Auxin Regulation of Flower Bud Formation in Tobacco Explants

Smulders, M. J. M, Janssen, G. F. K, Croes, A. F., Barendse,G. W. M., and Wullems, G. J. 1988. Auxin regulation of flowerbud formation in tobacco explants.—J. exp. Bot. 39: 451–459. The auxin 1-naphthaleneacetic acid (NAA) induces flower budformation in tobacco explants at concentrations between 0.1and 10 mmol m^–3. This regeneration process is completedin 14 d. The presence of NAA in the medium is required onlyduring the first 4 d of culture. In this period the explantstake up NAA at a very high rate, which is proportional to theconcentration in the medium. Due to the high uptake from a limitedvolume, the external concentration diminishes and, as a consequence,the uptake rate declines rapidly. A negative correlation wasfound between medium volume and the NAA concentration optimalfor bud formation. It is concluded that it is not the concentrationbut the dose of hormone taken up during the induction periodwhich determines the number of flower buds formed. Key words: Auxin, NAA, development, tissue culture     

Inhibition by Ethylene of Auxin-Promotion of Flower Bud Formation in Tobacco Explants Is Absent in Plants Transformed by Agrobacterium rhizogenes

The in vitro regeneration of flower buds was studied in pedicel explants from tobacco (Nicotiana tabacum L., cv Petit Havana) transformed with Agrobacterium rhizogenes, pRi 1855 (agropine type). At a low concentration (0.1 micromolar) of 1-naphthalene-acetic acid, pedicel strips from phenotypically aberrant plants regenerated two to three times more flower buds than explants from untransformed tobacco. Intermediate bud numbers were observed in transformants with a less extreme phenotype. The results can be explained by an increased sensitivity of the transformed explants to auxin with respect to flower bud regeneration. The effect of transformation on the auxin response is fully accounted for by the absence of a negative interaction of endogenous ethylene with 1-naphthaleneacetic acid, a phenomenon normally encountered in untransformed tissues. Three observations led to this conclusion. Application of 1 micromolar AgNO₃ to untransformed explants increased the number of flower buds to the level observed in transformed tissues but had no effect on transformed pedicel strips; exposure to 10 microliters per liter ethylene strongly reduced the response to auxin at all concentrations in untransformed explants but was almost ineffective in the transformed tissues; and endogenous ethylene synthesis occurred at the same rate in both types of explants.     

油茶花芽分化形态结构及内源激素变化

以普通油茶(Camellia oleifera) 4个无性系为材料,结合油茶成花的动态观察和花芽分化过程的石蜡切片形态观察,采用酶联免疫吸附分析法测定花芽中玉米素核苷(ZR)、脱落酸(ABA)、生长素(IAA)、赤霉素(GA) 4种内源激素含量,探讨油茶花芽分化与内源激素的关系。油茶花芽分化过程可分为6个时期：前分化期(10 d)、萼片形成期(20 d)、花瓣形成期(30 d)、雌雄蕊形成期(20 d)、子房与花药形成期(10 d)和雌雄蕊成熟期(20 d),历时3～4个月。油茶不同无性系的花芽分化时间略有不同。油茶花芽中ZR含量相对较低(5.102～16.412 ng·g–1 FW),ABA含量相对较高(76.815～137.648 ng·g–1 FW)。其中,粤华5号和湘林8号的ZR、ABA含量变化趋势一致,岑软3号和岑软2号含量变化趋势一致。油茶花芽中IAA含量相对较高,为49.072～135.622 ng·g–1 FW,随着花芽分化进程,IAA含量均呈先升后降再升的变化趋势。GA含量相对较低,为5.616～13.720 ng·g–1 FW,随时间变化,呈现出不断降低的趋势。其中,不同无性系的IAA、GA含量变化趋势一致,而ZR、ABA含量变化趋势有所差异。ZR有利于花器官形成;高浓度IAA促进油茶花芽分化,低浓度IAA有利于开花;花芽中IAA与ABA存在明显的颉颃作用;GA抑制花芽分化。     

