Identification of Genes Associated with Chlorophyll Accumulation in Flower Petals

Plants have an ability to prevent chlorophyll accumulation, which would mask the bright flower color, in their petals. In contrast, leaves contain substantial amounts of chlorophyll, as it is essential for photosynthesis. The mechanisms of organ-specific chlorophyll accumulation are unknown. To identify factors that determine the chlorophyll content in petals, we compared the expression of genes related to chlorophyll metabolism in different stages of non-green (red and white) petals (very low chlorophyll content), pale-green petals (low chlorophyll content), and leaves (high chlorophyll content) of carnation (Dianthus caryophyllus L.). The expression of many genes encoding chlorophyll biosynthesis enzymes, in particular Mg-chelatase, was lower in non-green petals than in leaves. Non-green petals also showed higher expression of genes involved in chlorophyll degradation, including STAY-GREEN gene and pheophytinase. These data suggest that the absence of chlorophylls in carnation petals may be caused by the low rate of chlorophyll biosynthesis and high rate of degradation. Similar results were obtained by the analysis of Arabidopsis microarray data. In carnation, most genes related to chlorophyll biosynthesis were expressed at similar levels in pale-green petals and leaves, whereas the expression of chlorophyll catabolic genes was higher in pale-green petals than in leaves. Therefore, we hypothesize that the difference in chlorophyll content between non-green and pale-green petals is due to different levels of chlorophyll biosynthesis. Our study provides a basis for future molecular and genetic studies on organ-specific chlorophyll accumulation.     

杂交兰花瓣类黄酮成分分析及其对花色的影响

为探究杂交兰花色形成的物质基础,以7个杂交兰品种为研究材料,采用显色反应、紫外 可见光谱方法初步判断花瓣中色素的类型;利用超高效液相色谱串联质谱(UPLC MS/MS)技术分析黄色系品种‘玉凤’和红色系品种‘福韵红霞’中代谢物组分及其含量差异,为探索杂交兰花色形成机理和兰花育种提供理论依据。结果表明：(1)7个品种均含有黄酮类化合物,不含类胡萝卜素,除‘玉凤’和‘双艺雅凤’外均含有花青素。(2)在代谢物中检测到160种差异代谢物,显著富集在花青素生物合成途径与黄酮和黄酮醇生物合成途径。(3)矢车菊素衍生物和芍药素衍生物是红色系杂交兰‘福韵红霞’的主要花色素成分;槲皮素类衍生物是黄色系‘玉凤’的主要花色素成分,推测花青素和黄酮醇生物合成的分流是杂交兰花色变化的主要原因。     

The Differentiation of Pigmentation in Flower Parts, IV. Flavonoid Elaborating Enzymes From Petals of Impatiens balsamina s

Extracts of the flower petals of Impatiens balsamina L. contain enzymes which catalyze the glycosylation of phenolic compounds. Enzymes have been extracted which glycosylate hydroquinone to arbutin and at least 3 different flavonols to the 3-monoglucoside. The hydroquinone glucosylating enzyme is similar to enzymes previously described except that it requires an unidentified low molecular weight cofactor. The glucosylation of flavonols follows normal enzyme kinetics; it requires a nucleotide diphosphate glucose donor for activity, and is made more evident by the presence of glucono-1:5-lactone, an inhibitor of endogenous glucosidases. It is suggested that the flavonol glucosylating enzyme acts naturally to glucosylate a precursor of both flavonols and anthocyanins to the 3-monoglucoside. The only elaboration of an anthocyanin observed with petal extracts was an acylation of pelargonidin-3-monoglucoside.     

