Disruption of the distylous syndrome in Primula veris

Background and Aims Distyly is a floral polymorphism characterized by the presence of two discrete morphs with reciprocal positioning of anthers and stigmas in flowers on different plants within the same population. Although reciprocal herkogamy and associated floral traits are generally thought to be discrete and strict polymorphisms, little is known about variation in floral traits related to the distylous syndrome within and among populations of a single species. In this study, variation in floral morphology and reciprocal positioning of the sexual organs in the distylous Primula veris (cowslip) is quantified.Methods Data were collected in ten populations occurring in two contrasting habitat types (grasslands and forests), and for each population the average level of reciprocity was assessed, the strength of the self-incompatibility system was determined, and seed production under natural conditions was quantified.Results In grassland populations, flowers showed clear distyly with low and symmetric reciprocity indices at both the lower and upper level. In forests, P. veris produced larger flowers that showed strong deviations in stigma–anther separation, especially in the L-morph. This deviation was mainly driven by variation in stigma height, resulting in high and asymmetric reciprocity indices and the occurrence of several short-styled homostylous plants. Self-incompatibility was, however, strict in both habitats, and morph ratios did not differ significantly from isoplethy. The observed shift in reciprocity in forest populations was associated with a significant reduction in seed production in the L-morph.Conclusions The results indicate that populations of P. veris show habitat-specific variation in flower morphology. Deviations from perfect reciprocal positioning of stigmas and anthers also translate into reduced seed production, suggesting that small changes in sexual organ reciprocity can have far-reaching ecological and evolutionary implications.     

Effects of population size and pollinator limitation on seed-set of Primula sieboldii populations in a fragmented landscape

Female reproductive success, fruit and seed-set and factors affecting them were measured for 3 years (1995–1997) for 24 populations of Primula sieboldii E. Morren, a threatened Japanese plant species in a highly fragmented landscape in southern Hokkaido. The smaller populations (genets <=; 3) set almost no seeds consistently during the study years. In larger populations (genets >= 7), proportions of failed flowers without damage from fungi or herbivores were relatively high, but fruit set per flower varied among the populations in 1995. Correlations of fruit (r = 0.589, P = 0.011) and seed- set (r = 0.688, P < 0.01) with population pollinator availability were highly significant. In 1995, pollination failure would be the most plausible reason for the variation in reproductive success among the populations. However, pollinator availability was relatively high in all populations in 1996 and 1997. In these years, higher proportions of fruit failed because of antagonistic biological interactions. In the populations with poor pollinator availability in 1995, mean fruit and seed-sets were greater in the long-styled morph than in the short-styled morph. Such between-morph differences are thought to be caused by partial self-compatibility of the long-styled morph and the results of hand pollination experiments support this hypothesis.     

Decrease of sexual organ reciprocity between heterostylous primrose species,with possible functional and evolutionary implications

Background and Aims

Heterostyly is a floral polymorphism that has fascinated evolutionary biologists since Darwin''s seminal studies on primroses. The main morphological characteristic of heterostyly is the reciprocal placement of anthers and stigmas in two distinct (distyly) floral morphs. Variation in the degree of intermorph sexual reciprocity is relatively common and known to affect patterns of pollen transfer within species. However, the partitioning of sexual organ reciprocity within and between closely related species remains unknown. This study aimed at testing whether intermorph sexual reciprocity differs within vs. between primrose species that hybridize in nature and whether the positions of sexual organs are correlated with other floral traits.

Methods

Six floral traits were measured in both floral morphs of 15 allopatric populations of Primula elatior, P. veris and P. vulgaris, and anther–stigma reciprocity was estimated within and between species. A combination of univariate and multivariate approaches was used to test whether positions of reproductive organs were less reciprocal between than within species, to assess correlations between sexual organ positions and other corolla traits, and to quantify differences between morphs and species.

Key Results

The three species were morphologically well differentiated in most floral traits, except that P. veris and P. vulgaris did not differ significantly in sexual organ positions. Overall, lower interspecific than intraspecific sexual organ reciprocity was detected. This decrease was marked between P. elatior and P. vulgaris, intermediate and variable between P. elatior and P. veris, but negligible between P. veris and P. vulgaris.

