DEVELOPMENT OF EXCISED FLORAL BUDS OF AQUILEGIA: THE COCONUT-MILK PROBLEM

In an attempt to define a medium and to investigate the effects of glycine, floral buds of Aquilegia were grown in culture on coconut milk and defined media containing either White's minerals or a new mineral formula, with and without glycine. Development obtained on the best defined medium was 66 to 76% of that achieved on the equivalent coconut-milk medium. The inability of buds to achieve development equal to that on the coconut-milk medium may be partially explained by the existence of a barrier to carpel formation and development. The effect of glycine on floral buds grown on a defined medium was dependent not only upon the minerals used but also on the stage of development of the bud. With White's minerals, glycine inhibited the development of buds in early and intermediate stages but was slightly promotive with buds in later stages. With the new minerals, glycine was slightly promotive with early and intermediate stages of development but was inhibitory with later stages. Further work is necessary before development on a defined medium will equal or exceed development on the coconut-milk medium.     

IN VITRO CULTURE OF FLORAL BUDS OF AQUILEGIA

Tepfer, S. S., R. I. Greyson, W. R. Craig, and J. L. Hindman. (U. Oregon, Eugene.) In vitro culture of floral buds of Aquilegia. Amer. Jour. Bot. 50(10): 1035–1045. Illus. 1963.—Floral buds at various stages of development, from early stages before sepal initiation to late stages with young carpel primordia present, were grown in culture on various agar media. A basic medium containing White's minerals, Nitsch's trace elements, coconut milk, sucrose, and assorted water-soluble vitamins was developed for growth of the buds. The addition of indoleacetic acid, gibberellic acid, and kinetin to the basic medium extended the developmental limits of buds at nearly all stages and decidedly improved the continued development of carpels. On this medium buds produce all of their organ primordia, growing from early stages to about the size of flowers at anthesis, but will not develop unless the sepals are removed. This inhibiting effect of sepals is not understood at this time. Stamen development is consistently poor beyond the point of differentiation of anther and filament, even with the addition of hormones. The development of buds in culture is illustrated and compared with development in intact plants. With further improvement of the medium, it is hoped that these buds may be used for experiments testing theories of floral morphogenesis.     

THE EFFECTS OF HORMONES UPON THE DEVELOPMENT OF EXCISED FLORAL BUDS OF AQUILEGIA

Young excised floral buds of Aquilegia were grown on defined medium containing kinetin, indoleacetic acid (IAA), or gibberellic acid (GA₃). Only when 10⁻⁶ or 10⁻⁷ m kinetin was added to the basal medium was there a significant increase in the number of initiated whorls of primordia. Buds on the basal medium or on medium with IAA or GA₃ failed to initiate carpels. On medium with 10⁻⁶ or 10⁻⁷ m kinetin, buds successfully initiated a normal whorl of five carpels. A high level of inorganic nitrogen was also required for the initiation of carpels. With 10⁻⁵ m kinetin, individual buds initiated from 6–18 carpels. Staminodial primordia of these buds were replaced with carpels, or the floral apex enlarged to accommodate a single whorl of many carpels. Kinetin did not support the further differentiation of the floral organs. Sepals, petals, and carpels did differentiate on medium with GA₃, but stamens aborted. However, on medium with GA₃ and kinetin, stamen primordia differentiated into short filaments and anthers. Further unknown growth factors appear to be required for the complete differentiation of floral primordia into mature organs.     

THE EFFECTS OF AMINO ACIDS UPON THE DEVELOPMENT OF EXCISED FLORAL BUDS OF AQUILEGIA

Young excised floral buds of Aquilegia were grown on a chemically defined medium containing various concentrations of single amino acids or mixtures of amino acids. γ-Amino butyric acid significantly promoted floral development through the initiation and differentiation of carpels. These floral organs were generally absent on the basal medium. Alanine, glutamic acid, and aspartic acid had no effect upon floral development. All other amino acids were either ineffective at lower concentrations and inhibitory at higher concentrations or were inhibitory at all concentrations. Casein hydrolysate and a mixture of amino acids found in coconut milk were ineffective. The addition of both γ-amino butyric acid and alanine to the basal medium promoted development approaching that achieved on the coconut-milk medium. However, further growth factors appear to be required before development on coconut-milk medium is equalled or exceeded.     

