Connecting plant–microbial interactions above and belowground: a fungal endophyte affects decomposition

Mutualisms can strongly affect the structure of communities, but their influence on ecosystem processes is not well resolved. Here we show that a plant–microbial mutualism affects the rate of leaf litter decomposition using the widespread interaction between tall fescue grass (Lolium arundinaceum) and the fungal endophyte Neotyphodium coenophialum. In grasses, fungal endophytes live symbiotically in the aboveground tissues, where the fungi gain protection and nutrients from their host and often protect host plants from biotic and abiotic stress. In a field experiment, decomposition rate depended on a complex interaction between the litter source (collected from endophyte-infected or endophyte-free plots), the decomposition microenvironment (endophyte-infected or endophyte-free plots), and the presence of mesoinvertebrates (manipulated by the mesh size of litter bags). Over all treatments, decomposition was slower for endophyte-infected fescue litter than for endophyte-free litter. When mesoinvertebrates were excluded using fine mesh and litter was placed in a microenvironment with the endophyte, the difference between endophyte-infected and endophyte-free litter was strongest. In the presence of mesoinvertebrates, endophyte-infected litter decomposed faster in microenvironments with the endophyte than in microenvironments lacking the endophyte, suggesting that plots differ in the detritivore assemblage. Indeed, the presence of the endophyte in plots shifted the composition of Collembola, with more Hypogastruridae in the presence of the endophyte and more Isotomidae in endophyte-free plots. In a separate outdoor pot experiment, we did not find strong effects of the litter source or the soil microbial/microinvertebrate community on decomposition, which may reflect differences between pot and field conditions or other differences in methodology. Our work is among the first to demonstrate an effect of plant–endophyte mutualisms on ecosystem processes under field conditions.     

Microtuberization of layered shoots and nodal cuttings of potato: The influence of growth regulators and incubation periods

A protocol is presented for the rapid induction of microtubers on micropropagated, layered potato shoots of Kennebec, Russet Burbank and Superior in medium devoid of growth regulators. Layered shoots microtuberized more rapidly and produced significantly larger microtubers compared with nodal cuttings. The addition of coumarin or (2-chloroethyl)-trimethylammonium chloride and benzyladenine to microtuberization medium, either had no effect or significantly reduced microtuber weight per shoots compared with medium containing only 80 g × 1^-1 sucrose and minimally affected the number of microtubers per shoot. Increasing the incubation period from 28 to 56 days did not affect the number but significantly increased the weight of microtubers per shoot and substantially increased the proportion, up to 20%, of microtubers heavier than 1 gram.Abbreviations Ba benzyladenine - ccc (2-chloroethyl) trimethylammonium chloride - coumarin 2h-1-benzopyran-2-one - ga₃ gibberellic acid     

Characterization of variability and relationships among components of partial resistance to leaf rust in CIMMYT bread wheats

Summary A study of spring bread wheat (Triticum aestivum) germ plasm developed at the International Maize and Wheat Improvement Center (CIMMYT) showed highly significant phenotypic variability for each component of partial resistance (namely, uredial appearance period, latency period, uredial number and uredial size) to Puccinia recondita f. sp. tritici. All of the wheat genotypes displayed longer uredial appearance and latency periods and decreased uredial number and uredial size when compared to the susceptible check cultivar Morocco. Positive correlations between uredial appearance period and latency period, and uredial number and uredial size, and negative correlations between uredial appearance and latency periods and uredial number and uredial size, inclusive, suggested that the components of partial resistance were either tightly linked or under pleiotropic genetic control. Compared to Morocco, all entries had slow disease progress in the field and variation occurred in the germ plasm for the area under the leaf rust progress curve. Disease progress was negatively correlated with uredial appearance and latency periods, whereas a positive correlation was observed with uredial number and uredial size. Certain genotypes displayed high levels of partial resistance resulting in low disease incidence in the field.     

