MORPHOGENESIS IN TRICHODERMA: AUTONOMOUS AND NONAUTONOMOUS PIGMENTATION IN HETEROKARYONS OF COLOR MUTANTS IN TRICHODERMA VIRIDE

The final step in the process of conidiation—conidial pigmentation—was studied in the fungus Trichoderma viride. Twenty-nine auxotrophic, color mutants, isolated from the same green wildtype strain, were paired to produce stable heterokaryons in all possible combinations and grouped according to their complementation behavior. No complementation (green pigmentation) was found in any of the heterokaryons formed by pairs of white (W) mutants. However, these mutants could be separated into two groups with respect to their behavior when paired with yellow (Y) and brown (Br) mutants. When W^c mutants were paired with any of the Y or Br mutants complementation took place. However, W^d mutants displayed this reaction with only one group of yellow mutants (Y^a) and not with the other (Y^b) nor with Br mutants. In noncomplementing heterokaryons such as Y^b/W^d, only yellow and white conidia were produced, pigmentation being autonomous. On the other hand, in heterokaryons in which complementation did take place, as for instance Y^a/W^d, green as well as white and yellow conidia were produced. Differential sensitivity to UV irradiation was used to show that the green conidia were of either W^d or F^a genotype, indicating a nonautonomous type of gene action. It is suggested that the genes W^c, Y^a, Y^b and Br have a sequential structural role in the biosynthesis of the green pigment, while W^d controls the activity of three (W^c, Y^b, Br) of these genes.     

BUZZ-POLLINATION AND PATTERNS IN SEXUAL TRAITS IN NORTH EUROPEAN PYROLACEAE

Flower biology and pollination of Moneses uniflora, Orlhilia secunda, Pyrola minor, P. rotundifolia, P. chlorantha, and Chimaphila umbellata are described and discussed in relation to patterns in sexual traits and possible evolution of buzz-pollination within the group. The large number of pollen grains are packed into units of monads in Orthilia, tetrads in Moneses and Pyrola, or polyads in Chimaphila. Pollen is the sole reward to visiting insects except in the nectar-producing O. secunda and C. umbellata. Correlations are present between several sexual traits. Pollen: ovule ratios are very low, indicating efficient pollen transfer. Use of polyads may reduce the number of siring males. This trend may be counteracted by larger stigmas as in C. umbellata. Increasing pollen unit size may enhance relatedness of seeds, reduce abortion, and improve seed set. This may explain the evolution, within the Pyrolaceae and other families, of higher ovule numbers with larger pollen units. The plants are mainly pollinated by bumblebees. Chimaphila umbellata is visited by nectar-collectors; Moneses uniflora, Pyrola minor, P. rotundifolia, and P. chlorantha are visited exclusively by pollen-buzzers; and Orthilia secunda is visited by both nectar-collectors and pollen-buzzers. Nectar secretion is suggested to be an ancient attractant in the group with C. umbellata having the least specialized pollination system, and buzz-pollination in M. uniflora and in the studied Pyrola species regarded as an advanced system.     

