贝各家系闭壳肌饱和脂肪酸(SFA)、单不饱和脂肪酸(MUFA)和多不饱和脂肪酸(PUFA)相对丰度比较;
图3是虾夷扇贝各家系闭壳肌脂肪酸EPA与DHA含量与ω-3/ω-6比值。
具体实施方式
下述非限定性实施例可以使本领域的普通技术人员更全面地理解本发明,但不以任何方式限制本发明。
以下结合实施例对本发明作具体的介绍。
实施例
一种虾夷扇贝高品质脂肪酸闭壳肌家系选育方法,包括以下步骤:
(1)、家系建立:在长海县大长山岛海区,每年的2~4月份海水水温在2~10℃左右,此时是虾夷扇贝的苗种繁育季节。根据动物育种学原理,建立虾夷扇贝全同胞家系40个。
(2)、幼贝育成:将上述40个全同胞家系采用常规的方法进行幼虫培育,然后将幼虫放至大长山岛海区进行中间暂养和海上育成,在当年的10月份,家系幼贝的壳长达到5~6 cm,接下来便可以对高脂肪酸闭壳肌家系进行筛选。
(3)、生长性状测定:当自然海区的水温在5-6℃时,从海上随机选取12个家系各30~ 50枚健康活泼幼贝,各家系用游标卡尺(精度为0.01mm)测量30枚壳长、壳宽和壳高,以电子天平(精度为0.01g)称量扇贝湿重;并剖取软体部于60℃烘干至恒重,以分析天平(精度为0.0001g)分别称量闭壳肌与软体部干重,并计算出柱率(出柱率=闭壳肌干重/软体部干重)。
脂肪酸组分测定:同一家系闭壳肌(干样)通过完全随机区组平均分成三个生物学重复,以IKA-A11基本型研磨机粉碎至60目网筛过滤,未通过的颗粒用玻璃研钵研磨直至全部过筛为止。称取粉碎后的闭壳肌2~3g,加入3%(质量体积比)的二丁基羟基甲苯(BHT)甲醇溶液,以改进的Folch法(Folch J,Lees M,Sloane-Stanley G H.A simple method forthe isolation and purification of total lipids from animal tissues[J].J BiolChem,1957,226(1): 497-509.)萃取脂质。所得脂质用0.5mol/L的KOH-甲醇溶液于70℃水浴中皂化1h,用三氟化硼(BF3)催化法(Metcalfe L D,Schmitz A A,Pelka J R.Rapidpreparation of fatty acid esters from lipids for gas chromatographic analysis[J].Analytical Chemistry,1966,38(3):514-515.)制取脂肪酸甲酯,并转移到石油醚中,取上清液用于气相色谱分析。采用37种脂肪酸甲酯标准样品(SIGMA公司)定性,通过面积归一化法定量。气相色谱分析条件:日本岛津GC—2010 型气相色谱仪配有抗氧化交联石英毛细管色谱柱FFAP(30m×0.25mm,0.3μm);检测器为FID(氢火焰离子化检测器);载气为高纯氮气;恒流为1mL/min;分流比为100:1;汽化室温度为260℃,色谱柱初温160℃,以2℃/min程序升温至230℃,并保持基线平稳。
获得高出柱率高品质脂肪酸闭壳肌家系:虾夷扇贝各家系生长性状参见图1,闭壳肌脂肪酸组成及评价指标见图2至图3及表1。筛选出柱率高且EPA与DHA含量高及ω-3/ω-6脂肪酸比值高的家系即为高出柱率高品质脂肪酸闭壳肌家系。
(4)、将步骤(3)所得的高出柱率高品质脂肪酸闭壳肌家系经2-3年培育成可繁育成体,作为培育F2代的亲本。
虾夷扇贝高出柱率高品质脂肪酸家系闭壳肌评定:
参照图1(图中家系编号B01-B12从左向右依次排列),随机选取的12个家系中,家系 B05的闭壳肌干重最重,与家系B04、B10差异不显著,三个家系显著重于其余各家系;在出柱率方面,家系B04最高,家系B10次之,二者差异不显著,且与家系B05、B06、B07、 B09差异不显著,但是显著高于其余各家系,在12个选育家系中,B04、B05和B10家系的出柱率优势最显著。
表1为虾夷扇贝各家系闭壳肌脂肪酸组成,参照表1、图2至3,各家系闭壳肌的EPA+DHA 含量占总脂肪酸总量的45%以上,家系B11的EPA与DHA含量最高,占脂肪酸总量的49.34%,与家系B06、B12、B07、B05、B10、B03差异不显著,家系B04的EPA与DHA含量最低(45.68%),显著低于其余各家系。各家系PUFA中ω-3/ω-6脂肪酸比值为10.94~12.69;家系B11的ω-3/ω-6脂肪酸比值最大(12.69),与家系B01、B02、B03、B04、B05、B07、B10、 B12差异不显著,家系B11、B10、B07和B04显著高于家系B06、B08、B09,且家系B08 的ω-3/ω-6脂肪酸比值最低(10.94),显著低于其余各家系。
结合家系的闭壳肌出柱率,综合分析B05和B10为虾夷扇贝闭壳肌高出柱率高品质脂肪酸家系,其闭壳肌出柱率高于家系平均值10%以上,脂肪酸的EPA+DHA含量高于家系平均值5%以上,ω-3/ω-6脂肪酸比值高于家系平均值5%以上。
本发明的方法,利用人工培育的虾夷扇贝全同胞家系,比较不同家系的出柱率和脂肪酸组成,筛选出的家系闭壳肌具有高出柱率、高品质脂肪酸等优势,出柱率高于平均值10%以上,EPA与DHA含量和ω-3/ω-6脂肪酸比值分别高于平均值5%以上。
表1虾夷扇贝各家系闭壳肌脂肪酸组成
Claims (4)
1.一种虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,其特征在于,包括以下步骤:
(1)、家系建立:在虾夷扇贝的苗种繁育季节,建立若干同胞或者半同胞家系;
(2)、幼贝育成:将步骤(1)所得的家系经常规的幼虫培育、中间暂养和海上育成环节后,获取壳长在5~6cm的扇贝,待用;
(3)、获得高出柱率高品质脂肪酸闭壳肌家系:在步骤(2)所得的扇贝中筛选闭壳肌出柱率高的家系,进一步筛选闭壳肌脂肪酸的EPA与DHA含量高并且ω-3/ω-6脂肪酸比值高的家系;
(4)、将步骤(3)所得的高出柱率高品质脂肪酸闭壳肌家系经2-3年培育成可繁育成体,作为培育F2代的亲本。
2.根据权利要求1所述的虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,其特征在于,所述步骤(3)中闭壳肌出柱率的值高于家系平均值10%以上。
3.根据权利要求1所述的虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,其特征在于,所述步骤(3)中闭壳肌脂肪酸的EPA与DHA含量高于家系平均值5%以上,ω-3/ω-6脂肪酸比值高于家系平均值5%以上。
4.根据权利要求1-3任意一项所述的虾夷扇贝高出柱率高品质脂肪酸闭壳肌家系选育方法,其特征在于,每个家系所用于分析的幼贝数不少于30枚。
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