Effect of GPAG Supplement on Porcine Oocyte Maturation and Its Following Embryonic Development by Parthenogenetic Activated

The aim of this study was to compare the effect of GPAG and commonly used FCS on porcine oocyte maturation and subsequent embryonic development after the fertilization. COCs were aspirated from follicles and cultured for 16, 24, 32, 40 and 48 h in TCM-199 medium either with GPAG or FCS. After 24 h with GPAG, 89.4% of oocytes reached M Ⅰ stage while in the medium supplemented with FCS, only 27.7% of oocytes reached the same stage （P〈0.05）. Prolonged incubation for up to 32 h clearly demonstrated that some of oocytes cultured in GPAG medium were at M Ⅱ stage （35.7%）, few of oocytes from FCS medium were at M Ⅱ stage （7.5%） （P〈0.05）. Both groups of oocytes reached the same stage of maturation within 48 h. After 48 h of culture, the oocytes with extruded polar bodies were inseminated. Fertilized oocytes were cultured in PZM3 medium supplemented with 3 mg.mL of BSA. After 7 days, the development and the quality of embryos were evaluated. The results showed that the maturation of oocytes in the presence of GPAG significantly increased their subsequent developmental ability when compared with FCS supplementation （29.2% ： 18.9% of blastocysts, P〈0.05）. However, differential staining revealed that once blastocysts were formed in either group, they had the same total cell number （39 ： 38） and the ICM/total cell ratio （0.26 ： 0.28）     

“胰岛素-转铁蛋白-硒钠”对山羊卵母细胞体外成熟及胚胎发育的影响(英文)

[Objective] The research aimed to enhance culture efficiencies of oocyte and embryo of goat in vitro and to explore serum-free culture system in vitro.[Method] At present,the conventional solutions of oocyte maturation and embryo development in vitro were always added into 1% ITS(Insulin-transferrin-selenium) or using 1% ITS to replace FBS in 2 kinds culture solutions for conducting in vitro cultures of goat oocyte and parthenogenetic embryo.The influences of ITS on their developments were detected.[Result] ITS in maturation liquid of oocytes could not increase oocytes maturation rate but significantly increased blastocyst rate (58.06% vs. 48.19%)of parthenogenetic embryo.If FBS in maturation liquid of oocytes was replaced by ITS, the maturation rate, cleavage rate and blastocyst rate were basically unchanged.Adding ITS into embryo medium could increase blastocyst rate (68.30% vs. 56.82%)of parthenogenetic embryo of goat.If FBS in embryo medium was replaced by ITS,the cleavage rate didn’t change basically,while the blastocyst rate in ITS was obviously lower than that in FBS group(42.33% vs.56.82%).[Conclusion] ITS could promote maturation of oocyte in vitro and early embryonic development, in addition,ITS could replace serum in maturation medium of oocyte as serum-free culture system for conducting relevant researches.     

Studies on Nuclear Transplantation in Mammalian Embryos

A series of experiments were conducted to study the major procedures in nuclear transplanation such as oocyte enucleation and activation,electrofusion and developent of the nuclear transplant embryos in the mouse,rabbits and sheep.The important results are as follows:1.In the mouse,only 35% of the oocytes collected 15-16h after hCG had a notable first polar body(FPb) and those without FPb were enucleated by removing cytoplasm from the PVS-wider side and the enucleation rate was similar to that in the oocytes with FPb,and the enucleation rate of removing 1/3 cytoplasm was remarkably higher than that of removing 1/4 cytoplasm.2.Among the three fusion media tested;mannitol and sucrose solutions produced better results than M2 in electrofusion of mouse 2-cell embryos.Under favorable pulse conditions,the osmotic pressure of fusion medium had no motable effect on electrofusion,but as the conditions became so unfavorable that some embryos began to lyse,the fusion rates in hypertonic mannitol solution were significantly higher than those in isotonic or hypotonic solutions.A wide range of pulse strengths (0.31-2.04kv/cm) and durations(10-1280us) were used and 100% of fusion were obtained in many cases.Optimal pulse durations were plotted for field strengths to obtain high fusion rates(96%-100%)in mouse2-cell embryos.3.With one pulse of 0.45kv/cm,satisfactory results of mouse oocyte activation were obatined only when the duration increased to 160us or longer,The activation rate increased as the oocytes got older.Some of the oocytes ar.rested at metaphaseⅢ after electrical stimulation and their proportion to the number of oocytes not activated increased with egg age.4.10% and 31% of the nuclear transplant embryos developed to morula or blastocyst stage in sheep and rabbits,respectively,with Chinese-made hormones and chemicals.     