CHROMOSOME HOMOLOGY IN SOME INTERCONTINENTAL HYBRIDS IN HIBISCUS SECT. FURCARIA

Ten kinds of interspecific hybrids were obtained involving the following species: H. surattensis L. (2x, genome constitution BB), H. sudanensis Hochr. (2x, GG), and H. rostellatus Guill. and Perr. (4x, GGHH) from Africa; H. furcatus Roxb. non Willd. (8x) from India and Ceylon; H. furcellatus Lam. and H. bifurcatus Cav. (both 4x, PPQQ) from South America; and H. heterophyllus Vent. (6x) from Australia. Chromosome pairing in pollen mother cells (PMC's) at metaphase I in the 4x hybrids H. bifurcatus-rostellatus and H. furcellatus-rostellatus indicated that the parents have one genome in common (Q = G or H). Hibiscus furcatus was shown earlier to have a B genome; hybrids of H. surattensis-sudanensis F₁ X furcatus were hexaploid, having received an unreduced gamete from their hybrid parent, and had approximately 36 II, 36 I in PMC's. The genome formula of H. furcatus may therefore be designated BBGGWWZZ. The hybrid H. rostellatus-furcatus (BGGHWZ) confirmed that H. furcatus has a G genome in common with H. rostellatus; pairing of the other three genomes was inconsistent, as was that in H. rostellatus-heterophyllus. Some samples of the latter approached 36 II, 36 I, expected if H. heterophyllus were GGHHJJ; other samples had less pairing. Hibiscus furcatus-heterophyllus hybrids apparently arose from unreduced gametes of H. heterophyllus and originated as decaploids rather than heptaploids; chromosome number was unstable in PMC's. Nevertheless, multivalents, especially trivalents, were frequent enough to suggest that H. furcatus and H. heterophyllus share G genomes. On the other hand, an 8x H. bifurcatus-furcatus hybrid, which apparently arose from an unreduced gamete of H. bifurcatus, had a low multivalent frequency. Hybrids were obtained of H. heterophyllus X sudanensis and H. surattensis-sudanensis X heterophyllus, but the plants were weak and were not analyzed cytologically. We suggest that the New World, African, Indian, and Australian genomes which retain a considerable degree of homology (G or H or both) were distributed by land prior to separation of the southern continents by continental drift.     

PHOTOPERIOD INDUCED CHANGE IN PHYLLOTAXIS IN XANTHIUM

Photoperiodic floral induction in Xanthium, achieved by subjecting the plants to two long nights, is accompanied by a transient change of the phyllotaxis from the (2, 3) contact parastichy pattern of vegetative plants, to a (3, 5) pattern during the transition. To specify the phyllotaxis, two parameters were estimated from transverse sections of apical buds of control and treated plants: the divergence angle, α, and the plastochron ratio, a. The plastochron ratio decreased progressively during transition from the vegetative to the reproductive state of growth, from a = 1.48 initially to a = 1.15 six days after the beginning of induction. The divergence angle was not altered during the transition. This change in phyllotaxis is interpreted as a change in the relative positioning of leaf primordia on the transitional apex. This transient change appears to be identical with the previously described long-term change of the phyllotaxis of Xanthium brought about by treatment of plants with gibberellic acid.     

COUNTERVAILING SELECTION IN DIFFERENT FITNESS COMPONENTS IN FEMALE RED DEER

In individually monitored red deer (Cervus elaphus) living in the North Block of the Isle of Rhum, Scotland, juvenile survival is related to the genotype at the enzyme loci Mpi and Idh-2 (each with two alleles, f and s). To establish whether other fitness components also are related to genetic differences, we examined whether age at first breeding, fecundity, and adult survival of females were related to genotype at the same loci. Fertility in females shot outside the study area was also analyzed in relation to Mpi and Idh-2 genotype. The analyses controlled for phenotypic and environmental factors affecting female reproductive performance. At Mpi, f-carrying females in the study area bred earlier than ss individuals and tended to be more fecund. However, no association was found between Mpi genotype and adult survival. In culled females, Mpi f-carriers were more likely to be pregnant than ss females. At Idh-2, homozygous females in the study area started breeding earlier than heterozygous females. Idh-2 fs and ss females were more fecund than ff females though this relationship was complicated by an interaction with spring temperature in the year of birth. When the population was at high density, adult survival of Idh-2 ss females was better than survival of ff females, which was, in turn, better than survival of fs females. No association was found between Idh-2 genotype and fertility in culled females. Overall, the associations found in female reproductive measures favor those genotypes that survive particularly badly over the first two years of life. This result supports the idea that countervailing selection in different fitness components (antagonistic pleiotropy) is a common and powerful force maintaining polymorphism in natural populations. It may also explain how fitness components can have large heritabilities while overall fitness may have a low heritability.     

UNUSUAL PHENOMENA IN THE LIFE HISTORIES OF FLORIDEAE IN CULTURE1

Several Florideae grown in natural seawater media under defined laboratory conditions have interesting and unusual life histories. Antithamnion occidentale males of one generation produced tetraspores that gave rise to nonsporangiate males. The functional females of A. pygmaeum developed spermatangia and tetrasporangia; the tetraspores formed new females. Antithamnion defectum tetrasporophytes of one generation bore spermatangia in addition to tetrasporangia; the tetraspores gave rise to typical gametophytes. Tetraspores from successive generations of Callitham-nion sp. developed into tetrasporophytes and males but no females were produced. Functional female gametophytes of Platythamnion sp. bore abortive tetrasporangia. Field-collected plants of two species of Fauchea produced tetraspores that yielded additional sporangiate plants: those of F. pygmaea being bispo-rangiate and tetrasporangiate, and those of F. lacini-ata being strictly tetrasporangiate. Male plants of Pleonosporium vancouverianum from a running seawater table bore spermatangia and polysporangia when collected. The same plants in unialgal culture produced only spermatangia.     

