Theobroxide induces tubers in potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) and flower buds in morning glory (<Emphasis Type="Italic">Pharbitis nil</Emphasis>) under non-inductive high temperatures

Induction of some plant organs including tubers and flower buds begins with sensing environmental cues, such as photoperiod and temperature in the leaves. Theobroxide has been shown to induce potato tuberization and flower-bud formation in morning glory under non-inductive photoperiodic conditions, stimulating the activity of lipoxygenase (LOX) and the synthesis of jasmonic acid (JA). In the present study, the ability of theobroxide to overcome the inhibitory effect of unfavorable high temperature on the induction of tubers in potato and flower buds in morning glory was examined. Both tuber induction and flower-bud formation under non-inductive high temperatures were promoted by the application of theobroxide at a high concentration. However, although theobroxide treatment resulted in an increase in fresh weight during potato tuber growth at 30°C, morning glory plants treated with theobroxide at 35°C failed to bloom, implying that theobroxide may assist only in flower-bud formation.     

Allene oxide cyclase is essential for theobroxide-induced jasmonic acid biosynthesis in Pharbitis nil

Theobroxide, a natural product, strongly stimulates the biosynthesis of jasmonic acid (JA) in Pharbitis nil. In this study, we investigated the accumulation of protein by the immunoblot analysis of lipoxygenase (LOX), allene oxide synthase (AOS), and allene oxide cyclase (AOC), key enzymes in JA biosynthesis, and how the endogenous levels of JA in P. nil are affected by theobroxide. The effect of JA on the accumulations of these proteins was monitored simultaneously. The results show that theobroxide treatment led to a high level accumulation of JA, which is due to high accumulations of LOX, AOS, and AOC proteins induced by theobroxide treatment both under short day (SD) and long day (LD) conditions. However, under SD conditions AOS and AOC proteins are not enhanced by JA treatment. Kinetic analysis of protein levels shows that a biphasic activation of AOC protein by theobroxide is displayed and the first activation of AOC protein together with elevated JA levels is observed within 30min after treatment. Meanwhile, AOS and LOX proteins are activated by theobroxide later than AOC protein, suggesting that AOC plays an essential role in the initial JA formation induced by theobroxide. Since theobroxide-increased JA levels also show a biphasic manner similar to AOC activation and AOS, LOX proteins are activated later than AOC, and thus we propose a positive JA feedback regulation. Interestingly, AOS protein, which is also the enzyme for the biosynthesis of 9,10-ketol-octadecadienoic acid (KODA, a flowering inducing factor), accumulates markedly due to the simultaneous involvement of theobroxide and SD conditions, suggesting that AOS probably plays a role in flower bud formation in P. nil.     

Effects of theobroxide, a natural product, on the level of endogenous jasmonoids

The natural potato microtuber inducing substance, theobroxide, strongly induces the formation of tuber of potato (Solanum tuberosum L.) and flower bud of morning glory (Pharbitis nil) plants under non-inducing conditions (long days) (Yoshihara et al., 2000). In the present study, theobroxide was evaluated for its effect on the level of endogenous jasmonoids in different tissues of such two plants. An in vitro bioassay using cultures of single-node segments of potato stems was performed with the supplement of theobroxide in the medium. The endogenous jasmonic acid (JA) and its analogue tuberonic acid (TA, 12-hydroxyjasmonic acid) in segments and microtubers were quantitatively analyzed. The increase in the endogenous JA level caused by theobroxide was observed in both segments and microtubers. Endogenous TA was only detected in segments, and the content increased with the concentration of theobroxide. As for morning glory, the whole plant was sprayed with theobroxide for 1 approximately 5 weeks under different photoperiods and endogenous JA in the leaves was quantitatively analyzed. Theobroxide spraying increased the level of endogenous JA in the leaves of the plants grown under both long and short days.     

Inhibition of stem elongation in spinach by theobroxide

In the current study, we investigated the influences of theobroxide on stem elongation in spinach (Spinacia oleracea). Our results showed that stem elongation and flower formation were inhibited by spraying spinach plants with theobroxide under inductive, long day conditions (16 h light/8 h dark), while application of exogenous applied GA3 prevented the effect of theobroxide. Quantitative analysis showed that theobroxide suppressed GA1 biosynthesis, whereas the endogenous content of jasmonic acid was unchanged. However, under short day conditions (10 h light/14 h dark), there were no differences in stem length between treated and untreated plants. These results suggest that the inhibition of stem elongation by theobroxide is probably due to the suppression of gibberellin biosynthesis.     