Artificial Induction of Twinning by an Active Immunization of Beef Cows Against Inhibin Partially Purified from Porcine Seminal Plasma

Two hundred and seventy multiparous Chinese Yellow cattle (beef) were selected at 1 to 3months postpartum and divided into three groups (90 cows for each). Animals were given both a primary and booster immunizations with a total dose of 3 mg (Group Th) or 1.5 mg (Group Tl) of seminal preparation containing inhibin activity, emulsified with Freund‘s complete adjuvant and incomplete adjuvant (for booster), at 3 or 4-week intervals. Other cows were treated with the same volume of seminal preparation without inhibin activity as procedures mentioned above to serve as a control (Group C). Artificial inseminations were given twice at 8 - 12 h intervals when the cow was in heat. Jugular venous blood samples were collected from each cow and used to assay the presence of antibody against seminal preparation by double-diffusion in agar precipitation test and to detect the titer of inhibin antibody by an ELISA method. Data from 247 cows showed that 83.9% (73/87) of cows were in estrus and ovulated 89 ova altogether, of which 19 cows ovulated twin ova and 15 cows produced twins in Group Th (n = 87). However, only 61.1% (44/72) of cows in Group Tl (n=72) and 62.5% (55/88) of cows in Group C were in estrus and ovulated 46 and 52 ova altogether respectively.The ovulation rate (1.27 + 0.03), calving rate ( 126.3% ) and twinning rate (26.3%) in Group Th were greater than those in Groups Tl or C (P＜0.01). Furthermore, the ovulation rate was associated with antibody titer in sera of immunized animals (r = 0.7507, P ＜0.01). These results indicate that active immunization of postpartum cows against inhibin purified from porcine seminal plasma may increase the ovulation rate and induce twinning, suggesting the potential to develop a method to improve fertility in cows.     

Effects of Organic Manure Application with Chemical Fertilizers on Nutrient Absorption and Yield of Rice in Hunan of Southern China

To evaluate the effect of organic manure application with chemical fertilizers on rice yield and soil fertility under long-term double-rice cropping system, a six year field experiment was conducted continually in the paddy soil derived from Quaternary red clay in Hunan Province of southern China. Four different treatments, i.e., no nitrogen with chemical P and K （PK）, swine manure only （M）, N, P and K chemical fertilizers only （NPK）, and half chemical fertilizers combined with half swine manure （NPKM） with four replications were included. Each N, P and K application rate was the same at all the treatments （except the N application rate at PK） and N application rate was 150 kg N ha^-1. All fertilizers were applied to soil tillage layer with once application as baseal fertilizers. The nutrients uptake rate, grain yield, nitrogen use efficiency, and soil organic matter content at each treatment were investigated. The NPKM treatment achieved the highest mean annual yield of 12.2 t ha^-1 （68% higher than that of PK）. Higher dry matter accumulation and nutrients absorption were observed during the middle-late growth period in the NPKM treatment, with higher panicle number per unit and filled-grain number per panicle. Its average nitrogen use efficiency was 36.3% and soil organic matter increased by 18.5% during the experimental period in the NPKM treatment, which were significantly higher than those in the NPK treatment. Organic manure application with chemical fertilizers increased the yield and nitrogen use efficiency of rice, reduced the risk of environmental pollution and improved soil fertility greatly. It could be a good practical technique that protects the environment and raises the rice yield in this region.     

