Ovule abortion in white clover (Trifolium repens L.)

Plants of white clover ( Trifolium repens L.) cv. Olwen were grown in an open glasshouse maintained at a mean temperature of 20^oC and ovule growth and seed production measured. Differences in the rate of growth of ovules within ovaries were observed as early as 2 days after pollination. Ovules reached a maximum size after 8 days with the smallest only half the size of the largest. After 8 days, the smallest ovules became flaccid and shrivelled. Ovule position within the ovary had little effect on the frequency of seed set and although there was an apparently higher probability that central ovules produced a seed than those nearer the peduncle or style this was not statistically significant. Inflorescence position and floret position on the inflorescence had a significant effect on the number of seeds per floret and seed weight; the first formed inflorescences and the first florets to be pollinated on each inflorescence had more seeds per floret and heavier seeds and fewer florets with no seed than later pollinated florets. There were also differences between florets within the same whorl. The role of a number of factors which may influence floret site utilisation are discussed.     

Tissue-specific oxylipin signature of tomato flowers: allene oxide cyclase is highly expressed in distinct flower organs and vascular bundles

A crucial step in the biosynthesis of jasmonic acid (JA) is the formation of its correct stereoisomeric precursor, cis(+)12-oxophytodienoic acid (OPDA). This step is catalysed by allene oxide cyclase (AOC), which has been recently cloned from tomato. In stems, young leaves and young flowers, AOC mRNA accumulates to a low level, contrasting with a high accumulation in flower buds, flower stalks and roots. The high levels of AOC mRNA and AOC protein in distinct flower organs correlate with high AOC activity, and with elevated levels of JA, OPDA and JA isoleucine conjugate. These compounds accumulate in flowers to levels of about 20 nmol g-1 fresh weight, which is two orders of magnitude higher than in leaves. In pistils, the level of OPDA is much higher than that of JA, whereas in flower stalks, the level of JA exceeds that of OPDA. In other flower tissues, the ratios among JA, OPDA and JA isoleucine conjugate differ remarkably, suggesting a tissue-specific oxylipin signature. Immunocytochemical analysis revealed the specific occurrence of the AOC protein in ovules, the transmission tissue of the style and in vascular bundles of receptacles, flower stalks, stems, petioles and roots. Based on the tissue-specific AOC expression and formation of JA, OPDA and JA amino acid conjugates, a possible role for these compounds in flower development is discussed in terms of their effect on sink-source relationships and plant defence reactions. Furthermore, the AOC expression in vascular bundles might play a role in the systemin-mediated wound response of tomato.     

Effect of plant growth regulators on in vitro fiber development from unfertilized and fertilized Egyptian cotton ovules

Unfertilized and fertilized ovules of Gossypium barbadense Giza 45 (extra long staple variety) were used to study the effect of plant growth substances (auxins, gibberellins and cytokinins) on in vitro fiber initiation and development. Kinetin, alone did not increase total fiber unit (TFU) of unfertilized ovules, while an increase in TFU value occurred when a constant level of IAA and GA₃ were used either separately or in combination in the liquid medium. GA₃ used alone, produced a higher TFU value than that produced by IAA, whilst, IAA with a constant level of GA₃ (5 M) produced the highest value of TFU. GA₃ with a constant level of IAA (5 M) produced a lower TFU value. Kinetin reduced the stimulatory effect of IAA and GA₃ on TFU value when used in combination with either substance. In fertilized ovules, the highest level of TFU was reached when IAA, with a constant level of GA₃, was added to the medium, whilst its lowest level was obtained when IAA was used alone. Estimation of in vitro fiber production, as well as the effect of growth substances used in different concentrations on in vitro fiber initiation and development from unfertilized and fertilized ovules of Egyptian cotton varieties Gossypium barbadense Giza 45 are discussed.     

