The tomato cytosolic fructokinase FRK1 is important for phloem fiber development

Tomato (Solanum lycopersicum) plants have four fructokinase genes, SlFRK1-4. The SlFRK4 is expressed only in pollen whereas the other three are expressed in all plant parts. While SlFRK2 and SlFRK3 are involved in vascular tissue development and affects the shape, size, and cell-wall width of xylem vessels and xylem fibers, the role of SlFRK1 has not been studied previously. The current work investigates the expression of SlFRK1 using transgenic tomato plants expressing the β-glucuronidase reporter gene under the SlFRK1 promoter, as well as the role of SlFRK1 using transgenic plants with antisense suppression of SlFRK1. The SlFRK1 promoter is expressed primarily in vascular tissues and specific suppression of SlFRK1 reduces water transport in stems, but has no other anatomical or phenotypic effects. Combined suppression of SlFRK1 and SlFRK2 severely inhibited plant growth and an anatomical analysis revealed a reduction in secondary xylem area and distorted phloem fibers characterized by thin cell walls and reduced lignification. The results suggest that SlFRK1 is involved in vascular tissue development and hydraulic conductivity in tomato plants and that SlFRK1 is important for normal phloem fiber development, together with SlFRK2.     

Deciphering the Mechanisms of Endophyte-Mediated Biofortification of Fe and Zn in Wheat

An investigation was carried out to understand the mechanism(s) underlying enhanced Fe or Zn uptake in low Fe–Zn accumulator wheat genotype 4HPYT-414, due to inoculation of siderophore-producing and zinc-solubilizing endophytes—Arthrobacter sulfonivorans DS-68 and Arthrobacter sp. DS-179. Root anatomical features, using transmission electron microscopy (TEM), qualitative and quantitative aspects of production of organic acids and sugars in root exudates, and expression of TaZIP genes were analysed to relate to endophyte-mediated higher concentrations of Fe and Zn in the roots and shoots of wheat plants. TEM studies revealed that the endodermis, cortical region, root hair extension, xylem and xylem vessels, pericycle and vascular bundles were more pronounced and thicker in inoculated treatments, as compared to control. The organic acid profile of root exudates revealed five types of organic acids, with citric acid being predominant. Inoculation of A. sulfonivorans and Arthrobacter sp. brought about 5- and eightfold increases in the amounts of acids, respectively, as compared to control, particularly citric acid, succinic acid and acetic acid. Among the four TaZIP genes targeted, expression was achieved only for TaZIP3 and TaZIP7 genes, which showed 1–2 fold increases in the inoculated treatments. The results clearly indicated that the endophyte-mediated overexpression of TaZIP3 and TaZIP7 genes in roots and shoots, and the observed anatomical and exudate changes were acting synergistically in facilitating higher Fe and Zn translocation in roots and shoots.     

Defence reactions in the apoplastic proteome of oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis> var. <Emphasis Type="Italic">napus</Emphasis>) attenuate <Emphasis Type="Italic">Verticillium longisporum</Emphasis>growth but not disease symptoms

Background

Verticillium longisporum is one of the most important pathogens of Brassicaceae that remains strictly in the xylem during most stages of its development. It has been suggested that disease symptoms are associated with clogging of xylem vessels. The aim of our study was to investigate extracellular defence reactions induced by V. longisporum in the xylem sap and leaf apoplast of Brassica napus var. napus in relation to the development of disease symptoms, photosynthesis and nutrient status.

Results

V. longisporum (strain VL43) did not overcome the hypocotyl barrier until 3 weeks after infection although the plants showed massive stunting of the stem and mild leaf chlorosis. During this initial infection phase photosynthetic carbon assimilation, transpiration rate and nutrient elements in leaves were not affected in VL43-infected compared to non-infected plants. Proteome analysis of the leaf apoplast revealed 170 spots after 2-D-protein separation, of which 12 were significantly enhanced in response to VL43-infection. LS-MS/MS analysis and data base searches revealed matches of VL43-responsive proteins to an endochitinase, a peroxidase, a PR-4 protein and a β-1,3-glucanase. In xylem sap three up-regulated proteins were found of which two were identified as PR-4 and β-1,3-glucanase. Xylem sap of infected plants inhibited the growth of V. longisporum.

