Influence of temperature and Artemia enriched with ω‐3 PUFAs on the early ontogenesis of Atlantic sturgeon,Acipenser oxyrinchus Mitchill, 1815

Successful breeding of fish species in aquaculture depends on several factors, among which the temperature and feed are the most significant ones. This study was designed to evaluate the effect of temperature in the range of 14–18°C on the rate of embryogenesis and duration of larva period and to estimate the efficiency of Artemia nauplii enriched with PUFAs in growing sturgeon larvae. The temperature of 16°C is the most suitable for both egg incubation and Atlantic sturgeon prelarvae maintenance under aquaculture conditions. Even minor temperature fluctuations of 1 degree up or down leads to increased loss both of eggs and prelarvae. Increased temperature shortens the incubation period but has a lesser impact on the duration of prelarvae onset of external feeding. The technology of Artemia nauplii bioencapsulation with a PUFA‐containing supplement in A. oxyrinchus rearing increases sturgeon larvae weight by 1.5 times at a constant survival dynamic.     

红姜花体细胞胚胎发生及植株再生的研究

以红姜花（Hedychium coccineum）的花丝和花药为外植体,通过体细胞胚胎发生途径建立了植株再生体系。结果表明：外植体在MS + 4 mg ? L-1 2,4-D + 4 mg ? L-1 NAA + 1 mg ? L-1 6-BA + 30 g ? L-1 蔗糖 + 7 g ? L-1 琼脂的培养基上经过120 d诱导出愈伤组织,愈伤组织在MS + 1 mg ? L-1 2,4-D + 0.25 mg ? L-1 NAA + 0.25 mg ? L-1 6-BA + 30 g ? L-1 蔗糖 + 7 g ? L-1琼脂的培养基上经过继代筛选,获得浅黄色松散易碎的胚性愈伤组织。胚性愈伤组织在MS无机盐 + B5维生素+ 100 mg ? L-1谷氨酰胺 + 230 mg ? L-1 脯氨酸 + 100 mg ? L-1麦芽提取物 + 0.02 mg ? L-1 NAA + 0.02 mg ? L-1 TDZ + 0.5 ~ 1.0 mg ? L-1 2,4-D + 45 g ? L-1蔗糖的培养基中通过3个月的悬浮培养,可得到均质稳定的胚性细胞悬浮系（ECS）。胚性悬浮细胞在SH无机盐 + B5维生素 + 100 mg ? L-1谷氨酰胺 + 230 mg ? L-1脯氨酸 + 100 mg ? L-1麦芽提取物 + 0.25 mg ? L-1 NAA + 0 ~ 0.20 mg ? L-1 TDZ + 45 g ? L-1蔗糖 + 7 g ? L-1琼脂的体胚诱导培养基中培养10 d,可见到白色半透明体胚发生,20 d后体胚发育成熟。当TDZ浓度为0.15 mg ? L-1时,体胚诱导率高达4 500个 ? mL-1 PCV ECS（PCV：细胞密实体积）。在SH无机盐 + B5维生素 + 0.20 mg ? L-1 IAA + 0.25 ~ 1.0 mg ? L-1 6-BA + 30 g ? L-1蔗糖 + 7 g ? L-1琼脂的体胚萌发培养基上,体胚萌发率高达100%。将萌发的体胚转移到1/2MS + 1 g ? L-1活性炭成苗培养基中,进一步发育成正常的再生植株,植株室外栽培成活率达90%。     

蜡梅胚胎晚期丰富蛋白基因CpLEA的克隆及表达分析

从蜡梅 （Chimonanthus praecox）花cDNA文库中获得了1个胚胎晚期丰富蛋白（LEA）基因的cDNA全长序列,命名为CpLEA（GenBank登录号：JF412788）,该基因cDNA含有一个273 bp编码91个氨基酸的开放阅读框,从蜡梅基因组DNA中扩增该基因发现,该基因含有两个大小分别为35 bp和707 bp的内含子。生物信息学分析显示,CpLEA的编码蛋白含有4个胚胎晚期丰富蛋白第3族LEA蛋白特有的11个氨基酸的基元重复序列,进化树分析表明该编码蛋白与榛子的LEA蛋白亲缘关系最近。通过原核表达获得了与预期大小一致的融合蛋白。实时荧光定量PCR分析表明,该基因在蜡梅成熟种子中表达量最高,在根中几乎不表达;在盛开期花的各个部位中又以雌蕊中的表达量最高;在花发育早期表达量较高,随后下降并保持相对稳定,在衰败期突增并达到最高;检测该基因在脱落酸（ABA 50 μmol · L^-1）、低温（4 ℃）、高温（42 ℃）、干旱（30% PEG6000）和高盐（NaCl 1 mol · L^-1）5种外源非生物胁迫因子作用下的表达特性。结果表明,该基因能够被ABA、低温、高温、PEG 和NaCl诱导表达,并在随后的时间内呈现出不同的表达模式。表明CpLEA可能在蜡梅花发育和多种非生物胁迫响应中发挥作用。     

系谱和聚类分析在橡胶树内珠被组织培养中应用研究

分析16种不同基因型橡胶树幼果内珠被愈伤组织诱导率、分化率;追踪参试基因型系谱;并基于25对EST-SSRs分子标记数据对其进行聚类分析,研究系谱、聚类结果与体胚发生的关系.结果表明:16种基因型的愈伤诱导率均较高,位于68.46％-100％,7个基因型愈伤组织能诱导出体细胞胚状体,其中3个基因型能够较大量诱导体细胞胚状体,其余9个基因型的愈伤组织完全不具备分化能力;具有RRIM600亲缘的基因型体胚分化能力强,具有IAN873、PB5/51、热研217亲缘的基因型体胚分化能力弱或无体胚分化能力;16种基因型的相似系数位于0.56～0.89间,在遗传距离0.73时分为5类.体胚发生结果与系谱分析结果基本吻合,与聚类分析结果吻合度不大;提示可以利用系谱分析提高橡胶树组织培养的培养基筛选效率.最后讨论分子标记应用于组织培养的研究思路.     

