Ca2+对黄瓜子叶离体培养物花芽分化的影响

黄瓜子叶离体培养物分化培养0～8d期间,添加Ca2+有利于花芽分化,花芽分化率可从Ca2+0mmol/L的(7.9±5.6)%上升到Ca2+6mmol/L的(31.7±4.0)%;0～2d和2～4d无钙脉冲处理不利于花芽分化,高钙脉冲处理的花芽分化率比对照略高;4～6d和6～8d高钙脉冲不利花芽分化,而无钙脉冲处理使花芽分化率上升很多。尤其是在4～6d,高钙处理使花芽分化率从(22±1.5)%下降到(15.7±3.5)%。而无钙处理使花芽分化率从(22.4±1.4)%上升到(43±3.5)%。表明0～8d期间不同时间段对Ca2+的需求是有差别的。相关性分析表明0～8d期间外源Ca2+影响花芽分化率与总芽中花芽比例极显著相关,提示Ca2+可能影响子叶向花芽或营养芽分化的趋势。本文结合已报道的黄瓜子叶培养物花原基形成的时程,分析了Ca2+对花原基形成和分化的影响。     

离体黄瓜子叶节花芽分化与内源激素及多胺的关系

用高效液相色谱法(HPLC)测定了黄瓜子叶节花芽分化期(0-6天)内源激素及多胺的变化。结果显示,子叶培养0-2天生长素(IAA)、赤霉素(GA_3)、玉米素(ZT)、脱落酸(ABA)等4种内源激素均明显下降,4-5天略有上升,表明0-2天IAA、GA_3和ABA的剧降有利于花原基形成,3-5天较高的ZT含量有利于花器官原基的形成。除腐胺(Put)外,精胺(Spm)、亚精胺(Spd)、尸胺(Cad)在0-1天均下降,1-4天上升,4-5天再下降,Put在0-1天急剧上升,而后持续下降,表明高水平的内源多胺总量和Put可能有利于花原基分化,2天后Spm含量上升有利于花器官原基分化,而Cad含量变化可能是区别花芽和营养芽分化的特征之一。     

离体黄瓜子叶节花芽分化与内源激素及多胺的关系

用高效液相色谱法(HPLC)测定了黄瓜子叶节花芽分化期(0—6天)内源激素及多胺的变化。结果显示,子叶培养0—2天生长素(IAA)、赤霉素(GA3)、玉米素(ZT)、脱落酸(ABA)等4种内源激素均明显下降,4—5天略有上升,表明0-2天IAA、GA3和ABA的剧降有利于花原基形成,3—5天较高的ZT含量有利于花器官原基的形成。除腐胺(Put)外,精胺(Spm)、亚精胺(Spd)、尸胺(Cad)在0—1天均下降,1—4天上升,4—5天再下降,Put在0—1天急剧上升,而后持续下降,表明高水平的内源多胺总量和Put可能有利于花原基分化,2天后Spm含量上升有利于花器官原基分化,而Cad含量变化可能是区别花芽和营养芽分化的特征之一。     

4种钙素调节剂对紫花擎天凤梨花芽分化及内源激素含量的影响

以组培苗移栽2年的紫花擎天凤梨为试材,研究了外源乙烯催花条件下4种钙素调节剂(A23187、W-7、TFP、EGTA)处理对其花芽分化及内源激素含量的影响.结果表明:(1)4种处理均能使花芽分化过程中生长素(IAA)、玉米素(ZR)含量不同程度增加,并以A23187处理的IAA、ZR含量增加幅度最大.(2)赤霉酸(GA3)和脱落酸(ABA)含量在花芽孕育阶段先后下降至低谷,在花芽形态分化期又先后上升并出现高峰;ABA含量在A23187处理中下降早于其他处理,于处理3 d时首先降到最低,而在EGTA处理中下降速度最慢,至处理5 d时才降到最低值;在花芽孕育阶段,处理A23187的GA3含量下降最慢,但整体下降幅度最大;在此后花芽发端期各处理的GA3含量均出现高峰,但A23187和乙烯处理上升幅度均高于其他3个处理.(3)(ZR+ABA)/GA3、(ZR+IAA)/GA3比值在花芽孕育阶段上升并出现高峰,在花芽形态分化期下降到低谷.(4)4种处理花芽分化完成时间不一致,紫花擎天凤梨花芽分化在Ca2+促进剂A23187处理下提前,分化时间缩短;而在钙离子专一性螯合剂EGTA、钙调素(CaM)拮抗剂W-7和TFP处理下均得到延缓或抑制.研究发现,Ca2+-CaM信号系统参与了乙烯诱导的紫花擎天凤梨花芽分化过程且起着重要的调控作用.     