金银花花形基因的发掘及生物信息学分析

金银花作为我国重要的中药材,具有消炎、抗菌、抗病毒、抗氧化、防癌等多种功效。随着金银花市场供需矛盾日益加剧,通过分子标记辅助选择育种方法来培育高产优质品种势在必行。通过NCBI的Blast工具扫描金银花蛋白组数据发掘花形候选基因,并执行候选基因的亲缘关系分析、结构域分析、表达模式分析、理化性质分析、蛋白质结构预测等一系列生物信息学分析。依据拟南芥调控花形的ABE类基因,通过NCBI-Blast工具扫描金银花氨基酸序列,筛选出包含MADS结构域的8个调控花形的金银花候选基因。经LjaFGD表达模式分析发现,金银花的花中GWHGAAZE016592和GWHGAAZE014905表达量显著高于其他部位,可能正向调控金银花花形。GWHGAAZE014905是一个包含MADS结构域的调控花器官发育的B类基因;GWHGAAZE016592是AP3同源基因。生物信息学分析发现,GWHGAAZE016592和GWHGAAZE014905均是稳定的亲水蛋白,属于非分泌蛋白,包括Motif1、Motif3、Motif4、Motif2、Motif6和Motif5,蛋白质三级结构模板为6byy.2.A和4ox0.2.C。GWHGAAZE014905被定位到细胞核上,而GWHGAAZE016592被定位到叶绿体上,且包含1个位于151~173 bp的跨膜螺旋区域,属于膜蛋白。研究结果为分子标记辅助选择方式培育道地高产优质金银花品种提供了基因资源和分子标记。     

Effects of Epidermal Cell Shape and Pigmentation on Optical Properties of Antirrhinum Petals at Visible and Ultraviolet Wavelengths

We used the Mixta+ and mixta- lines of Antirrhinum majus as a model system to investigate the effects of epidermal cell shape and pigmentation on tissue optical properties in the visible and ultraviolet (UV) spectral regions. Adaxial epidermal cells of Mixta+ flowers have a conical-papillate shape; in the mixta- line the cells are slightly domed. Mixta+ cells contained significantly more anthocyanin and other flavonoids than mixta- cells when plants were grown under either high- or low-UV conditions. Mixta+ cells focused light (3.5-4.7 times incident) within their pigmented interiors, whereas mixta- cells focused light (2.1-2.7 times incident) in the unpigmented mesophyll. UV light penetrated the epidermis (commonly 20-50% transmittance at 312 nm) mainly through the unpigmented peripheral regions of the cells that were similar for the two lines, so that overall penetration through Mixta+ and mixta- epidermises was equal. However, maximum UV absorption in the central region of epidermal cells was slightly greater in Mixta+ than mixta-, and intact Mixta+ flowers reflected less light in the spectral regions with intermediate flavonoid absorbance. In both cases, about 50 to 75% of the difference could be attributed to cell shape and resulting changes in the optical pathlength or focusing.     

The Abscission of Rose Petals

Petal abscission was studied in twelve hybrid tea rose (Rosahybrida L.) cultivars. At about 20 °C the time to petalabscission in uncut stems in greenhouses was the same as incut stems placed in water in the greenhouse or in a climate-controlledroom. The time between petal unfolding and abscission dependedon the cultivar, and varied between 12 and 35 d. The time topetal abscission of the cultivars was inversely correlated withtheir flower diameter at full bloom (linear regression, r² =0·82). In the cultivars with a relatively large flowerdiameter (10-18 cm) the petals fell without visible desiccationsymptoms, whereas in the group with a small diameter the petalswere partially or fully desiccated when shed. Fertilization occurred in some flowers of a few cultivars studied.In cultivars with a relatively large flower diameter (Papa Meilland,Cocktail, Dr. Verhage, Tineke) it had no effect on the timeto abscission in Motrea, Europa, and Carolien roses, which bearsmall flowers, the petals fell after fertilization, whereasin unfertilized flowers of the latter group of cultivars anabscission zone just above the uppermost node became activeand all parts above this node (pedicel and flower) turned brownand desiccated, though remained attached for more than a month. It is concluded that in the cultivars investigated: (a) thetime to petal abscission was inversely related to their flowerdiameter, (b) abscised petals were more desiccated in cultivarsin which the time to abscission was longer, (c) fertilizationhad little effect on the time to abscission in most cultivars,whereas the absence of fertilization prevented petal abscissionin a number of the small-diameter cultivars where it was replacedby flower abscission, and (d) cutting and placement in waterat 20 °C did not affect the time to abscission.Copyright1995, 1999 Academic Press Abscission, fertilization, flowers, petals, Rosa hybrida L., rose, water stress, carbohydrate stress     