Somatic embryogenesis from stigmas and styles of grapevine

Summary An in vitro protocol has been developed for callus indiction, somatic embryogenesis, and plant regeneration from stigma-style culture of grapevine. Four different grapevine cultivars (Vitis vinifera L.: cvs. ‘Bombino Nero’, ‘Greco di Tufo’, ‘Merlot’, and ‘Sangiovese’) were tested. Exlants were cultured on Nitsch and Nitsch medium (NN) supplemented with various combinations of 6-benzylaminopurine (BA: 4.5 and 9.0 μM) and β-naphthoxyacetic acid (NOA; 5.0 and 9.9 μM). Sucrose (88 mM) was used as the carbon source. Somatic embryogenesis was induced within 3–7 mo. after culture initiation. Even though explants of different origin (unfertilized ovules and anthers) regenerated somatic embryos, the higher embryogenic potential was observed in stigma and style explants, with the exception of ‘Merlot’, which regenerated somatic embryos only from unfertilized ovules. The percentages of stigma-style explants producing somatic embryos was 7% in ‘Bombino Nero’ (cultured on NN medium supplemented 9.0 μM BA and 9.9 μM NOA). 14% in ‘Greco di Tufo’ (4.5 μM BA and 9.9 μM NOA), and 8% in ‘Sangiovese’ (9.0 μM BA and 9.9 μM NOA). The presence of growth regulators (BA and NOA) in the medium was essential for induction of somatic embryogenesis. Plants were regenerated on hormone-free NN medium containing 88 mM sucrose.     

Variability in the developmental stage of apricot ovules at anthesis and its relationship with fruit set

The stage of ovule development at anthesis and its relationship with fruit set was studied in several apricot cultivars growing in Mediterranean climatic conditions. Although generally the ovule was immature at anthesis, a great variability was found in the stage of development of the ovules from different cultivars. Considering functional ovules to be those with an embryo sac with at least four nuclei at anthesis, the earliest flowering varieties frequently showed more than 50% of functional ovules. Though these results could suggest that there is an influence of the chilling requirement on the percentage of functional ovules at anthesis, data recorded from two cultivars, ‘Goldrich’ and ‘Colorao’, with high chilling requirements, contrast with this suggestion. These results indicate that in apricot flowers the development of the embryo sac at anthesis is genotype dependent. The fact that a high frequency of functional ovules at anthesis was found, in cultivars with more than 50% of fruit set, suggests that, to be fertilised, a certain level of development of the apricot ovules at this time is necessary.     

Comparative floral structure and systematics in Crossosomatales (Crossosomataceae, Stachyuraceae, Staphyleaceae, Aphloiaceae, Geissolomataceae, Ixerbaceae, Strasburgeriaceae)

Floral structure of all putative families of Crossosomatales as suggested by molecular studies was comparatively studied. The seven comprise Crossosomataceae, Stachyuraceae, Staphyleaceae, Aphloiaceae, Geissolomataceae, Ixerbaceae, and Strasburgeriaceae. The entire clade (1) is highly supported by floral structure, also the clades (in sequence of diminishing structural support): Ixerbaceae/Strasburgeriaceae (2), Geissolomataceae/Ixerbaceae/Strasburgeriaceae (3), Aphloiaceae/Geissolomataceae/Ixerbaceae/Strasburgeriaceae (4), and Crossosomataceae/Stachyuraceae/Staphyleaceae (5). Among the prominent floral features of Crossosomatales (1) are solitary flowers, presence of a floral cup, imbricate sepals with outermost smaller than inner, pollen grains with horizontally extended endoapertures, shortly stalked gynoecium, postgenitally united carpel tips forming a compitum, stigmatic papillae two‐ or more‐cellular, ovary locules tapering upwards, long integuments forming zigzag micropyles, cell clusters with bundles of long yellow crystals, mucilage cells, seeds with smooth, sclerified testa and without a differentiated tegmen. Clade (2) is characterized by large flowers, petals forming a tight, pointed cone in bud, stamens with long, stout filaments and sagittate anthers, streamlined, conical gynoecium, antitropous ovules, rudimentary aril, lignified, unicellular, T‐shaped hairs and idioblasts with striate mucilaginous cell walls. Clade (3) is characterized by alternisepalous carpels, punctiform stigma formed by postgenitally united and twisted carpel tips, synascidiate ovary, only one or two pendant ovules per carpel, nectary recesses between androecium and gynoecium. Clade (4) is characterized by pronounced ‘pollen buds’. Clade (5) is characterized by polygamous or functionally unisexual flowers, x‐shaped anthers, free and follicular carpels (not in Stachyuraceae). Crossosomataceae and Aphloiaceae, although not retrieved as a clade in molecular studies, share several special floral features: polystemonous androecium; basifixed anthers without a connective protrusion; stigma with two more or less decurrent crests; camplyotropous ovules and reniform seeds; simple, disc‐shaped nectaries and absence of hairs. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 147 , 1–46.     

