不同激素添加成分和受精液对山羊卵巢卵母细胞体外成熟与受精的影响

就成熟培养液中添加成分和不同受精液对山羊卵母细胞体外成熟与体外受精的影响进行了研究.结果表明,成熟培养液中添加HCG组的卵母细胞成熟率(85.7%)极显著高于PMSG组(50%)和FSH LH组(57.1%)(P<0.01),受精率比较则HCG组(62.2%)极显著高于PMSG组(34.7%)(P<0.01),显著的高于FSH LH组(45.2%)(P<0.05);添加了17β-雌二醇的成熟培养液对山羊卵母细胞的成熟率和受精率没有明显影响(P>0.05),而添加5%山羊卵泡液的成熟培养液则使卵母细胞的成熟率和受精率明显下降,卵裂率(7,7%)却显著高于对照组(0.0%)和17β-雌二醇组(0.0%) (P<0.05);分别以TALP液、BO液、M-199液为基础液组成的三种受精液(I液、II液、Ⅲ液)中II液的受精率(62.2%)显著高于Ⅲ液(44.7%)(P<0.05),而且II液的卵裂率(21.6%)显著高于I液(9.1%)(P<0.05),极显著高于Ⅲ液(0.0%)(P<0.01).     

山羊卵泡卵母细胞体外成熟体外受精研究

对山羊孵巢卵泡卵母细胞的采集方法、 PMSG处理山羊促进多卵泡发育及受精体系进行了研究。结果表明：切割法获可用卵数高于抽吸法， PMSG处理获卵总数及可用卵数高于对照组， BO液法的受精率高于 TALP液法，而 TALP液法的桑椹胚率高于 BO液法。     

Gamma Irradiation of Embryogenic Callus Cultures and In Vitro Selection for Salt Tolerance in Sugarcane (Saccharum officinarum L.)

Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane (Saccharum officinarum L.) cv.CoC-671.Embryogenic calli were gamma irradiated and exposed to different levels of NaCl (42.8,85.6,128.3,171.1,213.9,256.7,299.5,or 342.2 mM).The relative growth rate (RGR) decreased progressively with increasing salt stress and was the least with a salt stress of 256.7 mM (0.25±0.009),almost 10 fold lesser than the control.The RGR was significantly lower in 85.6 mM and higher salt stressed calli than the control.The survival percent also decreased,with an increase in NaC1 concentration.In case of 10 and 20 Gy irradiated calli,regeneration was observed up to 85.6 mM NaCl selection,medium,whereas,higher treatments (128.3 mM and beyond) exhibited browning initially.However,in the subsequent subcultures,regeneration was obtained in the case of 10 and 20 Gy irradiated calli on 128.3 and 171.1 mM NaCI selections.Higher dose of gamma irradiation (40 Gy) also showed regeneration,but only with 85.6 mM NaCI selection.The unirradiated calli regenerated the highest number of plantlets followed by 10 and 20 Gy irradiated calli on salt selection.A total of 147 plantlets were selected from different salt levels.The salt selected plants are being tested for their field performance.     

牛卵巢卵母细胞体外成熟的研究

研究了卵巢采卵方法以及卵巢保存温度和时间、性周期阶段、激素和培养基对牛卵母细胞体外成熟的影响。结果表明:从屠宰场获取的卵巢,先切剖或抽吸卵泡再切割卵巢,可显著提高可用卵母细胞数。卵巢保存时间在 2 h 以内最好,最长不超过 6 h。30~39 ℃的生理盐水保存卵巢,卵母细胞的成熟率(63.9 %)和卵裂率(34.4 %)均显著高于 2~8 ℃(16.7 %、0 %)和 20~29 ℃(54.1 %、31.7 %)保存的。采集卵母细胞时卵巢所处性周期阶段(卵泡期、黄体期)对卵母细胞体外成熟的影响不大(成熟率分别为 69.2 %、64.3 %)。M199 是牛卵母细胞体外成熟理想的培养基,M199 + 50 IU/mL LH+ 100 IU/mL FSH + 1μg/mL 17?-E2和 M199 + 50 IU/mL GnRH+1μg/mL 17?-E2都是牛卵母细胞体外成熟理想的培养系统(成熟率分别为 69.4 %、67.5 %,卵裂率分别为 35.4 %、42.7 %)。     

生发泡移植过程对小鼠卵母细胞体外受精的影响

为研究生发泡(GV)移植后的重组卵母细胞能充分在体外成熟和发育,本试验比较了GV移植过程及体外成熟对小鼠卵母细胞体外受精的影响。通过透明带切口及GV移植获得的重组GV期卵母细胞的成熟率为87.0%(147/169),体外受精后发育到原核期胚和2-细胞期胚的比率分别为76.2%(112/147)和42.2%(62/147),与仅做透明带切口但不进行GV移植的对照组相似(P>0.05),表明GV移植过程未对卵母细胞体外受精后的早期胚胎发育能力产生不利影响。但与体内成熟的卵母细胞相比,体外成熟卵母细胞的受精率和卵裂率都明显降低,表明其进一步发育的能力受到影响。     

