水稻重复序列RRD3在转基因植物中的启动子功能

来源于水稻（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）的一个８２０ｂｐ多拷贝重复序列ＲＲＤ３,含有植物启动子ＴＡＴＡ－ｂｏｘ、ＣＡＡＴ－ｂｏｘ等特征保守基元。用ＲＲＤ３取代Ｔｉ载体ｐＢ１１２１中的ＣａｍＶ ３５Ｓ启动子,通过植物转化鉴定ＲＲＤ３的启动子功能。组织化学分析表明,根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ） Ｃｏｎｎ）ＬＢＡ４４０４转化后     

抑制根癌土壤杆菌生长和转移的水稻信号分子的鉴定

从水稻（ＯｒｙｚａｓａｔｉｖａＬ．ｃｖ．ＩＲ７２）根中分离出一种高效抑制根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍｔｕｎｅｆａｃｉｅｎｓ（ＳｍｉｔｈｅｔＴｏｗｎｓｅｎｄ）Ｃｏｎｎ）生长及抑制根癌土壤杆菌在水稻细胞表面附着的水稻信号分子：乙基４－（邻硝基苯基）－３－硫代脲酸酯（Ｔ）,该化合物是一种新发现的天然的硫脲化合物,对根癌土壤杆菌生长和根癌土壤杆菌在水稻细胞表面附着具有较强抑制作用,水稻对根癌土     

水稻内生联合固氮细菌的筛选,鉴定及其分布特性

利用乙炔还原法和固定１５Ｎ２ 活性测定法对分离自水稻（ Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．）“越富”种子、根、茎和叶的内生细菌进行了筛选,获得２９ 株具有体外固氮能力的水稻内生联合固氮细菌。鉴定结果表明它们分属于根癌土壤杆菌（ Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ （Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ） Ｃｏｎｎ） ,放射土壤杆菌（ Ａ． ｒａｄｉｏｂａｃｔｅｒ （Ｂｅｉｊｅｒｉｎｃｋ ｅｔ ｖａｎ Ｄｅｌｄｅｎ） Ｃｏｎｎ） ;阴沟肠杆菌（ Ｅｎｔｅｒｏｂａｃｔｅｒ ｃｌｏａｃａｅ （Ｊｏｒｄａｎ） Ｈｏｒｍａｅｃｈｅ ｅｔ Ｅｄｗａｒｄｓ） ,成团肠杆菌（ Ｅ． ａｇｇｌｏｍｅｒａｎｓ （Ｂｅｉｊｅｒｉｎｃｋ） Ｅｗｉｎｇ ｅｔ Ｆｉｆｅ） ,坂崎肠杆菌（ Ｅ． ｓａｋａｚａｋｉｉ Ｆａｍｅｒ ｅｔ ａｌ．） ;皮氏产碱菌（ Ａｌｃａｌｉｇｅｎｅｓ ｐｉｅｃｈａｕｄｉｉ Ｋｉｒｅｄｊｉａｎ ｅｔ ａｌ．） ,反硝化产碱菌（ Ａ． ｄｅｎｉｔｒｉｆｉｃａｎｓ （Ｌｅｉｆｓｏｎ ｅｔ Ｈｕｇｈ） Ｒｕｇｅｒ ｅｔ Ｔａｎ） ;类产碱假单胞菌（ Ｐｓｅｕｄｏｍｏｎａｓ ｐｓｅｕｄｏａｌｃａｌｉｇｅｎｅｓ Ｓｔａｎｉｅｒ） ,产碱假单胞菌（ Ｐ． ａｌｃａｌ     

固氮相关的两个植物基因转化烟草及其表达

豆科植物凝集和血红蛋白分别在植物识别其相应的根瘤菌和在根瘤内降低氧分压保护固氮酶的共生固氮作用中起重要作用。将豌豆（Ｐｉｓｕｍ ｓａｔｉｖａ Ｌ．）凝集素基因（ｐｌ）和Ｐａｒａｑｓｐｏｎｉａ ａｎｄｅｒｓｏｎｉｉ血红蛋白基因（ｐｈｂ）构建到同一植物表达载体上,通过根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导法转化烟草（Ｎｉｃｓ     