EFFECTS OF GROWTH SUBSTANCES ON EXCISED FLORAL BUDS OF AQUILEGIA

Buds at various stages of development were grown for 3 weeks on solid media containing coconut milk, minerals, vitamins, sucrose, and varying quantities of gibberellic acid, indoleacetic acid, and kinetin. The best average growth was obtained on media containing GA at 2.0 mg/liter, IAA at 1.0 mg/liter, and kinetin at 0.05 mg/liter. When results were compared for buds explanted at very young stages and at older stages, however, young buds attained better average growth on media with 0.5 mg/liter of IAA. Evidence is presented for interaction between IAA and kinetin. With buds explanted at young stages, as the kinetin concentration was increased, optimum growth occurred on media with increasing concentrations of IAA. With older buds, on the other hand, as the kinetin concentration was increased, optimum growth occurred on decreasing concentrations of IAA. Bud growth was compared on media with growth substances sterilized by autoclaving with growth on similar media with these substances filter-sterilized. Better growth occurred generally on the media with filter-sterilized ingredients. A long-range objective of this research is the development of a system that would make possible quantitative measurements of floral development in vitro.     

耧斗菜属花部形态发生

用扫描电镜观察了无距耧斗菜（Ａｑｕｉｌｅｇｉａ ｅｃａｌｃａｒａｔａ Ｍａｘｉｍ ．）和北美耧斗菜（Ａ． ｃａｅｒｕｌｅａＪａｍ ｅｓ．）雄蕊群和雌蕊群的发生过程,并在光学显微镜下检查了同一雄蕊群不同轮雄蕊小孢子发育的顺序． 发现耧斗菜属雄蕊群的发生是向心发生,而小孢子发育和花药成熟顺序是离心的． 说明该属雄蕊群离心发育是次生现象;它在不同类群中出现,可能具有平行演化的性质,其系统学价值尚需重新评价     

大岩桐花萼切块离体培养直接再生花芽的形态观察(简报)

前文发现在基本培养基上添加赤霉素和细胞分裂素能促进离体培养的大岩桐花蕾直接再生花芽[1],后又发现细胞分裂素能协同赤霉素促进离体培养大岩桐花萼切块高频率直接再生花芽[2,3],所得结果提示,这可能是一个研究赤霉素和细胞分裂素在花分化发育中作用的良好实验系统。为了完善这一实验系统,明确离体培养的大岩桐花萼切块培养物直接再生花芽的起源以及花分化的形态进程是十分必要的。为此我们对离体培养大岩桐花萼切块的形态变化在体视显微镜下进行了系统观察,并进行了石蜡切片及电镜扫描观察。进行大岩桐(Sinningia speciosa Hiern)花萼切…     

离体植物花芽和花器官的发育研究进展

在近二十几年,尤其是近十年由于离体培养技术的完善和进一步向精确的层次的发展,已使得许多种植物花序、花芽和单独花器官以及部分的花器官在体外成功地进行了培养和尝试,并且对花芽及花器官的离体发育有了更深入的认识。不同植物的花发育需要不同的植物生长调节剂以及它们的不同配比,并且不同植物在其花发育所需要的营养因子也存在着相当大的差异性。这种差异性又随植物器官及花发育的不同阶段而受到加大或缩小。通过对正常花及突变体花进行的离体培养实验研究已经对一些花器官发生过程中的调节程序有了新的了解。利用离体培养技术,包括刚发展起来的薄层细胞培养技术在阐明花发育的机理、形态建成的分子机制以及成花梯度的物质基础等问题上具有广阔的潜力。     

THE POLLINATION ECOLOGY OF AQUILEGIA ELEGANTULA AND A. CAERULEA (RANUNCULACEAE) IN COLORADO

Aquilegia elegantula Greene and A. caerulea James occur in montane and subalpine habitats in the southern Rocky Mountains of western North America. The red and yellow flowers of A. elegantula are nodding, odorless, protogynous, and secrete a concentrated (44%) sucrose nectar in the floral spurs. Seed set in flowers under pollinator exclosures was 12% while seed set in open-pollinated flowers was 65%. The flowers of A. elegantula are pollinated primarily by the Broad-tailed Hummingbird (Selasphorus platycercus [Swainson]) and by at least three species of pollen-foraging bumblebees, of which Bombus occidentalis Greene is the most common. The blue and white flowers of A. caerulea are erect, mildly fragrant, protandrous, and secrete a 26% sucrose nectar. Seed set in caged flowers in the field averaged 39%. in uncaged flowers 54%. The most important pollinators of A. caerulea are the crepuscular hawkmoth, Hyles (=Celerio) lineata (Fabricius) and ten species of pollen-foraging Bombus. The most abundant bumblebee species, B. occidentalis, is also a frequent nectar thief. Differences in pollination systems alone probably do not constitute an effective anti-hydridization mechanism between A. elegantula and A. caerulea, but do serve to reinforce differences in habitat and flowering time that distinguish the two species.     

