Sucrose‐cleaving enzymes and carbohydrate pools in Lilium longiflorum floral organs

The activities of soluble invertase (EC 3.2.1.26), cell wall invertase (EC 3.2.1.26) and sucrose synthase (EC 2.4.1.13) were determined in Easter lily ( Lilium longiflorum Thunb. cv. Nellie White) floral organs during flower development. These enzyme activities were correlated with dry weight gains and carbohydrate pools to investigate the importance of their expression in maintaining sink strength of floral organs. In the early stages of flower bud development, anthers exhibited the highest rates of dry weight gain and activity of sucrolytic enzymes. Once anther growth was completed, the dry weight gain of tepal, filament, stigma and style increased with a concomitant increase in hexose concentrations and invertase activity. Although all three enzymes capable of catalyzing sucrose cleavage were present in every flower organ of L. longiflorum , soluble invertase was the predominant enzyme in all flower organs except stigma where cell wall invertase dominated. Soluble invertase activity was highly correlated with dry weight gain in most of the flower organs.     

The Effect of the Fruit and Exogenous Hormones on Leaf Expansion and Composition in Citrus

Sanz, A., Martinez Cortina, C. and Guardiola, J. L. 1987. Theeffect of the fruit and exogenous hormones on leaf expansionand composition in Citrus.—J. exp. Bot. 38: 2033-2042. The effect of the developing flowers and fruitlets on leaf expansionand composition has been determined in leafy inflorescencesof Citrus sinensis L. Osbeck. During leaf expansion the developingflowers do not compete with the leaves, and their early removaldoes not affect leaf size and composition. Competition for mineralelements is established after flower opening, once leaf expansionis complete. No effect of the fruit on metabolizable carbohydratesin the leaves was found up to day 22 after flower opening. The response to exogenously applied growth regulators suggeststhat the differences in weight and composition between inflorescenceleaves and leaves from vegetative sprouts may be due to hormonally-mediateddifferences in sink strength. Gibberellic acid enhances laminaexpansion and increases the sink strength of the leaves. Kinetinenhances lamina expansion without affecting sink strength. Leafgrowth is directly related to both soluble and wall-bound acidinvertase activities; however, the GA₃ effects on leaf growthand sink strength are unrelated to invertase activity and tothe rate of starch accumulation. Key words: Acid invertase, amylase, carbohydrates, Citrus, leaf growth     

Growth, sucrose synthase, and invertase activities of developing Phaseolus vulgaris L. fruits

Activities of the sucrose-cleaving enzymes, acid and neutral invertase and sucrose synthase, were measured in pods and seeds of developing snap bean (Phaseolus vulgaris L.) fruits, and compared with ¹⁴C-import, elongation and dry weight accumulation. During the first 10 d post-anthesis, pods elongated rapidly with pod dry weight increase lagging behind by several days. The temporal patterns of acid invertase activity and import coincided closely during the first part of pod development, consonant with a central role for this enzyme in converting imported sucrose during pod elongation and early dry weight accumulation. Later, sucrose synthase became the predominant enzyme of dry weight accumulation and was possibly associated with the development of phloem in pod walls. Sucrose synthase activity in seeds showed two peaks, corresponding to two phases of rapid import and dry weight accumulation; hence, sucrose synthase was associated with seed sink growth. Acid invertase activities in seeds were low and did not show a noticeable relationship with import or growth. All neutral invertase activities, during pod and seed development, were too low for it to have a dominant role in sucrose cleavage. Changes in activities of certain sucrose-cleaving enzymes appear to be correlated with certain sink functions, including import, storage of reserves, and biosynthetic activities. The data supports the association of specific sucrose-cleaving enzymes with the specific processes that occur in the developing pods and seeds of snap bean fruits; for example, acid invertase with pod elongation and sucrose synthase with fruit dry matter accumulation.     

Invertase Activity, Soluble Carbohydrates and Inflorescence Development in the Tomato (Lycopersicon esculentum Mill.)