在水稻花培后代中RFLP零等位的遗传分析

在构建水稻分子图谱的研究中,有４ 个ＲＦＬＰ探针揭示了一对籼粳稻之间的零等位现象．其中ＲＧ ２２９ 揭示了籼稻“圭６３０”（Ｏｒｙｚａ ｓａｔｉｖａ ｓｓｐ． ｉｎｄｉｃａ）两个零等位位点;ＲＧ ４１９,ＲＧ４２４ 及ＲＧ ３５３ 揭示了粳稻“０２４２８”（Ｏ． ｓａｔｉｖａ ｓｓｐ．ｊａｐｏｎｉｃａ）的零等位． 利用８１ 个圭６３０／０２４２８ 的花培植株（ＤＨ）进行的遗传分析结果表明,这些零等位位点与ＭｃＣｏｕｃｈ 发表的水稻分子图谱中其相邻分子标记间的连锁关系发生了变化．另外,用ＲＧ ６８４ 探针在一些ＤＨ植株中检测出有新的零等位出现,虽然双亲都具有与ＲＧ ６８４ 同源的序列． 零等位的出现可能与转座因子的转位有关     

转BmK IT4基因烟草的抗虫性

用酶切方法从「ｐＢＳ－ＢｍＫ ＩＴ４质粒中获得ＢｍＫ ＩＴ４基因片段,并构建ＣａＭＶ ３５Ｓ启动子下的ＢｍＫ ＩＴ４基因表达质粒ｐＥ３－ＢｍＫ ＩＴ４,以根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｒｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导的叶盘法转化云烟（Ｎｉｃｏ－ｔｉａｎａ ｔａｂａｃｕｍ Ｌ．）Ｋ３２６叶片,获得４５株抗卡那霉素的再生植株。用这些再生植株进行抗虫     

转基因烟草中Bt毒蛋白基因的表达行为

构建了高效植物表达载体ｐＢｉｎＭｏＢｃ,该载体携带超强表达复合启动子ＯＭ及Ω因子控制下的ＣｒｙＩＡ（ｃ）基因。采用根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导的方法转化烟草（Ｎｉｃｏｔｉａｎａ ｔａｂａｃｕｍ Ｌ．）,ＥＬＩＳＡ检测表明,大多数转基因烟草中ＣｒｙＩＡ（ｃ）基因表达量均超过０．１％,最高可达０．２５５％;转基因烟草     

毛白杨遗传转化系统优化的研究

研究了影响根癌土壤杆菌（Ａｇｒｏｂａｃｔｅｒｉｕｍ ｔｕｍｅｆａｃｉｅｎｓ（Ｓｍｉｔｈ ｅｔ Ｔｏｗｎｓｅｎｄ）Ｃｏｎｎ）介导的毛白杨（Ｐｏｐｕｌｕｓ ｔｏｍｅｎｔｏｓａ Ｃａｒｒ．）遗传转化的若干因素,建立了毛白简单,高铲的遗传转化系统,获得了大量正常转基因植株。在共培养培养基中附加２００μｍｏｌ／Ｌ的乙酰丁香酮的同时降低ｐＨ值可以显著提高毛白杨转化频率     

由RTS—barnase嵌合基因的表达导致的雄性不育水稻植株

用ＰＣ赍水稻品种ＩＲ３６的总ＤＮＡ中扩增了ＲＴＳ基因５’调控区（－１２８８＋２５＿顺序并与从解粉芽孢杆菌（Ｂａｃｉｌｌｕｓ ａｍｙｌｏｌｉｑｕｅｆａｃｉｅｎｓ）染色体ＤＮＡ中克隆的ＲＮａｓｅ（ｂａｒｎａｓｅ）基因编码区构建了嵌合基因,通过根癌农杆菌介导转化水稻品种ＺＨ１１,所获得转基因植株的主要性状与供体亲本无显著差异,但开花后药壁不开裂,并且大多数植株的花粉不被Ｉ－ＫＩ染色,自交条件下结实率为完     

