Exploiting wild population diversity and somaclonal variation in the salt marsh grass Distichlis spicata (Poaceae) for marsh creation and restoration

The salt marsh grass Distichlis spicata was regenerated from tissue culture and propagated in a greenhouse. Selected regenerants, along with selections from six wild populations, were grown for two years in a common garden flood-irrigated thrice weekly with tidal creek water. Selected wild and regenerated plants were also planted in a created salt marsh. Significant differences among regenerant and wild population selections were found in several functionally important salt marsh plant characteristics, including potential detritus production, belowground organic matter production, canopy structure, and decomposition rate. A combination of characteristics not found in the wild populations was evident in a regenerated line that exhibited both a high detritus production potential and a high decomposition rate. The amount of variation that occurred among regenerants from one parental line via somaclonal variation was similar to that which occurred among the wild population selections. Results of this study suggest that tissue culture may provide a means of producing marsh grasses with specific characteristics for directing the functional development of newly created salt marshes.     

Intrapopulation sex ratio variation in the salt grass Distichlis spicata

ABSTRACT In many dioecious plant populations, males and females appear to be spatially segregated, a pattern that is difficult to explain given its potentially high costs. However, in asexually propagating species, spatial segregation of the sexes may be indistinguishable from superficially similar patterns generated by random establishment of a few genets followed by extensive clonal spread and by gender-specific differences in rates of clonal spread. In populations where a significant fraction of individuals are not flowering and gender cannot be assigned to this fraction, apparent spatial segregation of the sexes may be due to differential flowering between the sexes. We confirm reports that flowering ramets of the clonal, perennial grass Distichlis spicata are spatially segregated by sex. We extend these studies in two fundamental ways and demonstrate that this species exhibits true spatial segregation of the sexes. First, using RAPD markers, we estimated that at least 50% of ramets in patches with biased sex ratios represent distinct genotypes. Second, we identified a RAPD marker linked to female phenotype (eliminating the possibility that gender is environmentally determined) and used it to show that the majority of patches exhibit significantly biased sex ratios for both ramets and genets, regardless of flowering status.     

Structural features of the salt glands of the leaf of Distichlis spicata 'Yensen 4a' (Poaceae)

The epidermal salt glands of the leaf of Distichlis spicata ‘Yensen 4a’ (Poaceae) have a direct contact with one or two water-storing parenchyma cells, which act as collecting cells. A vacuole occupying almost the whole volume of the collecting cell has a direct exit into the extracellular space (apoplast) through the invaginations of the parietal layer of the cytoplasm, which is interrupted in some areas so that the vacuolar-apoplastic continuum is separated only by a single thin membrane, which looks as a valve. On the basis of ultrastructural morphological data (two shapes of the extracellular channels, narrow and extended, are found in basal cells), the hypothesis on the mechanical nature of the salt pump in the basal cell of Distichlis leaf salt gland is proposed. According to the hypothesis, a driving force giving ordered motion to salt solution from the vacuole of the collecting cell through the basal cell of the salt gland to cap cell arises from the impulses of a mechanical compression–expansion of plasma membrane, which penetrates the basal cell in the form of extracellular channels. The acts of compression–expansion of these extracellular channels can be realized by numerous microtubules present in the basal cell cytoplasm.     

Microsatellite markers isolated from saltgrass (Distichlis spicata)

Twelve polymorphic microsatellite DNA loci were isolated from saltgrass (Distichlis spicata) and optimized for future studies of its breeding system. The loci were screened for variability among 24 individuals from two populations. The primers amplified loci with numbers of alleles ranging from four to 14 per locus and polymorphic information content from 0.481 to 0.951. Observed heterozygosity varied from 0.227 to 0.958.     

Communities of arbuscular mycorrhizal fungi in Stipa krylovii (Poaceae) in the Mongolian steppe

We examined arbuscular mycorrhizal (AM) fungi colonizing the roots of Stipa krylovii, a grass species dominating the grasslands of the steppe zone in Hustai and Uvurkhangai in Mongolia. The AM fungal communities of the collected S. krylovii roots were examined by molecular analysis based on the partial sequences of a small subunit of ribosomal RNA gene as well as AM fungal colonization rates. Almost all AM fungi detected were in Glomus-group A, and were divided into 10 phylotypes. Among them, one phylotype forming a clade with G. intraradices and G. irregulare was the most dominant. Furthermore, it was also found that most of the phylotypes include AM fungi previously detected in high altitude regions in the Eurasian Continent. Significant correlations were found among soil total N, total plant biomass and AM fungal colonization ratio, which suggested that higher plant biomass may be required for the proliferation of AM fungi in the environment. Meanwhile, redundancy analysis on AM fungal distribution and environmental variables suggested that the effect of plant biomass and most soil chemical properties on the AM fungal communities were not significant.     

