Formation of sperm cells inGalium mollugo (Rubiaceae),Trichodiadema setuliferum (Aizoaceae), andAvena sativa (Poaceae)

The formation of sperm cells has been examined ultrastructurally in the tricellular pollen grains ofGalium mollugo L. (Rubiaceae).Trichodiadema setuliferum Schwantes (Aizoaceae), andAvena sativa L. (Poaceae). After detachement from the intine the generative cell of all three species lies free within the vegetative cytoplasm. The two sperm cells are built inTrichodiadema andAvena by a single separating wall, while inGalium mollugo two independent walls are formed. However, both mechanisms separate the two male gametes completely.     

Subdivision ofPelargonium sect.Cortusina (Geraniaceae)

The sect.Cortusina (Dc.)Harv. s.l. is divided into the sectt.Cortusina (Dc.)Harv. s. str. andReniformia (Knuth)Dreyer, comb. nov.,P. cortusifolium L'Hérit.,P. crassicaule L'Hérit.,P. echinatum Curt.,P. magenteum J. J. A. Van der Walt, andP. sibthorpifolium Harv. remain in the sect.Cortusina s. str., while the speciesP. album J. J. A. Van der Walt,P. dichondrifolium Dc.,P. odoratissimum (L.)L'Hérit.,P. reniforme Curt., andP. sidoides Dc. are removed from the sect.Cortusina s.l. and included in the revised sect.Reniformia. Both sections are well characterized by morphological, anatomical, karyological and palynological features. The chromosomes ofCortusina s. str. are small and based upon x = 11; those ofReniformia are larger and based upon x = 8. The size and surface sculpture of the pollen grains differ markedly between the two sections. Flavones and hydrolysable tannins occur in sect.Reniformia, but are absent in sect.Cortusina s. str. After hydrolysis hydroxybenzoic acids, which are present in large amounts inReniformia, occur only in traces inCortusina. The two sections also occupy distinctly different geographical areas. A close relationship between sect.Reniformia and some species of sect.Ligularia (Sweet)Harv. is indicated by the occurrence of artificial hybrids.     

Pollen grain morphology ofCoris (Primulaceae)

Pollen grain morphology, sculpturing, and wall ultrastructure are investigated in two species ofCoris (Primulaceae),C. monspeliensis L. andC. hispanica Lange. The study includes both acetolysed and unacetolysed pollen. No evidence of any major palynological difference is recorded between these two species, apart from a somewhat larger pollen inC. monspeliensis. However,Coris can be distinguished from the remaining members of thePrimulaceae by the conjunction of relatively large pollen grains, prominent margo, and particular tectal pattern causing a reticulate surface with minute luminal perforations decreasing towards the colpi. From both these distinctive features, and others typically primulaceous, some evolutionary considerations are inferred. Finally, the higher proportion of irregular grains inC. hispanica is interpreted in light of environmental stress.     

The annual species ofAdonis (Ranunculaceae)—a polyploid complex

The five annual species ofAdonis L., sect.Adonis, growing in Israel, form a series of diploid, tetraploid and hexaploid species. Their somatic chromosome numbers are 2n = 16 inA. annua L.,A. dentata Del. andA. palaestina Boiss., 2n = 32 inA. microcarpa DC., 2n = 48 inA. aestivalis L.; counts forA. dentata, A. palaestina andA. microcarpa are new records. There are indications that alloploidization may have been involved in the process of speciation in sect.Adonis. A taxonomic survey of the 8 species of the section reveals that a higher ploidy level is usually combined with a larger distribution area.     

Neue Taxa vonTillandsia subgenusDiaphoranthema (Bromeliaceae) aus Bolivien und Argentinien

5 new taxa are described and illustrated; their position within the subgenus is discussed:T. hirta W. Till & L. Hromadnik,T. cotagaitensis L. Hromadnik,T. caliginosa W. Till, andT. gilliesii Baker subsp.polysticha W. Till & L. Hromadnik are members of a group includingT. myosura Grisebach exBaker,T. mandonii E. Morren exMez in DC.,T. retorta Grisebach exBaker em.Grisebach andT. andicola Gillies exBaker. T. brealitoensis L. Hromadnik is related toT. angulosa Mez in DC. but distinct and possibly of hybrid origin.