Conclusions

Differences in anther and stigma heights between the analysed primrose species were of the same magnitude or larger than intraspecific differences that altered pollen flow within other heterostylous systems. Therefore, it is possible to suggest that considerable reductions of sexual organ reciprocity between species may lower interspecific pollen flow, with potential effects on reproductive isolation.     

Characterization of agronomic traits and markers of recombinant inbred lines from intra- and interracial populations of Phaseolus vulgaris L.

The value of intra- and interracial populations in common bean (Phaseolus vulgaris L.) needs to be determined in order to create useful genetic variation for maximizing gains from selection, broadening the genetic base of commercial cultivars, and making efficient use of available resources. Five large-seeded parents of race Nueva Granada (N), two small-seeded race Mesoamerica (M), and one medium-seeded race Durango (D) were hybridized to produce one intraracial (N x N) and three interracial (two N x M and one N x D) populations. Seventy-nine F₂-derived F₆ lines randomly taken from each population along with their parents were evaluated for agronomic traits and markers at Palmira and Popayán, Colombia, in 1990 and 1991. Variation for agronomic traits and for morphological, protein, and isozyme markers was larger in interracial populations than in the intraracial population. Mean seed yield of all lines as well as yield of the highest yielding line from two interracial populations were significantly higher than that of the intraracial population. The highest ( 0.80±0.15) heritability was recorded for 100-seed weight. Values for seed yield varied from 0.19±0.17 to 0.50±0.16. Gains from selection (at 20% selection pressure) for seed yield ranged from 3.9% to 11.4%. Seed yield was positively associated with biomass yield, pods/m², and days to maturity, but harvest index showed negative correlations with these traits and a positive value with 100-seed weight. Polymorphism was recorded for phaseolin, lectins, protein Group-1 and protein Group-2 fractions, and six isozyme loci. Lines with indeterminate growth habit had significantly (P < 0.01) higher seed yield than lines with determinate growth habit in a Redkloud x MAM 4 population. Also, 23 other associations of markers with agronomic traits other than seed yield were recorded. Of these associations, lines with T phaseolin, the Diap1 ² allele, and lilac flower color tended to possess greater seed weight.     

Benzylaminopurine induces phenocopies of floral meristem and organ identity mutants in wild-typeArabidopsis plants

Arabidopsis thaliana (L.) Heynh. has been used as a model system to investigate the regulatory genes that control and coordinate the determination, differentiation and morphogenesis of the floral meristem and floral organs. We show here that benzylaminopurine (BAP), a cytokinin, influences flower development inArabidopsis and induces partial phenocopies of known floral homeotic mutants. Application of BAP to wild-type inflorescences at three developmental stages results in: (i) increase in floral organ number; (ii) formation of abnormal floral organs and (iii) induction of secondary floral buds in the axils of sepals. These abnormalities resemble the phenotypes of mutants,clv1 (increase in organ number),ap1,ap2,ap3 (abnormal floral organs) andap1 (secondary floral buds in the axils of first-whorl organs). In addition, BAP induces secondary floral buds in the axils of perianth members ofapt2-6, ap3-1 andag mutants, and accentuates the phenotype of theapt2-1 mutant to resemble theapt2-6 mutant. These observations suggest that exogenous BAP suppresses the normal functioning of the genes for floral meristem identity and thereby affects flower development and the later stages of floral organ differentiation.Abbreviations BAP N6-benzylaminopurine - CK cytokinin     

Changes in cotyledon mRNA during floral induction of Pharbitis nil CV. Violet

Floral induction in seedlings of Pharbitis nil Choisy cv. Violet, with one cotyledon removed, was manipulated by applying various photoperiodic treatments to the remaining cotyledon. Populations of polyadenylated RNA from treated cotyledons were examined to identify messages specifically involved in floral induction. The RNA was translated in vitro using a wheat-germ system, and the resulting translation products were analysed by two-dimensional polyacrylamide gel electrophoresis. Substantial qualitative and quantitative differences were found between mRNA from cotyledons of seedlings kept in continuous light (non-induced) and of seedlings given a 16-h dark period (induced). In contrast, inhibition of flowering with a night-break resulted only in one detectable, quantitative difference in mRNA.Abbreviations CL continuous light - kDa kilodalton - NB 16 h darkness+10 min red-light break, 8 h into the dark period - poly(A)⁺ RNA polyadenylated RNA (isolated by binding to a cellulose oligodeoxythymidine affinity column) - SD short day (16 h dark) - SDP short-day plant - SDS sodium dodecyl sulfate     