黄瓜离体子叶节花芽和营养芽分化中CFL基因的表达

CFL基因是从黄瓜中克隆到的拟南芥LEAFY(LFY)同源基因.以离体黄瓜子叶培养物成花为实验体系,利用mRNA原位杂交技术对CFL基因在花芽和营养芽分化过程中的时空表达进行了分析.结果如下:在花芽分化过程中,CFL基因在花原基形成、花器官原基分化及各轮花器官形成之初强表达,在花器官形成以后表达减弱或不表达;在营养芽分化过程中,CFL基因在分生组织、叶原基和幼叶中有明显表达,在成熟组织中不表达.结果说明CFL基因的表达在黄瓜子叶节花芽和营养芽分化中原基的分化形成是必需的.结果提示CFL基因可能参与细胞分裂调控和启动、营养性分生组织向花分生组织转变等过程.     

低温处理青花菜萌动种子对花芽分化的促进作用

低温(0～2℃)处理萌动种子可使青花菜的花芽分化提前.种子萌动时期经低温处理10 d(T10)和20d(T20)的青花菜植株花芽分化开始时的节位分别比对照(CK)低0.86个节位和1.03个节位.花芽分化临界期、第1级侧花茎原基分化期和第2～3级侧花茎原基分化期经低温处理10d和20 d均分别比对照提前5 d和6d.当进入花芽分化临界期时,叶片中GA3、可溶性蛋白质含量以及POD和转化酶活性均开始剧增,在第1级侧花茎原基分化期和第2～3级侧花茎原基分化期,叶片中POD、转化酶活性和GA3均有一高峰值.萌动种子经一段时间的低温处理后,首先诱导了GA3的合成,从而提高POD和转化酶等酶的活性,而这两种酶活性的升高对花芽分化有利.     

无外源激素条件下液体和固体培养基对黄瓜子叶培养器官分化的不同影响

在固体ＭＳ基本培养基上,黄瓜子叶能形成约０．８％的直接花芽,其分化高峰约为４５天左右,而在液体ＭＳ基本培养基上,黄瓜子叶花芽分化率为８％,其分化高峰期约在３０天左右．实验结果表明：液体培养基对黄瓜子叶花芽及根的分化效果明显好于固体培养基,而对于营养芽的分化固体培养基好于液培养基,花芽分化与子叶衰老存在着较密切的关系。     

罗汉果花芽分化过程中形态及其激素水平变化特征

采用石蜡切片和酶联免疫法(ELISA)对罗汉果雄性、雌性、两性花芽分化过程的形态和激素水平变化进行观测,为罗汉果开花调控和品种选育提供科学依据。结果表明:(1)罗汉果雄性、雌性、两性花的花芽分化过程均可分为花芽未分化期、花芽分化初期、花序分化期、萼片原基分化期、花瓣原基分化期、雄蕊原基分化期和雌蕊原基分化期7个阶段。雄蕊原基分化期前,3种花芽分化过程无明显差异,各时期形态特征均依次为:茎端呈圆锥状(花芽未分化期)→茎端经半球形变成扁平状(花芽分化初期)→距茎端5~7节位处分化出穗状花序(花序分化期)→小花原基周围形成5个萼片原基(萼片原基分化期)→萼片原基内侧形成5个花瓣原基(花瓣原基分化期)。雄蕊和雌蕊原基分化期,3种花芽分化过程存在明显差异,雄蕊原基内侧出现雌蕊原基后,雄花芽雄蕊原基继续发育成雄蕊,雌蕊原基停滞生长,退为一个小突起;雌花芽雌蕊原基继续发育成雌蕊,雄蕊原基生长缓慢,退化为小花丝;两性花芽雌蕊和雄蕊原基均继续发育,形成外观正常的雌蕊和雄蕊。(2)内源激素脱落酸(ABA)、赤霉素(GAs)和玉米素核苷(ZR)含量在3种花芽分化过程中变化规律相似,即ABA含量在花芽生理分化期降低,花芽形态分化期升高,而GAs和ZR含量则基本保持不变;吲哚乙酸(IAA)含量在3种花芽分化过程中变化存在明显差异,雌花芽IAA含量在花芽生理分化期升高,花芽形态分化期逐渐降低,而雄性和两性花芽的IAA含量则基本保持不变。ABA/GAs、ABA/IAA、ZR/IAA和ZR/GAs激素含量比值在3种花芽分化过程中变化规律相似,ABA/GAs在花芽生理分化期降低,花芽形态分化期升高,而BA/IAA、ZR/IAA和ZR/GAs则基本保持不变。研究认为,罗汉果花芽分化过程经历一个"两性期",高ABA含量和ABA/GAs比值有利于罗汉果花芽分化,IAA可能对罗汉果花性分化具有重要作用。     