Absence of the glutamine-synthetase-linked methylammonium (ammonium)-transport system in the cyanobiont of Cycas-cyanobacterial symbiosis

Using the ammonium analogue ¹⁴CH₃NH ₃ ⁺ , ammonium transport was studied in the cyanobiont cells freshly isolated from the root nodules of Cycas revoluta. An L-methionine-dl-sulphoximine (MSX)-insensitive ammonium-transport system, which was dependent on membrane potential (), was found in the cyanobiont. However, the cyanobiont was incapable of metabolizing exogenous ¹⁴CH₃NH ₃ ⁺ or NH ₄ ⁺ because of the absence of another ammonium-transport system responsible for the uptake of ammonium for assimilation via glutamine synthetase (EC 6.3.1.2). Such a modification seems to be the result of symbiosis because the free-living cultured isolate, Anabaena cycadeae, has been shown to possess both the ammonium-transport systems.Abbreviations and symbol ATS/ATSs ammonium transport system/systems - Chl chlorophyll - GS glutamine synthetase - MSX L-methionine-dl-sulphoximine - membrane potential     

Variable effects of endophytic fungus on seedling establishment of fine fescues

Seedborne systemic endophytic fungi of grasses are thought to be plant mutualists, because they have been shown to improve their host’s resistance against biotic and abiotic stresses. The interactions in plant–endophyte associations vary from mutualistic to parasitic with environmental conditions and the genotypes of interacting species. The possible pros and cons of endophytic fungi are expected to be most evident during the seedling establishment, where host fitness is most directly affected. If this holds true, endophytes may play a focal role in local adaptation of hosts to different environments. We examined if endophyte-infected and uninfected seeds and seedlings of two native grass species, Festuca rubra and F. ovina, differ in seed germination and seedling growth rates under greenhouse conditions. The germination of F. rubra seeds was also studied in the field. This is the first time that the effects of Epichloë endophyte on seedling establishment of fine fescues from natural populations have been experimentally evaluated. Mother plant (seed family) had a marked effect on many response variables in both grass species. Length and mean biomass of tillers of endophyte-infected (E+) F. ovina seedlings were lower, but root:shoot ratios were higher than in endophyte-free (E?) seedlings. In F. rubra, the effects of the endophyte were dependent on the habitat where the seeds were collected. The E+ seeds from river banks germinated faster than E+ seeds from meadows, and E+ seedlings from the river banks produced fewer but taller and heavier tillers than the other seedlings. Our data suggest that the effects of the endophyte infection on the seedling stage of fine fescues are dependent the species of grass, host genetic background and mother plant habitat. The germination strategy and growth form of E+ red fescue seedlings from river banks may be beneficial to surviving in the harsh conditions of that habitat.     

Effect of stalk damage on growth and yield of certain maize cultivars by the maize stalk borer Chilo partellus

The effects of different degrees of stem tunneling caused by different levels of artificial infestation by the larvae of Chilo partellus (Swinhoe) on growth and yield of 4 maize cultivars were studied. The susceptible Inbred A suffered much more tunneling than ICZ1-CM, ICZ2-CM and Katumani Composite B. This tunneling influenced significantly the plant growth of all the four cultivars. There was a significant negative correlation between tunneling and yield for the cultivars Inbred A and Katumani Composite B, but such a relationship was not significant for ICZ1-CM and ICZ2-CM. Such differences in the relationships between tunneling and yield among the 4 cultivars are due to different degrees of tunneling caused by C. partellus larvae to the plants of these cultivars.

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Résumé L'étude a porté sur la croissance et la production avec différents niveaux de minage des tiges de 4 cultivars de maïs, lors de divers degrés de contamination artificielle par des chenilles de C. partellus. Le cultivar sensible Inbred A souffre plus des mines de C. partellus que ICZ1-CM, ICZ2-CM et Katumani composite B. Les mines influent significativement sur la croissance des 4 cultivars. Il y a une corrélation négative significative entre le minage des tiges et la production pour les cultivars Inbred A et Katumani composite B, mais la relation n'est pas significative pour ICZ1-CM et ICZ2-CM. Les différences entre le minage des tiges et la récolte des 4 cultivars sont dues aux différents degrés de minage des tiges de ces cultivars par C. partellus.

