Effects of embryonic exposure to α‐lipoic acid or ascorbic acid on hatching rate and development of zebrafish (Danio rerio)

This study investigates the effects of embryonic exposure to two different antioxidants on growth and development in fish. Zebrafish (Danio rerio) embryos (100 per group) were exposed to lipoic acid (LA, 6–12 μM) or ascorbic acid (AA, 100–200 μM) and the hatching rate, standard lengths (SL) at hatching, development and growth post‐hatching monitored. The SLs at hatching were increased (P<0.05) in both antioxidant‐exposed groups relative to the controls, with no effect on yolk reserves. This enhanced development persisted up to 15 days post hatching. At hatching, cell proliferation rates (P<0.0005) and basic fibroblast growth factor (P<0.001), were greater in the antioxidant‐exposed fish than in the controls (0 μM antioxidant); no oxidative DNA damage was detected (P>0.05). Activity of the endogenous antioxidant enzyme superoxide dismutase was greater (P<0.001) in LA‐treated fish than in the controls. The results suggest that embryonic treatment of zebrafish with LA or AA during embryogenesis enhanced cell proliferation, leading to increased somatic growth in the larval stages, persisting into the juvenile stage. The findings support the treatment of embryonic fish with antioxidants for enhanced results in aquaculture.     

牙鲆胚胎及其初孵仔鱼的盐度耐受力

实验盐度设置为５,１０,１５,２０,２５,３０,３５,４０,４５,５０。当盐度为５～３５时,牙鲆（Ｐａｒａｌｉｃｈｔｈｙｓ ｏｌｉ－ｖａｃｅｕｓ）胚胎的孵化率都高于９５％;仔鱼体态正常,７２ｈ后的存活率高于９５％。刚孵化的仔鱼在低盐度下的体长较长,但７２ｈ后,没有明显差异（Ｐ〉０．０５）。自然条件（盐度为３２）孵化的仔鱼转移到不同盐度的溶液中,在盐度５～４５内,仔鱼的存活率高于５０％。实验初步表明     

液体培养过程中金花茶体细胞胚增殖优化

为获得大量性状一致的金花茶体细胞胚,试验采用单因素与完全随机设计,在液体培养过程中先后研究了碳源组合、脯氨酸浓度及培养基体积与接种量对其增殖的影响。结果表明：碳源组合对体细胞胚增殖无显著影响,但以40 g·L-1蔗糖与20 g·L-1山梨醇作为碳源时,低畸形胚的产生频率较低;脯氨酸浓度对体细胞胚增殖有极显著影响且适宜的浓度为2g·L-1;培养基体积、接种量及两者的交互作用均对体细胞胚增殖有极显著影响且其中影响最大的为培养基体积,最佳的培养基体积为45 mL,最佳的接种量为0.5或1.0 g。采用以上优化结果对体细胞胚进行液体培养,其增殖系数可达5.65且一致性良好。     

显微注射胚胎干细胞对长爪沙鼠胚胎发育的影响

将不同数量的成系的小鼠胚胎干细胞(ES细胞)显微注射注入长爪沙鼠(Merionesunguiculatus)的不同胚期的胚胎中,研究其对胚胎发育的影响,以期初步探索嵌合胚胎制作的条件。试验结果表明,显微注射2～5、6～10和11～15个ES细胞数量之间对早囊/囊胚期胚胎的成活率和孵化率的影响均不显著,但显微注射11～15个ES细胞的成活率和孵化率较低,分别为82.14%和64.29%;显微注射2～5个ES细胞的成活率和孵化率均较高,分别为90.90%和78.79%。显微注射11～15个ES细胞和未经显微注射的胚胎成活率分别为95.12%和82.14%,差异极显著(P<0.01);孵化率分别为90.48%和64.29%,差异极显著(P<0.01)。不经显微注射的桑椹/囊胚、经显微注射的8～16-细胞期胚胎和早囊/囊胚,其成活率分别为94.92%、91.53%和87.50%,差异均不显著(P>0.05);从孵化率来看,未经显微注射的桑椹/囊胚和经显微注射的8～16-细胞期胚胎其孵化率分别为89.83%和61.02%,差异极显著(P<0.01)。     

农杆菌介导法将Bt基因转入大豆的研究

以6个品种大豆体细胞胚为受体,以Bt基因为目的基因,应用根癌农杆茵介导法对大豆进行了遗传转化,经体细胞胚萌发、生根及壮苗培养和卡那霉素选择培养,获得了完整的抗性再生苗,经PCR、PCR-Southern和点杂交分析鉴定,初步证明外源基因Bt已整合到大豆的基因组中。     