根癌农杆菌介导的水稻转化及转基因R1代植株表型特征

用根癌农杆菌介导的方法对２个粳稻品种和４个籼稻品种进行了转化。粳稻成熟胚和籼稻幼胚来源的愈伤组织用携带质粒ｐＧＩＨ的农杆菌ＥＨＡ１０１感染,对所有品种均获得较高的愈伤转化频率（２０．８３％～６２．３２％）。粳稻“申香粳４号”植株转化频率为１７．３９％,“秋丰”为９．２１％。４个品种籼稻中仅“超丰早１号”获得１株转化植株。Ｓｏｕｔｈｅｒｎ杂交分析、ＧＵＳ染色、Ｔ－ＤＮＡ整合边界序列分析等结果表明外源     

Transfer of cultured rabbit embryos

Embryo transfer experiments were carried out to study the developmental capacity of cultured rabbit embryos when transferred to recipients of variable postovulatory maturity. Rabbit embryos were flushed from the oviduct at 26 hours postcoitum (pc) and cultured in a modified Ham's F-10 medium supplemented with bovine serum albumin (BSA) for a period of 70 hours. At 96 hours pc the cultured embryos, which ranged from the early morula to the expanding blastocyst stage, were transferred to pseudopregnant recipients mated to vasectomized males 36 to 96 hours prior to the transfer procedure. Greatest embryo survival occurred when transfers were made to either the oviducts or uterine horns of recipients at 48 hours pc. Intermediate results for both implantation rates and number of young born were obtained with recipients at 36, 60, 72, and 84 hours pc. Transferred embryos consistently failed to survive the uterine environment of recipients 96 hours pc at transfer although this group was synchronous with embryonic chronological age. Oviductal transfers were generally more successful than uterine transfers. Markedly higher rates of embryo survival resulted from embryos that were collected 60 and 72 hours pc and transferred directly to synchronous recipients without an interim period of culture. Dissimilarity of development for in vivo grown rabbit embryos and those cultured in synthetic medium is demonstrated.     

Utero‐tubal transfer of mouse embryos

Summary: A successful embryo transfer depends on the quality of the transferred embryos, recipients, and the transfer techniques. Among these, transfer techniques are often the limiting factor because transfer methodologies and personal skills vary. Suboptimal embryo transfer procedures can compromise transgenic experiments (pronuclear microinjection and gene targeting) and critical steps of mouse colony maintenance (embryo cryopreservation and mouse line rederivation). Here we present an efficient and simple procedure utilizing specific designs to improve the transfer quality. A 100% implantation rate is observed after the utero‐tubal embryo transfer, which indicates that the modified method successfully prevents the embryos from flowing out of the punctured hole during embryo transfer. We believe this alternative methodology is able to fulfill the need of high efficiency of animal production. genesis 30:77–81, 2001. © 2001 Wiley‐Liss, Inc.     

IVF-ET时影响胚胎着床率的因素分析

胚胎移植成功的标志性事件是胚胎着床。着床是一个高度协调的事件,影响胚胎着床的因素主要有胚胎质量和子宫内膜容受性两方面。     

水稻“双－3”多胚发生的研究

水稻ＭＩＶ（双－３、籼稻）传粉后可以有多个花粉管同时进入胚囊．大多数胚囊的合子发育为一个正常的胚,但是有少数合子胚发生裂生并分化形成双胚芽和一胚根．有些胚囊的助细胞和卵细胞同时受精后,分别发育为助细胞胚和合子胚;有些胚囊中的反足细胞团可直接发育为胚．可见“双－３”水稻除有正常合子胚外还存在助细胞胚和反足细胞匹的多胚现象．     

水稻“双－3”多胚发生的研究

叶秀,陈泽濂,黎垣庆 　　水稻ＭＩＶ（双－３、籼稻）传粉后可以有多个花粉管同时进入胚囊．大多数胚囊的合子发育为一个正常的胚,但是有少数合子胚发生裂生并分化形成双胚芽和一胚根．有些胚囊的助细胞和卵细胞同时受精后,分别发育为助细胞胚和合子胚;有些胚囊中的反足细胞团可直接发育为胚．可见“双－３”水稻除有正常合子胚外还存在助细胞胚和反足细胞匹的多胚现象．     