The concentration of reducing sugars in the developing firstinflorescence of the tomato (Lycopersicon esculentum Mill.)increased steadily between the macroscopic appearance of theflower buds and the initial stages of fruit expansion. Overthis period sucrose concentrations remained relatively constant.The rise in reducing sugar concentration was accompanied byan increase in the activity of an acid invertase. In individualflower buds invertase activity rose to a maximum shortly beforeanthesis and declined sharply as the anthers dehisced. Increased planting densities and removal of source leaves reducedthe rate of dry matter accumulation by the first inflorescenceand increased the incidence of flower bud abortion. These changeswere correlated with reductions in reducing sugar concentrations,in reducing sugar/sucrose ratios and in acid invertase levels.Removal of young leaves at the shoot apex significantly increasedthe relative growth rate of the inflorescence and led to a substantialincrease in its invertase content. These treatments had relativelylittle effect on sucrose concentration in the inflorescence. The data are consistent with the operation of an invertase-mediatedunloading mechanism for transported sucrose at sinks in theflower buds. It is suggested that the retarded development ofthe first inflorescence and the high incidence of flower budabortion observed under conditions of reduced photoassimilateavailability are causally related to the decline in invertaseproduction in the flower buds. Possible mechanisms for the regulationof invertase synthesis in the flowers are discussed. Lycopersicon esculentum Mill, tomato, inflorescence development, invertase, sink activity     

A novel role of BELL1-like homeobox genes, PENNYWISE and POUND-FOOLISH, in floral patterning

Flowers are determinate shoots comprised of perianth and reproductive organs displayed in a whorled phyllotactic pattern. Floral organ identity genes display region-specific expression patterns in the developing flower. In Arabidopsis, floral organ identity genes are activated by LEAFY (LFY), which functions with region-specific co-regulators, UNUSUAL FLORAL ORGANS (UFO) and WUSCHEL (WUS), to up-regulate homeotic genes in specific whorls of the flower. PENNYWISE (PNY) and POUND-FOOLISH (PNF) are redundant functioning BELL1-like homeodomain proteins that are expressed in shoot and floral meristems. During flower development, PNY functions with a co-repressor complex to down-regulate the homeotic gene, AGAMOUS (AG), in the outer whorls of the flower. However, the function of PNY as well as PNF in regulating floral organ identity in the central whorls of the flower is not known. In this report, we show that combining mutations in PNY and PNF enhance the floral patterning phenotypes of weak and strong alleles of lfy, indicating that these BELL1-like homeodomain proteins play a role in the specification of petals, stamens and carpels during flower development. Expression studies show that PNY and PNF positively regulate the homeotic genes, APETALA3 and AG, in the inner whorls of the flower. Moreover, PNY and PNF function in parallel with LFY, UFO and WUS to regulate homeotic gene expression. Since PNY and PNF interact with the KNOTTED1-like homeodomain proteins, SHOOTMERISTEMLESS (STM) and KNOTTED-LIKE from ARABIDOPSIS THALIANA2 (KNAT2) that regulate floral development, we propose that PNY/PNF-STM and PNY/PNF-KNAT2 complexes function in the inner whorls to regulate flower patterning events.     

Sucrose Metabolism at Three Leaf Development Stages in Bean Plants

Sucrose metabolism was studied at three leaf development stages in two Phaseolus vulgaris L. cultivars, Tacarigua and Montalban. The changes of enzyme activities involved in sucrose metabolism at the leaf development stages were: (1) Sink (9-11 % full leaf expansion, FLE): low total sucrose phosphate synthase (SPS) activity, and higher acid invertase (AI) activity accompanied by low sucrose synthase (SuSy) synthetic and sucrolytic activities. (2) Sink to source transition (40-47 % FLE): increase in total SPS and SuSy activities, decrease in AI activity. (3) Source (96-97 % FLE): high total SPS activity, increased SuSy activities, decreased AI activity. The hexose/sucrose ratio decreased from sink to source leaves in both bean cultivars. The neutral invertase activity was lower than that of AI; it showed an insignificant decrease during the sink-source transition. This revised version was published online in June 2006 with corrections to the Cover Date.     