     

Nuclear Overhauser effect investigation on GM1 ganglioside containingN-glycolyl-neuraminic acid (II3Neu5GcGgOse4Cer)

The conformational properties of the oligosaccharide chain of GM1 ganglioside containingN-glycolyl-neuraminic acid, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer, were studied through NMR nuclear Overhauser effect investigations on the monomeric ganglioside in dimethylsulfoxide, and on mixed micelles of ganglioside and dodecylphosphocholine in water. Several interresidual contacts for the trisaccharide core--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-were found to fix the relative orientitation of the three saccharides, while the glycosidic linkage of the terminal -Gal-was found to be quite mobile as the -Gal-(1-3)--GalNAc-disaccharide exists in different conformations. These results are similar to those found for two GM1 gangliosides containingN-acetyl-neuraminic acid and neuraminic acid [1].Abbreviations Ganglioside nomenclature is in accordance with Svennerholm [23] and the IUPAC-IUB Recommendations [24] GM3(Neu5Ac) II³Neu5AcLacCer, -Neu5Ac-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM3(Neu5Gc) II³Neu5GcLacCer, -Neu5Gc-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Ac) II³Neu5AcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Gc) II³Neu5GcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu) II³NeuGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu-(2-3)]--Glc-(1-1)-Cer - GD1a IV³Neu5AcII³Neu5AcGgOse₄Cer, -Neu5Ac-(2-3)--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GalNAc-GD1a IV⁴GalNAcIV³Neu5AcII³Neu5AcGgOse₄Cer, -GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - Neu neuraminic acid - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolyl-neuraminic acid - Cer ceramide     

Genetic and environmental considerations for evaluating crown position of wheat

Summary Crown position affects winter survival of fallsown wheat (Triticum aestivum L.) Direct or indirect selection for crown depth has been little practiced. Reports have suggested that short subcrown internode length was closely related to semidwarf plant height and that semidwarfism was related to poor emergence. This study determined the relationships among crown depth, plant height, and emergence rate index in three wheat populations. The efficiency of evaluating crown placement in the field was examined and additional information was obtained on its genetic control. The F₂-derived F₄ and F₅ lines from the crosses of female parents Daws, Nugaines, and Stephens with male parent Selection 7952 were planted at Central Ferry and Pullman, Washington, respectively. Correlations from each population indicated that crown depth and subcrown internode length were not closely associated with plant height and emergence rate index. Crown depth was a more reliable indicator of crown placement than subcrown internode length. Adjustment of the data for seed depth differences was essential for evaluating subcrown internode length but less important for evaluating crown depth. After adjustment for seed depth, narrow-sense h ² values for subcrown internode length and crown depth were 0.25–0.41. Crown depth and subcrown internode length were inherited as quantitative traits in phenotypes that expressed variable dominance. Modest gains due to selection for crown depth were achieved.Contribution from USDA-ARS and College of Agriculture and Home Economics Research Center, Washington State University, Scientific Paper No. 7795     

Temporal and spatial patterns of soil water following wildfire-induced changes in plant communities in the Great Basin in Nevada,USA

Infection of tall fescue (Festuca arundinacea Schreb.) with its endemicNeotyphodium coenophialum-endophyte (Morgan-Jones and Gams) Glenn, Bacon and Hanlin appears to reduce copper (Cu) concentrations in forage and serum of grazing animals, contributing to a range of immune-related disorders. A greenhouse experiment was conducted to identify effects of novel endophyte strains on Cu acquisition by tall fescue (Festuca arundinacea Schreb.) varieties Grasslands Flecha and Jesup infected with a novel, non ergot producing endophyte strain AR542, and two perennial ryegrass (Lolium perenne L.) varieties Aries and Quartet infected with a novel, non lolitrem B producing strain AR1, and their noninfected (E−) forms. Individual endophyte/grass associations were cultivated in nutrient solutions at 1.0 (P+) and 0.0 mM (P−) phosphorus concentrations. The Cu²⁺-binding activity of extracellular root exudates, and concentrations of Cu and other heavy metals in roots and shoots were measured. Extracellular root exudates of AR542-infected vs. E− tall fescue had higher Cu²⁺-binding activity only in P− nutrient solution as shown by lower concentration of free Cu²⁺ (0.096 vs. 0.188 mmol Cu²⁺ g⁻¹ root DM, respectively). The Cu²⁺-binding activity by root exudates of perennial ryegrass was not affected by endophyte infection, but was higher (i.e., lower concentration of free Cu²⁺) in P− vs. P+ nutrient solution (0.068 vs. 0.114 mmol Cu²⁺ g⁻¹ root DM). In this hydroponic experiment, Cu concentrations in shoots of both grasses were not a function of Cu²⁺-binding activity and endophyte effects on heavy metal concentrations in shoots and roots were specific for each variety. The Cu²⁺-binding activity of extracellular root exudates may affect Cu accumulation by field-grown, endophyte-infected tall fescue under P-limiting growth conditions and warrants verification by more specific methods.     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