     

Taxonomic and morphological studies inHypecoum sect.Hypecoum (Papaveraceae)

Hypecoum sect.Hypecoum comprises 8 species, one with 2 additional subspecies, in the Mediterranean area and the Middle East. Five of these are recognized for the first time, viz.Hypecoum procumbens L. subsp.fragrantissimum Å. E. Dahl,H. procumbens L. subsp.atropunctatum Å. E. Dahl,H. trullatum Å. E. Dahl,H. angustilobum Å. E. Dahl andH. torulosum Å. E. Dahl. H. dimidiatum Delile andH. pseudograndiflorum Petr., which have generally been included inH. imberbe Sm., are reestablished as distinct species. Morphology and variation patterns are described and discussed with special reference to mating systems. Self-incompatibility is dominating butH. procumbens subsp.procumbens andH. torulosum are self-compatible. The broad variation ranges of the former taxon in traits presumably related to mating system (petal, anther and stigma sizes as well as pollenovule ratios) indicate varying rates of outcrossing. SEM micrographs are given of petal and stigma surfaces and of pollen grains. Chromosome counts are presented for seven taxa. All are diploid with 2n = 16.     

Agouticarpa, a new neotropical genus of Tribe Gardenieae (Rubiaceae)

Agouticarpa, a new genus of the informalAlibertia group (Rubiaceae), is described and illustrated. It comprises six species, and occurs from Costa Rica to Bolivia. Three species are here described as new:Agouticarpa grandistipula, A. hirsuta, andA. velutina. Additionally, three new combinations are made:A. curvifolia andA. williamsii (previously inGenipa) andA. isernii (previously inAlibertia). Agouticarpa is characterized by being dioecious, having elliptic to abovate, membranaceous stipules, male flowers in a branched dichasial or thyrse-like inflorescence, a poorly developed cup-shaped calyx, pollen grains with 3–7 apertures, and large globose fruits.     

A functional interpretation ofLactuceae (Compositae) pollen

Pollen morphology and ultrastructure inLactuceae pollen is considered in relation to the accomodation of volume changes, pollination biology and exine-held substances. Echinate pollen grains, such as those ofCatananche, are shown to accomodate volume changes by folding along the colpi and possibly by volume changes in the cavea. The different patterns of echinolophate pollen respond in different ways. Folding along the colpi is important inScorzonera andTragopogon and to a limited extent inCichorium andEpilasia whilst inScolymus the colpi are almost immobilized. Movements of the lacunar floors take over the harmomegathic function to compensate for lack of colpus mobility. Bulging of the intine at the apertures and changes in the size of the cavea may account for part of the volume change accomodated in any pollen type. Echinolophate pollen is interpreted as being a superior means of regulating volume changes with the most economical and mechanically efficient use of wall material which has evolved independently in several tribes ofCompositae.     

Ribosomal RNA genes structure in somePopulus spp. (Salicaceae) and their hybrids

The tandemly repeated multigene families encoding 18S and 25S rRNAs were studied at the restriction enzyme level inPopulus alba L.,Populus deltoides Bartr. exMarsh.,Populus trichocarpa Torr. & Gray and in the hybrids between the last two mentioned species. The analysis of single and double digestion with EcoRI, BamHI, XbaI, and SstI endonucleases showed the presence of single repetitive unit types of 12.25 and 11.75kb inP. alba andP. trichocarpa, respectively.P. deltoides showed two rDNA gene types having the same length (12.25Kb) but different nucleotide sequence in the IGS. The rDNAs genes ofP. deltoides andP. triochocarpa are inherited codominantly in their hybrids.     

Pollen morphology inParis and its related genera

Pollen grains ofParis, Medeola, andScoliopus have been examined with light, scanning electron, and transmission electron microscopies.P. quadrifolia L.,P. verticillata Bieb.,P. delavayi Franch. andP. polyphylla Smith have monosulcate pollen characterized by psilate, foveolate or reticulate exine sculpture. In contrast to the former species,P. japonica (Fr. et Sav.) Franch. andP. tetraphylla A. Gray have monosulcate pollen with gemmate (rarely rugulate) exine.Medeola has monosulcate pollen with reticulate exine that is distinct from that ofParis. Scoliopus has monosulcate pollen characterized by a peculiar reticulate exine pattern. The palynological evidence suggests thatParis andTrillium are closely related to each other, andMedeola andScoliopus should be separated fromParis andTrillium.     