Floral organ growth and carbohydrate content during pollen development in Lilium

In order to understand floral sugar physiology, we correlated the growth of the organs with carbohydrate content in the flower of Lilium cv. “enchantment” during pollen development. In a previous work, we distinguished two phases in pollen ontogenesis: the anther growth phase, from the microspore mother cell until the vacuolated microspore, and the anther maturation phase, from the vacuolated microspore until anthesis. In the present work, we showed that during the growth phase, the anther underwent most of its size and dry weight growth, whereas the growth rate of nonanther organs was reduced. Anther and filament possessed the highest amounts of carbohydrates, which decreased progressively until the vacuolated microspore stage. During the maturation phase, sucrose and starch increased in all floral organs. Anther growth was completed at the Mi stage, whereas the nonanther organs began exponential growth. From these observations, we concluded that hierarchic nutritional correlations exist between the flower organs, in which the anther is the main actor: during the anther growth phase, the anther represents the highest sink strength floral organ, and mainly attracts assimilates through the filament. During the anther maturation phase, anther growth is achieved, its needs decrease, and assimilates are thus available for neighboring organs, which undergo intense growth until anthesis.     

A morphological study of flower initiation and development in strawberry (Fragaria x ananassa) using cryo-scanning electron microscopy

The morphological changes in the apex of strawberry (Fragaria x ananassa) cv. Elsanta during flower initiation and early development were studied by means of apical dissections and cryo-scanning electron microscopy. Characteristic stages of development were recorded from the earliest discernible evidence of floral initiation until anthesis. The results are discussed in relation to previous studies of floral development in strawberry.     

蝴蝶兰花发育的分子生物学研究进展

蝴蝶兰花非常独特且高度进化,如萼片瓣化、瓣片特化为唇瓣、雌雄蕊合生成合蕊柱及子房发育须由授粉启动等,是单子叶植物花发育研究的理想材料。近年来蝴蝶兰花发育分子生物学取得了重要进展。该文就近年来国内外有关蝴蝶兰开花转换及花器官发育相关基因研究以及B类基因与兰花花被的进化发育关系方面的研究进展进行综述。研究表明:MADS基因在蝴蝶兰开花转换及花器官发育过程中起重要作用,推测其中的DEF(DE-FICIENS)-like基因早期经过2轮复制,形成了4类不同的DEF-like基因,进而决定兰花花被属性。蝴蝶兰花发育分子生物学的深入研究,将极大地利于通过基因工程手段提高蝴蝶兰花品质如花色改良及花期调控等,推动分子育种进程。     

Floral development in bolting garlic

Garlic (Allium sativum L.) is a completely sterile plant, propagated only vegetatively. The aim of this research was to study the sequence of morphological processes occurring during floral initiation and development of a number of bolting garlic accessions from the Allium gene bank in Israel by using SEM. The garlic inflorescence is an umbel-like flower arrangement, the branches (flower clusters) of which arise from a common meristem. The numerous flowers have a distinct morphology typical of the genus Allium. Flower-stalk elongation precedes the swelling of the apical meristem and its subdivision into several centers of floral development. Within clusters, floral primordia develop unevenly. Differentiation of topsets begins after floral differentiation on the peripheral part of the apical surface, and their size, number and rate of development vary among genotypes. At least four morphological types differing in flower/topset ratio were distinguished among the 12 clones studied in this investigation. For further studies of flowering physiology and fertility restoration, only clones which can differentiate the greatest proportion of normal flowers and the least of topsets in the apical meristem should be selected. Received: 28 June 2000 / Revision accepted: 6 November 2000     

The Effect of Gibberellic Acid on Starch Breakdown in Sprouting Tubers of Solanum tuberosum L.