同株紫斑牡丹中单瓣与重瓣花芽分化及内源激素含量比较北大核心CSCD

该研究以紫斑牡丹‘关公红’为试验材料,通过调查‘关公红’的开花情况,采用光学显微镜、扫描电镜技术考察花芽分化和花瓣发育过程的结构特征,并测定同株‘关公红’单、重瓣花朵的花芽在不同分化阶段内源激素含量,分析单、重瓣花朵的花芽分化及内源激素含量的变化特征,为紫斑牡丹不同花型的培育奠定理论基础。结果表明:(1)‘关公红’单瓣花朵比重瓣花朵花期长,单瓣花朵花瓣表皮细胞光滑平整无明显褶皱,而重瓣花朵花瓣的表皮有明显的褶皱;重瓣花的花芽分化时间明显早于单瓣,花瓣原基的体积也明显增大,这可能与重瓣花中雄蕊瓣化有关系。(2)低浓度GA_(3)有利于‘关公红’单瓣花朵和重瓣花朵的花芽分化;高浓度的ABA促进重瓣花原基的形成;ZR含量增加有助于紫斑牡丹的花芽分化。(3)较高ZR/IAA、ZR/GA_(3)、ABA/GA_(3)、ABA/IAA比值有利于‘关公红’重瓣花朵的花芽分化;ABA/IAA、ZR/IAA比值的增加有利于‘关公红’单瓣花朵花芽分化;(GA_(3)+IAA+ZR)/ABA与(IAA+ZR)/GA_(3)比值减小分别促进重瓣花朵花瓣原基和雄蕊原基的形成。     

Ethylene Binding to Senescing Carnation Petals

Brown, J. H., Legge, R. L., Sisler, E. C, Baker, J E. and Thompson,J. E. 1986. Ethylene binding to senescing carnation petals.—J.exp. Bot 37: 526–534. Ethylene binding to carnation petals is significantly higheron a fresh weight basis for young fully expanded flowers thanfor older flowers showing petal-inrolling symptoms. The peakin ethylene binding precedes the climacteric-like rise in ethyleneproduction even when production of endogenous ethylene is inhibitedby incorporating amino-oxyacetic acid into the petals. FromScatchard analyses of ethylene binding, it has been estimatedthat petals from young fully expanded flowers have 11?10⁴ bindingsites per cell, whereas petals from senescent flowers showingextensive petal inrolling have 0.6?1010⁴ binding sites per cell.It is also apparent from the Scatchard analyses that the affinityof binding sites for ethylene decreases with advancing age ofthe flowers. The decline in number of binding sites with advancingage can be essentially accounted for by the extensive breakdownof membranes that accompanies senescence. However, the decreasedaffinity for ethylene in the older tissue suggests that thebinding sites become altered with advancing senescence. Key words: Carnation, ethylene binding, senescence     

The Complexity of Background Clutter Affects Nectar Bat Use of Flower Odor and Shape Cues

Given their small size and high metabolism, nectar bats need to be able to quickly locate flowers during foraging bouts. Chiropterophilous plants depend on these bats for their reproduction, thus they also benefit if their flowers can be easily located, and we would expect that floral traits such as odor and shape have evolved to maximize detection by bats. However, relatively little is known about the importance of different floral cues during foraging bouts. In the present study, we undertook a set of flight cage experiments with two species of nectar bats (Anoura caudifer and A. geoffroyi) and artificial flowers to compare the importance of shape and scent cues in locating flowers. In a training phase, a bat was presented an artificial flower with a given shape and scent, whose position was constantly shifted to prevent reliance on spatial memory. In the experimental phase, two flowers were presented, one with the training-flower scent and one with the training-flower shape. For each experimental repetition, we recorded which flower was located first, and then shifted flower positions. Additionally, experiments were repeated in a simple environment, without background clutter, or a complex environment, with a background of leaves and branches. Results demonstrate that bats visit either flower indiscriminately with simple backgrounds, with no significant difference in terms of whether they visit the training-flower odor or training-flower shape first. However, in a complex background olfaction was the most important cue; scented flowers were consistently located first. This suggests that for well-exposed flowers, without obstruction from clutter, vision and/or echolocation are sufficient in locating them. In more complex backgrounds, nectar bats depend more heavily on olfaction during foraging bouts.     