舞钢玉兰芽种类与成枝成花规律的研究

报道了舞钢玉兰芽的种类、分枝习性与成枝生长规律,拟花蕾、着生位置、解剖结构及其分化发育成花规律。从中发现：（1）当年生枝上有休眠芽、叶芽（侧叶芽和顶叶芽）、拟花蕾3种;（2）拟花蕾有缩台枝、芽鳞状托叶、雏枝、雏芽及雏蕾组成,因其外形似“花蕾“,称为“拟花蕾“;（3）缩台枝是枝与花着生的中间过滤枝变阶段,是由母枝顶端节间缩短、增粗的枝段和当年由雏枝生长的1次极短新枝所组成;（4）4-5月及7-8月前后两批形成的拟花蕾,均经过未分化发育期、花被分化发育期、雄蕊群分化发育期及离心皮雌蕊群分化发育期,各期均依次递后交错进行,但不逆转,也不能截然分开,直到翌春花分化发育全部结束,开花后才能结实;（5）芽鳞状托叶是托叶的变态,最外层薄革质,外面密被短柔毛,始落期6月中下旬,其余纸质--膜质,外面密被或疏被毛柔毛,翌春开花时脱落完毕;（6）雏蕾有雏梗、雏花及包被雏花的佛焰苞状托叶组成;（7）分枝习性与成枝生长规律与预生分枝及预生一同生分枝呈单阶无歧、单阶1歧生长规律,稀有单阶2歧生长规律。     

‘神马’菊花花芽分化与内源多胺的关系

用薄层层析-荧光法测定菊花'神马'花芽分化期间顶芽和叶片中多胺的动态变化,分析了菊花花芽分化与多胺的关系.结果表明,花芽分化起始期顶芽中的腐胺(Put)含量急剧下降,此后在低水平上波动;叶片内Put则于总苞鳞片分化初期大幅上升,其后各阶段处于较低的水平.顶芽中精胺(Spm)与亚精胺(Spd)含量呈平行波动上升趋势,顶芽中Spin在小花原基分化初期直到花冠分化中期处于优势地位,而顶芽中Spd并无明显变化.顶芽、叶片中的Spin变化趋势相反,顶芽中Spm、Spd的含量变化趋势十分相似,但叶片中却呈交替性变化.结果显示,菊花花芽分化过程中,Put含量的降低有利于启动菊花花芽分化,后期Spm的增加有利于小花的分化,叶片可能向顶芽提供Spm,顶芽和叶片中的Spd与小花原基分化有密切关系.     

桂花花芽分化期内源激素含量的变化

在桂花花芽分化期,采用酶联免疫吸附法(ELISA)测定桂花花芽内4种内源激素吲哚乙酸(IAA)、赤霉素(GA)、玉米素核苷(ZRs)和脱落酸(ABA)含量的动态变化.结果表明:在花芽生理分化期,GA和IAA含量有所增加,但在花芽形态分化开始以后GA的含量呈逐步下降趋势,直至分化结束;ZRs含量在花芽生理分化期呈上升趋势,在形态分化前期含量出现高峰,进入形态分化期后其含量下降;ABA含量在桂花花芽分化初期(苞片分化期)有逐步增加的趋势,在6月22日至7月2日达到最高水平,在花序分化以后至花芽形态分化结束,ABA的含量呈逐步下降趋势,但其变化相对平稳.研究认为桂花花芽分化期需要有高水平的ZRs、低水平的GA和IAA.     

The Initiation of an Endogenous Rhythm Affecting Flower Bud Formation in Pharbitis nil

When seedlings of Pharbitis nil are presented with an inductive dark period at varying times, they show a circadian fluctuation in the number of flower buds initiated. This study determines if this fluctuation is due to the plant's perception, at the time of the inductive dark period, of either a rhythmic, external, environmental stimulus or of an endogenous rhythm. Using experimental designs in which the time of planting, the time of seedling emergence from the soil, and the time at which the presentation of an inductive dark period are varied, this fluctuation in flower bud formation is shown to be due to an endogenous rhythm initiated or synchronized by some event associated with the emergence of the seedlings from the soil. The results are inconsistent with the hypothesis that the plants are responding to rhythmic external stimuli.     