不同条件对绒山羊体外培养初级毛囊生长的影响(英文)

[Objective] The aim of this study is to lay a foundation for illustrating the biological characteristics and growth regulation mechanism of hair follicles.[Method]Cashmere goat primary hair follicles were separated under aseptic condition and cultured in serum-free DMEM and serum-free Williams E media respectively;subsequently,the growth rate and morphological changes were observed under the inverted microscope.[Result]Hair follicles cultured in serum-free DMEM media showed a growth rate of 0.034 mm/d during the first 3 days,whose structure and morphological characteristics could maintian a stable status for a long time in the growth process.Hair follicles grew much faster in the serum-free Williams E media with a growth rate of 0.077 mm/d during the first 3 days.[Conclusion]There were significant differences(P<0.05)between the growth of cashmere goat hair follicles cultured in the 2 kinds of media.Serum-free Williams E medium was superior to serum-free DMEM medium.     

Culture of Eriocheir sinensis Larvae in Ponds in Greenhouse

A total of 15 pools were selected from a greenhouse, and they were randomly and evenly divided into three groups. In each group, one stocking density of Eriocheir sinensis was arranged. The results showed that after 45-d culture, the crab number per kilogram reached about 300 with survival rate of about 20%; the initial stocking density had significantly effect on the body size of E. sinensis on sel , instead on the survival rate of E. sinensis larvae. Therefore, it is feasible to conduct the culture of E. sinensis larvae in a pool in greenhouse.     

卵母细胞体外成熟时间对绵羊核移植效率的影响(英文)

[Objective] The study aimed to provide references for the time of oocyte maturation in vitro and enucleation in the course of sheep nuclear transfer(NT).[Method] Compared the effects of different maturation time of oocytes on enucleation efficiency and reconstructed embryo development by means of blind enucleation and fluorescence microscopy.[Result] Treatment of IVM(in vitro maturation)19-21 h was significantly higher than IVM 16-18 h treatment in oocyte maturation rate(P<0.05)and was significantly higher than IVM 22-24 h treatment in enucleation rate(P<0.05).Three treatments had no significant difference in cleavage rate and blastocyst rate(P>0.05),but IVM 19-21 h treatment was significantly higher than the other 2 treatments in average cell number of blastocysts(P<0.05).[Conclusion] The appropriate in vitro maturation time of oocytes was 19-21 h for sheep nuclear transfer,which could significantly improve the quality of blastocysts according to the cell number per blastocyst(P<0.05).     

卵母细胞体外成熟时间对绵羊核移植效率的影响(英文)

[Objective] The study aimed to provide references for the time of oocyte maturation in vitro and enucleation in the course of sheep nuclear transfer(NT).[Method] Compared the effects of different maturation time of oocytes on enucleation efficiency and reconstructed embryo development by means of blind enucleation and fluorescence microscopy.[Result] Treatment of IVM(in vitro maturation)19-21 h was significantly higher than IVM 16-18 h treatment in oocyte maturation rate(P<0.05)and was significantly higher than IVM 22-24 h treatment in enucleation rate(P<0.05).Three treatments had no significant difference in cleavage rate and blastocyst rate(P>0.05),but IVM 19-21 h treatment was significantly higher than the other 2 treatments in average cell number of blastocysts(P<0.05).[Conclusion] The appropriate in vitro maturation time of oocytes was 19-21 h for sheep nuclear transfer,which could significantly improve the quality of blastocysts according to the cell number per blastocyst(P<0.05).     

Analysis of Daylength and Temperature Characteristics of Fertility Conversion of 13 Newly Developed Genetic Male Sterile Lines of Rice

Based on 13 genetic male sterile lines of paddy rice newly-developed in academies of agricultural sciences of China, study was undertaken of the determination of ecological conditions for their fertility conversion in terms of the leveling of daylength and temperature and a fertility quantifying model in the context of observed bagged seedlings (in percentage) through self mating by pollen inside, daylength and temperature from the experiments with sowing by stage in three different latitude zones, i. e., at Sanya of Hainan Province, Guiyang of Guizhou and Wuhan of Hubei. Results show that five sterile lines are temperature-sensitive with the fertility conversion temperature below 24 ℃, two of the daylength-sensitive sterile lines have conversion daylength of ＞ 12. 4 h, suggesting their almost full sterility, and one of the photo- and temperature-sensitive lines needs shorter daylength either at high or at low temperature for fertility conversion. These eight sterile lines are of productive significance.     