Mapping of a chloroplast RFLP marker associated with the CMS cytoplasm of sugar beet (Beta vulgaris)

The Owen cytoplasm of male-sterile sugar beet is associated with several alterations of mitochondrial DNA and one additional HindIII site of chloroplast DNA. The region of this HindIII site has been cloned and sequenced. The site maps in a small reading frame (orf32) close to the ycf7 (orf31) gene in the petG-psbE region of chloroplast DNA. Possible functional implications of the results are discussed. The chloroplast RFLP marker described could be useful for studies on chloroplast-mitochondrial interactions, CMS of sugar beet, and the origin of the Owen cytoplasm.     

Salicylic acid-, but not cytokinin-induced, resistance to WClMV is associated with increased expression of SA-dependent resistance genes in Phaseolus vulgaris

Two-week-old Phaseolus vulgaris plants, wick-fed with 1 mmol/L salicylic acid (SA) or 50 nmol/L dihydrozeatin (DHZ), showed partial inhibition of the accumulation of white clover mosaic virus (WClMV) in infected primary leaves. This inhibition was measured as a decrease in the accumulation of both viral mRNA and viral coat protein, especially at the early stages of infection. Salicylic acid treatment resulted in moderately increased expression of phenylalanine ammonia lyase (PAL), NPR1, PR1 and HSP70 genes that participate in resistance to pathogens in plants. In contrast, DHZ treatments did not induce significant changes in expression of these genes. The expression of the P. vulgaris alternative oxidase (AOX) gene homolog, an enzyme implicated in plant resistance to viruses, showed low constitutive expression during the first 11 days post-infection and was not affected by either SA or DHZ. It appears that, while SA induced the NPR1-PR1 pathogen defense pathway genes, both SA and DHZ may use a different pathway to induce resistance to WClMV infection in P. vulgaris plants.     

Morphological and microsatellite diversity associated with ecological factors in natural populations of Medicago laciniata Mill. (Fabaceae)

Genetic variability in 10 natural Tunisian populations of Medicago laciniata were analysed using 19 quantitative traits and 12 polymorphic microsatellite loci. A large degree of genetic variability within-populations and among-populations was detected for both quantitative characters and molecular markers. High genetic differentiation among populations for quantitative traits was seen, with Q _ST = 0.47, and F _ST = 0.47 for microsatellite markers. Several quantitative traits displayed no statistical difference in the levels of Q _ST and F _ST . Further, significant correlations between quantitative traits and eco-geographical factors suggest that divergence in the traits among populations may track environmental differences. There was no significant correlation between genetic variability at quantitative traits and microsatellite markers within populations. The site-of-origin of eco-geographical factors explain between 18.13% and 23.40% of genetic variance among populations at quantitative traits and microsatellite markers, respectively. The environmental factors that most influence variation in measured traits among populations are assimilated phosphorus (P₂0₅) and mean annual rainfall, followed by climate and soil texture, altitude and organic matter. Significant associations between eco-geographical factors and gene diversity, H _e , were established in five-microsatellite loci suggesting that these simple sequence repeats (SSRs) are not necessarily biologically neutral.     

Plastid DNA diversity in natural populations of Beta maritima showing additional variation in sexual phenotype and mitochondrial DNA

Summary Plants of two natural populations of Beta maritima, characterized by high percentages of male-sterile plants, have been investigated for organelle DNA polymorphism. We confirm the two classes of mitochondrial DNA variation previously described: (i) mitochondrial DNA (mtDNA) type N is associated with male fertility, whereas mtDNA type S can cause cytoplasmic male sterility (CMS); (ii) the 10.4-kb linear plasmid is observed in both types of mitochondria and is not correlated with the cytoplasmic male sterility occurring in this plant material. A third polymorphism is now described for chloroplast DNA (ctDNA). This polymorphism occurs within single populations of Beta maritima. Three different ctDNA types have been identified by HindIII restriction analysis. Among the plants studied, ctDNA type 1 is associated with N mitochondria and type 2 with S mitochondria. Chloroplast DNA type 3 has been found both in a fertile N plant and in a sterile S plant. This finding suggests that the chloroplast DNA polymorphism reported is not involved in the expression of male sterility. A comparison with Beta vulgaris indicates that ctDNA type 3 of Beta maritima corresponds to the ctDNA of fertile sugar beet maintainer lines. The three types of Beta maritima ctDNA described in this study differ from the ctDNA of male-sterile sugar beet.     