Sources of variation in the interaction between three cereal aphids (Hemiptera: Aphididae) and wheat (Poaceae)

The relative contributions of host plant, herbivore species and clone to variation in the interaction between cereal aphids and wheat were investigated using five clones each of three species, Rhopalosiphum padi (Linnaeus), Sitobion avenae (Fabricius) and Schizaphis graminum (Rondani), on seedlings of two cultivars of Triticum aestivum L. and one cultivar of Triticum durum Desf. More individuals and biomass of R. padi than of the other two species were produced on seedlings. The three wheat cultivars lost similar amounts of biomass as a result of infestation by aphids, with the amount lost depending on aphid species: S. avenae caused the lowest loss in biomass. Variation in aphid biomass production was due mostly to differences among aphid species (70%), less to the interaction between wheat type and aphid species (7%), and least to aphid clone (1%). The specific impact of the aphids on the plants ranged from 1.7 to 3.7 mg of plant biomass lost per mg of aphid biomass gained, being lowest for R. padi and highest for S. graminum. Variation in plant biomass lost to herbivory was due mostly to unknown sources of error (95%), probably phenotypic differences among individual seedlings, with 3% due to aphid species and none attributable to aphid clone. For these aphid-plant interactions, differences among aphid clones within species contributed little to variation in aphid and plant productivity; therefore, a small sample of clones was adequate for quantifying the interactions. Furthermore, one clone of each species maintained in the laboratory for about 200 parthenogenetic generations was indistinguishable from clones collected recently from the field.     

Unusual chromosome pairing and B chromosomes inBriza spicata (Poaceae)

One plant from a population ofBriza spicata (Poaceae) was found to have highly irregular meiotic behaviour. It is characterized by having a reduced chiasma frequency, a large between cell variance in chiasma frequency and the formation of multivalents involving pairs of A chromosomes. The B chromosome present in this plant also forms multivalents with a pair of A chromosomes. It is suggested that the normal control of strict bivalent pairing has broken down and homoeologous chromosomes are associating as multivalents. Furthermore, the partial homology of the B chromosome with a pair of A chromosomes is revealed.     

丛枝菌根真菌和植物寄生线虫

本文综述了土壤微生物中丛枝菌根真菌和植物寄生线虫的互作关系及其互作机理,并阐述了丛枝菌根真菌在防治植物线虫病害方面的应用前景和实际操作中应注意的技术环节。     

Allozyme variation within and between populations inLolium (Poaceae)

Isozyme analysis was used to determine genetic variation within and between populations of sevenLolium species. All populations from the inbreeding species (L. temulentum, L. remotum, L. loliaceum, andL. persicum) were completely fixed for all enzymes scored. They also contained, for four of the five enzyme systems studied, exactly the same allelic variant. The three cross-breeding species showed large within-population variation and much less between-population variation. The great similarity of the allozymic variants found in all species, made the division of the genusLolium into species on basis of allozymic data difficult. It was not possible to separate the different inbreeding species from each other. Within the cross-breeding groupL. multiflorum andL. rigidum could be distinguished fromL. perenne. L. multiflorum, andL. rigidum could, with more difficulty, also be separated from each other. Allelic variation could have more relation with the provenance of the populations than with taxonomic classification.Genetic variation inLolium spp. II. For first part see Pl. Syst. Evol. 188: 87–99.     

