洋桔梗离体快繁技术研究

[目的]建立洋桔梗高效的快速繁殖体系,为工厂化生产提供可行的操作程序。[方法]以国外进口的洋桔梗F1代种子(品种名称为Polestar yellow)无菌苗的叶片为外植体,对不定芽的诱导、单芽增殖、无菌苗生根等进行了研究。[结果]结果表明:6-BA和IBA组合对洋桔梗叶片不定芽的诱导效果较6-BA和NAA组合好,叶片最佳的分化培养基为MS+6-BA0.6 mg/L+IBA0.04 mg/L,1个月内正常芽平均分化数为8.3;最佳的单芽增殖培养基为MS+6-BA0.8 mg/L+NAA0.04 mg/L,1个月内正常芽的增殖倍数为7.4;无菌苗在含有IBA0.25 mg/L的1/2 MS培养基上,生根率、平均每苗的根数均最大。[结论]该结果为洋桔梗的工厂化生产奠定了基础。     

潘那利番茄叶片组织培养及植株再生研究

研究以潘那利番茄的叶片为外植体,研究不同激素及其不同浓度组合对潘那利番茄愈伤组织的诱导、不定芽分化及其生根的影响.结果表明.愈伤组织诱导和芽分化的最适培养基为MS+6-BA 3.0 mg·L-1+IAA0.2 mg·L-1,生根的最适培养基为MS+IAA 0.1 mg·L-1,试管苗移栽后成活率为87%.该研究结果为潘...     

九个粳稻品种未传粉子房的培养

9个粳稻品种未传粉子房的培养试验表明,品种间愈伤组织的诱导率、分化率均存在差异。早粳优于晚梗。抽穗期早的品种具有较高的成苗率。外源激素对诱导分化具有十分重要的作用。MCPA(2-甲基-4-氯苯氧乙酸)优于2.4-D。     

新疆扁桃花粉萌发及花柱离体培养试验研究

对‘纸皮'、‘双果'、‘克西'、‘麻壳'、‘267'共5个扁桃品种进行花粉萌发试验,结果表明‘纸皮'花粉萌发率最高,为84.83%。‘克西'的萌发率最低,为0.65%。通过对5个品种自交和杂交授粉后的花柱进行离体培养可以看出,自交时,‘双果'ב双果'的自交组合优势最为明显;杂交时,‘双果'ב纸皮'的授粉组合优势最为明显。     

不同浓度硫酸盐对水稻土中异化铁还原过程的影响

研究不同电子受体之间的竞争关系对揭示厌氧水稻土中微生物作用导致的氧化还原过程变化机理具有重要的理论意义。本研究采用土壤泥浆厌氧培养、人工合成氧化铁体系接种土壤浸提液厌氧培养及接种铁还原菌纯培养等试验方法,通过向培养体系中添加SO24-,探讨了硫酸盐作为竞争电子受体对不同铁还原体系中Fe（Ⅲ）还原的影响。结果表明,在2种水稻土的泥浆培养过程中,Fe（Ⅲ）还原速率均随着SO24-浓度增加而降低,但Fe（Ⅱ）的最终累积量却较对照处理有明显的增加。添加硫酸盐对Fe（Ⅲ）还原速率（k）的影响表现为：石灰性水稻土〉酸性水稻土;而最终Fe（Ⅱ）累积增加率则为：酸性水稻土〉石灰性水稻土。由接种不同水稻土浸提液的培养试验看出,添加SO24-后Fe（Ⅲ）还原受到显著的抑制,但随着培养时间延长Fe（Ⅲ）还原反应依然可以进行,并且Fe（Ⅱ）累积量最终达到与CK相同的水平。在接种铁还原菌的纯培养试验中,添加SO24-对供试的4株铁还原菌的Fe（Ⅲ）还原过程并未产生抑制效应,表明铁还原菌本身并不受硫酸盐的影响。     

奶牛乳腺上皮细胞的不同培养方法比较及激素和细胞因子对β-酪蛋白mRNA表达的诱导

本试验旨在建立一种高效的奶牛乳腺上皮细胞培养方法,并研究不同激素和细胞因子对其表达β-酪蛋白mRNA的诱导作用.分别采用组织块培养法和机械破碎法培养奶牛乳腺上皮细胞,利用成纤维细胞和乳腺上皮细胞对胰酶的敏感性不同,对获得的细胞进行纯化.通过细胞计数法测定纯化细胞的生长曲线,通过免疫荧光组织化学染色法检测角蛋白18的表达,通过实时定量PCR法检测8种不同激素和细胞因子组合培养液诱导细胞表达β-酪蛋白mRNA的效果.结果表明:通过机械破碎法可以分离得到增殖旺盛的奶牛乳腺上皮细胞,与组织块培养法相比,具有细胞迁出速度快等优点.细胞生长曲线呈典型的“S”型.在纯化的乳腺上皮细胞及第20代乳腺上皮细胞中都检测到角蛋白18的表达.100 ng/mL类胰岛素生长因子Ⅰ(IGF-Ⅰ)显著提高了乳腺上皮细胞中β-酪蛋白mRNA的表达(P＜0.05).由结果可知,本试验采用的机械破碎法是一种高效的奶牛乳腺上皮细胞培养方法,100 ng/mL IGF-Ⅰ对细胞中β-酪蛋白mRNA表达的诱导效果最好.     

