Genetic and genomic relationships in Leymus Hochst

Genetic and genomic relationships among three taxonomically related species of Leymus, northern European L. arenarius (octoploid, 2n = 56), northern American/Pacific L. mollis (tetraploid, 2n = 28) and central Eurasian L. racemosus (tetraploid, 2n = 28), were examined using molecular and cytogenetic methods. The amplified fragment length polymorphism (AFLP) analysis clearly differentiated Icelandic populations of L. arenarius from Alaskan populations of L. mollis. The former group is more genetically homogeneous than the latter. Leymus arenarius in Iceland has a common gene pool and a relatively recent origin. The Alaskan L. mollis, on the other hand, is probably a glacial survival that has accumulated high level of genetic variation and has differentiated into subspecies. Analysis of the 18S-26S ribosomal genes, by restriction fragment length polymorphism (RFLP) and fluorescence in situ hybridization (FISH), revealed a very close relationship between the octoploid northern European L. arenarius and the tetraploid Eurasian L. racemosus, such that the former could have originated from the latter, probably via interspecific hybridization. Leymus-specific DNA sequences were isolated and used for analyzing genetic relatedness among five Leymus species and four Psathyrostachys species. The RFLP analysis of retrotransposon sequence pLm44 and ribosomal clone pTa71 clearly revealed a close relationship between these two genera, i.e. higher variation was found within genera than between them. The results support the previous notion that Leymus is autopolyploid having all genomes being designated Ns as in Psathyrostachys, but a major taxonomic revision of this group would require analysis of more species.     

Molecular variation in Leymus species and populations

Icelandic populations of European lymegrass [Leymus arenarius (L.) Hochst.] were examined using amplified fragment length polymorphism (AFLP) and restriction fragment length polymorphism (RFLP) of the major ribosomal genes (18S-5.8S-26S rDNA), in comparison with Alaskan populations of its closely related species L. mollis (Trin.) Pilger. The AFLP profiles emerged as two distinct entities, clearly separating the two species, and based on species-specific bands it was simple to distinguish these two morphologically similar species. The rDNA-RFLPs also differentiated the species. Within species, the Icelandic L. arenarius was more homogeneous than the Alaskan L. mollis, and its variation was dispersed over geographically different populations, suggesting a common gene pool. The variation among the Alaskan L. mollis was more extensive and its interrupted pattern may be the result of gene introgression at subspecies level. Within a 40-year-old population of L. mollis established in Iceland from Alaskan material, the molecular profiles separated old and new genotypes. Both AFLP and rDNA revealed the new genotypes to be extremely similar. This rapid change in allele frequency is thought to be the result of adaptation to a new environment.     

Genomic and genetic relationships among species of Leymus (Poaceae: Triticeae) inferred from 18S-26S ribosomal genes

The 18S-26S ribosomal genes in three closely related species of Leymus (Poaceae: Triticeae) were examined using fluorescence in situ hybridization (FISH) and restriction fragment length polymorphism (RFLP). Both approaches revealed a close relationship between L. arenarius (8x = 56, northern European) and L. racemosus (4x = 28, central Eurasian), whereas L. mollis (4x = 28, northern American/Pacific) was distinct. Each species had three homologous pairs of major rDNA loci: a1, a2, and a3 for L. arenarius; m1, m2, and m3 for L. mollis; and r1, r2, and r3 for L. racemosus. Leymus arenarius had in addition three minor loci, a4, a5, and a6. The major loci of L. arenarius and L. racemosus were identical, indicating that the former species could have originated from the latter, via interspecific hybridization and/or polyploidy. The rDNA-RFLPs further indicated relationships of these species to other species of Leymus (L. karellini, 8x = 56 and L. angustus, 12x = 84) and Psathyrostachys (P. fragilis, P. huashanica, P. juncea, and P. lanuginosa, which are all diploids). A phenogram constructed from 20 BamHI, EcoRI, and DraI rDNA fragments revealed closer relationship between the two genera, Leymus and Psathyrostachys, than that among species within a genus.     

Absence of the J genome in Leymus species (Poaceae: Triticeae): evidence from DNA hybridization and meiotic pairing.