Floral determination in the terminal bud of the short-day plant Pharbitis nil

Temporal and spatial aspects of floral determination in seedling terminal buds of the qualitative short-day plant Pharbitis nil were examined using a grafting assay. Floral determination in the terminal buds of 6-day-old P. nil seedlings is rapid; by 9 hr after the end of a 14-hr inductive dark period more than 50% of the induced terminal buds grafted onto uninduced stock plants produced a full complement of flower buds. When grafted at early times after the end of the dark period the terminal buds of induced plants produced three discrete populations of plants: plants with no flowers, plants with two axillary flowers at nodes 3 and 4 and a vegetative terminal shoot apex, and plants with five to seven flowers including a terminal flower. The temporal relationship among these populations of plants produced by apices grafted at different times indicates that under our conditions, the region of the terminal bud that will form the axillary buds at nodes 3 and 4 becomes florally determined prior to floral determination of the region of the terminal bud giving rise to the nodes above node 4.     

Relation Between Environmental Factors and the LOX Activities Upon Potato Tuber Formation and Flower-bud Formation in Morning Glory

Life cycles of plants including tuberization and flowering are strongly related to environmental factors such as photoperiod and temperature. Theobroxide induces potato tuber formation and flower bud formation of morning glory under non-inductive conditions and stimulates the activity of lipoxygenase (LOX). In this study, to understand the LOX activity more systematically, the relationships between LOX activity and light and temperature, which effects potato tuber and flower-bud formation, have been investigated. The results showed that LOX activity in morning glory was greatly enhanced up to 30 min and then declined after switching from the light to the dark condition, while the activity did not vary when switching from the dark to the light condition. In addition, the temperature profile of measured LOX activity in the potato and morning glory plants was nearly consistent with the time taken to form potato tubers and flower buds in morning glory, respectively, at different growing temperatures. These results strongly suggest that LOX activity is directly connected with light and temperature to regulate the formation of tubers and flower-buds.     

Inhibitory Effect of Methyl Jasmonate on Flowering and Elongation Growth in Pharbitis nil

The effect of methyl jasmonate (JA-Me) on the floral bud formation and elongation growth in the short-day plant Pharbitis nil was investigated. The placing of 4-day-old seedlings of P. nil in a solution of JA-Me for a period of 24 h before an inductive (16 h or 14 h of darkness) night led to a dramatic reduction in the number of flower buds formed by the plant. Plants treated with JA-Me also totally lost their capacity to form a generative terminal bud. JA-Me applied after photoinduction does not inhibit flowering. Gibberellic acid (GA3) partly reverses the inhibitory effect of JA-Me. Plants treated simultaneously with JA-Me and GA3 formed about 3 flower buds more than plants treated with JA-Me only. JA-Me at a concentration of 10-7 M stimulates slightly, but at higher concentrations it inhibits root growth and shoot growth. A distinct lack of correlation between the effect of JA-Me on inhibition of flowering and shoot and root growth was noted. This indicates the independent action of JA-Me in controlling both processes.     

Inhibitory Effect of Methyl Jasmonate on Flowering and Elongation Growth in Pharbitis nil

The effect of methyl jasmonate (JA-Me) on the floral bud formation and elongation growth in the short-day plant Pharbitis nil was investigated. The placing of 4-day-old seedlings of P. nil in a solution of JA-Me for a period of 24 h before an inductive (16 h or 14 h of darkness) night led to a dramatic reduction in the number of flower buds formed by the plant. Plants treated with JA-Me also totally lost their capacity to form a generative terminal bud. JA-Me applied after photoinduction does not inhibit flowering. Gibberellic acid (GA3) partly reverses the inhibitory effect of JA-Me. Plants treated simultaneously with JA-Me and GA3 formed about 3 flower buds more than plants treated with JA-Me only. JA-Me at a concentration of 10-7 M stimulates slightly, but at higher concentrations it inhibits root growth and shoot growth. A distinct lack of correlation between the effect of JA-Me on inhibition of flowering and shoot and root growth was noted. This indicates the independent action of JA-Me in controlling both processes.     