马铃薯花芽分化与内源激素动态变化的关系

以马铃薯2个开花品种YS205和HZ88以及未开花品种YS304为材料,采用石蜡切片法观察花芽分化的解剖学特征,并用酶联免疫吸附法(ELISA)测定开花与未开花品种不同时期叶内4种内源激素——生长素(IAA)、赤霉素(GA3)、玉米核苷素(ZR)、脱落酸(ABA)含量的动态变化,探讨成花过程中叶片内源激素含量和比值的变化与成花的关系,为马铃薯花期调控、栽培和杂交育种提供理论依据。结果显示:(1)马铃薯花芽内部发育与外部形态之间有相对稳定的时序性对应关系,从花芽分化初期生长锥逐渐向上凸起,到雌雄蕊原基形成,完成整个花芽分化仅用了1周左右的时间,此阶段YS205和HZ88的花芽分化过程无明显差异,而YS304未成花。(2)随着花器官的不断发育,HZ88子房形成2室,每室多个胚珠,而YS205的子房在发育过程中发生了变异,形成完整的3室;YS205和HZ88都先后经历了授粉受精过程,HZ88具有一定的自交结实率,YS205无坐果率。(3)马铃薯3个品种的IAA、GA3、ZR和ABA含量在花芽分化前期均较低,且无明显差异性;随后,3个品种的IAA和GA3含量均呈先升后降的单峰变化曲线,但YS304上升和下降的速度较2个开花品种快,而且YS205和HZ88的ZR和ABA含量则始终高于YS304。(4)开花品种YS205和HZ88的ABA/IAA、ABA/GA3、ZR/IAA、ZR/GA3比值均高于未开花品种YS304,且随生育期变化趋势不同。研究表明,较低水平的IAA、GA3和高水平的ABA、ZR均有利于促进马铃薯花芽分化,反之则抑制花原基形态的建成。     

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.     

Changes in Cytokinins before and during Early Flower Bud Differentiation in Lychee (Litchi chinensis Sonn.)

Lychee (Litchi chinensis) has been analyzed for cytokinins in buds before and after flower bud differentiation, using reversephase high performance liquid chromatography in combination with Amaranthus bioassay and gas chromatography-mass spectrometry-selected ion monitoring. Four cytokinins, zeatin, zeatin riboside, N⁶-(δ²-isopentenyl)adenine, and N⁶-(δ⁶-isopentenyl) adenine riboside, were detected in buds. There was an increase of cytokinin activity in the buds during flower bud differentiation. In dormant buds, the endogenous cytokinin content was low, and the buds did not respond to exogenous cytokinin application. Application of kinetin promotes flower bud differentiation significantly after bud dormancy. These results are interpreted as an indication that the increase in endogenous cytokinin levels during flower bud differentiation may be correlative rather than the cause of flower bud initiation.     

光皮树花芽分化过程中内源激素含量变化的研究

以4年生光皮树嫁接苗为材料,采用酶联免疫吸附法（ELISA）测定同龄开花光皮树与不开花光皮树叶内4种内源激素生长素（IAA）、赤霉素（GA3）、玉米核苷素（ZR）、脱落酸（ABA）含量的动态变化,研究成花过程中叶片内源激素含量的变化与成花的关系。研究结果表明：在芽分化时期开花光皮树和未开花光皮树叶内4种内源激素含量动态变化存在显著差异。有花芽分化的光皮树保持相对较低和稳定IAA、GA3含量,ABA含量高且变化幅度较大,ZR含量相对较低,但随着花的形成含量逐渐升高;而在相同的生长季内,无花光皮树IAA、GA3含量先升高后降低,ABA含量先逐渐升高,然后下降,而ZR含量呈现出先逐渐升高然后降低,再升高再下降的变化趋势。因此,4种内源激素含量的动态变化影响光皮树的成花过程。     

无花果花芽分化过程中内源激素含量的变化

无花果花芽分化过程中的花托和小花形成阶段,新梢第7或第8节位芽中,脱落酸（ABA）和玉米素核苷（ZRs）含量先升高,后保持高水平：生长素（IAA）和赤霉素（GA1＋3）含量先下降,后保持低水平。     

香荚兰花芽分化至萌发期内源激素的变化

以香荚兰 (Vanillafragrans)为材料 ,研究不同栽培条件下花芽分化和萌发期内源激素变化 ,分析和探讨内源激素在花芽分化和萌发中的作用 ,香荚兰花芽分化时期茎里的激素含量降低 ,芽里激素含量升高 ,其中相对高的ZR和ZR ABA有利于分化 ,IAA和IAA ABA的一定增加也利于分化 ,过高或没有IAA的增加则不利于花芽分化。大多数花芽形成于倒垂茎蔓上 ,花芽分化期 (11～ 12月 ) ,倒垂茎蔓的茎里生长类激素含量降低大于竖立茎蔓 ,芽的激素含量增高则多于竖立茎蔓 ,倒垂茎蔓的这种变化可能是有利于花芽分化。香荚兰生长中顶端优势明显 ,去顶后侧芽里ZR、GA、IAA增高 ,这与 11～ 12月去顶促进倒垂茎蔓开花可能有关。     