Restriction fragment length polymorphism analysis of loci associated with disease resistance genes and developmental traits in Pisum sativum L.

An F₂ population of pea (Pisum sativum L.) consisting of 174 plants was analysed by restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. Ascochyta pisi race C resistance, plant height, flowering earliness and number of nodes were measured in order to map the genes responsible for their variation. We have constructed a partial linkage map including 3 morphological character genes, 4 disease resistance genes, 56 RFLP loci, 4 microsatellite loci and 2 RAPD loci. Molecular markers linked to each resistance gene were found: Fusarium wilt (6 cM from Fw), powdery mildew (11 cM from er) and pea common Mosaic virus (15 cM from mo). QTLs (quantitative traits loci) for Ascochyta pisi race C resistance were mapped, with most of the variation explained by only three chromosomal regions. The QTL with the largest effect, on chromosome 4, was also mapped using a qualitative, Mendelian approach. Another QTL displayed a transgressive segregation, i.e. the parental line that was susceptible to Ascochyta blight had a resistance allele at this QTL. Analysis of correlations between developmental traits in terms of QTL effects and positions suggested a common genetic control of the number of nodes and earliness, and a loose relationship between these traits and height.     

Induction and floral determination in the terminal bud of Nicotiana tabacum L. cv. Maryland Mammoth,a short-day plant

Floral determination in the terminal bud of the short-day plant Nicotiana tabacum cv. Maryland Mammoth has been investigated. Plants grown continuously in short days flowered after producing 31.4±1.6 (SD) nodes while plants grown continuously in long days did not flower and produced 172.5±9.5 nodes after one year. At various ages, expressed as number of leaves that were at least 1.0 cm in length above the most basal 10-cm leaf, one of three treatments was performed on plants grown from seed in short days: 1) whole plants were shifted from short days to long days, 2) the terminal bud was removed and then rooted and grown in long days, and 3) the terminal bud was removed and then rooted and grown in short days. Whole plants flowered only when shifted from short days to long days at age 15 or later. Only rooted terminal buds from plants at age 15 or older produced plants that flowered when grown in long days. Only terminal buds from plants at age 15 or older that were rooted and grown in short days produced the same number of nodes as they would have produced in their original locations while buds from younger plants produced more nodes than they would have in their original locations. Thus, determination for floral development in the terminal bud, as assayed by rooting, is simultaneous with the commitment to flowering as assayed by shifting whole plants to non-inductive conditions.Abbreviations LD long day(s) - SD short day(s) - DN dayneutral     

浙江省野生蜡梅花部形态变异及其与环境因子的相关性

以浙江省蜡梅自然分布区的5个居群为研究对象,采用巢式方差分析、主成分分析、相关分析、聚类分析等多种分析方法,探究蜡梅花部的形态变异及其与环境因子相关性以及居群间和居群内的表型多样性。结果表明:蜡梅花部性状中除雄蕊长度、雌蕊数、雌蕊长度、内被片数以外,其他12个性状在居群间均有极显著差异。胚珠数的平均变异系数最大,花冠内径的平均变异系数最小,各性状的平均变异系数为15.03%(10.81%—23.38%)。五尖山的表型多样性最丰富,碧东山居群最小,种群间平均表型分化系数为44.38%(1.57%—89.62%),种群内变异大于种群间变异,种群内变异是花部变异的主要来源。主成分分析显示花冠直径、花冠内径、花筒深度和中被片长对种群变异起主要贡献作用。花部多数性状间存在显著或极显著的相关关系,生态因子中花部性状与土壤酸度相关性最大,土壤中花部性状与大量元素相关性最大,微量元素次之,中量元素最小。通过UPGMA聚类分析可以将5个种群分成两组。浙江省野生蜡梅花部形态存在丰富的变异和多样性,花部部分性状与土壤中钙、铁、铜含量和海拔有显著或极显著相关关系。     