The quality after culture in vitro or in vivo of porcine oocytes matured and fertilized in vitro and their ability to develop to term

The quality of porcine blastocysts produced in vitro is poor in comparison with those that develop in vivo. We examined the quality of in vitro‐matured and fertilized (IVM/IVF) oocytes, their abilities to develop to blastocysts under in vivo and in vitro conditions, and the potential of the embryos to develop to term after transfer. IVM/IVF oocytes were either transferred and the embryos recovered on Days 5 and 6 (100% and 87.5%, respectively) (‘ET‐vivo’ embryos), or cultured in vitro for 5 or 6 days (‘IVC’ embryos). The proportion of blastocysts differed significantly between the two groups on Day 5 (20.6% and 8.0%, respectively), but not on Day 6 (23.8% and 21.2%, respectively). The mean number of cells in ET‐vivo blastocysts on Days 5 or 6 was significantly higher (72.8 and 78.7, respectively) than that in IVC blastocysts (22.1 and 39.7, respectively). When IVM/IVF oocytes and IVC blastocysts on Day 6 were transferred, all (three and three, respectively) developed to piglets (16 and 16, respectively), without any difference in the rates of development to term (2.1% and 2.6%, respectively). These data suggest that, although blastocyst production differs between the two culture conditions, IVM/IVF oocytes possess the same ability to develop to term.     

鳜下丘脑神经元细胞培养

为了建立可行、高效的下丘脑神经元细胞技术,本研究通过分离鳜下丘脑组织,用Ⅰ型胶原酶将组织消化成单细胞悬液,并用完全培养液进行培养,在培养的第3、4、5和6天观察细胞的形态。结果显示,培养第3天时细胞贴壁量少,胞体小并且呈单个分布;在培养第4天,细胞的数量显著增多,且具有典型的神经元的形态,胞体饱满;第5天神经元胞体的融合度进一步增大,突起增长增粗,许多分支互相交错连接而形成密集的神经纤维网络;第6天细胞活性减弱,开始凋亡。使用CCK-8法检测细胞活性,结果显示,在培养第5天,细胞活性最高;采用荧光定量PCR(RT-PCR)法和免疫荧光鉴定法对神经元细胞进行鉴定,经RT-PCR检测发现,培养的下丘脑细胞可以表达神经元特异基因noggin,经免疫荧光鉴定,下丘脑神经元细胞纯度为95.9%。研究表明,通过此培养方法能够获得纯度较高的鳜下丘脑神经元细胞,为进一步在细胞水平研究鳜的摄食与能量代谢相关机理奠定基础。     

Binding and bioactivity of insulin in primary cultures of carp (Cyprinus carpio) hepatocytes

Isolated carp hepatocytes cultured in serum-free, chemically defined medium were used to investigate within the same cell preparation characteristics of the binding of insulin as well as effects of insulin on cellular metabolism. The binding of human [¹²⁵I]-insulin to carp hepatocytes was studied in kinetic, saturation and displacement experiments. A dependency of insulin binding on the collagenase used for cell isolation was demonstrated. Insulin binding decreased during the first 12h of culture but remained constant during the following 12h. The kinetic experiments revealed that [¹²⁵I]-insulin binding reached a steady state within 20–30 min of incubation. The mathematical analysis of the saturation experiments demonstrated the existence of two populations of binding sites, one with high affinity (K_d1 = 5.5 pM) and low capacity (B_max1 = 0.14 fmol/mg protein or 77 binding sites/cell) and one with low affinity (K_d2 = 2.4 nM) and high capacity (B_max2 = 17.6 fmol/mg protein or 9623 binding sites/cell). In competition experiments, 312 pM [¹²⁵I]-insulin was displaced by cold insulin, IGF-I and IGF-II with IC₅₀ values of 2.2, 7.9 and 20.3 nM, respectively. Glucagon was without effect. Binding of insulin to carp hepatocytes resulted in a significant reduction of glucose release and a significant increase of protein synthesis as of de novo fatty acid synthesis. dedicated to Prof. Dr. W. Hanke on the occasion of his 65^th birthday.     

Growth and production of the Amazonian tambaqui in fixed cages under different feeding regimes

Experimental culture of the native Amazonian fish tambaqui, Colossoma macropomum, in fixed cages was carried out over a period of 8 months, in Lake Urubu (Rio Grande do Norte, Brazil), to assess the viability of fixed cage culture of tambaqui and to test the influence of diet on growth rates. Nine synthetic net cages (1 m³) were each stocked with 45-day-old fish (mean weight 3 g; mean total body length 51 mm) at a density of 34 fry m^–3. During the first 2 months of culture, fish were fed a balanced formulated feed on an as-fed basis at the rate of 5% body weight day^–1. During months 3–8 this continued for fish in treatment 1 while those in treatment 2 were fed tropical regional fruits, on a wet weight basis at the rate of 5% body wt day^–1. Fish in treatment 3 were given no supplementary feed. Monthly biometric measurements were made on all fish. Fixed cage fish culture was shown to be a viable and simple technique. Survival in all treatments was 100%. With balanced supplementary feed, production was 14.4 kg m^–3, compared with 4.9 kg m^–3 and 2.1 kg m^–3, respectively, in the treatments where fish were fed with fruits and were not given any supplementary feed.