石斛离体培养中ABA对诱导花芽形成的影响

由兰科植物铁皮石斛（Ｄｅｎｄｒｏｂｉｕｍ ｃａｎｄｉｄｕｍ Ｗａｌｌ．ｅｘ Ｌｉｎｄｌ．）种子诱导形成的愈伤组织,在光照下置于ＭＳ附加０．３ ｍ ｇ／ＬＮＡＡ 的培养基上繁殖,可以形成原球茎。将原球茎转入ＭＳ含２ ｍ ｇ／Ｌ ６－ＢＡ 和０．５ ｍ ｇ／ＬＮＡＡ 的培养基上,花芽形成频率为２７．０％ 。原球茎先在０．５ ｍ ｇ／ＬＡＢＡ的培养基上预培养１５ ｄ,再转入含２ ｍ ｇ／Ｌ６－ＢＡ 的ＭＳ培养基上培养,花芽形成频率明显提高,可达８４．４％ ,而且每株植株花的数目增加;但是在仅有ＡＢＡ 的ＭＳ培养基上培养的原球茎再生的植株未见花芽形成     

FLORAL DEVELOPMENT IN MYRISTICA (MYRISTICACEAE)

Myristica fragrans and M. malabarica are dioecious. Both staminate and pistillate plants produce axillary flowering structures. Each pistillate flower is solitary, borne terminally on a short, second-order shoot that bears a pair of ephemeral bracts. Each staminate inflorescence similarly produces a terminal flower and, usually, a third-order, racemose axis in the axil of each pair of bracts. Each flower on these indeterminate axes is in the axil of a bract. On the abaxial side immediately below the perianth, each flower has a bracteole, which is produced by the floral apex. Three tepal primordia are initiated on the margins of the floral apex in an acyclic pattern. Subsequent intercalary growth produces a perianth tube. Alternate with the tepals, three anther primordia arise on the margins of a broadened floral apex in an acyclic or helical pattern. Usually two more anther primordia arise adjacent to each of the first three primordia, producing a total of nine primordia. At this stage the floral apex begins to lose its meristematic appearance, but the residuum persists. Intercalary growth below the floral apex produces a columnar receptacle. The anther primordia remain adnate to the receptacle and grow longitudinally as the receptacle elongates. Each primordium develops into an anther with two pairs of septate, elongate microsporangia. In pistillate flowers, a carpel primordium encircles the floral apex eventually producing an ascidiate carpel with a cleft on the oblique apex and upper adaxial wall. The floral ontogeny supports the morphological interpretation of myristicaceous flowers as trimerous with either four-sporangiate anthers or monocarpellate pistils.     

FLORAL DEVELOPMENT IN CAESALPINIA (LEGUMINOSAE)

Utilizing scanning electron microscopy, we studied the early floral ontogeny of three species of Caesalpinia (Leguminosae: Caesalpinioideae): C. cassioides, C. pulcherrima, and C. vesicaria. Interspecific differences among the three are minor at early and middle stages of floral development. Members of the calyx, corolla, first stamen whorl, and second stamen whorl appear in acropetal order, except that the carpel is present before appearance of the last three inner stamens. Sepals are formed in generally unidirectional succession, beginning with one on the abaxial side next to the subtending bracts, followed by the two lateral sepals and adaxial sepal, then lastly the other adaxial sepal. In one flower of C. vesicaria, sepals were helically initiated. In the calyx, the first-initiated sepal maintains a size advantage over the other four sepals and eventually becomes cucullate, enveloping the remaining parts of the flower. The cucullate abaxial sepal is found in the majority of species of the genus Caesalpinia. Petals, outer stamens, and inner stamens are formed unidirectionally in each whorl from the abaxial to the adaxial sides of the flower. Abaxial stamens are present before the last petals are visible as mounds on the adaxial side, so that the floral apex is engaged in initiation of different categories of floral organs at the same time.     

FLORAL DEVELOPMENT IN GYMNOTHECA CHINENSIS (SAURURACEAE)

The development of the inflorescence and flowers are described for Gymnotheca chinensis Decaisne (Saururaceae), which is native only to southeast China. The inflorescence is a short terminal spike of about 50–70 flowers, each subtended by a small bract. There are no showy involucral bracts. The bracts are initiated before the flowers, in acropetal order. Flowers tend to be initiated in whorls of three which alternate with the previous whorl members. No perianth is present. The flower contains six stamens, and four carpels fused in an inferior ovary containing 40–60 ovules on four parietal placentae. Floral symmetry is dorsiventral from inception and throughout organ initiation. Floral organs are initiated in the following order: 1) median adaxial stamen, 2) a pair of lateral common primordia which bifurcate radially to produce two stamen primordia each, 3) median abaxial stamen, 4) a pair of lateral carpel primordia, 5) median adaxial carpel, 6) median abaxial carpel. This order of initiation differs from that of any other Saururaceae previously investigated. The inferior ovary results from intercalary growth below the level of stamen attachment; the style elongates by intercalary growth, and the four stigmas remain free. The floral structure of Gymnotheca is relatively advanced compared to Saururus, but its assemblage of specializations differs from that of either Anemopsis or Houttuynia, the other derived genera in the Saururaceae.