INBREEDING DEPRESSION IN TWO MIMULUS TAXA MEASURED BY MULTIGENERATIONAL CHANGES IN THE INBREEDING COEFFICIENT

In mixed-mating plant populations, one can estimate the relative fitness of selfed progeny w by measuring the inbreeding coefficient F and selfing rate s of adults of one generation, together with F of adults in the following generation (after selection). In the first application of this multigenerational method, we estimated F and s for adults over three consecutive generations in adjacent populations of two annual Mimulus taxa: the outbreeding M. guttatus and the inbreeding M. platycalyx. This gave estimates of w for the last two generations. Although average multilocus selfing rates were high in both taxa (0.63 in M. guttatus; 0.84 in M. platycalyx), the relative fitness of selfed progeny averaged only 0.19 in M. guttatus and 0.32 in M. platycalyx. An alternative estimator for w that incorporates biparental inbreeding gave even lower estimates of w. These values are significantly below the 0.5 threshold thought to favor selfing, and show that partially selfing populations can harbor substantial genetic load. In accordance with the purging hypothesis, the more highly selfing M. platycalyx showed marginally lower inbreeding depression than M. guttatus in both years (P = 0.08). Inbreeding depression and selfing rates also varied among years in concert among taxa. Several sources of bias are discussed, but computer simulations indicate it is unlikely that w is biased downwards by linkage of marker loci to load loci.     

MORNING-EVENING ADMINISTRATION TIME DIFFERENCES IN DIGOXIN KINETICS IN HEALTHY YOUNG SUBJECTS

Digoxin, frequently used in the treatment of congestive heart failure, has a very narrow therapeutic index. We studied the differences in digoxin pharmacokinetics when ingested in the morning versus evening. A single digoxin (0.25 mg) dose was given orally to the same group of 10 diurnally active healthy (6 male and 4 female) volunteers in the morning at 08:00 and evening at 20:00 in separate experiments scheduled 2 weeks apart. Blood samples were collected at specific times for 48h after each timed dose; digoxin was determined by radioimmunoassay (RIA). Maximum plasma concentration C_max; T_max, the time to reach C_max; area under plasma concentration curve AUC; and elimination half-time T_1/2 of digoxin were determined. T_max was statistically significantly shorter (54 min) following 08:00 dosing compared to 20:00 dosing (96 min). Although the C_max was higher after morning than evening dosing, it was not significantly so. No other parameter of digoxin pharmacokinetics except T_max exhibited administration time dependency. (Chronobiology International, 18(5), 841–849, 2001)     

中国的被孢霉种类

被孢霉属(Mortierella Coemans)是接合菌纲(Zygomycetes)、毛霉目(Mucorales)、被孢霉科(Mortierellaceae)中的一个大属,目前已知约有90种;主要存在于土壤、植物残体、动物粪便等基物中。我国过去对被孢霉的研究不多,在《中国真菌总汇》(1979)中记录了8个种。本研究从全国22个省、市、自治区采集的2000多号样品中,分离到约220个被孢霉菌株。本研究主要采用Gams (1970, 1977)的分类系统进行分类鉴定,并对该系统进行了修改。在属下分3个亚属(Micromucor, Mortierella和Gamsiella), 8个组(Actinomortierella, Alpina,Hygrophila, Mortierella, Schmuckii, Simplex, Spinosa和Stylospora),单囊霉(Haplosporangium)被承认为独立的一个属。本研究鉴定出22个种和3个变种,包括一个新种(武夷山被孢霉Mortierella wuyishanensis sp. nov.)和一个新变种(极细无色被孢霉Mortierella hyalina(Harz) W. Gams var. subtilissima var. nov.), 14个中国新纪录。这14个新纪录为:产芽胞被孢霉(Mortierella. gemmifera M. Ellis)、园圃被孢霉(M. horticola Linnem.)、矮小被孢霉(M. humilis Linnem.)、无色被孢霉(M. hyalina(Harz) W. Gams)、印度被孢霉(M. indica B.S. Mehrotra)、英杜被孢霉(M. indohii C.Y. Chien),詹金氏被孢霉(M. jenkinii (A.L. Sm.) Naumov)、可疑极小被孢霉(M. minutissima Tiegh. var. dubia Linnem.)、易变被孢霉(M. mutabilis Linnem.)、微孢被孢霉(M. parvispora Linnem.)、角胞拉曼被孢霉(M. ramanniana(Moller) Linnem. var. angulispora (Naumov) Linnem.)、网孢被孢霉(M. reticulata Tiegh.& G. Le Monn.)、多疣被孢霉(M. verrucosa Linnem.)、轮枝被孢霉(M. verticillata Linnem.)。文中讨论和评价了一些分类性状,还列出分亚属、分组、分种和变种的检索表.每个分类单元都有描述和讨论以及线条图、并列出分布地区。     