Research on streptococci in the UK

Pleurotus florida produced high amounts of laccase (4.60 U/ml) in malt extract broth after 12 days' growth under stationary conditions. The production of laccase was semi-constitutive. Hyperlaccase mutants ofP. florida were obtained through mutagenesis of mycelial protoplasts usingN-methyl-N-nitro-N-nitrosoguanidine (50 g/ml) for 2 min. Three hyperlaccase mutants were selected showing growth and enzyme production responses similar to the parent.     

Two-dimensional 1H NMR study of two cyclobutane type photodimers of thymidylyl-(3′→5′)-thymidine

2D NMR spectroscopy and J coupling constant analysis are applied to resolve the structure of two photoproducts of thymidylyl-(35)-thymidine. These products are cyclobutane type thymine dimers possessing the cis-syn (the predominant one) and trans-syn geometry. The cis-syn is formed in an ANTI-ANTI conformation about the N-glycosyl linkages and resembles the normal base-stacked configuration. The glycosidic conformation in solution of the 5 terminal fragment differs from the crystal in which the less common SYN conformation is observed. In this isomer only the sugar pucker of the 3 terminal fragment is changed substantially with respect to the dinucleotide. The trans-syn isomer is formed in a SYN-ANTI glycosidic conformation. In this isomer the sugar puckers of both deoxyribose rings are affected and a preference for a pure 2-endo conformation is observed.Abbreviations dTpdT 2-deoxythymidylyl-(35)-2-deoxythymidine - dTp[]dT cyclobutane type photodimers of dTpdT - dTp- and dTp[]- their 5' terminal fragments (fragment A) - -pdT and-[]pdT their 3 terminal fragments (fragment B) - RP-HPLC reversed-phase high-performance liquid chromatography - COSY two-dimensional correlated spectroscopy - 2D NOE two-dimensional nuclear Overhauser spectroscopy     

An improved selection method for λlacZ − phages based on galactose sensitivity

The determination of thelacZ mutant frequency in gt10lacZ phage vectors isolated from the transgenic mouse strain 40.6 (MutaMouse), requires the screening of large numbers of phages on -galactosidase activity. Existing methods rely on distinguishing a few white plaques on X-gal containing plates amongst a multide of blue ones which is both time-consuming and expensive. The new screening method described here employs the galactose sensitiveEscherichia coli C lacZ recA galE strain into which a multicopy plasmid has been introduced, which results in over-expression of thegalK andgalT genes. In the presence of phenyl--d-galactopyranoside, a substrate for -galactosidase, this leads to the suppression of lacZ ⁺ phage propagation without affecting the ability of lacZ ^– phages to form plaques. With this method it is possible to screen 1.5×10⁶ phages on a single 9-cm Petri dish. Furthermore, the need for blue/white screening has been eliminated.     

Effects of fungal endophytes on the seed and seedling biology of Lolium perenne and Festuca arundinacea

Summary Many grasses are infected by endophytic fungi that grow intercellularly in leaves, stems, and flowers and are transmitted maternally by hyphal growth into ovules and seeds. The seed biology and seedling growth of endophyte-infected and uninfected perennial ryegrass (Lolium perenne) and tall fescue (Festuca arundinacea) were investigated under controlled environmental conditions. The percentage of filled seeds produced by infected tall fescue was over twice of uninfected tall fescue; infected and uninfected perennial reegrass had similar percentages. Weights of seeds from infected and uninfected plants were similar in both species. Seeds from infected plants of both species exhibited a higher rate of germination than seeds from uninfected plants. Shoot growth in the greenhouse was compared by making three sequential harvests of above-ground plant parts from infected and uninfected plants of both species. Infected perennial ryegrass plants produced significantly more biomass and tillers than uninfected plants after 6 and 10 weeks of growth and significantly more biomass after 14 weeks of growth. Infected tall fescue plants produced significantly more biomass and tillers than uninfected plants after 10 and 14 weeks of growth. The physiological mechanism of enhancement of growth is not known. The results of this study suggest that infected plants may have a selective advantage in populations with uninfected members.     