AE油佐剂灭活疫苗对鸡胚和仔鸡的保护力试验

三批禽脑脊髓炎(AE)油佐剂灭活疫苗免疫开产前种鸡,种鸡免疫2~3个月后所产种蛋对禽脑脊髓炎病毒(AEV)鸡胚易感性试验的受保护率达100%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEV VR株攻毒的受保护率亦达100%; 种鸡免疫6~7个月后所产种蛋对AEV VR鸡胚易感性试验的受保护率达75%~80%,而同期所产种蛋孵出的仔代雏鸡,在出雏后3周内对AEVVR株攻毒的受保护率达70%~90%。试验结果表明,AE灭活疫苗AEV鸡胚易感性试验的鸡胚保护率与同期雏鸡对AEV VR肌肉途径攻毒的保护率基本相同。     

根癌农杆菌介导转化籼稻影响因素的研究

选用我国籼型杂交稻中的几个重要亲本 ,以携带有双元载体 p CAMBIA1 3 0 1的根癌农杆菌 EHA1 0 5为载体 ,以GUS基因的瞬间表达为依据 ,研究根癌农杆菌介导转化籼稻的影响因素 ,确立对转化频率有重要影响的几个条件。结果表明 :经短期预培养 (4～ 5d)的幼胚是较为理想的转化受体材料 ;在共培养基中添加较高浓度的乙酰丁香酮 (3 0 0～ 4 0 0μmol/ L)可提高 GUS基因的瞬间表达频率 ;以 CC培养基作为共培养后抗性愈伤筛选的基本培养基有利于抗性愈伤组织的筛选培养。按此确立的条件 ,在特青、龙特甫 (B)、珍汕 97(B)、盐恢 559等籼稻品种或亲本中的 GUS瞬间表达率高达 80 % ,抗性愈伤组织转化率高达 70 %     

提高N6培养基钙浓度对玉米幼胚培养的影响

筛选试验结果表明 ,将 N6 培养基 Ca2 + 浓度从 1.13mmol/ L提高到 5 m mol/ L ,对玉米幼胚胚性愈伤组织的诱导、继代和分化具有促进作用。在一次继代过程中 ,Ca2 +浓度为 5 m mol/ L 的培养基上培养的胚性愈伤组织相对生长量比 1.13m mol/ L 的原培养基增加 0 .8倍。 Ca2 +浓度提高以后 ,在培养基中再添加钙调蛋白 (Ca M)抑制剂盐酸氯丙嗪 (CPZ) ,可抑制 Ca2 +的促进作用 ,说明钙增加后对玉米胚性愈伤组织诱导和继代的促进作用受Ca2 + / Ca M信息系统控制。用改进后的 N6 培养基 ,可提高生产上大面积推广的玉米自交系胚性愈伤组织的诱导率和继代生长量     

根据雏鸡血清酶活性对蛋鸡进行早期选种的研究

本研究测定了147只8、12周龄S220母雏血清碱性磷酸酶(AKP)、谷草转氨酶和乳酸脱氢酶活性,估测其遗传力及其与产蛋性能间的表型相关和遗传相关。结果表明,8周龄血清AKP活性遗传力为0.62,300日龄产蛋量遗传力为0.20,两者间的遗传相关为0.8245。利用8周龄血清AKP活性作为辅助性状对300日龄产蛋量进行间接选择,其间接选择效率为个体选择的1.4519倍,为家系选择的1.4837倍,为合并选择的1.1887倍。因此,8周龄血清AKP活性可作为蛋鸡早期选种血液生化指标之一。     

Lipid-based transfection as a method for gene delivery in zebrafish (Danio rerio) embryos

A major challenge to the widespread production of transgenic, knockout and knockdown zebrafish has been the absence of a simple and effective procedure for introducing macromolecules into the fertilized egg. None of the existing techniques for gene transfer in fish embryos has proven to be a major advance over cytoplasm microinjection, which is a technically demanding and time‐consuming procedure. This report addresses this need, considering that the development of protocols for lipid‐based transfection with fish embryos would considerably simplify gene transfer in this complex biological model. In this study, lipid‐based transfection with two different reporter vectors was carried out in zebrafish embryos at different developmental stages. The parameters tested included different plasmid/transfection reagent ratios as well as the influence of an added transfection enhancer reagent. When embryos were transfected in the blastula stage with a pEGFP‐N1 vector, more than 35% successfully incorporated the plasmid and expressed the fluorescent protein 24 h after transfection. The transfection enhancer did not show any significant effect in our experiments. This work presents an approach to implement this technique as a faster, cheaper and more practical alternative than microinjection.