POLYMORPHISM IN RINGED PLOVERS

The rather similar ringed plovers Charadrius hiaticula and C. semipalmatus represent essentially allopatric, circumpolar populations which overlap in the breeding season only in a narrow zone in the eastern Canadian Arctic. In eight fiords on eastern Baffin Island, semipalmatus was the more numerous by a factor of almost three. Both plovers occurred together, although not in equal numbers, in the same wide range of habitats. Pair formation in both plovers apparently took place at the same time, but for more than half of the individuals, not in the same geographic area. The majority of the hiaticula individuals arrived after most semipalmatut, and the late arriving hiaticula appeared to be paired upon reaching Baffin Island. There was an overlap in their arrival times of about five days. Mixed hiaticula-semipalmatus courtship groups occurred during this period. Pair forming activities of hiaticula and semipalmatus appeared similar but were not studied closely. Analysis of the non-overlapping territories of two populations suggested that the plovers did discriminate one type from the other for there was a tendency for unlike pairs to nest side by side. The significance of this odd pattern is not known. Analysis of the mensural and plumage characteristics of adult plovers from the overlap zone seemed to indicate that gene flow between the two was absent and that they were behaving as distinct species. However, mixed pairs occurred in five out of eight fiords. Considering the populations of all fiords, the number of mixed pairs found (20) was significantly greater than theoretical expectation, which allowed for an occasional mixed mating since a choice of mate was not always possible. All mixed matings produced mixed broods. The young were either hiaticula or semipalmatus. No intermediates occurred. Moreover, 10 pairs in which both birds were pheno-typically “semipalmatus” produced both semipalmatus and hiaticula-type chicks. No intermediates were detected. Egg and chick mortality probably prevented the detection of more such cases. All hiaticula x hiaticula matings produced only hiaticula-type chicks. The inheritance data indicated that hiaticula and semipalmatus are morphs of a single polymorphic species. If a switch gene is operating, it may have originally controlled only toe-webbing, and the other differences may have accumulated as the result of geographic isolation. That differences in plumage pattern and body size are greatest where separation was probably longest supports this hypothesis. One possible model considered semipalmatus-type individuals to be either SS or Ss and hiaticula-types to be ss. Accordingly, all semipalmatus-types involved in mixed matings with hiaticula- types would have been Ss individuals because all such matings produced chicks of both types. Circumstantial evidence suggested that during the last 80 years, the relative number of the two types in the overlap area has shifted in favour of the semipalmatus- type. This might have been the result of selective factors intrinsic to the polymorphism.     

LEAF GAS EXCHANGE IN QUERCUS MACROCARPA (FAGACEAE): RAPID STOMATAL RESPONSES TO VARIABILITY IN SUNLIGHT IN A TREE GROWTH FORM