To test the presence of a J genome in the type species of Leymus, L. arenarius, its total genomic DNA and that of tetraploids L. mollis, L. salinus ssp. salmonis, L. ambiguus, L. chinensis, L. secalinus, L. alaicus ssp. karataviensis, and L. innovatus were probed with the 277-bp insert of pLeUCD2, which can hybridize with the J, S, and P but not with the N, R, V, Q, I, T, and ABD genomes. The DNA probe hybridized with PalI- or TaqI-digested total DNAs from Thinopyrum elongatum (JeJe diploid) and T. elongatum x Psathyrostachys juncea (JeN hybrid), but not with those from L. arenarius (NNNNXXXX octoploid) and all tetraploid Leymus species (NNXX). Attempts to cross diploid Thinopyrum and tetraploid Leymus species yielded only one triploid hybrid, T. elongatum x L. salinus ssp. salmonis. Meiotic chromosome associations at metaphase I of pollen mother cells in the triploid hybrid averaged 19.69 univalents, 0.64 bivalents, and 0.01 trivalents per cell. Chromosome pairings in the tetraploid hybrids of L. mollis x L. salinus ssp. salmonis, and the reciprocal cross, indicate that L. mollis and L. salinus ssp. salmonis shae the same genomic constitution. Both the DNA probe and genome analysis results confirm the absence of the J genome in the seven additional Leymus species tested. Meiotic data indicated that tetraploid Leymus species could not have the genome formula N1N1N2N2; thus their genome formulas should remain as NNXX until the source of X is identified.     

Isolation, characterization, and analysis of Leymus-specific DNA sequences.

Genomic Southern hybridization using labeled total genomic DNA of Leymus mollis as probe showed intense hybridization signals on all restriction enzyme digested DNA from five species of Leymus Hochst., and four species of Psathyrostachys Nevski. Experiments using the same L. mollis probe, but with unlabeled blocking DNA from Psathyrostachys, showed no hybridization at all. These two genera evidently had the same genomic content. Southern hybridization without blocking allowed identification of DNA fragments abundant in Leymus and Psathyrostachys. Fragments potentially specific to Leymus were cloned. Five repetitive DNA clones from L. mollis and L. arenarius were characterized: pLmIs1, pLmIs44, pLmIs51, pLmIs53, and pLaIs56. These clones hybridized to both Leymus and Psathyrostachys on Southern blots - no clone hybridized to only one of these genera. Both Southern blot and fluorescence in situ hybridization (FISH) experiments showed that all the clones contained dispersed repetitive sequences. They painted all and whole chromosomes uniformly except at centromeres, telomeres, and nucleolar organiser regions. Three of these clones, i.e., pLmIs1, pLmIs44, and pLmIs53, were essentially specific to Leymus and Psathyrostachys - little or no hybridization was detected in other genera such as Triticum, Hordeum, Thinopyrum, or Elymus. Sequence analysis further revealed that the clones were part of retroelements. In particular, the clone pLmIs44 produced hybridization profiles suitable for analysis of genetic relatedness among species. The present study shows that Leymus and Psathyrostachys share the same basic genome, Ns, and therefore provides strong evidence for combining these two genera.     

Arbuscular Mycorrhizae of Leymus arenarius on Coastal Sands and Reclamation Sites in Iceland and Response to Inoculation

The occurrence of arbuscular mycorrhizae fungi (AMF) was examined in natural and reclaimed sand dunes in Iceland. On the coastal sand‐plain of Myrdalssandur no spores of AMF were found on barren sand, and very few were found in 1‐ and 5‐year‐old reclamation sites of Leymus arenarius (lymegrass). A significantly higher number of AMF spores and root colonization were found in a 10‐year‐old reclamation site and in a natural old dune system of L. arenarius. AMF spores showed seasonal variation with higher occurrence in the fall than in the spring. On the volcanic island Surtsey, no AMF spores or root colonization were found on L. arenarius in 6‐ and 10‐year‐old dunes. However, AMF spores and root colonization were found in the 22‐year‐old Leymus dune. On Surtsey, a colonization pattern was found where AMF non‐dependent plants were the first colonizers followed by AMF facultative plants, then by AMF‐dependent plants. Today AMF facultative plants have the highest number of species but AMF non‐dependent plants have the largest population size. Three different AMF inocula were tested on L. arenarius; growth of seedlings was improved significantly by an indigenous AMF inoculum compared with commercial inocula. Inoculation of nursery grown seedlings of L. arenarius followed by transplantation into barren sand seeded with L. arenarius is recommended because this method will ensure establishment of the plant and enhance succession.     

