The Effect of Vitrification on Mouse Oocyte Apoptosis by Cryotop Method

Background : Oocyte cryopreservation is one of the most important topics in the field of assisted reproductive technology to preserve women fertility, but relationship between cryopreservation and apoptosis is still a matter of debate. The present study was aimed to investigate the effects of vitrification on apoptosis in mouse oocytes by Cryotop method. Method: A total of 200 germinal vesicle (GV) and 200 metaphase II (MII) oocytes were obtained from ovaries and fallopian tubes of NMRI mice, respectively and divided into control and experimental groups. Oocytes in experimental group were vitrified by Cryotop using vitrification medium and were kept in liquid nitrogen for one month. The survival rate of oocytes was evaluated after 2 hour incubation time. Then, the oocyte apoptosis was evaluated by TUNEL technique and compared with those in control group. The data was compared statistically using SPSS software and chi-square test. Results: The survival rates of vitrified GV (93%) and MII oocytes (88%) showed a significant decrease compared with the control group (P<0.05), but there was no significant difference in survival rate of both vitrified oocyte groups. The incidence of apoptosis in vitrified and control GV oocytes showed no significant difference (13% vs. 7%), but the rate of apoptosis in vitrified MII oocytes increased significantly not only in comparison with MII control group (25% vs. 5%) but also with vitrified GV oocytes (P<0.05). Conclusion: The results indicate that vitrification increases apoptosis in mouse MII oocytes and apoptosis may play a role in MII oocyte injury after vitrification. Key Words: Vitrification, Apoptosis, Oocytes     

剑麻组培过程中玻璃化问题的探讨

本文对剑麻组培过程中导致玻璃化的因素进行了试验，结合相关的研究报道，阐述了这些影响因子与玻璃化之间的关系并提出了一些控制玻璃化的方法。     

剑麻组培过程中玻璃化问题的探讨

本文对剑麻组培过程中导致玻璃化的因素进行了试验,结合相关的研究报道,阐述了这些影响因子与玻璃化之间的关系并提出了一些控制玻璃化的方法.     

影响柑桔球形合子胚离体培养的若干因素

通过对柑桔球形合子胚不同发育期的分离培养、不同基本培养基及附加物、蔗糖浓度等因素对柑桔合子胚离体培养影响的研,结果表明:授粉后50d是柑桔合子胚离体培养的最适时期;Whiet基本培养基比MS基本培养基更适于柑桔球形合子胚离体培养。适当增加培养基的渗透压和添加水解乳蛋白,特别是袖子胚乳 ,可提高幼胚的诱导率,有利于柑桔球形胚的发育。比较适宜的蔗糖浓度为10%一 15%,水解乳蛋白浓度为400mg/L     

氯溴异氰尿酸在马铃薯试管苗培养中的作用

以中薯 2号和Favorita两个品种为试验材料 ,将MS培养基给以不同浓度的氯溴异氰尿酸处理 ,来取代高压灭菌过程 ,研究氯溴异氰尿酸的杀菌效果和对试管苗的影响。结果表明 :在培养基中添加氯溴异氰尿酸处理的浓度为 0 70～ 0 4 0 g/L ,均有良好的杀菌效果 ;处理浓度为 0 70 g/L的培养基 ,能够诱使试管苗单节茎段产生丛生芽 ,0 5 7～ 0 4 0 g/L的处理浓度 ,可以有效地缩短试管苗的节间长度 ,增加单位高度内的叶片数 ,提高试管苗的移栽成活率。     

钼对茶树体内无机元素含量及分布的影响

用水培、土施和喷施方法,研究了 Mo 对茶树体内无机元素吸收和分布的影响,结果表明,茶树对 Mo 有很强的吸收能力,并积累在叶、根中。土施以盆土表面积每平方厘米50—200μg效果较好,但喷施易造成叶中 Mo 过剩。水培 Mo 处理能有效地提高叶中 B、Na 的含量,促进 K 的吸收,减少体内 S 的含量,P、Ca、Cu、Fe 在茎中积累,Mg、Mn 在叶中有所增加。Mo 对 Al、Zn 含量无影响。施肥方式不同,Mo 效应也有区别,土施 Mo 肥能增加新梢中的大部分元素含量,喷施Mo 肥则使新梢中的大部分元素含量趋于下降。     

