Gibberellic acid impairs fertilization in Clementine mandarin under cross-pollination conditions

We investigated the effect of gibberellic acid (GA₃) in the fertilization of Clementine mandarin cv. ‘Clemenules’ (Citrus clementina Hort. ex Tan.), a parthenocarpic variety that produces seedless fruit due to its self-incompatible nature, but yields seedy fruits when grown under cross-pollination conditions.Experiments were conducted with on-tree ‘Clemenules’ flowers and ‘Fortune’ mandarin pollen (C. clementina Hort. ex Tan. × C. reticulata Blanco), which is sexually compatible with the former. Preanthesis treatment at −2 days after anthesis (−2 DAA) enhanced ovule abortion in both unpollinated and cross-pollinated (at +2 DAA) flowers. In the latter, the number of pollen tubes reaching the ovules was significantly reduced although pollen grains were not treated; thus, fertilization was partially avoided and seed set was reduced. When GA₃ was applied at anthesis (0 DAA) at the time of pollination, ovule abortion was again enhanced, and pollen tube growth was completely arrested; thus, fertilization was prevented and seed set was impeded. When GA₃ was applied 24 h after pollination (+1 DAA in flowers pollinated at anthesis), pollen tube growth was impaired but not arrested and ovule abortion was enhanced; therefore, fertilization was not prevented but impaired.We conclude that, when applied the days around anthesis, GA₃ (10 mg l⁻¹) impairs fertilization by either enhancing ovule abortion or reducing pollen tube growth, in ‘Clemenules’ flowers under cross-pollination conditions. The intensity of the response depends on the physiological flower state at the moment of treatment.     

Regeneration of the Digestive Tube in the Holothurian Apostichopus japonicusafter Evisceration

We performed a TEM study of regeneration of the intestine in the Far Eastern trepang, the holothurian Apostichopus japonicus, after evisceration. The following stages were distinguished in the restoration process of the digestive tube: the growth of connective tissue along the margin of the mesenterium, in the place of rupture; dedifferentiation of cells and their migration and proliferation; the rooting of the esophagus lining into the connective tissue anlage; and the transformation of esophagus cells into cells of the middle part of the intestine. The migration of epithelium into the area of regeneration takes place through a solid cellular layer, without breaking of the cell contacts. The mitotic activity was registered in all stages of restoration; the dividing cells were located chaotically, without the development of a blastema.     

Actin and pollen tube growth

Summary Actin microfilaments (MFs) are essential for the growth of the pollen tube. Although it is well known that MFs, together with myosin, deliver the vesicles required for cell elongation, it is becoming evident that the polymerization of new actin MFs, in a process that is independent of actomyosin-dependent vesicle translocation, is also necessary for cell elongation. Herein we review the recent literature that focuses on this subject, including brief discussions of the actin-binding proteins in pollen, and their possible role in regulating actin MF activity. We promote the view that polymerization of new actin MFs polarizes the cytoplasm at the apex of the tube. This process is regulated in part by the apical calcium gradient and by different actin-binding proteins. For example, profilin binds actin monomers and gives the cell control over the initiation of polymerization. A more recently discovered actin-binding protein, villin, stimulates the formation of unipolar bundles of MFs. Villin may also respond to the apical calcium gradient, fragmenting MFs, and thus locally facilitating actin remodeling. While much remains to be discovered, it is nevertheless apparent that actin MFs play a fundamental role in controlling apical cell growth in pollen tubes.Dedicated to Professor Brian E. S. Gunning on the occasion of his 65th birthday     

花粉萌发和花粉管生长发育的信号转导

显花植物授粉是一个复杂的发育过程。从花粉落在柱头上开始,经过粘附、识别、水合、萌发,花粉管在花柱内生长,直至到达子房发生双受精作用,整个过程发生在雌、雄两性细胞和组织之间,受到严格的遗传控制和细胞控制。一方面雌雄配子的基因型决定两者是否亲和,另一方面雌雄两性细胞间发生复杂的相互作用,细胞外信号分子是这些过程的主要调控因子。当花粉或花粉管细胞感知外部信号后,必然通过信号转导级联反应,达到控制萌发、调整花粉管生长方向等目的。这一系列动力学的细胞事件,关系到受精的成败。因此研究此过程中的信号及其转换机…     

Inhibition of the growth of apple pollen tubes by EDTA, surfactants and fungicides