Responses in net photosynthesis (A), stomatal conductance to water vapor (g), and leaf xylem pressure potential (ψ) were measured in the deciduous tree Quercus macrocarpa during alternating periods of sun (photosynthetic photon flux, PPF > 1,500 μmol m^-2 sec^-1) and shade (ca. 350 μmol m^-2 sec^-1 simulating cloud cover). Measurements were made on trees growing at the gallery forest-prairie edge on the Konza Prairie Research Natural Area in northeast Kansas. The region is near the westernmost extension of the range of Q. macrocarpa where this species experiences significant seasonal water stress (minimum ψ < -2.9 MPa). Quercus macrocarpa was chosen for study because it has relatively high A (15 μmol m^-2 sec^-1) and g (300 mmol m^-2 sec^-1) in contrast to the deciduous and evergreen subalpine trees previously studied. Both trees and large saplings of Q. macrocarpa responded to alternating several minute periods of sun and shade with relatively rapid changes in A and g. Reductions in g (110 mmol m^-2 sec^-1) during shade periods lowered transpirational water losses (E) by 13% (and reduced A by 5%) relative to estimates of A and E made assuming g remained constant. Partial stomatal closure during shade was correlated with moderate enhancement in ψ (0.31 MPa) in Q. macrocarpa. However, greater increases in ψ were measured in adjacent prairie grasses exposed to similar periods of shade (0.72 MPa in Andropogon gerardii, 0.61 MPa in Sorghastrum nutans). Reduced variability in ψ in tree growth forms may reflect greater buffering of water relations associated with the large size of trees, the amount of tissue devoted to water storage, and differences in hydraulic resistance relative to herbs. Nonetheless, the gas exchange and water relations responses in Q. macrocarpa were much more similar to those previously measured in herbaceous subalpine and grassland species than to those documented for subalpine trees. Thus, rapid gas exchange responses to variable PPF may also occur in tree growth forms.     

THE PROLIFERATION KINETICS OF NHIK 1922 CELLS IN VITRO AND IN SOLID TUMOURS IN ATHYMIC MICE

The proliferation kinetics of cells of the line NHIK 1922 grown in vitro and as solid tumours in the athymic mutant nude mouse has been studied. In vitro, growth curves were determined for exponentially growing populations and for populations synchronized by mitotic selection. The phase durations for these populations were determined by flow cytofluorometric measurements of DNA-histograms and pulsed incorporation of [³H]TdR respectively. The generation time and the phase durations for synchronized populations were found to be about equal to those for exponentially growing populations. The duration of the phases G₁, S and G₂+ M was found to be 8·5–9·5, 11·0–12·0 and 6·0–6·5 hr respectively, i.e. the generation time was 26·5–27·0 hr. The proliferation kinetics in vivo were studied by flow cytofluorometry and by the technique of percentage labelled mitoses. The median duration of S-phase and (G₂+ M)-phase in vivo was found to be approximately the same as that observed in vitro, while the median duration of G₁-phase was found to be approximately 5 hr longer in vivo than under the present in vitro growth conditions. The growth fraction in vivo was estimated to be approximately 50%. The non-proliferative compartment of the tumour cells was found to consist mainly of cells with the DNA-content of cells in G₁-phase. It is concluded that the reduced rate of proliferation of NHIK 1922 cells in vivo is correlated with alterations in the duration of G₁-phase and, hence, the proportion of cells in G₁-phase.     

PHYTOCHROME LEVELS DETECTABLE BY IN VIVO SPECTROPHOTOMETRY IN PLANT PARTS GROWN OR STORED IN THE LIGHT

Tissues of fruits and vegetables from local markets were tested for reversible optical density changes at 660 and 730 m μ as a phytochrome assay. Over one-third showed activity, which bore no obvious relationship to any morphological or taxonomic category. Reports of high activity in Brassica oleracea botrytis (cauliflower) were confirmed, but the highest activities were in tissues from roots of Pastinaca (parsnip), receptacles of Cynara (globe artichoke) and vegetative buds of Brassica oleracea gemmifera (Brussels sprouts). Activity in Pastinaca and Cynara appears light-stable, with the far-red-absorbing form reverting to the red-absorbing form more or less quantitatively in darkness. Activity in B. o. gemmifera, by contrast, is light-labile, decaying without any apparent reversion; the high level of phytochrome initially present is due to light exclusion by the surrounding green tissues. Several of the materials described should prove valuable for further investigations on the nature and role of phytochrome.     