大白菜雄性不育系的育性温敏特性

[目的]探明大白菜雄性不育的育性转换特性。[方法]以大白菜温敏不育系A和小白菜可育品系"小青口"为试材,进行育性观察和花粉生活力测定。[结果]供试的大白菜不育材料属高温不育类型,诱导雄性稳定不育的温度阈值为日均温24℃以上,诱导雄性可育的温区为16℃以下,日均温16~24℃为育性转换温区;在我国北方春季大白菜繁种期间,花期日均温度大多在16℃以上,即使有少量花粉产生,授粉竞争力远不及父本正常可育花粉,利用温敏不育系杂交制种安全可靠;16℃以下的温度条件,在北方冬季或早春利用温室等保护地设施能轻易达到,大白菜温敏不育系自交繁殖的可操作性强。[结论]该研究为大白菜两系法杂交制种提供了依据。     

高温对滇型杂交水稻雄性不育系育性的影响

 本文在对滇型杂交水稻雄性不育性研究的基础上,提出了温度敏感雄性不育的概念。发现滇Ⅰ型粳稻不育系滇寻1号A是一个典型的温度敏感不育系,并提出利用这一特性来实现滇型杂交水稻种子生产“两系化”的设想。滇型杂交水稻雄性不育系异地种植的结果表明,除滇Ⅰ型滇寻1号A外,在不同的气侯条件下一般表现完全不育。滇寻1号A在较高温度地区种植时则表现出育性恢复的现象。进一步研究表明,其育性转换的主导因子是温度。育性转换的临界发育时期为花粉母细胞形成期至双核花粉发育期。育性转换的临界日平均温度为22.0℃。低于此温度时表现为完全不育。而高于此温度时,在一定范围内可育性随温度增高而增加。对此不育系自交结实后代的观察发现,自交结实后代仍表现为完全不育,亦具有温敏的特性。恢保关系与滇寻1号A本身完全相同。研究还表明:这是一个对温度敏感的不育系,可以通过异地播种选择在适宜温度条件下种植来繁殖不育系,或配制杂种而利用杂种优势,还可作生理遗传的研究材料。     

5460不育系育性转变敏感期试验初报

在人工气候室条件下,采用16h 长光照高温35℃,16h 长光照常温25℃,12h 短光照高温35℃,12h 短光照常温25℃的设计。研究了5460不育系在不同生长时期不同温光组合处理的花粉育性反应.结果表明:(1)5460不育系的育性转变敏感期是在减数分裂期,离抽穗前11d 左右.在育性转变敏感期间,当温度为35℃时,无论光照是12h 还是16h,其不育株率和花粉的不育度均达到100%;而温度为25℃时,无论光照长短,其花粉的育性正常,花粉可染色率80%以上,可育株率100%.(2)播种后至减数分裂期前,不但光照长短对花粉的育性未见有效应,而且在高温和常温下亦未发现对育性有影响.     

4种光温敏核不育水稻在南昌的育性表现

[目的]考查自然条件下4种不育系水稻在南昌的育性情况。[方法]2010年通过分期播种的方法分别对4个不同类型的实用光温敏核不育系准S、Y58S、P88S和培矮64S的育性转换特性进行系统研究。[结果]准S的敏感期为开花前9～10 d,58S的敏感期为开花前11～13 d,培矮64S的敏感期为开花前16～19 d,P88S没有明显的敏感期。从育性转换临界温度来看,准S最低,为27.2℃;培矮64S次之,为27.5℃;58S转换温度最高,为29.3℃。[结论]在南昌地区,培矮64S的育性波动较小,稳定期较长,较其他几个不育系好。     

不同来源培矮64S育性转换光温反应特性的分析

【目的】研究不同来源的5份培矮64S的育性转换光温反应特性。【方法】利用人工气候箱对5份培矮64S的自交结实率和花粉可育度进行研究，并分析育性转换发生变化的原因。【结果】在长日高温条件（14.5h/28℃）下，5份材料几乎完全不育，但在14.5h/24℃，除2份材料仍然保持较好的不育性外，其它材料都有一定的结实，其中4号材料的结实率达到9.05%，花粉可育度达到27.83%，与其它4份材料的差异达到极显著水平。另外，对历年培矮64S在气候箱内鉴定的自交结实率的结果进行分析比较，发现不同年度间繁育的培矮64S光温反应特性也存在差异。【结论】培矮64S表现出明显的地区间和年度间的差异。4号材料的不育起点温度发生变化，不符合不育系的光温反应鉴定标准。     