RcAGL61基因调控雄蕊和花瓣之间转变影响月季花瓣数量

【目的】为了探究月季重瓣性状的调控机制,课题组前期筛选到一个与花发育相关的AG同源基因RcAGL61,本研究对该基因的功能进行了分析。【方法】利用荧光定量PCR（qRT-PCR）对该基因在‘窄叶藤本月季花’ב月月粉’杂交群体中重瓣株系和单瓣株系花芽5个发育时期的表达模式进行了分析,以重瓣株系和单瓣株系为材料,克隆RcAGL61,并进行生物信息学分析、亚细胞定位及VIGS实验。【结果】（1）该基因表达水平在单瓣株系的五个发育时期均显著高于重瓣株系,在单瓣株系花发育的S4-S5期比S1-S3期表达量明显升高。（2）RcAGL61编码区序列在单瓣株系和重瓣株系中一致,长度为495 bp,与RcAG基因序列相似度为30.75%,编码164个氨基酸,含有一个MADS-Box保守结构域,属于MADS-Box基因家族。（3）亚细胞定位发现RcAGL61蛋白定位于烟草表皮细胞的细胞核。（4）沉默该基因后,瓣化雄蕊数量增加,雄蕊数量减少,花瓣数量增加,萼片数量和雌蕊数量无显著变化。【结论】RcAGL61参与调控了雄蕊原基和花瓣原基间的转变,影响了月季的花瓣数量。     

Physiological Responses of Chrysanthemum Petals during Senescence

Flower senescence was studied in three cultivars of Chrysanthemum coronarium L.: Snowball White, Yellow Chandrama (standard type flowers) and Spray Button (spray type flowers). Spray button flowers exhibited least respiration rate, less efflux of ions, minimum protease activity and less decline in fresh mass, relative water content and total soluble protein content with the progression of senescence. The Snowball White flowers showed highest respiration rate, great efflux of ions, maximum protease activity, high activity of hydrolytic as well as proteolytic enzymes, and high decline in fresh mass, relative water content, and total soluble protein content. Yellow Chandrama flowers showed responses similar to Snowball White flowers.     

An Effect of Ethylene on the Distribution of 14C-sucrose from the Petals to Other Flower Parts in the Senescent Cut Inflorescence of Dianthus caryophyllus

The distribution of carbon-14 in the flower parts of the cutcarnation inflorescence after feeding ¹⁴C-sucrose through thepetals was studied during natural ageing and after ethylenetreatment. Levels of ethylene which caused irreversible wiltingof petals also promoted an accelerated transfer of the radioactivesucrose to the nectar, gynaecium and stem. Since the nectarreceived a relatively large proportion of the radioactive carbon,the composition of the sugars in the nectar and the vascularizationof the nectary were investigated. Sucrose comprised about 85per cent of the nectar sugars and the balance was glucose andfructose. The vascular tissue closest to the nectary consistedof phloem elements; tracheary elements terminated deeper inthe receptacle and were surrounded by a ring of phloem. Thepercentage of solutes in the nectar was about 18 per cent andincreased when the flowering stems were placed in sucrose solutions;the solutes in the nectar were principally sugars. Taken togetherthe results show that the nectary can act as a sink for sucroseand, in the flower at least, that translocation of sucrose takesplace in the phloem. The results provide further evidence forthe hypothesis that ethylene promotes mobilization of substrateand an efflux of material from petals to the gynaecium, nectarand stem.     

The Nature of the White Colour of Petals

Whiteness in flowers has been ascribed to reflection of lightor to the presence of flavones in the tissue. Light reflectionfrom the surface of chrysanthemum petals and the pigment contentof the petals were determined before and after various treatments.After squashing or infiltration of the petals with water, lightreflection was reduced to half or less, whereas flavone contentremained almost unimpaired. Further reductions in light reflectionwere obtained by removing the cuticle. An albino Antirrhinumwas white in colour, yet contained no flavone. These resultsare discussed and it is concluded that whiteness is due to reflectionof light, particularly by the surfaces round air spaces.