Influence of endophyte infection, plant age and harvest interval on Rhopalosiphum padi survival and its relation to quantity of N-formyl and N-acetyl loline in tall fescue

Rhopalosiphum padi L. (Homoptera: Aphididae) is sensitive to loline alkaloids present in tall fescue, Festuca arundinacea Shreb., infected with the endophytic fungus, Acremonium coenophialum Morgan-Jones & Gams. Aphid survival was higher on endophyte-free plants regardless of plant age after germination or age of regrowth tissue after clipping. Survival of aphids on endophyte-infected grass was lower on young tissue but increased as plants aged, although it never reached the same level on endophyte-free plants. Both N-formyl and N-acetyl loline increased as uncut or regrowth tissue aged; however, this was influenced by the age of the plant at the initial cut and the clipping frequency. Although even small amounts of loline cause high aphid mortality, the aphids are able to survive on endophyte-infected plants if the tillers have senescing leaves which contain lower amounts of loline. Preference for senescing leaves may help R. padi avoid plant parts containing high amounts of toxic allelochemicals, thus contributing to higher numbers of aphids on older, endophyte-infected plants.     

Relationships of Agropyron intermedium chromosomes determined by chromosome pairing and alcohol dehydrogenase isozymes in common wheat background

Summary The relationships of Agropyron intermedium chromosomes in two wheat-Agropyron addition series were determined. Chromosome pairing behaviour revealed that the alien chromosome in lines TAF-2 and L7 of Vilmorin-A. intermedium set are homologous to the alien chromosomes in lines P and C of the Caribo-A. intermedium set respectively. Localization of alcohol dehydrogenase isozyme genes in Vilmorin-Agropyron addition line L4 and in Caribo-Agropyron line O indicated relationships with wheat chromosomes of homoeologous group 4.     

Fungal endophytes of grasses and their effects on an insect herbivore

Summary The effects of endophytic fungi (Tribe Balansiae, Clavicipitaceae, Ascomycetes) of grasses on an insect herbivore were studied by feeding paired groups of larvae of the fall armyworm (Spodoptera frugiperda, Noctuidae, Lepidoptera) leaves from either infected or uninfected individuals. Perennial ryegrass infected by the Lolium endophyte, tall fescue infected by Epichloe typhina, dallisgrass infected by Myriogenospora atramentosa, Texas wintergrass infected by Atkinsonella hypoxylon, and sandbur infected by Balansia obtecta were utilized. The endophytes of ryegrass and fescue previously have been shown to be toxic to mammalian herbivores and to deter feeding of some insect herbivores. In this study we extend the antiherbivore properties of those endophytes to the fall armyworm and demonstrate that fungal endophytes in three other genera have similar antiherbivore properties. For most grasses, survival and weights of fall armyworm larvae fed infected leaves were significantly lower and larval duration was significantly longer compared to larvae fed uninfected leaves. Resistance to herbivores may provide a selective advantage to endophyte-infected grasses in natural populations.     

Association of a fungal endophyte in perennial ryegrass with antibiosis to larvae of the southern armyworm, Spodoptera eridania

Larvae of the Southern armyworm Spodoptera eridania Cramer (Lepidoptera:Noctuidae), feeding on perennial ryegrasses (Lolium perenne L.) infected with an endophytic fungus (Acremonium loliae Latch, Christensen and Samuels), had a much lower survival rate (7–13%) than larvae feeding on endophyte-free ryegrasses (82–90%). Death of the larvae on endophyte-infected entries occurred rapidly between 144 h and 168 h of feeding. This corresponded with armyworms feeding on the base of the plant, where endophyte concentration is highest. Twenty-four hours after the start of the bioassay the larval mass and rate of larval development were significantly higher on endophytic entries. From 48–144 h no differences were seen, but after 144 h the mass of larvae on endophyte-infected grasses sharply declined. Observations from this bioassay further substantiate the association between A. loliae-infected ryegrass and antibiosis to several lepidopterous and coleopterous insect pests.