温光条件对胞质不育系V太911289a雄性育性影响的初步研究

[目的]研究温光条件对小麦胞质不育系V太911289a雄性育性的影响,探讨胞质不育系雄性育性的转换机制。[方法]于2003~2005年在湛江进行了分期播种试验和人工定温定光试验。[结果]在一定的温光条件下,胞质不育系V太911289a的雄性育性可以发生转换。温度是影响其自交种雄性育性的主要原因之一。当育性敏感期日平均温度低于19℃时,表现稳定不育;日平均温度为20~22℃时,则表现部分可育。光照时间是对其雄性育性也有一定的影响,长日照利于雄性发育。[结论]胞质不育系用于生产杂交种安全性远高于温敏不育系。     

温光条件对胞质不育系V_(太911289a)雄性育性影响的初步研究(英文)

[Objective] The study aimed to reveal the effect of photoperiod and temperature on male fertility of cytoplasmic male sterile line Vtai911289a in wheat and discuss the mechanism of male fertility alteration. [Method] The sowing-date tests and designed conditions were conducted during 2003-2005. [Result] Fertility of Vtai911289acould alter under specific photoperiod and temperature conditions. Temperature is one of the main factors influencing male fertility of the male sterile lines. Vtai911289a showed stable sterility under the condition of the mean of daily temperature at fertility sensitive stage lower than 19 ℃ and presented partial fertility when the mean of daily temperature at fertility sensitive stage lower than 20-22 ℃. Photoperiod to some extent affects the male fertility of Vtai911289a,long-day condition is helpful for the male fertility of the sterile line. [Conclusion] The application of photoperiod temperature-sensitive cytoplasmic male sterile line in production has a higher safety than that of temperature sensitive sterile line.     

不同类型光敏核不育水稻育性转换特点及应用研究

经1986～1988年.先后对十八套不同亚种类型的光敏感核不育系,在江西南昌,连续三年进行分期播种试验,已得到如下主要研究结果:1.基本探明了供试的十八套不同粳、籼亚种类型的光敏感核不育系,在南昌(28°41′N)的育性转换时期、阶段及稳定性.经育性转换及杂一代优势观察,已筛选出育性转换、开花习性及经济性状较好的粳型双9—2S、28006S和籼型W6417S、W6154S、赣ⅢS等光敏感核不育系.2.不同亚种类型及同一亚种不同品种的光敏感核不育系,其育性转换特点有明显差异.一般规律:籼型核不育系,普遍存在不育期较长,转育较晚,同时可育期易受低温影响结实的特点;而粳型核不育系,则普遍较籼型核不育系的不育期为短,育性转换较早,不育期因高温影响,常有部分散粉自交结实现象,但可育期因低温影响结实不明显.3.鉴于不同亚种类型光敏核不育系的育性转换特点不同,结合其开花习性差异对杂交制种的影响,从育种应用出发,研究提出:我国长江中下游稻作区,水稻光敏核不育系的选育,应重点主攻籼亚种,首先突破早熟早籼为宜.在选材上优先考虑矮秆、抗病、优质以及开花习性好,柱头外露率高等性状.     

生态条件对光（温）敏核不育水稻育性的影响

本文用Ｗ６１５４５、７００１５等一批灿粳核不育系为材料，通过分期播种、高海拔种植和人工控制光温处理试验，研究了光温条件对不同类型核不育系育性的影响。结果表明：以Ｗ６１５４５为代表的灿型核不育系的育性主要受温度影响，开花当时的低温和抽穗前的低温对育性同样有影响。以７００１Ｓ为代表的枝型核不育系的育性主要受光照影响，低温影响小。抽穗当时的低温对育性没有影响；抽穗前遇低温，１５～２０天后才有反映。生育前期的光温条件对育性有积累效应，同时抽穗但不同播期的不育株育性有差异。在岳西县高海拔长日低温条件下种植的核不育系不育期相对缩短，不育性降低，特别适用于从Ｆ２代中选光敏适宜温度范围宽的核不育株。