茶悬浮培养细胞玻璃化超低温保存研究

为保存具有稳定代谢能力的优良茶细胞系,本文对茶叶悬浮培养细胞玻璃化法超低温保存进行了初步研究。结果表明:预培养以4d最佳,60%PVS2冰浴装载以20min最好,100%PVS2冰浴脱水处理60min最优,40℃复温解冻细胞存活率最高,采用这些参数进行完整试验的细胞存活率达到76%,初步建立了茶悬浮培养细胞玻璃化超低温保存的冷冻程序。     

玉米幼胚培养及植株再生的研究

以不同玉米自交系幼胚为外植体,研究不同2,4-D浓度对愈伤组织诱导、6-BA浓度对愈伤组织分化和6-BA与NAA配合使用对试管苗生根的影响。结果表明:不同2,4-D浓度对愈伤组织诱导率影响差异不明显,但对胚性率诱导有明显影响,其诱导最佳培养基为MS+2,4-D 2 mg/L+水解酪蛋白500 mg/L+脯氨酸500 mg/L;愈伤组织分化的最佳培养基是MS+6-BA0.5 mg/L+水解酪蛋白500 mg/L+谷氨酰胺200 mg/L+PP₃₃₃ 2 mg/L;试管苗生根的最佳培养基是1/2 MS+6-BA0.5 mg/L+NAA2.0 mg/L+PP₃₃₃ 2.0 mg/L+活性炭0.2%。     

不同培养基及激素配比对小麦幼胚离体培养的影响

为了建立一个适于优良小麦品种基因枪转化的高效组织培养体系,以2个冬性小麦品种京841和中麦9号为供试材料,探讨了不同培养基、不同激素配比对小麦幼胚离体培养的影响。结果表明,在MS、N6B5MS、B5和L34种培养基中,L3培养基的出愈率及分化率最高,两品种平均出愈率达到88.9%,分化率达到36.6%;以L3为基本分化培养基,同时添加ZT3mg/L和NAA1mg/L配比对愈伤组织诱导分化的效果最好,分化率平均达到55.9%。     

二倍体马铃薯试管苗的培养及对叶肉原生质体融合的影响

以35份二倍体马铃薯试管苗为试验材料,对二倍体马铃薯试管苗培养的影响因素进行研究,目的是使试管苗的叶片增大,叶面积增加,并有利于进行叶肉原生质体的融合。结果表明:含有AgNO3的培养基(MS+0.5 mg/L NAA+1 mg/L AgNO3)适合22份供试二倍体马铃薯试管苗的培养;添加AgNO3比不添加AgNO3的试管苗的叶片明显增大,叶面积显著增加,添加2%蔗糖和2 mg/L AgNO3的培养基的试管苗叶面积最大;以二倍体野生种S.chacoense和二倍体材料DY4-5-10为亲本进行电融合,以添加2%蔗糖和1 mg/L AgNO3的培养基培养其试管苗,获得的融合产物可再生出完整植株,该培养基适合用于分离原生质体的二倍体马铃薯试管苗的培养。     

马铃薯试管苗不同培养方式中添加抑菌中草药萃取液的作用

以马铃薯品种东农305的脱毒试管苗为供试材料,采用固体培养和液体培养两种方式,在试管苗繁殖培养基(M S+3%白糖+6.5 g.L-1琼脂,pH约5.8)中加入0.05%的中草药萃取液,以常规的试管苗繁殖培养基为对照,进行试管苗生长状况的比较。试验采用二因素完全随机试验设计,3次重复,接种1周后开始取样调查,每周调查1次,固体培养调查4次,液体培养调查3次。调查苗高、茎粗、茎节数、根数和根长,对所有性状进行方差分析和差异显著性测验(SSR法)。试验结果表明:在试管苗繁殖培养基中添加中草药萃取液,对试管苗的苗高、茎粗和茎节数的增加均有一定的促进作用,只是在不同培养方式下的效果存在一定差异。对试管苗根部的作用在于增加根长,而不是增加根的数量。在本试验中,固体培养方式下添加中草药萃取液的处理效果最好,成苗时试管苗的苗高达7.3 cm,茎节数达7.1个,繁殖倍数高于其它处理。     