Ethylene and ABA interactions in the regulation of flower induction in Pharbitis nil

Hormones are included in the essential elements that control the induction of flowering. Ethylene is thought to be a strong inhibitor of flowering in short day plants (SDPs), whereas the involvement of abscisic acid (ABA) in the regulation of flowering of plants is not well understood. The dual role of ABA in the photoperiodic flower induction of the SDP Pharbitis nil and the interaction between ABA and ethylene were examined in the present experiments. Application of ABA on the cotyledons during the inductive 16-h-long night inhibited flowering. However, ABA application on the cotyledons or the shoot apices during the subinductive 12-h-long night resulted in slight stimulation of flowering. Application of ABA also resulted in enhanced ethylene production. Whereas nordihydroguaiaretic acid (NDGA) - an ABA biosynthesis inhibitor - applied on the cotyledons of 5-d-old seedlings during the inductive night inhibited both the formation of axillary and of terminal flower buds, application of 2-aminoethoxyvinylglycine (AVG) and 2,5-norbornadiene (NBD) - inhibitors of ethylene action - reversed the inhibitory effect of ABA on flowering. ABA levels in the cotyledons of seedlings exposed to a 16-h-long inductive night markedly increased. Such an effect was not observed when the inductive night was interrupted with a 15-min-long red light pulse or when seedlings were treated at the same time with gaseous ethylene during the dark period. Lower levels of ABA were observed in seedlings treated with NDGA during the inductive night. These results may suggest that ABA plays an important role in the photoperiodic induction of flowering in P. nil seedlings, and that the inhibitory effect of ethylene on P. nil flowering inhibition may depend on its influence on the ABA level. A reversal of the inhibitory effect of ethylene on flower induction through a simultaneous treatment of induced seedlings with both ethylene and ABA strongly supports this hypothesis.     

The role of ethylene in the flowering response of bulbous plants

Ethylene induces the flower formation and stimulates the flower-bud development of some bulbous plants exposed to the gas when the apex is in the vegetative state. For iris bulbs cv. Ideal maximum responses have been found after exposure to 5 ppm for 8 h; lower concentrations, shorter exposure periods and, depending on seasonal conditions, low temperatures during gas treatment, gave intermediate responses. The effects are opposite to the ethylene induced flower-bud blasting which occurs when bulbous plants are exposed to the gas after completion of the flower formation. Dry storage of the bulbs in atmospheres containing 5% CO₂ reduces the temperature-enhanced flower formation, suggesting a possible effect of endogenous ethylene. Presented at the International Symposium “Plant Growth Regulators” held on June 18–22 1984 at Liblice, Czechoslovakia.     

多胺在植物花发育中的作用

多胺是植物体内一类生理活性物质,其代谢变化与高等植物的花芽形成和发育关系密切。该论文综述了多胺在植物体内的形态和分布,以及成花诱导、花芽形成,尤其是雄性育性的发生和性别分化过程中的作用,并对今后的研究方向进行了讨论。     

Flower bud differentiation in Salix pentandra as affected by photoperiod, temperature and growth regulators

Flower bud initiation in seedlings and vegetatively propagated plants of Salix pentandra from different locations has been studied under controlled conditions. In mature plants flower bud formation was induced by 2-chloroethyltrimethylammoniumchloride (CCC) and by short day treatment. The effect of CCC was antagonized by GA₃. The critical photoperiod for flower bud formation was about 18 h for a southern clone (from 49°48'N), but cuttings of a northern ecotype (from 69°39'N) formed flower buds even at 24 h photoperiod. Generally, flower bud formation occurred simultaneously with apical growth cessation. However, apical growth cessation was not a prerequisite for floral initiation and flower buds were also found in elongating plants. Seedlings up to 60 days old did not form flower buds in growth chamber studies. The length of the juvenile phase has not been studied in detail, but cuttings taken from seedlings approximately 20 cm high and 60 days old were able to develop flower buds when treated with CCC. A gradual transition from the juvenile to the mature phase was obtained by repeated pruning of seedlings grown at 18°C and 24 h photoperiod.     