A COMPARISON OF IN VIVO AND IN VITRO STATHMOKINETIC METHODS FOR THE STUDY OF CELL PROLIFERATION IN HAMSTER ORAL EPITHELIA

Mitotic indices (MI) expressed as numbers of metaphase figures per 100 basal cells in the cheek pouch and palatal epithelium of the Syrian hamster following metaphase arrest with vinblastine sulphate (VLB) were compared using in vivo and in vitro techniques. The MI in vivo 4 1/2 hr after intraperitoneal injection of 4 mg VLB/kg body weight was 2·69 ± 0·37 for cheek pouch and 12·08 ± 1·09 for palate. MI in vitro was measured using small tissue explants cultured for 4 hr in medium supplemented with VLB at concentrations ranging from 6-600 μg/ml. The maximum MI for cheek pouch epithelium in vitro (2·7) did not differ significantly from that observed in vivo (P > 0·50) and was obtained in the presence of 12–30 μg VLB/ml, a concentration comparable with that used in vivo. In contrast, the maximum MI for palate epithelium in culture (5·6) was significantly lower than that in vivo (P < 0·001) and was only achieved in the presence of extremely high concentrations of VLB. Possible reasons are discussed for the discrepancy between the MI for palatal epithelium in vivo and in vitro.     

HYBRIDIZATION STUDIES IN PERENNIAL ZINNIAS

Results of hybridization studies among the 6 species of subgenus Diplothrix (Zinnia-compositae) are reported. Also included are analyses of chiasmata frequency in the parental species and 2 diploid hybrids. Pairing relationships and chiasmata frequency of the chromosomes in the hybrids of the diploid species indicate their genomes are homologous. Analyses of F₂'s of Z. juni-perifolia X acerosa (2n) show that production of pigment in ray flowers, C, from Z. juniperifolia segregates as a simple dominant over colorless, cc, from Z. acerosa. Morphological studies of the hybrids produced to date indicate that the origin of the polyploid taxa has been more circuitous than simple hybridization followed by chromosome doubling. The genomic constitution of Z. juniperifolia is designated A₁A₁, that of Z. acerosa A₂A₂, and that of Z. oligantha A₃A₃. Morphological data and chromosomal pairing in the polyploid hybrids suggest that the tetraploid species should be tentatively assigned genomic formulae as follows: A₁A₁A₂A₂ or A₁A₁A₃A₃ for both Z. citrea, and Z. grandiflora, and A₂A₂A₃A₃ for 4n Z. acerosa.     

KINETICS OF CELL PROLIFERATION IN THE PRE-IMPLANTED MOUSE EMBRYO IN VIVO AND IN VITRO

The cell proliferation of pre-implanted mouse embryos was investigated after development in vivo and in vitro. The studies were started at the pronuclear stage, 2 h post conception (p.c.) and continued until the hatching of blastocysts, 120–144 h p.c. The number of cell nuclei, the DNA content of each nucleus, the mitotic index and the labelling index were determined. From these data it was possible to calculate the length of the cell generation cycle and its various phases. With the exception of the first cell cycle the S-phase was constant. The G₁- as well as the G₂-phase varied in length during the different cell cycles. From 31–72 h p.c. the increase in cell number was exponential. After cultivation in vitro this increase was smaller than in vivo. At later periods the proliferation rate decreased with proceeding development. In late blastocysts most of the cells were in the G₁-phase. The development of the embryos was somewhat faster in vivo than in vitro. But in principle conditions were comparable.     

SEASONAL VARIATION IN HEART RATE VARIABILITY IN ASTHMATIC CHILDREN

Asthma is a “seasonal disease” with symptoms either aggravated by environmental changes during specific seasons or prevalent at certain times of the year for other reasons. We examined whether the heart rate variability (HRV) of asthmatic children changes by season. The HRV during a portion of one night (00:00–04:00) and day (12:00–16:00) and the entire 24h period (00:00–24:00) during each of the four seasons was analyzed. The data of 95 children with asthma and 106 healthy children, as controls, were assessed. In children with asthma during the 24h period, seasonal variation in the low-frequency (LF) band (0.04–0.15 Hz) and the high-frequency (HF) band (0.15–0.4 Hz) were detected (HF: F = 6.81, p =.0003; LF: F = 4.18, p =.008. The HF value in the summer was significantly higher than in autumn and spring (Scheffe test: autumn vs. summer, s = 4.46, p<.001; spring vs. summer, s = 2.86, p<.05), while the LF value in autumn was significantly lower than in summer (s = 3.42, p <.01). In the control group, no seasonal variation in HF, LF, or LF/HF was detected. The findings infer the HRV, a surrogate measure of autonomic nervous system function, of asthmatic children is more susceptible to seasonal changes brought about by either endogenous annual rhythms or environmental weather phenomena. (Chronobiology International, 17(4), 503–511, 2000)     