碳氮供给对小麦离体穗培养粒重、微量元素及蛋白质含量的影响

为了解碳氮供给与小麦粒重、Fe、Zn、Mn、Cu等微量元素以及蛋白质含量的关系,在离体穗培养条件下研究了灌浆初期和灌浆中期不同浓度C(蔗糖)、N(硝酸铵)供给对小麦粒重、微量元素(Fe、Zn、Mn、Cu)和蛋白质含量的影响.结果表明,随着培养基糖浓度的增大,粒重和Fe、Mn含量都表现为先升高后降低的趋势,在4%糖浓度时,均达到最高值;籽粒Zn、Cu和蛋白质含量表现为随糖浓度增大而持续降低.随着培养基N浓度的增加,粒重和Fe、Zn、Mn、Cu含量多表现为先升高后降低的趋势,且多在N浓度为0.07%时达到最高值,尤以灌浆中期开始的培养表现明显;籽粒蛋白质含量表现为随N浓度增大而持续增加.由此可见,外源C、N供给对粒重、微量元素和蛋白质含量有明显的调控效应.一定的糖供给可同时提高粒重和Fe、Mn含量,一定的N供给可同时提高粒重及微量元素和蛋白质含量.     

The Effects of Fibroblast Co-Culture and Activin A on in vitro Growth of Mouse Preantral Follicles

Background: This study was conducted to evaluate fibroblast co-culture and Activin A on in vitro maturation and fertilization of mouse preantral follicles. Methods: The ovaries from 12-14-day-old mice were dissected, and 120-150 μm preantral follicles were cultured individually in α-MEM as based medium for 12 days. A total number of 456 follicles were cultured in four conditions: (i) base medium as control group (n = 113), (ii) base medium supplemented with 30 ng/ml Activin A (n = 115), (iii) base medium co-cultured with mouse embryonic fibroblast (n = 113), and (iv) base medium supplemented with 30 ng/ml Activin A and co-cultured with fibroblast (n = 115). Rate of growth, survivability, antrum formation, ovulation, embryonic development and steroid production were evaluated. Analysis of Variance and Duncan test were applied for analyzing. Results: Both co-culture and co-culture + Activin A groups showed significant difference (P<0.05) in growth (on days 4, 6, and 8 of culture period) and survival rates. However, there was no significant difference in antrum formation, ovulation rate, and embryonic development of ovulated oocytes. There were significant differences (P<0.05) in the estradiol production on days 8, 10, and 12 between co-culture + Activin A and the control group. Progesterone production also was significant (P<0.05) in co-culture + Activin A group on days 6, 8, 10, and 12 compared to control group. Conclusion: Fibroblast co-culture and Activin A promoted growth and survivability of preantral follicles. However, simultaneous use of them was more efficient. Key Words: Fibroblast, Activin A, Follicle     

钙对非洲凤仙铅累积特性的影响

通过水培实验,研究钙对非洲凤仙铅耐性和累积特性的影响。结果表明,无钙处理下,随着铅处理浓度增加,非洲凤仙生物量逐渐降低,根系和叶片铅含量逐渐升高,铅处理浓度高于100 mg/L时,非洲凤仙各器官铅含量在10~250 mg/kg之间;与无钙处理相比,外源钙显著促进50、100和300 mg/L铅处理下非洲凤仙的生长;钙对非洲凤仙各器官铅含量影响随铅处理浓度不同而不同,与无钙处理相比,钙显著促进50和100 mg/L铅处理下根系铅累积,显著抑制300和600 mg/L铅处理下根系对铅的吸收累积,钙显著降低100和300 mg/L铅处理非洲凤仙地上部分铅含量;铅处理浓度为100 mg/L时,钙显著降低铅在根系和叶片中活性,也降低根向地上部分转运。由此可知,非洲凤仙对铅具有较高耐性,是Pb富集植物;外源钙能够缓解一定铅胁迫对非洲凤仙的毒害作用,推测缓解效应与钙降低地上部分铅累积含量和降低铅在植株体内活性有关。     

小麦成熟胚培养方法的优化及其在小麦遗传转化中的应用

为了优化小麦成熟胚培养体系,在前人报道的小麦成熟胚培养方法的基础上对小麦成熟种子处理及取胚方法进行了改进,发现胚切伤略带胚乳的取胚方法及浸泡过夜后4C低温处理法可以提高小麦成熟胚的再生效率;从19个小麦品种(系)中筛选出5个在此培养体系下具有较高再生率的基因型,分别为农大408、农大3787、保丰104、农大3965和京花1号,其再生率分别为73.1 1％、68.30％、65.30％、63.40％和57.60％.进一步利用基因枪轰击法对小麦品种辽春18和保丰104成熟胚诱导的愈伤组织进行了转化,分别获得转GUS基因和转Gcn5RNAi基因的小麦T代植株,经分子检测其转化率分别达到1.35％和1.80％.证明优化后的小麦成熟胚培养技术可以应用于小麦遗传转化研究.     