Comparative floral structure and systematics in Celastrales (Celastraceae, Parnassiaceae, Lepidobotryaceae)

Floral morphology, anatomy and histology in the newly circumscribed order Celastrales, comprising Celastraceae, Parnassiaceae and Lepidobotryaceae are studied comparatively. Several genera of Celastraceae and Lepidobotrys (Lepidobotryaceae) were studied for the first time in this respect. Celastraceae are well supported as a group by floral structure (including genera that were in separate families in earlier classifications); they have dorsally bulged‐up locules (and thus apical septa) and contain oxalate druses in their floral tissues. The group of Celastraceae and Parnassiaceae is also well supported. They share completely syncarpous gynoecia with commissural stigmatic lobes (and strong concomitant development of the commissural vascular bundles but weak median carpel bundles), only weakly crassinucellar or incompletely tenuinucellar ovules with an endothelium, partly fringed sepals and petals, protandry in bisexual flowers combined with herkogamy by the movement of stamens and anther abscission, and stamens fused with the ovary. In contrast, Lepidobotryaceae are more distant from the other two families, sharing only a handful of features with Celastraceae (not Parnassiaceae), such as pseudohermaphroditic flowers, united stamen bases forming a collar around the gynoecium and seeds with a conspicuous aril. However, all three families together are also somewhat supported as a group and share petals that are not retarded in late floral bud development, 3‐carpellate gynoecia, ventral slits of carpels closed by long interlocking epidermal cells and pollen tube transmitting tissue encompassing several cell layers, both integuments usually more than two cell layers thick, and only weak or lacking floral indumentum. In some molecular analyses Celastrales form an unsupported clade with Malpighiales and Oxalidales. This association is supported by floral structure, especially between Celastrales and Malpighiales. Among Celastrales, Lepidobotryaceae especially share special features with Malpighiales, including a diplostemonous androecium with ten fertile stamens, epitropous ovules with an obturator and strong vascularization around the chalaza. © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society, 2005, 149 , 129–194.     

Calcium in the micropylar secretion and receptivity of explanted Crocus ovules to intra- and interspecific pollen

The micropylar secrete of the ovules of Crocus vernus ssp. vernus was analyzed for the Ca⁺² content by atomic absorption, and its capacity to germinate and attract pollen was tested by pollinating explanted ovules, and incubating in absence of culture medium. The results display a Ca⁺² concentration of 28.9 mM in the micropylar secrete. On this secrete both compatible- and incompatible pollen germinates with a mean percentage of 53.7%, and their pollen tubes enter the micropylar canal with percentages of 32.3% to 21.0%. In situ the ovules fail to attract tubes of incompatible pollen. The results are discussed in relation to the ovule receptivity and the guided growth of pollen tubes, substantiating the model of the tropic growth towards increasing calcium concentrations.     

Carpeloidy in flower evolution and diversification: a comparative study in Carica papaya and Arabidopsis thaliana

Background and Aims

Bisexual flowers of Carica papaya range from highly regular flowers to morphs with various fusions of stamens to the ovary. Arabidopsis thaliana sup1 mutants have carpels replaced by chimeric carpel–stamen structures. Comparative analysis of stamen to carpel conversions in the two different plant systems was used to understand the stage and origin of carpeloidy when derived from stamen tissues, and consequently to understand how carpeloidy contributes to innovations in flower evolution.

Methods

Floral development of bisexual flowers of Carica was studied by scanning electron microscopy and was compared with teratological sup mutants of A. thaliana.

Key Results

In Carica development of bisexual flowers was similar to wild (unisexual) forms up to locule initiation. Feminization ranges from fusion of stamen tissue to the gynoecium to complete carpeloidy of antepetalous stamens. In A. thaliana, partial stamen feminization occurs exclusively at the flower apex, with normal stamens forming at the periphery. Such transformations take place relatively late in development, indicating strong developmental plasticity of most stamen tissues. These results are compared with evo-devo theories on flower bisexuality, as derived from unisexual ancestors. The Arabidopsis data highlight possible early evolutionary events in the acquisition of bisexuality by a patchy transformation of stamen parts into female parts linked to a flower axis-position effect. The Carica results highlight tissue-fusion mechanisms in angiosperms leading to carpeloidy once bisexual flowers have evolved.

Conclusions

We show two different developmental routes leading to stamen to carpel conversions by late re-specification. The process may be a fundamental aspect of flower development that is hidden in most instances by developmental homeostasis.