青海云杉花芽分化期内源激素含量的变化特征

采用酶联免疫法测定并分析了中国青藏高原东北边缘特有树种青海云杉花芽分化过程中内源激素的变化,以期为调控青海云杉花期调控提供理论依据。结果表明:(1)内源激素吲哚乙酸(IAA)、赤霉素(GAs)、玉米素核苷(ZR)和脱落酸(ABA)含量的变化存在一定的相似性,它们分别在青海云杉花芽生理分化前期和形态分化前期出现高峰;青海云杉叶片ZR/GAs、ZR/IAA之值在花芽生理分化期达到峰值,而ABA/GAs值在花芽生理分化期总体呈递增趋势。(2)青海云杉花芽生理分化期,其顶芽、侧芽中可溶性糖及蛋白质皆出现高峰;形态分化前期,顶芽及侧芽中蛋白质含量下降,但可溶性糖含量持续上升;而花芽生理分化期间,叶片中核酸含量总体皆呈递增趋势。研究认为,较高的ZR/GAs、ZR/IAA有利于青海云杉花芽生理分化,但对维持花芽形态分化可能不是必须的,而高的ABA/GAs、ABA/IAA可能是花芽形态分化能够顺利完成所不可缺少的;可溶性糖、蛋白质、核酸等结构物质和能量物质的积累有利于青海云杉花芽生理分化的完成。     

Endogenous Levels of Gibberellins, IAA and Cytokinins in Tobacco Crown Gall Tissues of Different Morphologies

Endogenous levels of gibberellin (GA) as well as IAA and cytokininsin teratomas and unorganized crown gall tissues of tobacco wereexamined by GC-SIM (for GA and cytokinin) or HPLC with a fluorescencedetector (for IAA). Two different types of crown gall inducedby octopine-type and nopaline-type Ti-plasmids were used. Inboth types, GA contents were higher in shoot-forming teratomasthan in unorganized calluses, while IAA contents were higherin unorganized calluses. But cytokinin contents in octopine-typecells were higher in unorganized calluses than in teratomas,whereas the contents in nopaline-type cells were higher in teratomas.Our results suggest that there is not always a relationshipbetween the cytokinin/IAA balance and tobacco crown gall morphology,but GA production in tobacco tissues is closely related to itsdifferentiation. ⁴ Present address: Agency for the Assessment and Applicationof Technology (BPPT), Jakarta Pusat, Indonesia. (Received September 1, 1986; Accepted February 16, 1987)     

Regulation by Cytokinins of Endogenous Levels of Jasmonic and Salicylic Acids in Mechanically Wounded Tobacco Plants

Plants respond differentially to wounding and pathogens usingdistinct signaling pathways, so that wound signals are transmittedto jasmonic acid (JA) which induces basic pathogenesis-related(PR) proteins, whereas pathogenic signals cause, in additionto JA, accumulation of salicylic acid (SA) which stimulatesproduction of acidic PR proteins. Transgenic tobacco plantsexpressing a gene for a small GTP binding protein respond abnormallyto mechanical wounding to produce SA and consequently acidicPR proteins, suggesting that wound signals cross with pathogensignaling pathways [Sano et al. (1994) Proc. Natl. Acad. Sci.USA 91: 10556]. This unusual signal crossing is associated witha highly sensitive wound-response of transgenic plants which,upon wounding, produce JA at least eighteen hours earlier thanwild-type plants. When wildtype plants are wounded in the presenceof the synthetic cytokinin, benzylaminopurine, production ofJA begins six hours earlier than in untreated samples, and alsoSA begins to accumulate. The cytokinin antagonist, 2-chloro-4-cyclohexylamino-6-ethylamino-s-triazine,erases these effects. Because transgenic plants constitutivelyproduce four-to six-fold higher amounts of endogenous cytokininsthan wild-type plants, it is concluded that cytokinins are indispensablefor control of endogenous levels of SA and JA. (Received April 23, 1996; Accepted June 10, 1996)     