Human Ovarian Tissue Vitrification/Warming Has Minor Effect on the Expression of Apoptosis-Related Genes

Background: In this study, we evaluated the incidence of apoptosis at the ultrastructural levels and expression of some apoptosis-related genes in vitrified human ovarian tissue just after warming. Methods: Human ovarian tissue biopsies from 23 women after caesarean section were transported to the laboratory within 2 hours, and then they were cut into small pieces. Some pieces were vitrified and warmed and the other samples were considered as control. Apoptosis was assessed by a transmission electron microscope and also by molecular analysis of pro-apoptotic (Fas, FasL, Bax, p53, caspase8, and caspase3) and antiapoptotic (Bcl-2 and BIRC5) genem RNA levels using real-time RT-PCR before and after vitrification. Results: No sign of apoptosis was shown ultrastructurally in vitrified samples. The level of FasL, Bcl-2, Bax, p53, and caspase3 mRNA and Bax:Bcl-2 ratio were similar in non-vitrified and vitrified groups; however, the expression of Fas and caspase8 genes was higher and BIRC5 was lower in vitrified samples compared to non-vitrified group (P<0.05). Conclusion: The fine structure of human vitrified ovarian tissue was well preserved; moreover, vitrification was shown to affect the expression of some apoptosis-related genes. However, additional study is needed to confirm this observation.Key Words: Vitrification, Apoptosis, Gene expression, Ovary, Humans     

杨桃草酸的分布及其果实草酸含量变化的研究

采用离子色谱法对新加坡甜杨桃和广州本地酸杨桃的叶、叶柄、根和不同发育时期的果实及其不同部位进行了草酸含量的测定,同时对不同施肥处理、热水处理的甜杨桃果实分别进行了草酸含量的测定。结果表明,杨桃果实草酸含量先迅速上升,然后逐渐下降,谢花80d左右处于低峰值;杨桃叶片的草酸含量最高,果肉的草酸含量最低;施有机肥可有效降低甜杨桃果实水溶性草酸含量;热水处理3 min可降低果肉水溶性草酸含量42%以上。     

夏玉米叶片叶绿素含量的时空动态及其与植株含氮率关系的研究

在不同施氮水平下,研究了夏玉米郑单958叶片叶绿素含量(SPAD值)的时空动态变化及其与植株含氮率的关系。结果表明,夏玉米子粒生长期棒三叶SPAD值与棒三叶、全株总叶片及整株含氮率均有极显著线性相关关系。不同施氮水平子粒生长期叶片叶绿素含量的空间分布均以穗位叶为中心呈现出中间高两边低的趋势,SPAD值依次为中部叶>上部叶>下部叶。施氮可调节各部位叶片SPAD值或氮素含量,但并未改变其空间分布模式。不同施氮水平子粒生长期上部和中部叶片叶绿素含量随时间的动态变化总体上呈现先增大后减小的趋势,下部叶片的叶绿素含量则呈持续下降的趋势。夏玉米不同叶位叶片对氮肥的反应均表现在一定范围内(N≤207 kg/hm²),子粒生长期各叶片叶绿素含量随施氮量的增加而增加,当总施氮量增为276 kg/hm²时,各叶片叶绿素含量不再明显增加,相对来说中部叶比上部和下部叶对氮肥的反应更为敏感。     

TDZ对尾细桉叶片离体再生的影响

为合理利用生物质炭改良热带地区土壤提供理论依据,通过培养试验,研究不同水分培养条件下添加生物质炭对砖红壤pH及养分含量的影响。结果表明,生物质炭能显著提高土壤pH,增加土壤有机碳及全N、全P和全K的含量;淹水显著提高添加了生物质炭的土壤碱解N含量,但极大地降低了其速效P和速效K含量。说明利用生物质炭改良热带土壤时,要根据改良目的合理进行水分管理。     

生物质炭对砖红壤pH及养分含量影响研究

为提高软木地板表面的物理性能,采用弹性漆对软木地板表面进行涂饰处理,研究不同弹性漆的涂漆量、油漆固化温度对软木地板表面耐磨、耐冲击性、表面色度学特性的影响。结果表明：与对照相比,软木地板表面漆膜具有优良的耐磨性、表面磨耗值随着弹性漆涂漆量的增加而降低,表面磨耗值为0.015 6～0.031 0 g/100 r;弹性漆对软木地板表面抗冲击改善幅度较大;油漆固化温度和涂漆量对软木地板表面颜色影响较小。