小鼠腔前卵泡体外发育和体外排卵的研究

为探明小鼠腔前卵泡体外发育及体外排卵的特征和规律性 ,为利用卵巢资源奠定基础 ,采用机械方法分离未成熟小鼠腔前卵泡进行体外培养 .以 α- MEM加亚硒酸钠、维生素 C,L-谷氨酰胺、丙酮酸钠为基础培养液 ,分组添加 10 %的 2种不同来源的血清和 1μg/ m L FSH.在多卵泡培养体系中 ,培养 9～ 10 d,每天观察其体外发育情况 ,测定卵泡和卵母细胞大小 .培养结束时 ,用 h CG进行体外诱导排卵 .结果表明 :体外培养过程中 ,腔前卵泡贴附培养皿底 ,相互聚集粘附形成团状结构 ,在细胞铺层微滴培养中 ,卵泡呈单个生长 ,并发育形成卵泡腔或腔样结构 ;经9d体外培养 ,小鼠腔前卵泡达到排卵前大小 (>40 0 μm) ,卵母细胞从开始时的 5 3.2 μm生长到 6 8.0～ 70 .8μm,总存活率为 6 1.0 % ;卵母细胞的大小及存活率在不同的血清添加组间差异不显著 ;用 h CG可诱导体外排卵 ,发生卵泡破裂 ,卵母细胞排出 ,卵丘细胞扩展 ,卵母细胞生化泡破裂 ,放出第一极体 .总排卵率达 5 9.5 % .因此 ,采用多卵泡培养体系 ,小鼠腔前卵泡能够发育至成熟排卵 .     

Gamma Irradiation of Embryogenic Callus Cultures and In vitro Selection for Salt Tolerance in Sugarcane （Saccharum officinarum L.）

Radiation induced mutagenesis followed by in vitro selection was employed for salt tolerance in popular Indian sugarcane （Saccharum officinarum L.） cv. CoC-671. Embryogenic calli were gamma irradiated and exposed to different levels of NaCl （42.8, 85.6, 128.3, 171.1,213.9, 256.7, 299.5, or 342.2 mM）. The relative growth rate （RGR） decreased progressively with increasing salt stress and was the least with a salt stress of 256.7 mM （0.25±0.009）, almost 10 fold lesser than the control. The RGR was significantly lower in 85.6 mM and higher salt stressed calli than the control. The survival percent also decreased, with an increase in NaCl concentration. In case of 10 and 20 Gy irradiated calli, regeneration was observed up to 85.6 mM NaCl selection, medium, whereas, higher treatments （128.3 mM and beyond） exhibited browning initially. However, in the subsequent subcultures, regeneration was obtained in the case of 10 and 20 Gy irradiated calli on 128.3 and 171.1 mM NaCl selections. Higher dose of gamma irradiation （40 Gy） also showed regeneration, but only with 85.6 mM NaCI selection. The unirradiated calli regenerated the highest number of plantlets followed by 10 and 20 Gy irradiated calli on salt selection. A total of 147 plantlets were selected from different salt levels. The salt selected plants are being tested for their field performance.     

奶牛卵丘细胞和共培养细胞对体外受精胚胎发育的影响

对奶牛卵母细胞进行体外受精,研究了卵丘细胞和共培养细胞对体外受精胚胎发育的影响.结果表明,奶牛卵丘细胞有助于卵母细胞的体外受精,能够显著提高体外受精的卵裂率(70.7%)和囊胚率(22.8%),而在B2胚胎培养液中添加Vero作为共培养细胞能显著提高体外受精后胚胎的囊胚率(22.7%).     

兰花试管开花研究进展

阐述兰科植物试管开花研究的一些进展,着重介绍实生苗、组培苗以及环境、营养条件、培养基附加成分等因素对开花的影响。     

培养条件对绵羊卵巢卵母细胞体外成熟及受精的影响

对绵羊卵巢卵母细胞体外成熟培养及体外受精培养条件进行筛选,结果表明:在基础培养基中添加FCS可以显著提高卵母细胞的体外成熟率,而添加10%、15%和20%FCS的组间,卵母细胞的成熟率差异不明显,表明在基础培养液中添加10%FCS即可使绵羊卵母细胞的体外成熟率达到一个比较高的水平;体外培养时间不同对绵羊卵母细胞的成熟有较大的影响,培养24 h和26 h的卵母细胞成熟率(分别为73.3%和77.5%)显著高于培养18 h和20 h的卵母细胞成熟率(分别为62.5%和65.0%);对活力较低的绵羊精子而言,采用直接上浮法要比离心洗涤法更有利于选取活力较高的精子,并可提高受精卵的卵裂率.