无花果花芽分化与内源激素含量的关系

在‘布兰瑞克’无花果花芽分化形态学研究的基础上,对花芽分化期无花果新梢第7或第8节位花芽中的玉米素核苷(ZRs)、脱落酸(ABA)、赤霉素(GA1 3)、生长素(IAA)4种内源激素含量的变化进行了探讨。结果表明,在无花果花芽分化阶段,GA1 3和IAA初期含量较高,后快速下降,后期稳定在较低水平;ZRs和ABA在初期含量较低,后大幅提高,后期稳定在较高水平。可见,较高水平的内源ZRs、ABA和较低水平的内源GA1 3、IAA,以及较高的ABA/IAA、ABA/GA1 3、ZRs/GA1 3和ZRs/IAA比值有利于无花果花芽分化。     

无花果花芽分化与植物激素含量的关系

在对布兰瑞克无花果花芽分化形态学研究的基础上,对花芽分化期无花果新梢第7或第8节位芽中植物激素的玉米素核苷（ZR）、脱落酸（ABA）、赤霉素（GA1＋3）、生长素（IAA）含量的变化进行了研究。结果表明,在花托和小花形成阶段,芽内需要较高水平的ZR、ABA和较低水平的GA1＋3、IAA。在花托和小花的形成阶段,ABA/IAA,ABA/GA1＋3,ZR/GA1＋3和ZR/IAA都有较高的比值。     

Dynamics of Endogenous Cytokinins during the Growth Cycle of a Hormone-Autotrophic Genetic Tumor Line of Tobacco

The profile of endogenous cytokinins in a genetic tumor line of tobacco, namely, Nicotiana glauca (Grah.) × Nicotiana langsdorffii (Weinm.), following 1 to 10 weeks of growth on solid medium was determined by radioimmunoassay. ³H-labeled cytokinins of high specific activity were added during tissue extraction to correct for the purification losses. Following subculture (of 4-week-old tissues when their cytokinin content is high) onto fresh medium the total cytokinin content continued to be high during the first week (1470 picomoles per gram fresh weight) when the tissue fresh weight remained essentially unchanged (lag phase). The cytokinin levels then declined by about half in 2- and 3-week-old tissues (626 and 675 picomoles per gram fresh weight, respectively), a period when rapid increase in tissue fresh weight was recorded. Increments of 840% and 2780% over initial fresh weight were obtained in 2- and 3-week-old cultures, respectively. The cytokinin content then increased to initial high levels in 4-week-old tissues (1384 picomoles per gram fresh weight) after which it gradually declined with tissue age. The lowest cytokinin levels (432 picomoles per gram fresh weight) were observed in 10-week-old tissues. Maximal tissue fresh weight (4030% increase over initial fresh weight) was recorded in 5-week-old cultures after which it decreased slowly to 77.5% of the highest tissue fresh weight in 10-week-old cultures. Zeatin appeared to be the dominant endogenous cytokinin in tissues of all ages. Other cytokinins quantified were dihydrozeatin, zeatin riboside, and dihydrozeatin riboside; the values may include contributions from aglucones derived from the hydrolysis of corresponding O-glucosides, since the entire basic fraction was treated with β-glucosidase before analysis. In addition the levels of isopentenyladenine, isopentenyladenosine, and the nucleotides of zeatin riboside, dihydrozeatin riboside, and isopentenyladenosine were also determined.     

Flower Bud Atrophy in Baccara Roses

Changes in gibberellin content and of the production of ethylene in the two upper shoots of roses were measured as affected by decreasing temperature and light intensity, factors which encourage flower atrophy. Decreases in temperature or light intensity to the whole plant reduce the endogenous content of gibberellin in the leaves of the two shoots. The decrease is more rapid and occurs earlier in leaves of the second shoot from the top, where “blindness” is more common. There was also a drop in the gibberellin content, when individual shoots were shaded, although the decrease was more moderate. A lowered light intensity reduced the production of endogenous ethylene by the two shoots, but in the second shoot this production increased after a number of days, before atrophy of the flower bud took place. It is suggested that gibberellin participates in the endogenous control of rose flower development, and the possibility is discussed that it acts by directing the translocation of metabolites to the flower bud.