Pollen diameter and fertility in nine species ofPuccinellia (Poaceae)

Diameter and fertility of pollen grains were surveyed in nine species ofPuccinellia Parl. (Poaceae), mainly from the Iberian Peninsula, allowing information about reproductive behaviour and biological characteristics of pollen of these taxa. Maximum diameter was measured with an image processor and fertility was evaluated by staining the grains with Müntzing's mixture. Significant differences in both characters were found between the taxa. In consequence, maximum diameter and fertility proved to have some taxonomic relevance inPuccinellia.     

Relationships between species ofLeymus,Psathyrostachys, andHordeum (Poaceae,Triticeae) inferred from Southern hybridization of genomic and cloned DNA probes

We have used total genomic DNA as a probe to size-fractionated restriction enzyme digests of genomic DNA from a range ofTriticeae species from the generaLeymus Hochst.,Psathyrostachys Nevski, andHordeum L., and hybrids betweenHordeum andLeymus to investigate their taxonomic relationships. Genomic Southern hybridization was found to be an effective and simple way to assess the distribution and diversity of essentially species-specific and common, repetitive DNA sequences, and is hence especially useful in evolutionary studies. The DNA sequences ofH. vulgare seem to diverge substantially from those ofH. brachyantherum, H. lechleri, H. procerum, andH. depressum. The genome ofThinopyron bessarabicum shows little homology to those of theLeymus species investigated, confirming thatT. bessarabicum is not an ancestral genome inLeymus. Although the genomes ofLeymus andPsathyrostachys share substantial proportions of DNA sequences, they include divergent repeated sequences as well. Hybridization with a ribosomal DNA probe (pTa 71) showed that the coding regions containing structural genes encoding the 18 S, 5.8 S, and 26 S ribosomal RNA were conserved among the species investigated, whereas the intergenic spacer region was more variable, presenting different sizes of restriction fragments and enabling a classification of the species. The rye heterochromatin probe pSc 119.2 hybridized to DNA fromH. lechleri andT. bessarabicum, but not to DNA from the other species investigated.     

Pollen evolution and systematics inAnnonaceae with special reference to the disulcate Australian endemic genera

All genera ofAnnonaceae endemic in Australia (Ancana, Fitzalania, Haplostichanthus) show almost exactly the same type of disulcate (disulculate) pollen with intact exine extending over the sulci. Tetrad stages inHaplostichanthus andAncana reveal a latudinal subequatiorial orientation of the two sulci at the proximal hemisphere. Sometimes they fuse into a ±zonosulcate aperture.Fissistigma pollen grains are ±globose and have a flattened pole with a central elevation and a concentric groove, covered by a somewhat reduced exine. This palynological characters give further support for separating the generaAncana andFissistigma. Germination was observed inHaplostichanthus where the pollen tube emerges at one of the two sulci and inFissistigma where the flattened part breaks up during germination. The aperture types described here are obviously transitional stages between aperturate and inaperturate pollen grains and are discussed in regard to pollen evolution.     

Pubic symphyseal synostosis and sexual dimorphism of the pelvis inPresbytis cristata andPresbytis rubicunda

This study examines the relationship between public symphyseal synostosis and sexual dimorphism of the pelvis in two sympatric species ofPresbytis—P. cristata andP. rubicunda. Whereas no specimen ofP. cristata shows fusion of the interpubic joint, a high percentage of female (43.8%) and male (83.3%)P. rubicunda have a fused public symphysis. As females of both species are similar in body size, they are predicted to give birth to similarly sized newborns. Based on comparison with other anthropoids, the percentage dimorphism in the ischiopubic index inP. cristata andP. rubicuda suggests selection on pelvic capacity in relation to obstetrics. In species characterized by cephalopelvic constriction (i.e., the size of the fetal cranium closely approximates the capacity of the maternal birth canal), successful birth seems possible only by a hormonally induced increase in pelvic joint mobility during delivery. However, fusion of the interpubic joint obviates pelvic joint mobility. Consequently, this study tests the hypothesis thatP. rubicunda shows obstetric adaptations of the pelvis that are not found inP. cristata. The results show that pelvic capacity is larger in females than males in bothP. cristata andP. rubicunda; the sexual difference is most pronounced at the inlet. Moreover, the pattern of pelvic dimorphism is nearly identical between the species. When females of the two species are compared,P. rubicunda evidences a shorter distance between the sacroiliac and hip joints and a wider bituberous diameter. The former is related to interspecific differences in locomotion, and the latter is associated with obstetrics.     