Patterns of population genetic structure for springtails and mites in southern Victoria Land, Antarctica

We sequenced the mitochondrial (mt) DNA cytochrome c oxidase subunit I gene to examine comparative phylogeographic patterns for the springtail Gomphiocephalus hodgsoni and the mite Stereotydeus mollis throughout their ranges in southern Victoria Land, Antarctica. Our aim was to extend previous genetic work to encompass a large ice-free area in the Dry Valleys. In particular, we sought to determine if this new region harboured high levels of genetic diversity and if patterns of genetic structure were congruent across taxa. Phylogenetic and nested clade analyses for G. hodgsoni and S. mollis showed similar patterns of population sub-structuring among locations and highlighted several potential refugia that may have existed during glacial maxima. We identified greater levels of genetic divergence in S. mollis and suggest that there is a nucleotide substitution (mutation) rate difference between S. mollis and G. hodgsoni, and/or that S. mollis has had a longer association with the Antarctic landscape.     

我国特有珍稀植物翅果油树濒危原因分析

翅果油树是我国特有种,为国家2级珍稀濒危保护植物,主要分布于山西南部的低山和丘陵区以及陕西秦岭北麓的户县。翅果油树分布区的生态环境特征是：热量充足,降水量较少,立地条件较差。翅果油树濒危的主要原因有（1）果实形态结构特殊,不利于传播和萌发,（2）果实结实率较低,（3）种子寿命短,发芽率低,（4）翅果油树幼苗缺乏竞争力和（5）人力破坏严重。此外,针对翅果油树的濒危现状和原因,还提出了科学保护对策。     

Host plant dispersion, leaf hopper movement and disease transmission

Abstract.

• 1 The plant-to-plant movement of the corn leafhopper, Dalbulus maidis Delong & Wolcott, and the spread of the leafhopper-borne maize rayado fino virus were investigated in four patterns of maize (Zea mays) dispersion.
• 2 D. maidis was less abundant and the spread of the virus was slower in dense stands of maize than in sparse stands.
• 3 When plant density was held constant, leafhoppers were more abundant in maize stands with relatively equidistant plant spacing (uniform dispersion) than in stands with densely-sown rows (linear dispersion) or double-sown hills (clumped dispersion), but there was no difference in virus incidence among these plant dispersion patterns.
• 4 Leafhoppers were less likely to move to adjacent plants in uniform plant dispersion patterns than in either linear or clumped dispersion patterns. This result may explain the lack of higher virus incidence in uniform stands, despite higher leafhopper abundance.
• 5 Leafhopper movement was consistent with a simple rule: the shorter the distance to the next adjacent plant, the more likely a leafhopper is to move between plants.
• 6 These results demonstrate that host plant dispersion can affect the abundance and behaviour of highly mobile herbivorous insects even when plant density is constant.

     

Chemical Composition and Recrystallization of Epicuticular Waxes: Coiled Rodlets and Tubules

Abstract: Coiled rodlets characterize several non-related taxa within the angiosperms. They often occur together with tubules but sometimes also with platelets or transitional forms between them. The ultrastructure chemistry, and recrystallization of epicuticular waxes of three species were investigated by high-resolution scanning electron microscopy, gas chromatography, and mass spectrometry. Whereas Buxus sempervirens (Buxaceae) and Chrysanthemum segetum (Asteraceae) show coiled rodlets in combination with tubules, Leymus arenarius (Poaceae) exhibits tubules but no coiled rodlets. Chemical analyses reveal that the predominating β-diketones of all species differ completely in their molecular structure. Those of the former two species are mainly substituted in carbon atom positions up to 12. In contrast, the wax of L. arenarius contains only hentriacontane-14,16-dione and 25-hydroxy-hentriacontane-14,16-dione. Standard solutions of the total waxes from B. sempervirens, C. segetum and L. arenarius, the purified β-diketone fraction from C. segetum and hentriacontane-14,16-dione from Secale cereale were taken for recrystallization experiments under different conditions in relation to solvent and crystallization velocity. It was demonstrated that coiled rodlets grew exclusively from total waxes of B. sempervirens and C. segetum, and its β-diketone fraction but never from L. arenarius wax or pure hentriacontane-14,16-dione. The recrystallization experiments pointed out that conditions, such as the chemical environment and physical factors, strongly influence the formation of coiled rodlets and tubules. It is concluded that coiled rodlets are formed by self-assembly in close dependence on the position of β-diketo substitution. The future role of β-diketones in the classification of coiled rodlets within wax crystals is discussed.     