Survival rate to term of chimeric morulae produced by aggregation of five to nine embryos in the mouse, Mus musculus

Chimeric morulae from five to nine embryos were produced by aggregation after removal of the zonae pellucidae and brief incubation in phytohemagglutinin-P. After 28 hr of in vitro culture, these aggregates were transferred to the left uterine horn of pseudopregnant recipient female mice. Each recipient also received control embryos in the right uterine horn. Genetic markers were included so that aggregate-derived offspring could be distinguished from control offspring. Aggregate-derived embryos survived to term, but at a much reduced rate. In the 25 recipients that produced litters, 6.2% (N=211) of the aggregated embryos developed into liveborn young. Survival-to-term for control embryos was 38.8% (N=387). Survival-to-term was independent of the number of embryos aggregated. The aggregate-derived embryos were apparently undergoing size regulation following implantation.     

Comparative study of somatic embryogenesis from immature and mature embryos and organogenesis from leaf-base of Triticale

Immature and mature zygotic embryos of hexaploid, Triticale var. DT-46 formed an embryogenic callus, with subsequent somatic embryo formation upon subculture to MS (Murashige and Skoog, 1962) or N₆ (Chu et al., 1975) nutrient medium supplemented with various concentrations (9.0–22.5 M) of 2,4-dichlorophenoxyacetic acid (2,4-D). Of the two types of explants, embryogenic tissue from immature embryos responded at a higher frequency, to form somatic embryos over the callus surface. Leaf-base segment cultured on to 2,4-D-containing medium formed a tissue which did not form somatic embryos and instead differentiated into shoot-buds. N₆ medium proved to be more effective than MS in support of somatic embryogenesis or shoot-bud formation. Regeneration of plantlets occurred on 2,4-D-free basal medium. These in vitro-formed plantlets were successfully transferred to soil and set seed.     

Deep and intermediate complex morphophysiological dormancy in seeds of Ferula gummosa (Apiaceae)

We tested the hypothesis that seeds of the monocarpic perennial Ferula gummosa from the Mediterranean area and central Asia have deep complex morphophysiological dormancy. We determined the water permeability of seeds, embryo morphology, temperature requirements for embryo growth and seed germination and responses of seeds to warm and cold stratification and to different concentrations of GA₃. The embryo has differentiated organs, but it is small (underdeveloped) and must grow inside the seed, reaching a critical embryo length, seed length ratio of 0.65–0.7, before the seed can germinate. Seeds required 9 weeks of cold stratification at <10°C for embryo growth, dormancy break and germination to occur. Thus, seeds have morphophysiological dormancy (MPD). Furthermore, GA₃ improved the germination percentage and rate at 5°C and promoted 20 and 5% germination of seeds incubated at 15 and 20°C, respectively. Thus, about 20% of the seeds had intermediate complex MPD. For the other seeds in the seed lot, cold stratification (5°C) was the only requirement for dormancy break and germination and GA₃ could not substitute for cold stratification. Thus, about 80% of the seeds had deep complex MPD.     

被子植物胚柄研究进展

胚柄在胚胎发育过程中是一个暂时性的结构, 但却起着为胚体提供营养和生长调节因子的作用。相对胚体而言, 胚柄具有细胞类型单一、结构相对独立及发育时间短等特点, 其在胚胎发育的基因调控和细胞命运决定研究中具有独特的优势。该文从胚柄的形成及特点、胚柄在胚胎发育中的作用、信号传递系统对胚柄的影响以及与胚柄相关基因的功能等方面进行综述, 以期为研究胚柄在胚胎发育过程中所起的作用以及胚柄的命运决定提供参考信息。     

鸡胚内抗氧化物质的分布与变化

生物体内存在多种内源性抗氧化物质,在生命过程中发挥着基本的防御功能,是人们十分关注的研究领域。本文综述了近年来鸡胚内抗氧化物质的形成与来源等研究成果,分析了鸡胚孵育过程中维生素(A、C、E)、类胡萝卜素、硒、过氧化氢酶和超氧化物歧化酶等主要抗氧化物质的分布与变化,及内源性抗氧化系统的形成,旨在为今后的研究提供有益的科学依据。