Pollen morphology ofSonchus and related genera,and a general discussion

The pollen morphology of the taxa belonging to the generaAetheorhiza Cass.,Launaea Cass.,Reichardia Roth andSonchus L. in the Iberian Peninsula has been studied with light and electron microscopy. The pollen is 3(-4)-zonocolporate and echinolophate (without polar lacunae, but in general with prelacunae), with equatorial ridges and 15–20 lacunae: 3–4 poral, 6–8 abporal and 6–8 paraporal. Small to medium size, P × E = 19–36 × 23–42 µm; sometimes two different sizes have been found. Exine 3–9 µm thick and ornamentation microreticulate and echinate. The results clearly show the relationships between genera. Moreno-Socías, E., Mejías, J. A., Díez, M. J., 1994: Morfología polínica deLactuceae (Asteraceae) en la Península Ibérica, I.Lactuca y géneros relacionados. — Acta Bot. Malacitana.19: 103–113.     

Weitere Untersuchungen über Proteinkörper in Zellkernen und ihre taxonomische Bedeutung

In 83 species of the familiesMonotropaceae, Apocynaceae, Oleaceae, Scrophulariaceae, Lentibulariaceae, Bignoniaceae, Martyniaceae, Myoporaceae, Verbenaceae, Lamiaceae, Campanulaceae, andLiliaceae, protein bodies in the cell nuclei have been found, in 68 of these species for the first time. On the basis of their structure in accordance with morphological characters the generaBurgsdorfia, Hesiodia, Olisia, andPhlomoides of theLamiaceae are accepted;Lamium is divided intoLamium, Lamiastrum andOrvala; two new combinations are established:Kickxia campyloceras (Rech. fil. &Esfandiari)Speta andEtornotus papilionaceus (Burm. in L.)Speta. Deviating shape or lack of protein bodies corroborate former taxonomic decisions, e.g. the transfer ofMonotropa toMonotropaceae or the separation ofGaleopsis andLadanum; Veronica schmidtiana should not be included inPseudolysimachion. Systematic affinities are discussed primarily withinScrophulariaceae because nuclear protein bodies have been studied already in many species of this family. ForCampanula patula two 2 x populations are reported.

Herrn Professor Dr. L.Geitler zum 80. Geburtstag gewidmet.     

Pollen morphology of the cypripedioid orchids

Pollen morphology of the four traditional genera (Paphiopedilum, Selenipedium, Cypripedium, Phragmipedium) comprising the cypripedioid orchids indicates that the monads are sulcate, more or less smooth-surfaced, and covered by a non-acetolysis resistant layer called elastoviscin. Evidence from ultrathin sections of pollen grains shows that typical exine layers are present only inSelenipedium, modified inPhragmipedium and absent inPaphiopedilum and most species ofCypripedium; that a small, inconspicuous portion of the grain surface is constructed as a sulcus; and that the pollen grain wall acts as a sponge in rapidly absorbing water. Several instances of parallelisms between non-related families and among different groups of orchids are reported and new ideas on the evolution of theCypripedioideae are presented.Dedicated to Prof. DrE. Tschermak-Woess on the occasion of her 70th birthday.Reprint requests toM. Hesse.     