Past and future range shifts and loss of diversity in dwarf willow (Salix herbacea L.) inferred from genetics, fossils and modelling

Aim Climate change may cause loss of genetic diversity. Here we explore how a multidisciplinary approach can be used to infer effects of past climate change on species distribution and genetic diversity and also to predict loss of diversity due to future climate change. We use the arctic‐alpine plant Salix herbacea L. as a model. Location Europe, Greenland and eastern North America. Methods We analysed 399 samples from 41 populations for amplified fragment length polymorphism (AFLP) to identify current patterns of genetic structure and diversity and likely historical dispersal routes. Macrofossil records were compiled to infer past distribution, and species distribution models were used to predict the Last Glacial Maximum (LGM) and future distribution of climatically suitable areas. Results We found strong genetic differentiation between the populations from Europe/East Greenland and those from Canada/West Greenland, indicating a split probably predating the LGM. Much less differentiation was observed among the four genetic groups identified in Europe, and diversity was high in the Scandinavian as well as in southern alpine populations. Continuous distribution in Central Europe during the last glaciation was inferred based on the fossil records and distribution modelling. A 46–57% reduction in suitable areas was predicted in 2080 compared to present. However, mainly southern alpine populations may go extinct, causing a loss of about 5% of the genetic diversity in the species. Main conclusions From a continuous range in Central Europe during the last glaciation, northward colonization probably occurred as a broad front maintaining diversity as the climate warmed. This explains why potential extinction of southern populations by 2080 will cause a comparatively low loss of the genetic diversity in S. herbacea. For other species with different glacial histories, however, the expected climate‐change induced regional extinction may cause a more severe loss of genetic diversity. We conclude that our multidisciplinary approach may be a useful tool for assessing impact of climate change on loss of genetic diversity.     

The inter- and intraspecific relationships of Holcus lanutus L. and H. mollis L. sensu lato (Gramineae)

Holcus lanatus and H. mollis were initially considered to be diploids, but their hybrids proved male sterile. Subsequent cytological studies showed H. mollis to be made up of a polyploid series with 2 n = 28, 35, 42 and 49. The planned hybridizations which then became possible showed the races to be fertile in varying degrees with each other and with H. lanatus. Self-fertile plants were found among all groups. Spikelets with two hermaphrodite functional florets were present within the panicles of many plants in the mollis groups and also in H. lanatus. Rhizomes were normal in H. mollis but never developed in H. lanatus. The chromosome races of H. mollis represent 'Cryptic' species since they cannot be separated on morphological criteria. A diploid H. mollis has been reported from Portugal.     

Trypsin from Greenland cod, Gadus ogac. Isolation and comparative properties

Trypsin(ogen) was isolated from the pyloric ceca of Greenland cod. Greenland cod trypsin catalyzed hydrolysis of N alpha-benzoyl-DL-arginine p-nitroanilide, tosyl arginine methyl ester and protein and was inhibited by the serine protease inhibitor PMSF and other well-known trypsin inhibitors. Greenland cod trypsin was more stable at alkaline pH than at acid pH; and was inactivated by relatively low thermal treatment. Like other trypsins, the enzyme was rich in potential acidic amino acid residues but poor in basic amino acid residues and had a molecular weight of 23,500; but it had less potential disulfide pairs, less alpha-helix and a lower H phi ave than other trypsins previously characterized. Reactions catalyzed by Greenland cod trypsin were not very responsive to temperature change, such that specific activity was relatively high at low reaction temperature.     

Ecophysiological differences among Leymus mollis populations across a subarctic dune system caused by environmental, not genetic, factors

Plant species that persist during succession, from the colonization to the stabilization stages, face major environmental changes. Such changes are believed to have significant effects on species performance. In subarctic coastal dune systems, Leymus mollis colonizes the embryo dunes, on the upper limit of the beach. It reaches its maximum density on the foredune, but also grows on older, stabilized ridges. This paper reports on the phenotypic variations of some ecophysiological traits associated with the persistence of L. mollis on a dune system on the east coast of Hudson Bay (northern Quebec). Leymus mollis ramets tend to have a lower net carbon assimilation rate and water use efficiency, and a higher substomatal CO₂ concentration on the stabilized dune than on the foredune. However, these physiological differences cannot be explained by differences in leaf morphology or nitrogen content. Under controlled conditions, ecophysiological differences observed in the field disappear, suggesting that these are not genetic but determined by environmental changes along the foredune-stabilized dune gradient. We propose that higher net carbon assimilation rate on the foredune might be related to higher sink strength in relation to the growth-stimulating effect of sand burial.     