Growth inhibition by exogenous proline and its metabolism in saltgrass (Distichlis spicata) suspension cultures

The growth of Distichlis spicata suspension cultures in LS medium without NaCl was inhibited 54% by 2 mM proline. In medium containing 260 mM NaCl, 10 mM proline inhibited growth by only 22%. The uptake and metabolism of 10 mM L-[1-¹³C] proline was followed by ¹³C NMR and ninhydrin analyses of suspensions cultured in the presence of 0 or 260 mM NaCl. Uptake of 85 to 92% of the exogenous proline occurred within 72 h in all media. In 10 mM proline and no NaCl, cellular proline reached a maximm of 51.5 moles/g FW compared to 1.9 moles/g FW in suspensions not grown on proline. In medium containing 260 mM NaCl and proline, cellular proline reached 59–65 moles/g FW compared to 30–40 moles/g FW in controls grown without proline. The ¹³C-label in the proline-C₁ was either retained in proline or disappeared, presumably released as carbon dioxide, by catabolism through the TCA cycle. Since no metabolite of ¹³C-proline was detected by NMR, proline was considered to be the molecule which inhibited the suspension culture growth.Abbreviations LS Linsmaier and Skoog medium - FW fresh weight - DW dry weight - P5C 1-pyrroline-5-carboxylate - TCA tricarboxylic acid cycle - FID free-induction-decay - NMR nuclear magnetic resonance spectroscopy - T₁ spin-lattice relaxation time - NOE Nuclear Overhauser Effect.     

Xyloglucanases in the interaction between saprobe fungi and the arbuscular mycorrhizal fungus Glomus mosseae

We studied the production of xyloglucanase enzymes of pea and lettuce roots in the presence of saprobe and arbuscular mycorrhizal (AM) fungi. The AM fungus Glomus mosseae and the saprobe fungi Fusarium graminearum, Fusarium oxysporum-126, Trichoderma harzianum, Penicillium chrysogenum, Pleurotus ostreatus and Aspergillus niger were used. G. mosseae increased the shoot and root dry weight of pea but not of lettuce. Most of the saprobe fungi increased the level of mycorrhization of pea and lettuce, but only P. chrysogenum and T. harzianum inoculated together with G. mosseae increased the dry weight of pea and lettuce respectively. The AM and saprobe fungi increased the production of xyloglucanases by plant roots. The level of xyloglucanase activities and the number of xyloglucanolytic isozymes in plants inoculated with G. mosseae and most of the saprobe fungi tested were higher than when both microorganisms were inoculated separately. The possible relationship between xylogucanase activities and the ability of AM and saprobe fungi to improve the dry weight and AM root colonization of plants was discussed.     

Sex‐specific interaction between arbuscular mycorrhizal and dark septate fungi in the dioecious plant Antennaria dioica (Asteraceae)

Male and female plants of dioecious species often differ in their resource demands and this has been linked to secondary sexual dimorphism, including sex‐specific interactions with other organisms such as herbivores and pollinators. However, little is known about the interaction between dioecious plants and fungal root endophytes. Plants may be simultaneously colonised by arbuscular mycorrhizal (AM) and dark septate (DS) fungi. While it is well established that AM mutualism involves reciprocal transfer of photosynthates and mineral nutrients between roots of host plants and these fungi, the role of DS fungi remains controversial. Here, we report the temporal and spatial variation in AM and DS fungi in female, male and non‐reproductive Antennaria dioica plants in three natural populations in Finland during flowering and after seed production. Females had higher colonisation by AM fungi, but lower colonisation by DS fungi than male and non‐reproductive plants. The higher AM colonisation was observed during flowering, and this difference varied among populations. Our results suggest that females and males of A. dioica interact with AM and DS fungi differently and that this relationship is dependent on soil fertility.     

Chloroplast DNA variation in the genusSecale (Poaceae)

The chloroplast genomes of 44 accessions ofSecale were surveyed for restriction site polymorphisms. The accessions were chosen to represent the geographic as well as taxonomic range of the genus. Using 12 restriction enzymes a total of 348 sites were detected. Twenty-nine mutation sites were phylogenetically informative and used in a cladistic analysis. Further, a 0.1 kb insertion separatedSecale from the outgroup species. Only the annual speciesS. sylvestre was distinct from the rest of the taxa. Cultivated rye together with both wild annual and wild perennial accessions were mixed among each other. Sequence divergence (p) among taxa ofSecale was low, varying from 0.000 to 0.005, suggesting a rather recent origin of the genus.     