The Involvement of Cyclic ADPR in Photoperiodic Flower Induction of Pharbitis nil

Cyclic adenosine diphosphate ribose (cADPR) is a potent endogenous calcium-mobilizing agent synthesized from NAD⁺ by ADP-ribosyl cyclases described for several animal cells. Pharmacological studies suggest that cADPR is an endogenous modulator of Ca²⁺-induced Ca²⁺ release channels. There is also information about the sub-micromolar concentration of cADPR in plant cells. Whether cADPR can act as a Ca²⁺-mobilizing intracellular messenger in plant tissue is an unresolved question. Despite the obvious importance of monitoring cADPR cellular levels under various physiological conditions in plants, its measurement has been technically difficult and requires specialized reagents. In the present study a widely applicable sensitivity assay for cADPR is described. We show that Pharbitis nil tissue from cotyledons contains a certain cADPR level. To explain the possible roles of this second messenger in photoperiodic flower induction, some physiological experiments were also performed. The exogenous applications of cADPR to Pharbitis nil plants, which were exposed to a 12-h-long subinductive night, significantly increased flowering response. Nevertheless 8-Br-cADPR inhibited flowering when these compounds were applied during a 16-h-long inductive night. The effect of ruthenium red, a calcium channel blocker and ryanodine, a calcium channel stimulator, on the photoperiodic induction of flowering was also studied. Ruthenium red, when applied before and during an inductive 16-h dark period, slightly inhibited flowering, whereas ryanodine, when applied before and during a 12-h long subinductive night, stimulated flower bud formation. We also confirmed evidence that Ca²⁺ ions are involved in the photoperiodic induction of flowering. Thus, the obtained results may suggest the involvement of cyclic ADPR-activated Ca²⁺ mobilization in the photoperiodic flower induction process in Pharbitis nil.     

Involvement of signal transduction pathway components in photoperiodic flower induction in Pharbitis nil

The possible participation of several major components of the signal transduction pathway in photoperiodic flower induction was examined in Pharbitis cotyledons. Exogenous applications of GTP-γ-S (1–10 μ M ) or of the phorbol ester, phorbol 12-myristate-13-acetate (PMA, 0.1–5.0 μ M ) to Pharbitis plants held under a marginal inductive period (11.5 h dark) significantly increased their flowering response. Membrane lipid fluidity, GTP-binding and protein kinase activity were increased following a single flowering-inducing dark period of 16 h; however, a light-break of 10 min that abolished flower induction failed to reverse the dark-induced increase in these processes. Photo-inductive dark conditions significantly increased the content of diacylglycerol (DAG) and phosphoinositides in the cotyledon membranes, together with the activities of their kinases, and a light break decreased them to control levels and below. In addition, a single spraying with GTP-γ-S or PMA at 1 μ M significantly increased both the lipid content and the kinase activities. These compounds also enhanced the kinase activities in vitro. It is concluded that DAG and phosphoinositide metabolism play a role in the linking of the photoperiodic induction of the phytochrome with the flowering response in Pharbitis nil .     

Photoperiod and temperature effects on in vitro growth and flowering of P. pusilla, an epiphytic orchid.

Psygmorchis pusilla Dodson and Dressler, an epiphytic orchid, has been shown to be an interesting model to study in vitro flower formation. In the present study, the effects of photoperiod and temperature on vegetative and reproductive development were investigated. Although photoperiod had limited effects on leaf number, an etiolating process was verified in darkness and a higher growth was detected under long days. A positive relationship was observed between long days and floral spike formation. However, plant incubation under 20 h photoperiod or longer days negatively affected floral bud development, inhibiting anthesis and reducing flower longevity. Higher soluble sugar and starch levels were detected in plants cultivated under long days, while chlorophyll and carotenoids contents were negatively affected under these conditions. Plants showed great sensitivity to temperature variations; 27 degrees C being the most adequate for growth, leaf and floral spike formation. Temperatures of 22 and 32 degrees C were not appropriate for in vitro development of P. pusilla.     