Behavior of organelle nuclei (nucleoids) in generative and vegetative cells during maturation of pollen inLilium longiflorum andPelargonium zonale

Summary The behavior of organelle nuclei during maturation of the male gametes ofLilium longiflorum andPelargonium zonale was examined by fluorescence microscopy after staining with 4,6-diamidino-2-phenylindole (DAPI) and Southern hybridization. The organelle nuclei in both generative and vegetative cells inL. longiflorum were preferentially degraded during the maturation of the male gametes. In the mature pollen grains ofL. longiflorum, there were absolutely no organelle nuclei visible in the cytoplasm of the generative cells. In the vegetative cells, almost all the organelle nuclei were degraded. However, in contrast to the situation in generative cells, the last vestiges of organelle nuclei in vegetative cells did not disappear completely. They remained in evidence in the vegetative cells during germination of the pollen tubes. InP. zonale, however, no evidence of degradation of organelle nuclei was ever observed. As a result, a very large number of organelle nuclei remained in the sperm cells during maturation of the pollen grains. When the total DNA isolated from the pollen or pollen tubes was analyzed by Southern hybridization with a probe that contained therbc L gene, for detection of the plastid DNA and a probe that contained thecox I gene, for detection of the mitochondrial DNA, the same results were obtained. Therefore, the maternal inheritance of the organelle genes inL. longiflorum is caused by the degradation of the organelle DNA in the generative cells while the biparental inheritance of the organelle genes inP. zonale is the result of the preservation of the organelle DNA in the generative and sperm cells. To characterize the degradation of the organelle nuclei, nucleolytic activities in mature pollen were analyzed by an in situ assay on an SDS-DNA-gel after electrophoresis. The results revealed that a 40kDa Ca²⁺-dependent nuclease and a 23 kDa Zn²⁺ -dependent nuclease were present specifically among the pollen proteins ofL. longiflorum. By contrast, no nucleolytic activity was detected in a similar analysis of pollen proteins ofP. zonale.     

Improvement of cotton fiber quality by transforming the<Emphasis Type="Italic"> acsA</Emphasis> and<Emphasis Type="Italic"> acsB</Emphasis> genes into<Emphasis Type="Italic"> Gossypium hirsutum</Emphasis> L. by means of vacuum infiltration

A novel method for the genetic transformation of cotton pollen by means of vacuum infiltration and Agrobacterium-mediated transformation is reported. The acsA and acsB genes, which are involved in cellulose synthesis in Acetobacter xylinum, were transferred into pollen grains of brown cotton with the aim of improving its fiber quality by incorporating useful prokaryotic features into the colored cotton plants. Transformation was carried out in cotton pollen-germinating medium, and transformation was mediated by vector pCAMBIA1301, which contains a reporter gene -glucuronidase (GUS), a selectable marker gene, hpt, for hygromycin resistance and the genes of interest, acsA and acsB. The integration and expression of acsA, acsB and GUS in the genome of transgenic plants were analyzed with Southern blot hybridization, PCR, histochemical GUS assay and Northern blot hybridization. We found that following pollination on the cotton stigma transformed pollen retained its capability of double-fertilization and that normal cotton seeds were produced in the cotton ovary. Of 1,039 seeds from 312 bolls pollinated with transformed pollen grains, 17 were able to germinate and grow into seedlings for more than 3 weeks in a nutrient medium containing 50 mg/l hygromycin; eight of these were transgenic plants integrated with acsA and acsB, yielding a 0.77% transformation rate. Fiber strength and length from the most positive transformants was 15% greater than those of the control (non-transformed), a significant difference, as was cellulose content between the transformed and control plants. Our study suggests that transformation through vacuum infiltration and Agrobacterium mediated transformation can be an efficient way to introduce foreign genes into the cotton pollen grain and that cotton fiber quality can be improved with the incorporation of the prokaryotic genes acsA and acsB.Communicated by D. Bartels     

Dimorphic exine sculpturing in three distylous species ofLinum (Linaceae)

DistylousLinum grandiflorum, L. mucronatum, andL. pubescens have dimorphic pollen grains. In short-styled flowers the exine has monomorphic processes each with a ring of papillae. In long-styled flowers the exine has two types of processes: small processes terminating in a spinule and larger processes with a central spinule surrounded by a ring of spinuels, papillae or buttresses. The distylousLinum species of four different sections, in which morph-specific differences in the wall structure of stigmatic papillae have been reported, also exhibit exine dimorphism. The patterns of these dimorphisms are similar in all four sections in which distyly is present.