茶条槭自然种群种子和果实表型多样性研究

为探析茶条槭(Acer ginnala)种子和果实表型差异程度及变异规律,该研究采用巢式方差分析、主成分分析、聚类分析等方法,对茶条槭主要分布区内7个种群种子和果实的12个表型性状进行比较研究,分析茶条槭种群间和种群内的表型多样性以及与地理生态因子的关系。结果表明:除种子长/宽(SLW)外,其它11个表型性状在茶条槭种群间和种群内均具有显著差异;各性状平均变异系数(CV)为13.90%,变异幅度为8.14%～32.08%;种群间翅果性状的平均变异系数(15.63%)高于种子性状的平均变异系数(8.71%),表明种子性状的稳定性高于果实性状。主成份分析中,果实形态特征对茶条槭种群表型的贡献率大于种子贡献率。种群间表型分化系数为35.47%,说明种群内变异(64.53%)大于种群间变异(35.47%),暗示变异主要来源于种群内变异。茶条槭种子和果实表型性状受地理生态因子影响较小,主要受自身遗传因素的影响。基于种群间欧式距离的聚类分析将茶条槭7个种群分为2大支,没有严格按照地理距离而聚类,表明茶条槭种群表型性状变异的不连续性。茶条槭不同种群种子和果实具有较高的表型多样性,与其分布范围和生物学特性有关。     

Promotion of elongation and acid invertase activity in Phaseolus vulgaris L. internode segments by phenylacetic acid

Phenylacetic acid (PAA) significantly stimulated the elongation of isolated Phaseolus vulgaris internodal segments and prevented the decline in acid invertase specific activity observed in segments incubated in the absence of growth substances. Unlike IAA, which stimulated both elongation and invertase activity over a very wide range of concentrations (<10^-4 - 1 mol.m^-3; optimum 10^-2 mol.m^-3), the response to PAA was restricted to a much narrower range of concentrations (3 × 10^-2 - 1 mol.m^-3; optimum ca. 1–2 × 10^-1mol.m^-3). At the optimum concentration of PAA, the stimulation of both responses was about 63–75% of that induced by the optimum concentration of IAA. The differences in the concentration range and magnitude of the responses to IAA and PAA were not due to differences in uptake of the two compounds. The stimulation of elongation by both compounds was prevented by 3.6 × 10^-2mol.m^-3 cycloheximide (CH), and acid invertase activites were greatly reduced compared with samples treated with growth substances alone. A saturating concentration of the specific auxin efflux carrier inhibitor N-1-naphthylphthalamic acid (NPA) slightly promoted the growth of control segments, probably by reducing the loss of residual endogenous auxin to the incubation medium. The elongation induced by PAA at its optimum concentration was considerably greater than the elongation induced by NPA, indicating that PAA did not cause growth by preventing the loss of endogenous auxin from the segments. Elongation responses to combinations of IAA and PAA suggested that the compounds were acting additively and that they were affecting growth by the same mechanism.     

Quantitative trait loci for plant height in four maize populations and their associations with qualitative genetic loci

Summary We report that plant height quantitative trait loci (QTLs) identified in a given small population are not consistent with QTLs identified in other small populations, and that most QTLs are in close proximity to mapped qualitative genetic loci. These observations provide evidence to support the hypothesis that qualitative genetic loci are the same loci that affect quantitative traits, and affirm that these modest experiments probably identify real QTLs.     