抗松针褐斑病湿地松体细胞胚胎发生与植株再生

为将抗松针褐斑病湿地松(Pinus elliottii Engelm.)良种快速推广,以未成熟合子胚为外植体,研究了合子胚发育阶段对胚性愈伤组织诱导以及ABA与PEG8000对体细胞胚成熟的影响,并探讨了体细胞胚质量及干化条件对体细胞胚萌发的影响。结果表明:发育至2~4阶段的合子胚最利于胚性愈伤组织的诱导,LP+16 mg·L~(-1)ABA+180 g·L~(-1)PEG8000+2 g·L~(-1)AC适合于体细胞胚的成熟,高质量体细胞胚经"滤纸容器法"干化2周后萌发,萌发的体细胞胚于1/4WPM+0.3 mg·L~(-1)NAA+1 mg·L~(-1)IBA上培养70 d左右根部发育,转入1/2WPM中,30 d左右侧根产生发育完整的再生植株。     

Yolk mobilization in perch, Perca fluviatilis L., embryos

During embryogenesis of the perch, Perca fluviatilis L., a gradual proteolysis of yolk proteins was revealed by SDS-PAGE and immunoblotting and was comparable to other teleosts. A 118 kDa polypeptide was processed in two 96 and 18 kDa descendants; further proteolysis produced two peptides of 64 and 31 kDa. Ultrastructure and immunocytochemistry were used to compare electrophoresis patterns with morphological changes in the developing embryo and to localise yolk proteins in embryonic cell layers. In contrast to most other freshwater teleost, in spawned eggs of the perch yolk spheres fused together forming a homogenous yolk mass; lipid droplets formed a single oil drop on top of the egg. Combination of electrophoresis data and light and electron microscopical observations suggested a specific and parallel mobilization of yolk and lipid. In addition, yolk absorption by the yolk syncytial layer was observed. Numerous yolk platelets were found in the yolk syncytial layer and, in later stages of embryogenesis, also in the inner cell mass, but never in the cells of the enveloping layer.     

豆科牧草沙打旺抗乙硫氨酸变异系筛选

以NaN3诱变处理的沙打旺胚性愈伤组织为材料,用甲硫氨酸类似物—乙硫 氨酸为选择剂,筛选到1株抗性稳定且能再生植株的抗0.6 mmol L－1乙硫氨酸 变异系。该变异系细胞对乙硫氨酸的抗性是野生型细胞的8倍。当把变异系愈伤组织传入 体细胞 胚胎诱导培养基上,平均体细胞胚胎发生频率为13.9%,平均每g鲜重愈伤组织可产生21 个体     

龙眼胚性培养物APX同工酶的分析方法建立及其在龙眼体胚发生过程中的变化

以龙眼胚性培养物为材料,进行抗坏血酸过氧化物酶(APX)同工酶的聚丙烯酰胺凝胶电泳分析方法研究,建立一套适合分离龙眼胚性培养物中不同类型APX同工酶的垂直板聚丙烯酰胺凝胶电泳方法;进一步研究了APX同工酶在龙眼体胚发生过程中的变化,发现在胚性愈伤组织、球形胚、子叶形胚3个阶段APX同工酶存在差异.     

Histological Study on Soybean Somatic Embryogenesis

Soybean somatic cell could induce the development of embryoid which was similar to embryo morphologically and structurally. Somatic embryogenesis system of soybean was used to conduct genetic transformation of soybean because of its several advantages such as higher transformational efficiency,beetter synchronism and fewer plant chimeras among transgenic plants. After infected with agrobacterium tumefaciens,the initiation,differentiation and development of young cotyledon embryogenic cell of soybean which was cultured on selective culture medium with kanamycin were investigated through histological study. The result showed that somatic embryo was differentiated in non-bud differentiation way. The embryogenic cells were differentiated from epidermis of explant or cells in 1 layer or 2 layers,with the division of embryogenic cells and degradation and disorganization of surrounding cells,the embryogenic cells would form embryoid with analogous suspensor structure.Later,globular embryoid would extrude from epidermis then developed into heart-shape embryo.The experiment was expected to provide theoretical reference for the construction of high transformational system of using plant somatic embryogenesis induced by young cotyledon of soybean.     

长江和黄河流域棉区棉花品种体细胞胚胎发生和植株再生比较研究

选用我国长江和黄河流域棉区各10个棉花品种以及珂字201和YZ1共22个基因型,研究和优化棉花体细胞胚胎发生的相关条件参数。在此基础上,比较了两生态区棉花品种的体细胞再生能力。结果表明, 在IBA+KT的激素组合下,多数基因型有较强再生能力;在愈伤组织诱导时期,铵态氮是必需的,而在愈伤组织继代分化时期,一定浓度的硝态氮则能促进分化;没有铁盐不能诱导出愈伤组织,而铁盐浓度为56 mg L^-1时有利于胚分化;长江流域和黄河流域棉花品种的植株再生潜力没有明显差别,但长江流域的品种体细胞胚胎发生所需的时间长。本文首次获得了鄂抗棉3号、5号、鄂棉20、鄂棉23、豫棉9号、豫早73、豫棉12、豫棉1221等8个品种的体细胞胚胎发生和植株再生。