The treatment of potato tubers with 150 µmol dm^–3gibberellic acid (GA₃) stimulated starch breakdown and hexoseaccumulation in tuber tissues and the transfer of dry matterto stems. These effects could not be accounted for by enhancedactivities of starch phosphorylase, amylase and acid invertase.Indeed enzyme activities either declined or remained relativelyconstant as starch degradation and hexose accumulation proceeded.Changes in the rate of starch depletion were related to changesin sink strength and sink type, the onset of tuber initiationin controls causing the rate of starch degradation to exceedthat in GA₃-treated tissues, in which tuberization was inhibited. Solanum tuberosum L., gibberellic acid, starch breakdown     

Large flowers tend to be short-lived in Mediterranean ecosystems: Insights from three Cistus species

Larger and longer lived flowers receive more pollinators, but may also involve increased water maintenance costs under hot, dry environments. Hence, smaller and/or short-lived flowers may buffer such costs. We surveyed floral longevity in three large-flowered Mediterranean Cistus species. We hypothesize that: (1) in Cistus, floral longevity decreases with increasing air temperature and flower size; (2) in C. ladanifer, flower size and longevity increase along an altitudinal gradient; (3) floral longevity is differentially affected by temperature rather than flower size along the gradient; (4) under similar temperature, floral longevity decreases with flower size. For each species, we evaluated the effects of flower size and air temperature on floral longevity. Specifically, floral longevity was surveyed along an altitudinal gradient in the largest flowered species Cistusladanifer. Floral longevity in Cistus species lasted < 1 d and was affected by air temperature, independently of flower size. In C. ladanifer, flower size increased along the gradient but floral longevity decreased. Still, floral longevity decreased with increasing air temperature and, to a lesser extent, with flower size. Together, our findings show a triangular relationship among air temperature, flower size and floral longevity with margins for plasticity to accommodate pollinator attraction with the costs of large-flowered Mediterranean plants.     

Roles of gibberellins in increasing sink demand in Japanese pear fruit during rapid fruit growth

Our previous work demonstrated that exogenous gibberellins (GAs) applications during rapid fruit growth significantly increases sink demand and results in a larger fruit in Japanese pear. In an attempt to unravel the mechanism of increased sink demand by applied GAs, the histology, cell wall components of the flesh, and carbon accumulation in the fruit were assessed for Japanese pear (Pyrus pyrifolia, cultivar ‘Kousui’), as were the activities of sucrose- and sorbitol-cleaving enzymes. Our results show that most vascular tissues occurred in core tissue with very little vascular tissue in the flesh. Application of a mixture of GA₃ + GA₄ in lanolin paste significantly increased the amount of ethanol-insoluble solids, e.g., total pectins, hemicellulose, and cellulose in the cell walls. There was a significantly increased sink demand (assessed by ¹³C accumulation in the fruit) by the applied GAs, and this increased sink strength was closely related to increased activities of cell wall-bound invertase in the core, neutral invertase and NAD-dependent sorbitol dehydrogenase in the flesh during rapid fruit growth. As well, concentrations of sorbitol and sucrose in the flesh were decreased by GA application, while glucose concentration increased. Most importantly, the fact that sink activity can be increased by GA application implies that endogenous GAs are likely to be important modulators for sugar metabolism. Hence, selecting for genotypes with elevated GA production in the growing fruit and increased activities of key enzymes for sugar metabolism could result in increased fruit size.     

Partitioning of [14C]sucrose and Acid Invertase Activity in Reproductive Organs of Pepper Plants in Relation to Their Abscission Under Heat Stress

The effect of heat stress on processes related to carbohydratepartitioning was investigated in young bell pepper (Capsicumannum L. cv. Maor) plants in relation to abscission of theirreproductive organs at different stages of development. None of the reproductive organs abscised after 5 d in a normalday/night temperature regime (25/18°C). With a temperatureregime of 35°C day, 25°C night, abscission occurredin only a small portion of the flower buds and none of the flowersand fruitlets. However, when temperatures in the day and nightwere reversed (25/35°C, day/night) all the buds and someof the flowers abscised during that time period. The young fruitat the first node did not abscise under any temperature regime.The abscission rate of the flower buds was reduced under heatstress if the developing fruit at the first node had been removed. High temperature during either the light or dark periods reducedthe export of [¹⁴C]sucrose from the source leaf (fed for 48h with [¹⁴C]sucrose). Both heat stress and fruit presence reduced the relative amountof [¹⁴C]sucrose which was exported to the flower buds, flowersand roots. Likewise, these treatments reduced the concentrationof reducing sugars in the reproductive organs. Concomitantly,the heat stress and fruit presence on the first node reducedthe activity of soluble acid invertase in the flower buds andthe roots, but not in young leaves. Overall, the results show that heat stress causes alternationin sucrose distribution in the plant, but may also have specificeffects on metabolic activities related to sucrose import andutilization in flower buds and flowers which in turn may enhancetheir abscission. Bell pepper, (Capsicum annuum L. cv. Maor), abscission, acidinvertase, heat stress, reproductive organs, sink leaves. Bell pepper, (Capsicum annuum L. cv. Maor), abscission, acid invertase, heat stress, reproductive organs, sink leaves.     