超早熟栽培草莓花芽分化进程的扫描电镜观察

于2011年7月23日至8月24日,对草莓品种‘红颜’进行低温(夜温为17～20℃变温)、短日照(黑暗时间为16 h)处理,从处理之始到花序现蕾,约每3～5 d取样1次,实体解剖镜下解离草莓顶芽,材料均用FAA固定,临界点干燥,扫描电镜观察,以确定保护地栽培草莓的花芽开始分化始期,为生产中草莓尽早适期定植提供依据.根据扫描观察的微形态特征,将草莓花芽分化的全过程分为花芽未分化期、花芽分化始期、生殖顶端膨大期、花序分化期、顶花花萼与花瓣形成期、雄蕊形成期和雌蕊形成期共7个时期.花芽未分化期,顶芽一直处于营养生长阶段;花芽分化始期,草莓顶芽逐步由尖锐、狭窄的营养顶端向平坦、宽大的生殖顶端发育;生殖顶端膨大期,生殖顶端隆起而膨大;花序分化期,顶花序逐步分化出顶花原基和侧花原基;顶花花萼、花瓣形成期,花萼(包括副萼)、花瓣原基相继分化完成;雄蕊形成期,两轮雄蕊原基相继分化形成;雌蕊形成期,大量雌蕊原基逐步隆起,心皮原基逐步卷合、完成花芽分化.研究认为,草莓的定植期以生殖顶端充分膨大期为宜,即低温短日照处理15～20 d后即可定植大田.     

Amiloride和Ni2+抑制缺血/复灌引起的大鼠心肌细胞内钙的增加

本文旨在研究Na+/H+交换以及Na+/Ca2 +交换对模拟缺血 /复灌引起的大鼠心肌细胞内游离钙水平变化的调节作用。分别利用模拟缺血液和正常台氏液对大鼠心肌细胞进行缺血 /复灌处理 ,在缺血期间分别应用Na+/H+交换抑制剂阿米洛利 (amiloride)、Na+/Ca2 +交换抑制剂NiCl2 以及无钙液 ,观察它们对细胞内游离Ca2 +浓度变化的影响。利用Zeiss LSM 5 10激光共聚焦显微镜检测、采集细胞内游离Ca2 +的指示剂Fluo 3 AM的荧光信号 ,计算出相对于正常(缺血前 )的相对荧光强度 ,以表示胞内游离Ca2 +浓度的变化。结果显示 ,模拟缺血引起大鼠心肌细胞内游离Ca2 +持续上升 ,缺血前的相对荧光强度值为 10 0 % ,模拟缺血 5min后为 140 3± 13 0 % (P <0 0 5 ) ,复灌 15min后为 142 8±15 5 % (P <0 0 5 )。经 10 0 μmol/Lamiloride、5mmol/LNiCl2 和无钙液分别预处理 ,模拟缺血 5min后的相对荧光强度分别为 10 1 4± 16 3 % (P <0 0 5 )、110 4± 11 1% (P <0 0 5 )和 10 7 1± 10 8(P <0 0 5 ) ;复灌 15min后则分别为 97 8±14 3 % (P <0 0 5 )、10 6 2± 14 5 % (P <0 0 5 )和 10 6 6± 15 7(P <0 0 5 )。另外 ,与对照组细胞相比 ,再灌注期间NiCl2和无钙液处理的细胞钙振荡的产生幅度明显减弱 ,amilorid     

黄瓜子叶节离体培养物花分化Ca^2＋敏感期的研究（简报）

Cotyledonary nodes of cucumber cultured on calcium-free medium for 0, 1, 2, 3, 4, 5, 6d respectively, were transferred to medium with 6.0 mmol/L CaCl2 for 24h, then returned to calcium-free medium. Cotyledonary nodes cultured on calcium-free or 6.0 mmol/L CaCl2 medium for all time, were taken as controls. Results showed that cotyledonary nodes were transferred to 6.0 mmol/L CaCl2 medium for 24h during 0-3d after the beginning of culture, percentage of floral bud formation at cotyledonary nodes was increased significantly. Transferring cotyledonary nodes on the 3d day after the beginning of culture was achieved best effect, percentage of floral bud formation was up to 34.3%. We deduced that the calcium sensitive period during floral differentiation of cucumber cotyleddonary node cultured in vitro may be 0-4d after the beginning of culture.     