The effects of EDTA (ethylene diamine tetraacetic acid), Kresoxim‐methyl, Wettable Sulphur 80 WP and the surfactants Triton X‐100, Triton X‐114, Dehydol TA 5 and Dehydol TA 29 on pollen tube growth of Malus domestica, Borkh. cvs ‘Golden Delicious’ and ‘Mondial Gala’ were determined by image analysis. The ED₅₀ ‐ and ED₉₀ ‐ values (effective doses giving 50% and 90% reduction of growth) of the chemicals, except for the surfactants, were determined. The selected test substances showed an inhibiting effect on pollen tube growth, with Kresoxim‐methyl being the most active. Surfactants are components of pesticides and they also caused inhibition of pollen tube growth but only at application rates more then 10‐fold higher than typically recommended. Pollen of both apple cultivars were almost equally sensitive to the chemicals.     

Efficient cloning of plant genomes into bacterial artificial chromosome (BAC) libraries with larger and more uniform insert size

The construction of bacterial artificial chromosome (BAC) libraries remains relatively complex and laborious, such that any technological improvement is considered to be highly advantageous. In this study, we addressed several aspects that improved the quality and efficiency of cloning of plant genomes into BACs. We set the 'single tube vector' preparation method with no precipitation or gel electrophoresis steps, which resulted in less vector DNA damage and a remarkable two- to threefold higher transformation efficiency compared with other known vector preparation methods. We used a reduced amount of DNA for partial digestion (up to 5 microg), which resulted in less BAC clones with small inserts. We performed electrophoresis in 0.25 x TBE (Tris, boric acid, ethylenediaminetetraacetic acid) buffer instead of 0.5 x TBE, which resulted in larger and more uniformly sized BAC inserts and, surprisingly, a two- to threefold higher transformation efficiency, probably due to less contamination with borate ions. We adopted a triple size selection that resulted in an increased mean insert size of up to 70 kb and a transformation efficiency comparable with that of double size selection. Overall, the improved protocol presented in this study resulted in a five- to sixfold higher cloning efficiency and larger and more uniformly sized BAC inserts. BAC libraries with the desired mean insert size (up to 200 kb) were constructed from several plant species, including hexaploid wheat. The improved protocol will render the construction of BAC libraries more available in plants and will greatly enhance genome analysis, gene mapping and cloning.     

Histological study of post-pollination events in Spathodea campanulata beauv. (Bignoniaceae), a species with late-acting self-incompatibility

The reproductive biology of Spathodea campanulata was investigated by means of hand-pollination experiments, observations of pollen tube growth using fluorescence microscopy, and serial sections of ovules in selfed and crossed pistils. Only cross-pollinated flowers developed fruits, and all selfed flowers abscised within 3-4 d. However, self pollen tubes grew successfully to the ovary, penetrating and fertilizing the majority of ovules by 48 h, indicating that S. campanulata is a species with late-acting self-incompatibility. The incidences of ovule penetration, fertilization and endosperm initiation were all significantly slower in selfed vs. crossed pistils, although no other signs of malfunctioning were detected. The possible role of such slow self pollen tube effectiveness as a recognition event is discussed within the context of the slow but not entirely suppressed self pollen tube growth reported for some species with conventional homomorphic self-incompatibility.     

Behavior of storage lipids during development and germination of olive ( Olea europaea L.) pollen

Summary.  The presence of abundant oil bodies in the mature olive pollen grain has led us to focus on the behavior of these lipid bodies during pollen development and in vitro pollen germination. The appearance, increase, and accumulation of lipid bodies have been determined by following the sequential development of the pollen grain. Semithin slices of anthers and pollen grains were stained with Sudan Black B in order to identify neutral lipids. Ultrastructural studies were also carried out. Our results show a notable increase in lipid bodies between the young-pollen-grain stage and the mature-pollen-grain stage. Substantial polarization of lipid bodies was observed after 1 or 2 h of pollen incubation in germination medium. During pollen tube growth, the lipid bodies are located near the germinative aperture after 3 h of incubation, as well as inside the pollen tube, thus suggesting that the lipid bodies move from the pollen grain to the pollen tube. After 7 h of germination the presence of lipid bodies inside the pollen tube is no longer substantial. Our results support the idea that lipid bodies are involved in pollen germination, stigma penetration, and pollen tube growth. These results are discussed in connection with their implications for the pollen germination process. Received June 4, 2002; accepted October 29, 2002; published online April 8, 2003 RID="*" ID="*" Correspondence and reprints: Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, Consejo Superior de Investigaciones Científicas, Profesor Albareda 1, 18008 Granada, Spain.