AgNO3处理下银杏幼胚愈伤组织的形成和激素变化

采用石蜡切片技术和酶联免疫吸附法(ELISA),以银杏幼胚为外植体,添加0,1和5 mg/L AgNO3作处理,观察21 d内愈伤组织的形成过程,测定内源激素吲哚乙酸(IAA)、保幼激素类似物(ZR)、赤霉素(GA)和脱落酸(ABA)的变化,分析AgNO3对银杏幼胚愈伤组织分化潜能的影响。结果表明:根据解剖结构愈伤形成可分为4个阶段。第1阶段(0~7 d),IAA升高,ABA降低,表皮细胞启动脱分化。第2阶段(≥7~11 d),4种激素浓度均升高,愈伤组织和细胞团初步形成。第3阶段(≥11~16 d),GA、ABA大幅降低,细胞团大量形成。第4阶段(≥16~21 d),ABA骤降、GA迅速上升,愈伤组织和细胞团增殖。1 mg/L AgNO3处理下分生细胞团种类最多,其次为5mg/L处理,0 mg/L处理最少。研究认为,银杏幼胚愈伤组织具有较强的分化潜能,AgNO3能影响其内源激素的平衡,调控分化方向。     

四种鸢尾属植物种子休眠和萌发研究

燕子花(Iris laeviga ta)、玉蝉花(Iris ensa ta)、溪荪(Iris sanguinea)、黄菖蒲(Iris pseudacorus)是鸢尾科4种喜水湿植物。经萌发实验证明,除溪荪种子没有休眠外,其它3种种子都有休眠。经去除种皮、低温沙藏、赤霉素处理和胚培养4种处理,证明燕子花、玉蝉花、黄菖蒲中抑制种子萌发的主要因素是胚乳。不同种打破休眠的有效方法不同,其中50 mg/L GA3处理可提高黄菖蒲种子的发芽率,达到84.4%,而低温沙藏30 d可使玉蝉花种子的发芽率达到94.4%,低温沙藏90 d则可提高燕子花的发芽率达到87.8%。     

萝卜与大头菜属间杂种胚的离体培养

用萝卜雄性不育系DH303A作母本,以大头菜为父本进行属间远缘杂交,杂交后代的初期结荚率为64%~88%,横径第7天停止增加,荚长第8天停止生长,授粉以后第7天开始败育,而且越往后败育得越快。在杂种胚发育后5～8 d进行胚抢救,用生芽培养基（1/2 MS＋1.0 mg/L 6-BA＋0.2 mg/L NAA＋0.5%活性炭）和生根培养基（1/2 MS＋0.2 mg/L 6-BA＋1.0 mg/L NAA＋0.5%活性炭）进行离体培养,能成功获得幼苗。     

野牛草成熟胚植株再生及其影响因素研究

以野牛草Buchloe dactyloides成熟胚为外植体,对外植体灭菌方式及影响成熟胚再生的因素进行了研究.结果表明:70%酒精处理60 s,75% NaClO溶液(原溶液含有效氯为7%～10%)处理30 min,污染率最低为0.59%.6-BA 0.1 mg/L,2,4-D浓度增至3 mg/L时,愈伤组织诱导率达最高,为80.27%,但愈伤组织结构疏松、呈水渍状.附加5 mg/L硝酸银(AgNO3)或硫代硫酸银(STS)时,愈伤组织诱导率略有降低,愈伤组织结构紧密且表面有颗粒状突起,当AgNO3或 STS浓度为10 mg/L以上时,均不利于愈伤组织的诱导.愈伤组织继代培养过程中,3/4倍MS大量元素用量、聚乙烯吡咯烷酮(PVP) 200 mg/L、维生素C(Vc )200 mg/L及水解酪蛋白(CH) 1 000 mg/L可减轻愈伤组织褐化程度.不同诱导培养基所获得的愈伤组织与其植株再生能力有关,以不加任何植物生长调节剂的MS培养基(MS0)为分化培养基,仅在附加STS的诱导培养基中所得的愈伤组织能够形成再生植株,再生率为10%.     