黄瓜子叶节离体培养物花分化Ca2+敏感期的研究

植物开花是一个非常复杂的过程,对其机理的研究有着重要的实践意义和理论意义。近百年来进行了广泛的生理生化研究,提出过多种假设。自90年代初成功分离了花器官分化和发育基因以来,使这一研究领域进入了分子水平。钙不仅是植物的矿质营养元素之一,而且Ca~(2+)作为植物的第二信使,参与细胞内多种生理生化活动。许多研究结果表明,Ca~(2+)在成花诱导、花芽形成和分化过程中起重要作用,我们自建立黄瓜子叶培养物离     

大岩桐花瓣切块离体培养高频率花芽再生

该文报道了大岩桐花瓣切块离体培养再生花现象,花瓣切块再生花有两种方式：一种是仅再生花芽（命名为BF）;另一种是既再生花芽也再生营养芽（命名为BF＋V）。花芽再生的能力与光照、花芽大小及培养基中赤霉素和细胞分裂素浓度紧密相关。当培养基中含有1.0 mg/L GA3时,BA的添加会显著增加总花芽（BF＋BF＋V）的形成率,添加0.5 mg/L BA时,总花芽形成率达100%。在暗中培养时,BF达93.4%。不同大小花芽的花瓣再生花的能力不同,7 mm直径花芽的BF最高,达86.7%。同时,对花芽再生过程中花瓣切块的组织结构形态变化也进行了观察。     

Changes of Endogenous Hormone Contents During Floral Bud and Vegetative Bud Differentiation in Thin Cell Layer Culture of Cichorium intybus L. Explant

The sectioned thin cell layers (TCL) of flower stalk of Cichorium intybus L. were cultured in MS medium supplemented with NAA and BA or IAA and BA where floral and vegetative buds were developed from the explant. Endogenous IAA, DHZ+DHZR, iPA increased significantly during the floral bud formation, while Z+ZR remained changed. The levels of cytokinins, DHZ +DHZR, iPA, and Z-f-ZR all increased significantly during the vegetative bud formation, however IAA level was reduced during the first 7 days of culture and increased to two-thirds of initial values on the day when the bud primordia were formed. The results suggested that the initiation of floral buds was associated with a high IAA/CTK ratio, whereas the induction of vegetative bud differentiation was related to a low IAA/CTK ratio.     

喷施烯效唑对苹果顶芽激素水平和花芽分化的影响

用烯效唑(uniconazol,S3307)1 g/L喷洒“红富士“苹果树降低了顶芽IAA、GA1,3,4,7含量,提高了ZR、ABA含量,从而提高了ZR/IAA、ZR/GA1,3,4,7、ABA/IAA和ABA/GA1,3,4,7比值.烯效唑处理增加了花芽形成百分率,加速了花芽分化的进程,缩短了花芽形成的延续时期,但对花芽生理孕育临界时期长短没有影响.烯效唑处理对花芽的节位数没有影响,但使叶芽节位数增加了1节.     

Winter bud content according to position in 3-year-old branching systems of 'Granny Smith' apple

An investigation was made of the number of preformed organs in winter buds of 3-year-old reiterated complexes of the 'Granny Smith' cultivar. Winter bud content was studied with respect to bud position: terminal buds were compared on both long shoots and spurs according to branching order and shoot age, while axillary buds were compared between three zones (distal, median and proximal) along 1-year-old annual shoots in order 1. The percentage of winter buds that differentiated into inflorescences was determined and the flowers in each bud were counted for each bud category. The other organ categories considered were scales and leaf primordia. The results confirmed that a certain number of organs must be initiated before floral differentiation occurred. The minimum limit was estimated at about 15 organs on average, including scales. Total number of lateral organs formed was shown to vary with both bud position and meristem age, increasing from newly formed meristems to 1- and 2-year-old meristems on different shoot types. These differences in bud organogenesis depending on bud position, were consistent with the morphogenetic gradients observed in apple tree architecture. Axillary buds did not contain more than 15 organs on average and this low organogenetic activity of the meristems was related to a low number of flowers per bud. In contrast, the other bud categories contained more than 15 differentiated organs on average and a trade-off was observed between leaf and flower primordia. The ratio between the number of leaf and flower primordia per bud varied with shoot type. When the terminal buds on long shoots and spurs were compared, those on long shoots showed more flowers and a higher ratio of leaf to flower primordia.     