Sucrose and starch catabolism in the anther of <Emphasis Type="Italic">Lilium</Emphasis> during its development: a comparative study among the anther wall,locular fluid and microspore/pollen fractions

In order to better understand the various pathways of sucrose and starch catabolism in the anther of lily (Lilium hybrida var. “Enchantment”), invertase (EC 3.2.1.26) and amylase (EC 3.2.1.1, EC 3.2.1.2) activities were measured separately in different fractions (anther wall, locular fluid and microspore/pollen) and correlated with the sugar content during anther development. Our findings showed significant differences among the fractions analyzed, suggesting that the regulation of sucrose and starch catabolism could follow distinct pathways in each fraction. Glucose and fructose amounts progressively decreased from anther wall to fluid and from fluid to microspore/pollen. Thus, the developing pollen could act as a sink for the carbohydrates that reach the anther. In this sense, cell wall-bound invertases seem to play a major role in soluble sugar partitioning in the different fractions of the anther. Sucrose concentration was found to be substantially higher in the locular fluid than in the other fractions, indicating a probable site for storage. On the other hand, the anther wall tissues could have a buffering function, storing nutrient surplus in starch grains and thus regulating the availability of soluble sugars in the whole anther. All these results proved the advantages of the experimental model proposed here, as well as its usefulness to investigate sugar metabolism in Lilium anthers.     

Post-translational inhibitory regulation of acid invertase induced by fructose and glucose in developing apple fruit

Acid invertase (EC 3.2.1.26) is one of the key enzymes involved in the carbohydrate sinkorgan development and the sink strength modulation in crops. The experiment conducted with ‘Starkrimson’ apple (Malus domestica Borkh) fruit showed that, during the fruit development, the activity of acid invertase gradually declined concomitantly with the progressive accumulation of fructose, glucose and sucrose, while Western blotting assay of acid invertase detected a 30 ku peptide of which the immuno-signal intensity increased during the fruit development. The immunolocalization via immunogold electron microscopy showed that, on the one hand, acid invertase was mainly located on the flesh cell wall with numbers of the immunosignals present in the vacuole at the late stage of fruit development; and on the other hand, the amount of acid invertase increased during fruit development, which was consistent with the results of Western blotting. The in vivo preincubation of fruit discs with soluble sugars showed that the activity of extractible acid invertase was inhibited by fructose or glucose, while Western blotting did not detect any changes in apparent quantity of the enzyme nor other peptides than 30 ku one. So it is considered that fructose and glucose induced the post-translational or translocational inhibitory regulation of acid invertase in developing apple fruit. The mechanism of the post-translational inhibition was shown different from both the two previously reported ones that proposed either the inhibition by hexose products in the in vitro chemical reaction equilibrium system or the inhibition by the proteinaceous inhibitors. It was hypothesized that fructose and glucose might induce acid invertase inhibition by modulating the expression of some inhibition-related genes or some structural modification of acid invertase.     

Carbon allocation during fruiting in Rubus chamaemorus

Background and Aims

Rubus chamaemorus (cloudberry) is a herbaceous clonal peatland plant that produces an extensive underground rhizome system with distant ramets. Most of these ramets are non-floral. The main objectives of this study were to determine: (a) if plant growth was source limited in cloudberry; (b) if the non-floral ramets translocated carbon (C) to the fruit; and (c) if there was competition between fruit, leaves and rhizomes for C during fruit development.