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Résumé Le ray-grass vivace (Lolium perenne), contaminé par le champignon endophyte Acremonium loliae Latch, Christensen & Samuels, a présenté une augmentation de la resistance à de nombreux coléoptères et lépidoptères nuisibles. Cette note examine les réactions de Spodoptera eridania Cramer (Lépido., Noctuidae) alimenté sur trois lignées de ray-grass contaminées par le champignon et trois lignées saines. Après 168 h d'alimentation sur ray-grass contaminé, les chenilles presentent une très forte mortalité; la survie n'est que de 7 à 13% contre 82 à 90% pour le ray-grass sain. Le décès brutal des chenilles correspond à leur alimentation sur la base de la plante ou la concentration du champignon est la plus forte. Les chenilles consomment constamment, broutant plus des 2/3 du feuillage du ray-grass; les broutements des six séries ne différaient pas significativement.Au bout de 24 h, la nombre de chenilles passées du 3ème au 4ème stade, et l'augmentation de poids sont plus élevés pour les séries sur plante contaminée, ce qui suggère un avantage initial pour les chenilles en présence de champignon endophyte, l'analyse en poids sec a montré que l'augmentation de poids initial est réel. Entre 48 et 144 h, cependant, le nombre de 4ème stade et le poids des chenilles sont les mêmes dans les deux séries. Après 144 h, le poids des chenilles sur ray-grass contaminé diminue significativement; aucune n'était parvenue au 5ème stade, contre 11% sur ray-grass sain. Nous n'avons pas observé de signes apparents de neurotoxicité. Au lieu de cela, il ya a eu interaction avec un processus physiologique fondamental, ce qui a provoqué une forte perte de poids larvaire et la mort, indiquant l'intervention d'inhibiteurs métaboliques.

     

Peculiar properties of the PsaF photosystem I protein from the green alga Chlamydomonas reinhardtii: presequence independent import of the PsaF protein into both chloroplasts and mitochondria

It has previously been shown that presequences of nuclear-encoded chloroplast proteins from the green alga Chlamydomonas reinhardtii contain a region that may form an amphiphilic -helix, a structure characteristic of mitochondrial presequences. We have tested two precursors of chloroplast proteins (the PsaF and PsaK photosystem I subunits) from C. reinhardtii for the ability to be imported into spinach leaf mitochondria in vitro. Both precursors bound to spinach mitochondria. The PsaF protein was converted into a protease-protected form with high efficiency in a membrane potential-dependent manner, indicating that the protein had been imported, whereas the PsaK protein was not protease protected. The protease protection of PsaF was not inhibited by a synthetic peptide derived from the presequence of the N. plumbaginifolia mitochondrial F₁ subunit. Furthermore, if the presequence of PsaF was truncated or deleted by in vitro mutagenesis, the protein was still protease-protected with approximately the same efficiency as the full-length precursor. These results indicate that PsaF can be imported by spinach mitochondria in a presequence-independent manner. However, even in the absence of the presequence, this process was membrane potential-dependent. Interestingly, the presequence-truncated PsaF proteins were also protease-protected upon incubation with C. reinhardtii chloroplasts. Our results indicate that the C. reinhardtii chloroplast PsaF protein has peculiar properties and may be imported not only into chloroplasts but also into higher-plant mitochondria. This finding indicates that additional control mechanisms in the cytosol that are independent of the presequence are required to achieve sorting between chloroplasts and mitochondria in vivo.Abbreviations cTP chloroplast transit peptide - mTP mitochondrial targeting peptide - Rubisco ribulose-1,5-bisphosphate carboxylase/oxygenase - pF₁(1,25) a synthetic peptide derived from the first 25 residues of the Nicotiana plumbaginifolia mitochondrial ATP synthase F₁ subunit - PsaF(2–30) and PsaF(2–61) mutant proteins lacking regions corresponding to residues 2–30 and 2–61 in the PsaF precursor protein, respectively     

Mechanisms of the filtering area adaptation in Daphnia

The changes of selected parameters of the filtering comb of the third thoracic limb were studied in a natural population of Daphnia pulicaria Forbes, as well as in experimental enclosures and in lab cultures, including individual life history. Two hypotheses were tested: 1. either these changes are related to the succession of clones coexisting within one population, or 2. the size of the filtering area changes gradually as an individual adaptation during the moulting. No evidence supporting the clonal hypothesis was found. On the contrary, the adaptability of the filtering comb is the same in a natural population as it is in a clone and in individuals.