The Exogenous Hormornal Control of the Development of Regenerated Flower Buds in Hyacinthus orientalis

The critical role of exogenous hormone on inducing the initiation of different floral organs in the regenerated flower bud and controlling their numbers was further evidenced. The initiation of the flower buds was first induced from the perianth explants of Hyacinthus orientalis L. cv. White pearl by a combination of 2 mg/L 6-BA and 0.1 mg/L 2,4-D, and then a continuous initiation of over 100 tepals (a flower bud of H. orientalis in situ has only 6 tepals) was successfully controlled by maintenance of such a hormone concentration. However, a change of hormonal concentration (2 mg/L 6-BA and 0-0.000 1 mg/L 2,4-D) caused cessation of continuous initiation of the tepals but gave rise to induction of stamen initiation. Keeping the changed hormone concentrations could successfully control the continuous initiation of over 20 stamens (a flower bud of H. orientalis in situ has only 6 stamens). The experiment showed that the number of identical floral organs in the regenerated flower buds can be controlled by certain defined concentrations of the exogenous hormones, and the amount of the induced identical floral organs has no effect on the differentiation sequence of the different floral organs in the regenerated flower bud. Based on a systematic research on controlling the differentiation of the floral organs from both the perianth explants and the regenerated flower buds by the exogenous hormones in H. orientalis over the past decade, the authors put forward here a new idea on the role of phytohormone in controlling the automatic and sequential differentiation of the different floral organs in flower development. The main points are as follows: 1. the development of flower bud in plant is a process in which all of the floral organs are automatically and sequentially differentiated from the flower meristem. 2. Experiments in vitro showed that the effect of exogenous hormones in controlling the initiation of different floral organs is strictly concentration dependent, i.e., one kind of the floral organ can continuously and repeatedly initiate from the flower meristem as long as it is maintained in that specific concentration of the exogenous hormone which is suitable for the initiation of that particular kind of floral organ. 3. It shows that the flower buds in situ must be automatically able to adjust the endogenous hormonal concentrations just after the completion of the differentiation of one whorl of floral organ to suit the differentiation of the next whorl. Thus, the phytohormone in different concentrations takes after many change-over switches of the organ differentiation and plays a connective and regulatory role between the differentiation of every two whorls of the floral organ. In other words, these change-over switches play the roles of inhibiting the expression of the genes which control the initiation of the floral organs in the first whorl, meanwhile, activating the expression of the genes which control the initiation of the floral organs in the second whorl during the successive initiation of the different floral organs from the flower bud. It results in the automatic and sequential initiation of the various floral organs from the floral meristem.      

赤霉素对大岩桐花蕾离体培养直接再生花芽的影响（简报）

植物经过一定时期的营养生长(或感受外界信号)后,就能产生成花刺激物。成花刺激物被运输到茎尖,诱导发生一系列的反应。随后其分生组织在一定时期内处于一个相对稳定的状态,即成花决定态。植物成花决定态建立的过程称为成花决定。对成花决定的研究进行了许多年,但是其确切的机理仍不清楚.     

Effect of ABA on the in Vitro Induction of Floral Buds of Dendrobium candidu Wall.ex Lindl.

An in vitro flowering system of Dendrobium candidurn Wall. ex Lindl., a wild species of orchid, was established. Callus was induced from seeds and protocorms formed on MS agar medium supplemented with 0.3 mg/L NAA under diffused light. Floral buds were induced when protocorms were cultured on MS agar medium supplemented with 2 mg/L 6-BA and 0.5 mg/L NAA, the frequency of floral bud induction being 27.0%. When protocorms were precultured on MS medium supplemented with 0. 5 mg/L ABA for 15 days and then transfered onto MS medium supplemented with 2 mg/L 6-BA, the frequency of floral bud induction increased greatly reach 84.0 % during a period of 5 months. Meanwhile the number of branches and floral buds also increased and in some instances inflorescence appeared. Never the less, no floral bud was formed if protocorms continued to be cultured on the ABA-containing MS medium.