How reproductive allocation and flowering probability of individuals in plant populations are affected by position in stand size hierarchy,plant size and CO2 regime

Aims We investigate the effect of position within a size-structured population on the reproductive allocation (RA) and flowering probability of individual plants of Sinapis arvensis. We also assess the effects of plant size and changing level of CO₂ on both responses.Methods Sinapis arvensis L., (field mustard), an annual agricultural weed, was grown in monoculture at six densities under ambient and elevated CO₂ in a study with 84 stands. Individual aboveground biomass and reproductive biomass were measured. Varying density produced a wide range of mean plant sizes across stands and size hierarchies within stands. Many (~40%) individuals had zero reproductive biomass. Employing a novel modelling approach, we analysed the joint effects of position in stand size hierarchy, plant size and CO₂ on RA and flowering probability of individuals.Important findings We found a strong effect of position within the size hierarchy of individuals in a population: for an individual of a given size, greater size relative to neighbours substantially increased RA and flowering probability at a single harvest time. There was no other effect of plant size on RA. We found a positive effect of elevated CO₂ on RA regardless of position within the size hierarchy. These observed patterns could impact doubly on the reproductive biomass (R) of small individuals. First, because RA is not affected by size, smaller plants will have smaller R than larger plants; and second, for smaller plants lower down in a population size hierarchy, their RA and hence R will be further reduced. These results suggest that size relative to neighbours may be independent of and more important than direct abiotic effects in determining RA. Further studies are required to evaluate how these observed patterns generalize to other populations in non-experimental conditions.     

Biomass, morphology and nutrient contents of fine roots in four Norway spruce stands

Fine root systems may respond to soil chemical conditions, but contrasting results have been obtained from field studies in non-manipulated forests with distinct soil chemical properties. We investigated biomass, necromass, live/dead ratios, morphology and nutrient concentrations of fine roots (<2 mm) in four mature Norway spruce (Picea abies [L.] Karst.) stands of south-east Germany, encompassing variations in soil chemical properties and climate. All stands were established on acidic soils (pH (CaCl₂) range 2.8–3.8 in the humus layer), two of the four stands had molar ratios in soil solution below 1 and one of the four stands had received a liming treatment 22 years before the study. Soil cores down to 40 cm mineral soil depth were taken in autumn and separated into four fractions: humus layer, 0–10 cm, 10–20 cm and 20–40 cm. We found no indications of negative effects of N availability on fine root properties despite large variations in inorganic N seepage fluxes (4–34 kg N ha⁻¹ yr⁻¹), suggesting that the variation in N deposition between 17 and 26 kg N ha⁻¹ yr⁻¹ does not affect the fine root system of Norway spruce. Fine root biomass was largest in the humus layer and increased with the amount of organic matter stored in the humus layer, indicating that the vertical pattern of fine roots is largely affected by the thickness of this horizon. Only two stands showed significant differences in fine root biomass of the mineral soil which can be explained by differences in soil chemical conditions. The stand with the lowest total biomass had the lowest Ca/Al ratio of 0.1 in seepage, however, Al, Ca, Mg and K concentrations of fine roots were not different among the stands. The Ca/Al ratio in seepage might be a less reliable stress parameter because another stand also had Ca/Al ratios in seepage far below the critical value of 1.0 without any signs of fine root damages. Large differences in the live/dead ratio were positively correlated with the Mn concentration of live fine roots from the mineral soil. This relationship was attributed to faster decay of dead fine roots because Mn is known as an essential element of lignin degrading enzymes. It is questionable if the live/dead ratio can be used as a vitality parameter of fine roots since both longevity of fine roots and decay of root litter may affect this parameter. Morphological properties were different in the humus layer of one stand that was limed in 1983, indicating that a single lime dose of 3–4 Mg ha⁻¹ has a long-lasting effect on fine root architecture of Norway spruce. Almost no differences were found in morphological properties in the mineral soil among the stands, but vertical patterns were apparently different. Two stands with high base saturation in the subsoil showed a vertical decrease in specific root length and specific root tip density whereas the other two stands showed an opposite pattern or no effect. Our results suggest that proliferation of fine roots increased with decreasing base saturation in the subsoil of Norway spruce stands.     