Methods

Floral and non-floral ramet activities were monitored during the period of flower and fruit development using three approaches: gas exchange measurements, ¹⁴CO₂ labelling and dry mass accumulation in the different organs. Source and sink activity were manipulated by eliminating leaves or flowers or by reducing rhizome length.

Key Results

Photosynthetic rates were lower in floral than in deflowered ramets. Autoradiographs and ¹⁴C labelling data clearly indicated that fruit is a very strong sink for the floral ramet, whereas non-floral ramets translocated C toward the rhizome but not toward floral ramets. Nevertheless, rhizomes received some C from the floral ramet throughout the fruiting period. Ramets with shorter rhizomes produced smaller leaves and smaller fruits, and defoliated ramets produced very small fruits.

Conclusions

Plant growth appears to be source-limited in cloudberry since a reduction in sink strength did not induce a reduction in photosynthetic activity. Non-floral ramets did not participate directly to fruit development. Developing leaves appear to compete with the developing fruit but the intensity of this competition could vary with the specific timing of the two organs. The rhizome appears to act both as a source but also potentially as a sink during fruit development. Further studies are needed to characterize better the complex role played by the rhizome in fruit C nutrition.Key words: Allocation pattern, ¹⁴C labelling, carbon translocation, carbon reserves, cloudberry, defoliation, fruit production, gas exchange, Rubus chamaemorus, source–sink relationship, flowering     

The developmental and organ specific expression of sucrose cleaving enzymes in sugar beet suggests a transition between apoplasmic and symplasmic phloem unloading in the tap roots

Sucrose utilisation in sink tissues depend on its cleavage and is mediated by two different classes of enzymes, invertase and sucrose synthase, which determine the mechanism of phloem unloading. Cloning of two extracellular (BIN35 and BIN46) and one vacuolar invertase (BIN44) provided the basis for a detailed molecular analysis of the relative contribution of the sucrose cleaving enzymes to the sink metabolism of sugar beets (Beta vulgaris) during development. The determination of the steady state levels of mRNAs has been complemented by the analysis of the corresponding enzyme activities. The present study demonstrates an inverse regulation of extracellular invertase and sucrose synthase during tap root development indicating a transition between functional unloading pathways. Extracellular cleavage by invertase is the dominating mechanism to supply hexoses via an apoplasmic pathway at early stages of storage root development. Only at later stages sucrose synthase takes over the function of the key sink enzyme to contribute to the sink strength of the tap root via symplasmic phloem unloading. Whereas mRNAs for both extracellular invertase BIN35 and sucrose synthase were shown to be induced by mechanical wounding of mature leaves of adult plants, only sucrose synthase mRNA was metabolically induced by glucose in this source organ supporting the metabolic flexibility of this species.     

Variability of Flavonol Contents during Floral Morphogenesis in the Poppy Papaver somniferum L.

We studied the contents of flavonols (kaempferol and quercetin) in the meristem of vegetative and generative apices of the main plant shoot in floral Papaver somniferum L. mutants, as well as in the normal plants at successive stages of flower development. Five stages of flower development were distinguished. Flavonols (kaempferol and quercetin) were present in all flower organs at all stages of floral morphogenesis we studied. However, their contents and distribution in different organs and at different stages of flower development markedly varied. No significant differences were found in the contents of flavonols in the meristems of vegetative and generative apices of the main shoot in the lines of floral mutants, as well as between the lines with different amounts of vegetative phytomeres. In the plants with normal flower structure, the contents of flavonols (kaempferol + quercetin) sharply increased with the beginning of differentiation of flower organs, i.e. from stage 3, to reach a maximum in the open flower, when gametogenesis is terminated and fertilization takes place. The level of flavonol contents in the petals (upper part) and stamen was at a maximum at all stages of flower development, while that in the gynaecium was at a minimum. The kaempferol : quercetin ratio was shifted towards quercetin at successive stages of flower development, most significantly in the stamens. The involvement of flavonols in the regulation of floral morphogenesis at stages of flower organs differentiation and functioning is discussed.