INTERSPECIFIC CROSSES IN KALMIA

Interspecific crosses among Kalmia species are reported for the first time. All possible crosses between K. angustifolia, K. cuneata, K. latifolia, K. hirsuta, K. polifolia var. rosmarinifolia and K. p. var. microphylla were attempted. Several crosses yielded no hybrids, many produced albino or yellow-green inviable seedlings, and six combinations produced at least a few seedlings with near-normal growth habit. Stylar inhibition of pollen tube growth prevented some species from hybridizing; reciprocal differences in crossability and pollen tube growth were noted. Kalmia species are highly heterozygous, but no evidence was found within a species for geographic variation of combining ability except for the two varieties of K. polifolia which had different crossing patterns with the other species. The anthocyaninless form of K. angustifolia, in contrast to the normal, has unique combining ability with K. latifolia. No genetic evidence was found to support the placement of K. hirsuta in another genus (Kalmiella) or to designate K. angustifolia var. caroliniana as a distinct species. Use of all the species to breed improved ornamentals is limited by genetic barriers and sterility of F₁ hybrids.     

IMMUNOHISTOCHEMICAL LOCALIZATION OF TYPE I AND II COLLAGENS IN THE INVOLUTING CHICK NOTOCHORDS IN VIVO AND IN VITRO

Embryonic chick notochords were studied during their metabolically active and involuting periods for the expression of collagen type I and II. The staining was carried out on notochords in vivo at stage 20 and stage 35 and on mesenchyme-contaminated and mesenchyme-free notochords at stage 20, which were cultured in vitro for 6 days. The results show that type II collagen is demonstrable in the notochords, at all the examined stages, both in vivo and in vitro. However, the expression of type I collagen was stage-dependent in vivo and in vitro. At stage 20, the perinotochordal sheath is positively immunostained for collagen type I, but the notochord itself is negative. At stage 35, the perinotochordal sheath as well as the notochord are positively immunostained for collagen type I. The mesenchyme-contaminated and the mesenchyme-free notochords and their sheaths are also positively immunostained for the type I collagen after6 days in vitro. The current results, at late developmental stages, indicate that the involuting notochords express collagen type I, which seems not to be altered by changing the micro-environment in vivo.     

α-METHYLTRYPTOPHAN: EFFECTS ON CEREBRAL MONOOXYGENASES IN VITRO AND IN VIVO

Following administration of x-methyltryptophan (AMTP) (50 mg/kg) there was a time dependent decrease of serotonin and a concomitant increase of α-methyl-5-hydroxy-tryptamine (AM-5-HT) in most cerebral areas. AMTP is hydroxylated to α-methyl-5-hydroxytryptophan (AM-5-HTP) by cerebral tryptophan hydroxylase in vitro and in vivo. Hydroxylation of AMTP in vitro and in vivo was markedly inhibited in p-chlorophenylalanine (p-CP) treated rats. After administration of AMTP, the conversion in vivo of tyrosine to norepinephrine was inhibited. This inhibition was not apparent in p-CP pretreated animals. p-Chloroamphetamine (p-CA) (10 mg/kg) lowered serotonin and AM-5-HT levels in some areas of the brain, but unlike p-CP, alone or in combination with AMTP it did not significantly inhibit hydroxylation of tryptophan (Trp). AMTP, as substrate of tryptophan hydroxylase, has a K_m of 1.08 × 10^-4 M (using 6-MPH₄, as cofactor) and as competitive inhibitor, a K₁ of 2.09 × 10^-4M with L-Trp as substrate. AMTP becomes an uncompetitive inhibitor when its concentration is equal to or exceeds that of L-Trp. D-AMTP is neither a substrate nor an inhibitor of tryptophan hydroxylase. DL-AM-5-HTP (K₁, 1.5 × 10^-5 M) and AM-5-HT (K¹ 4.0 × 10^-5 M) are competitive inhibitors.