不同外植体对多年生黑麦草愈伤组织诱导的影响

对多年生黑麦草的3个品种进行愈伤组织诱导,选取成熟种子、成熟胚、胚轴、胚根4种外植体。结果表明,胚轴、胚根诱导效果较差,成熟胚在愈伤组织的诱导率、鲜重及愈伤质量方面均优于成熟种子。2,4-D浓度在2~4mg/L,6-BA浓度为0.025mg/L时有利于成熟胚诱导产生愈伤组织;成熟种子为外植体时,2,4-D浓度在5~8mg/L,6-BA浓度在0.025mg/L或0.2mg/L有利于成熟种子诱导产生愈伤组织。2,4-D浓度为2mg/L,6-BA浓度为0.5mg/L时成熟胚和成熟种子诱导的愈伤组织鲜重均达到最大。特拉华在愈伤组织的诱导率、增殖速度及愈伤质量方面表现较优,托亚次之,尤文图斯较差。AC和LH有利于植株的再生。     

小麦幼胚愈伤组织辐照后代的遗传变异分析

以３个普通小麦品种为材料，用＾６０Ｃｏγ射线辐照小麦幼胚愈伤组织，在ＭＲ１、ＭＲ２代中才观察到广泛变异，其后代在育性、株高、抽穗期、千粒重、株型、蛋白质含量等方面者发生了明显变异，这些变异较单纯用组织培养得到的无性系的后代的变异频率更大，变异范围也更为广泛，并且从ＭＲ２中得到了几个有较高利用价值的变异株系。     

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     

In-vitro Fertilization of Ovules of Some Species of Brassicaceae*

The method of in-vitro fertilization of ovules can be successfully applied to various species of Brassincaceae. Mature embryos and plants were obtained after in-vitro pollinating the ovules of Arabis caucasica, Brassica napus, B. oleracea var. sabellica (kale), B. oleracea var. italica (broccoli), Diplotaxis tenuifolia, Moricandia arvensis and Sisymbrium Loeselli. In the case of Sinapis alba fertilization and embryo development did not occur. The same method has been successfully used for obtaining hybrid immature embryos at different stages of development from crosses between B. napus X D. tenuifolia, B. napus X M. arvensis, B. oleracea var. italica X D. tenuifolia, D. tenuifolia x B. napus, D. tenuifolia. X M. arvensis and D. tenuifolia X S. Loeselli. The present findings show that in-vitro pollination of ovules of various species of Brassicaceae makes it possible to (a) perform the whole process of fertilization and embryogenesis; and (b) obtain intergeneric hybrid embryos.     

Egg quality determination based on the shape of the lipid vesicle in common dentex, Dentex dentex

The shape of the lipid vesicle (the ratio between the maximal and the minimal diameter and the shape coefficient) of the eggs of Diplodus puntazzo and Sparus aurata is correlated with larval survival rate (%) and can be used as an egg quality marker as reported previously by Lahnsteiner and Patarnello (2005) . In the present study, this method for egg quality determination has also been validated for common dentex, Dentex dentex, after introducing several modifications in the calculation and evaluation procedure in order to obtain higher sensitivity. In the present study, it could be demonstrated that the shape of the lipid vesicle is not only correlated with the hatching rate but also with the survival rate of 1, 2 and 3 days post hatch larvae and therefore it can also be used as an indicator of larval viability at early stages of development.     

Ammonia dynamics in relation to hatching in Atlantic halibut (Hippoglossus hippoglossus L.)

Total ammonia content (T_Amm) and excretion (J_Amm), and ionic content (K⁺, Na⁺, Cl_-) have been studied in embryos and yolk-sac larvae of Atlantic halibut (Hippoglossus hippoglossus L.) maintained in darkness (6 °C and 34% S) or in light (light-arrested, nonon-hatching embryos). The T_Amm of the eggs increased from 250-300 nmol ind^-1 during the first 5-10 days post fertilization (dPF) to 400-500 nmol ind^-1 at the time of natural hatching in darkness. T_Amm did not start to decrease before 2-5 days after hatch, and yolk-sac larvae had a higher T_Amm compared to the light-arrested embryos of the same age and batch. J_Amm increased exponentially rom about 0.05 nmol ind^-1 h^-1 at 10 dPF to about 3 nmol ind^-1 h^-1 at hatch. Embryos undergoing hatching showed a significantly higher J_Amm than non-hatching, light-arrested embryos of the same age. The content of K+ in distilled water rinsed eggs declined from about 0.85 µmol ind^-1 at 2-4 dPF to about 0.15 µmol ind^-1 at hatch, while that of Cl^- remained constant at about t 0.85 µmol in^-1, and that of Na⁺ increased slowly from 0.10 to 0.20 µmol ind^-1 . Based on the measured ions, there was an anionic deficit in the egg, especially during the first week of development. When exposed to increased ambient total ammonia (0-27 mM NH4Cl), no mortality occurred, and no significant increase was found in the T_Amm of the Atlantic halibut egg before 9.2 dPF despite an estimated large inward diffusion gradient for the un-ionized ammonia species (NH₃). The delayed release of T_Amm comp ared to embryos and yolk-sac larvae of other marine teleosts, may relate to the deep-water spawning of the Atlantic halibut, and the buoyancy regulation of the yolk-sac larvae.