Genetic variation in the response of the weed Ruellia nudiflora (Acanthaceae) to arbuscular mycorrhizal fungi

The main goal of this work was to test for plant genetic variation in the phenotypic plasticity response of the weed Ruellia nudiflora to arbuscular mycorrhizal (AM) fungi inoculation. We collected plants in the field, kept them under homogeneous conditions inside a nursery, and then collected seeds from these parent plants to generate five inbred lines (i.e., genetic families). Half of the plants of each inbred line were inoculated with AM fungi while the other half were not (controls); a fully crossed experimental design was then used to test for the effects of treatment (with or without AM fungi inoculation) and inbred line (genetic family). For each plant, we recorded the number of leaves produced and the number of days it survived during a 2-month period. Results showed a strong positive treatment effect (plastic response to AM fungi inoculation) for leaf production and survival. Moreover, in terms of survival, the treatment effect differed between genetic families (significant genetic family by treatment interaction). These findings indicate that the positive effect of AM fungi on plant survival (and potentially also growth) differs across plant genotypes and that such condition may contribute to R. nudiflora’s capacity to colonize new environments.     

I. Pathogenic variation in fungi and bacteria and mycorrhizal compatibility: Intraspecific variation in a mycorrhizal association

Laboratory studies with combinations of the fly agaric, Amanita muscaria (L. ex Fries) Hooker and birch (Betula pendula (Roth.)) suggest that the genetical control of mycorrhizal formation has features in common with the legume/Rhizobium symbiosis. There are at least four factors that can be genetically controlled by either the host or the fungus: (a) mycorrhizal formation; (b) the extent of mycorrhizal development; (c) the pattern of mycorrhizal branching, and (d) the shape and size of the host root system.     

Geographic variation in the dichanthelium aciculare complex (Poaceae)

Geographic variation in theDicanthelium aciculare complex was studied by principal components analysis. Few of the attributes analyzed show high correlations with one another, but pubescence patterns are of value in the discrimination of taxa. Sharp differences between phenetic clusters are restricted to one or two geographical regions. When viewed as a whole the complex forms a mosaic of variation patterns and the boundaries between phenetic clusters fade. The complex is reduced to two species,Dichanthelium aciculare (Desv. ex Poir.) Gould & Clark andD. consanguineum (Kunth) Gould & Clark.     

Interactions between vesicular-arbuscular mycorrhizal fungi and asparagus

Summary The vesicular-arbuscular mycorrhizal (VAM) fungus,Glomus versiforme increased significantly the growth ofAsparagus officinalis under controlled conditions using Turface as the growth medium. The growth responses, including increases in root fresh weight, numbers of shoots, shoot dry weight, and shoot height follow a pattern similar to other mycorrhizal systems. Indigenous VAM fungi appeared to have negative effects on average shoot fresh and dry weight, number of shoots per pot and average shoot height on one year oldA. officinalis seedlings obtained from the field and grown under controlled conditions. These results may be due either to the high levels of soluble phosphate present in the soil or the ineffectiveness of the particular indigenous fungi as mycorrhizal fungi in asparagus. Indigenous mycorrhizal fungi overwinter in asparagus root crown as vesicles and as external and internal hyphae. Soil obtained from the same fields as the one year old crowns was a good source of mycorrhizal inoculum for sterile seedlings.     

Effects of nitrous oxide on mitochondrial and cell respiration and growth in Distichlis spicata suspension cultures

The reversible inhibition of respiratory activity could provide a novel approach to the preservation of traditionally hard to store plant germplasm such as clonal materials and recalcitrant seed. The gaseous anesthetic nitrous oxide caused a reversible, dose-dependent, partial inhibition of dioxygen utilization in mitochondrial particles isolated from cell suspension cultures of the salt-tolerant marsh grass Distichlis spicata, with maximal inhibition of 33% after 30 minutes exposure to an atmosphere of 80% nitrous oxide plus 20% oxygen. Respiration of whole cells required longer time to be affected by the anesthetic, and was reversibly inhibited an average of 19% when measured using a differential respirometer. Exposure to 80% nitrous oxide plus 20% oxygen for up to 10 days caused no measurable effect on cell growth.Abbreviations PCV packed cell volume - EDTA sodium ethylenediaminetetraacetic acid - BSA bovine serum albumin - MOPS 3(N-morpholino) propanesulfonic acid - TMPD N,N,N',N'-tetramethyl-p-phenylene diamine - STP standard temperature and pressure