Immimohistochemical localization of corticotropin-releasing factor in selected brain areas of the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia)

Summary Corticotropin-releasing factor (CRF) was localized in the brains of two passerine species, the European starling (Sturnus vulgaris) and the song sparrow (Melospiza melodia), by means of immunohistochemistry. The hypothalamic distribution of this peptide in these species includes a complex of immunoreactive perikarya observed in the paraventricular nucleus (PVN), in both its medial and lateral divisions. Nerve fibers were also seen running from these areas to the anterior median eminence (AME) where a terminal field is apparent. A wide variety of extra-hypothalamic nuclei containing CRF-immunoreactive cells and fibers were identified. An apparent CRF terminal field can be visualized in the lateral septum. A dense fiber plexus is present in the nucleus accumbens (Ac) and more caudally in the nucleus of the stria terminalis (nST). In colchicinepretreated animals, it was revealed that these areas also contain CRF-stained perikarya. The pattern of CRF immunoreactivity in the Ac-nST complex is continuous, with no distinction apparent between the nuclei. The medial preoptic area (mPOA) and the adjacent diagonal band of Broca contain CRF-fibers, while cells are apparent in the mPOA. In the mesencephalon, cells were visualized in the midbrain central gray; a terminal field and scattered positively stained perikarya were found in areas more ventral to the central grey that are adjacent to the third cranial nerve. Scattered cells were also seen at the border of the nucleus intercollicularis-nucleus mesencephalicus lateralis, pars dorsalis complex. In contrast to mammalian studies, no immunoreactive nerve fibers or perikarya were observed in telencephalic areas homologous to the mammalian neocortex. These studies confirm the presence of a CRF path-way regulating pituitary function and suggest a broad role played by CRF as a neuromodulator or neurotransmitter in autonomic and possibly behavioral activities in these species.     

Ectomycorrhizas associated with a relict population of Dryas octopetala in the Burren, western Ireland II. Composition, structure and temporal variation in the ectomycorrhizal community

The composition, structure and temporal variation of ectomycorrhizal (EM) communities associated with mountain avens (Dryas octopetala) in grass heaths of the Burren, western Ireland were assessed by using soil core sampling in two permanent plots and 30 other sites (196 cores in total). Of the 34 different EM types observed, 11 were common and constituted over 80% of the EM biomass. Four EM types, Craterellus lutescens, Tomentella sp., Dryadirhiza fulgens and Cenococcum geophilum were the most abundant as measured by EM length and frequency of occurrence in cores. The species profile and relative abundances were very similar in cores from the permanent plots and different sites in the Burren, indicating that they were all representative of the same EM community. The below-ground EM community in both plots was compared with production of basidiomes, and the latter was found to be an unreliable indicator of EM community structure. Temporal variation in the EM community was assessed by repeated core sampling of the two permanent plots over a 14-month period (between March 1998 and May 1999). No statistically significant shifts in EM abundance were found between sampling dates, probably as a consequence of the large variation in EM abundance between core samples over the sampling period. No significant relationship was found between rainfall, soil moisture or soil temperature and fluctuations in EM abundance. Patterns of total EM abundance and fluctuations in EM diversity were strongly correlated between the two permanent plots over the sampling period. Temporal fluctuations in the dominant EM type, Craterellus lutescens, were similar in both plots with respect to mycorrhizal length, biomass and relative abundance, and the patterns between both plots were positively correlated. EM diversity was negatively correlated with biomass of ectomycorrhizas of Craterellus lutescens in both plots, but it was significant only in plot 1.     

Low genetic variation in Swedish populations of the rare speciesVicia pisiformis (Fabaceae) revealed with rflp (rDNA) and RAPD

Nine Swedish populations, 1–5 individuals/population, and one cultivated individual of the rare speciesVicia pisiformis were investigated for genetic variation. In hybridizations with two rDNA probes using 8 restriction enzymes, only two individuals belonging to one population were polymorphic. A map of the rDNA gene cluster was constructed for four of the restriction enzymes used. Two of the polymorphic sites were mapped and were found to be located outside regions coding for rRNA, presumably caused by single point mutations or small deletions. The repeat length of the rDNA region was c. 10,000 bp, which corresponds well with the size found for other species belonging toFabaceae. No length polymorphism was found in the intergenic spacer, contrary to the situation found for most other plant species investigated for rDNA variation. The haplotype diversity for the species (Hsp Shannon) was very low (0.055). Within-population values (Hpop) was 0 for all populations except the variable one, which had 0.301. PCR amplification with 6 random primers also revealed very low levels of genetic diversity. A polymorphism was observed in a limited number of individuals for four populations. Hsp was 0.065 and was 0.050. The average D value (Wetton) for the PCR haplotypes was 0.99.