沙棘对中国亚湿润干旱区的杨树人工林生长与生产力的影响(英文)

通过将沙棘(Hippophae rhamnoides L．)与3种杨树品种(小黑杨(Populus cv.“Xiaohei”),昭林6号杨(P．cv．“Zhaolin06”)和欧美杨64号(P．euramericane cv．“N3016”))的人工林分别按株混和行混两种方式进行混交实验,研究了固氮植物沙棘对亚湿润干旱区的杨树人工林生长和生产力的影响。研究结果表明：无论哪一种杨树品种或混交方式,沙棘与杨树混交后能显著地增加杨树人工林的生长量,林分平均胸径增加6％～38％,林分平均高增加8％～23％。在株混方式中,杨树地上部生物量大于杨树纯林的地上部生物量。但是在行混方式中,呈现相反的规律,这是由于行混方式中单位面积的杨树株数少。无论哪一种杨树品种或混交方式,杨树与沙棘混交林的地上部净生产力大于杨树纯林的地上部生产力。在株混和行混两种方式中,沙棘占总地上部净生产力的比例分别为20％和41％,但草本植物所占的比重很小。     

Studies of Seedling Inoculation of Frankia in Elaeagnus mollis Diels

The research on the preparation of Frankia solution, seedling culture and inoculation was carried out. Results indicated that the collective amount of cells may be increased when the logarithm phase cells were used as the inoculum and regularly homogenizing and transplanting inoculum by the method of magnetic stirring were adopted during the cultural process of Frankia. The inoculative tests verified that the strain with the highest ability to infect Elaeagnus mollis Diels is SIB 1301118 isolated from Elaeagnus angustifolia L. other than SIB 1332281 isolated from Elaeagnus mollis Diels. The effectivities of inoculation for germinating seeds and the seedlings to be transplanted were seed soaking and root soaking in Frankia solution preparel respectively. Early inoculation was beneficial to the growth and nodulation of seedings. The fefectivity of late inoculation was bad. Sand-soil mixture (2:1) was properly suitable to the seed germination and seedling growth. The seedling growth and nodulative rate could be promoted when 1/4 Sileris-Yong nutrient solution was suppplimented regularly.     

Ectomycorrhizal fungal communities in late-successional Swedish boreal forests, and their composition following wildfire

This study was conducted to evaluate the effects of wildfires on ectomycorrhizal (EM) fungal communities in Scots pine ( Pinus sylvestris ) stands. Below- and above-ground communities were analysed in terms of species richness and evenness by examining mycorrhizas and sporocarps in a chronosequence of burned stands in comparison with adjacent unburned late-successional stands. The internal transcribed spacer (ITS)-region (rDNA) of mycobionts from single mycorrhizas was digested with three restriction enzymes and compared with an ITS–restriction fragment length polymorphism (RFLP) reference database of EM sporocarps. Spatial variation seemed to be more prominent than the effects of fire on the EM fungal species composition. Most of the common species tended to be found in all sites, suggesting that EM fungal communities show a high degree of continuity following low-intensity wildfires. Species richness was not affected by fire, whereas the evenness of species distributions of mycorrhizas was lower in the burned stands. The diversity of EM fungi was relatively high considering that there were only three EM tree species present in the stands. In total, 135 EM taxa were identified on the basis of their RFLP patterns; 66 species were recorded as sporocarps, but only 11 of these were also recorded as mycorrhizas. The species composition of the below-ground community of EM fungi did not reflect that of the sporocarps produced. EM fungal species present in our ITS–RFLP reference database accounted for 54–99% of the total sporocarp production in the stands, but only 0–32% of the mycorrhizal abundance.     

Accurate gene diversity estimates from amplified fragment length polymorphism (AFLP) markers

Three procedures for the estimation of null allele frequencies and gene diversity from dominant multilocus data were empirically tested in natural populations of the outcrossing angiosperm Persoonia mollis (Proteaceae). The three procedures were the square root transform of the null homozygote frequency, the Lynch & Milligan procedure, and the Bayesian method. Genotypes for each of 116 polymorphic loci generated by amplified fragment length polymorphism (AFLP) were inferred from segregation patterns in progeny arrays. Therefore, for the plus phenotype (band present), heterozygotes were distinguished from homozygotes. In contrast to previous studies, all three procedures produced very similar mean estimates of heterozygosity, which were in turn accurate estimators of the direct value (HO = 0.28). A second population of P. mollis displayed markedly lower levels of heterozygosity (HO = 0.20) but approximately twice as many polymorphic loci (284). These AFLP results show that biases in estimates of average null allele frequency and heterozygosity are largely eliminated in highly polymorphic dominant marker data sets displaying a J-shaped beta distribution with a high percentage of loci containing more than three null homozygotes and relatively few loci with no null homozygotes. This distribution may be typical of outcrossing angiosperms.