Tatjana Prebeg

Endogenous gibberellin profile during Christmas rose (Helleborus niger L.) flower and fruit development.

Gibberellins (GAs) were identified and quantified during flower and fruit development in the Christmas rose (Helleborus niger L.), a native of southeastern Europe with a long international horticultural tradition. Physiologically, the plant differs from popular model species in two major respects: (1) following anthesis, the initially white or rose perianth (formed in this species by the sepals) turns green and persists until fruit ripening, and (2) the seed is shed with an immature embryo, a miniature endosperm, and a prominent perisperm as the main storage tissue. GA1 and GA4 were identified by full-scan mass spectra as the major bioactive GAs in sepals and fruit. LC-MS/MS system in accord with previously verified protocols also afforded analytical data on 12 precursors and metabolites of GAs. In the fruit, GA4 peaked during rapid pericarp growth and embryo development and GA1 peaked during the subsequent period of rapid nutrient accumulation in the seeds and continued pericarp enlargement. In the sepals, the flux through the GA biosynthetic pathway was highest prior to the light green stage when the photosynthetic system was induced. Unfertilized, depistillated, and deseeded flowers became less green than the seed-bearing controls; chlorophyll accumulation could be restored by applying GA1, GA4, and, less efficiently, GA3 to the deseeded fruit. The sepals of unfertilized and depistillated flowers indeed contained very low levels of GA4 and gradually decreasing levels of GA1. However, the concentrations of their precursors and metabolites were less affected. These data suggest that a signal(s) from the fruit stimulates GA biosynthesis in the sepals resulting in greening. The fruit-derived GAs appear to be mainly involved in pericarp growth and seed development.

Ultrastructural characterization of the reversible differentiation of chloroplasts in cucumber fruit

The changes in plastid ultrastructure in the pericarp of cucumber (Cucumis sativus L) fruit were studied during fruit yellowing (which accompanied maturation) and regreening. In the course of fruit maturation, the thylakoid system was progressively reduced, and only a small number of membranes remained in the plastids of mature fruit. At the same time, the plastoglobules increased in size, often remaining in close proximity to the degrading thylakoids. In pericarp tissue which turned green again, the thylakoid network in the plastids was gradually reconstituted. Morphological similarities between the plastids in mature and regreening fruit indicated that the chloroplasts in regreened tissue were redifferentiated from the plastids of mature fruit. Reconstitution of the thylakoid system appeared to start from two morphologically distinct types of membranes: from double membranes which resembled thylakoids and from membrane-bound bodies (MBBs). The latter appeared to form thylakoids by two mechanisms: by detachment of extensions from their surfaces and by fragmentation. The plastoglobules remained in the plastids during thylakoid system reconstitution and were often observed in close proximity to developing thylakoids. In the course of chloroplast redifferentiation, several types of membraneous structures were found to be associated with the plastid envelope: (i) vesicles which appeared to separate from the envelope and to fuse subsequently with the developing thylakoids, (ii) tubules, and (iii) double-membrane sheets which appeared asde novo forming thylakoids.

Endogenous Auxin Profile in the Christmas Rose (Helleborus niger L.) Flower and Fruit: Free and Amide Conjugated IAA

The reproductive development of the Christmas rose (Helleborus niger L.) is characterized by an uncommon feature in the world of flowering plants: after fertilization the white perianth becomes green and photosynthetically active and persists during fruit development. In the flowers in which fertilization was prevented by emasculation (unfertilized) or entire reproductive organs were removed (depistillated), the elongation of the peduncle was reduced by 20–30%, and vascular development, particularly lignin deposition in sclerenchyma, was arrested. Chlorophyll accumulation in sepals and their photosynthetic efficacy were up to 80% lower in comparison to fertilized flowers. Endogenous auxins were investigated in floral and fruit tissues and their potential roles in these processes are discussed. Analytical data of free indole-3-acetic acid, indole-3-ethanol (IEt), and seven amino acid conjugates were afforded by LC-MS/MS in floral tissues of fertilized as well as unfertilized and depistillated flowers. Among amino acid conjugates, novel ones with Val, Gly, and Phe were identified and quantified in the anthers, and in the fruit during development. Reproductive organs before fertilization followed by developing fruit at post-anthesis were the main source of auxin. Tissues of unfertilized and depistillated flowers accumulated significantly lower levels of auxin. Upon depistillation, auxin content in the peduncle and sepal was decreased to 4 and 45%, respectively, in comparison to fruit-bearing flowers. This study suggests that auxin arising in developing fruit may participate, in part, in the coordination of the Christmas rose peduncle elongation and its vascular development.

DIFFERENTIATION OF CHROMOPLASTS IN CUCUMIS SATIVUS PETALS

The development of fibrillar‐type chromoplasts, with particular emphasis on the morphogenesis and structural characteristics of the fibrils, was studied in Cucumis sativus petals by light and electron microscopy. Chromoplast differentiation was characterized by the progressive accumulation of fibrils that arranged in a parallel fashion to form birefringent bundles. Immediately before flower opening, the fibrillar bundles in the petals lost their compact structure, and at anthesis, the majority of them partially or completely disorganized. In the plastids of senescing petals, most fibrils were arranged in parallel again. Ultrastructural studies indicated that the formation of at least some of the fibrils started within plastoglobules. The outer diameter of most fibrils ranged from 13 to 18 nm. When transversely sectioned, they were polygonal in shape, often resembling a pentagon or hexagon. In addition to these most common fibrils, different forms that appeared to be composed of two to several “incompletely separated” single fibrils were also frequently present. When standard preparation methods were used, the coat of the fibrils did not appear as a uniformly stained layer but consisted of darkly stained roundish subunits interposed with more lightly stained areas. The morphological observations are discussed with regard to the fine structural changes of chromoplasts during flower development, as well as to the structure of fibrils and the mechanisms of their formation.