野生胭脂花(Primula maximowiczii)生长发育和花芽分化研究

以北京东灵山胭脂花自然分布地野生群体中2年生以上植株为研究对象,于2008～2010年连续3年观测了野生胭脂花的年生长发育进程,并采用石蜡切片法制片,通过普通光学显微镜和体视显微镜观察了胭脂花花芽分化和花序发育的过程,以探明野生胭脂花生长发育和花芽分化的规律,为人工栽培胭脂花提供依据。结果显示:(1)胭脂花的年生长发育进程可分为萌芽期、营养生长期、开花期、果实发育期、果熟期、花芽分化期和休眠期等7个阶段;5～9月为胭脂花的生长季,生长环境凉爽,日平均气温为5℃～20℃。(2)胭脂花花芽分化期为7月中下旬～9月上旬,历时约2个月,整个过程包括未分化期、花芽原基分化期、小花原基分化期和小花分化期;花序上的小花由外向内逐渐形成并发育,雌雄蕊的发育从8月中下旬开始到9月上中旬结束;花序发育完全的胭脂花植株进入休眠期,经过当年10月份至来年4月份的低温阶段翌年开花。胭脂花花芽分化进程和外部形态密切相关,可根据植株的外部形态特征快速判定花序发育状况。     

The involvement of cyclic GMP in the photoperiodic flower induction of Pharbitis nil

The involvement of cGMP in the regulation of the flowering of Pharbitis nil was investigated through exogenous applications of cGMP and chemicals that are able to change the cGMP level and analyses of endogenous cGMP level. Exogenous applications of cGMP and 8-pCPT-cGMP (a cyclic GMP non hydrolyzed analog) to P. nil plants, which were exposed to a 12-h-long subinductive night, significantly increased flowering response. NS-2028 (guanylyl cyclase inhibitor) inhibited flowering when that compound was applied during a 16-h-long inductive night, whereas SNP (guanylyl cyclase activator) increased the flowering when plants were subjected to a 12-h-long subinductive night. The inhibitors of cyclic nucleotides phosphodiesterase (isobutyl-methylxanthine and dipyridamole), which increase the cytosolic cGMP level, promoted the flowering and allowed the length of the dark period necessary for induction of flowering to be reduced. The endogenous cGMP level was also measured after the treatment of P. nil seedlings with those chemicals. Results have clearly shown that compounds that were used in physiological experiments modulated endogenous cGMP level. There was a significant difference in the cyclic GMP level between 16-h-long night conditions and a long night with a night-break. During a long inductive night the oscillation of cGMP was observed with four main peaks in 4, 7, 11, 14 h, whereas a 10 min flash of red light in the middle of the night was able to modify these rhythmical changes in the second half of the long night. These results have shown that there are oscillations in the concentration of cGMP in the night and the biosynthesis and/or deactivation of cGMP is affected by light treatment and therefore it may be involved in the regulation of photoinduction processes in cotyledons. From these combined results, we propose a hypothesis that cGMP is involved in the control of photoperiodic flower induction in Pharbitis nil.     

不同激素对锦绣杜鹃的催花作用

在加温、补光条件下,研究赤霉素(GA3)、萘乙酸(NAA)和吲哚乙酸(IAA)处理对锦绣杜鹃花蕾发育、开花期及开花质量等的影响。结果表明:锦绣杜鹃花蕾经0～3000mg.L-1GA3涂抹后,其催花效果随处理浓度增大呈先升后降趋势,并于2000mg.L-1浓度下达到最佳,表现在花蕾发育快、开花期提前及开花质量高等;而0～3000mg.L-1NAA和IAA对锦绣杜鹃催花效果均不明显。锦绣杜鹃花蕾大小(包括长度和宽度)与开花期提前时间和花径之间均呈极显著正相关,与花蕾期则呈极显著负相关;花蕾期与开花期提前时间之间呈极显著负相关(P<0.01)。     

苹果花芽孕育机理的探讨

花芽孕育期,以环剥、多效唑＋环剥和ＧＡ３处理“红富士”苹果树,成花效应以多效唑＋环剥处理为最高（＋６８４％）。花芽孕育起动时,促花处理的短枝顶芽内ＩＡＡ、ＺＲ含量和ＩＡＡ／ＧＡｓ、ＺＲ／ＧＡｓ的比值均明显提高。在花芽孕育完成过程中,促花处理的短枝叶内淀粉含量、Ｃ／Ｎ比,芽内ＺＲ、ＩＡＡ含量和ＺＲ／ＧＡｓ明显提高,ＧＡｓ则明显下降。