Conclusion

V. longisporum infection did not result in drought stress or nutrient limitations. Stunting and mild chlorosis were, therefore, not consequences of insufficient water and nutrient supply due to VL43-caused xylem obstruction. A distinct array of extracellular PR-proteins was activated that might have limited Verticillium spreading above the hypocotyl. In silico analysis suggested that ethylene was involved in up-regulating VL43-responsive proteins.

     

Genetic structure of Sakhalin spruce (<Emphasis Type="Italic">Picea glehnii</Emphasis>) in northern Japan and adjacent regions revealed by nuclear microsatellites and mitochondrial gene sequences

The genetic structure of Sakhalin spruce (Picea glehnii) was studied across the natural range of the species, including two small isolated populations in south Sakhalin and Hayachine, by using six microsatellite loci and maternally inherited mitochondrial gene sequences. We also analyzed P. jezoensis, a sympatric spruce in the range. Genetic diversity of P. glehnii was higher in central Hokkaido and the lowest in the Hayachine. Bayesian clustering and principal coordinate analysis by using the microsatellites indicated that the Hayachine was clearly distinct from other populations, implying that it had undergone strong genetic drift since the last glacial period. P. glehnii harbored four mitochondrial haplotypes, two of which were shared with P. jezoensis. One of the two was observed without geographical concentration, suggesting its derivation from ancestral polymorphism. Another was observed in south Sakhalin and in P. jezoensis across Sakhalin. The Bayesian clustering—by using four microsatellite loci, including P. jezoensis populations—indicated unambiguous species delimitation, but with possible admixture of P. jezoensis genes into P. glehnii in south Sakhalin, where P. glehnii is abundantly overwhelmed by P. jezoensis; this might explain the occurrence of introgression of the haplotype of P. jezoensis into P. glehnii.     

Jasmonic acid modulates xylem development by controlling polar auxin transport in vascular tissues

Development of xylem cells is affected by environmental stresses such as drought and oxidative stress, and recent findings suggested that jasmonic acid (JA) mediates this process through interaction with other phytohormones such as cytokinin. In this study, we showed that polar auxin transport regulated by PIN3 and PIN7 is involved in the JA-mediated xylem development in vascular tissues. The mutant plants that lack the activity of PIN3 and PIN7 responsible for the auxin transport developed extra xylems in vascular tissues such as the JA-treated wild-type plants. Visualization of auxin response and xylem development in the roots treated with NPA, an inhibitor of polar auxin transport, suggested that disruption of polar auxin transport is involved in the xylem phenotype of pin3 pin7 double mutants. We also found that cytokinin increases expressions of PIN3 and PIN7 responsible for the auxin transport while JA decreases only PIN7. These suggested that PIN7-mediated polar auxin transport system modulates xylem development in response to JA. The finding that JA affects auxin distribution in root vascular tissues further supported this. Collectively, these suggest that JA promotes xylem development by disrupting auxin transport in vascular tissues, and the auxin efflux genes, more especially PIN7 whose expression is suppressed by JA mediates this process.     

Xylem anatomy and calculated hydraulic conductance of four <Emphasis Type="Italic">Nothofagus</Emphasis> species with contrasting distribution in South-Central Chile

Nothofagus obliqua, N. dombeyi, N. alpina and N. antarctica are characteristic tree species of the temperate forests on the western slopes of the Andes with centres of distribution that differ in their temperature and moisture regimes. We tested branch wood from co-occurring specimens of these species for the inherent differences in xylem anatomy and theoretical hydraulic conductance to evaluate their resistance to drought or frost. The hydraulic conductivity of the xylem was calculated using a modified Hagen–Poiseuille equation and related to wood density. Conduit dimensions were used to predict the water potential that would cause 50 % loss of hydraulic conductivity (Ψ ₅₀). Nothofagus alpina, which mainly grows at sites with low frost frequency, exhibited the largest conduits and the highest mean values for conduit area, fraction of conduit area in the cross-section and hydraulic conductivity, but the lowest wood density. Opposite relationships were found in the plastic N. antarctica, whose xylem seems to be least vulnerable to freezing-induced, but also to drought-induced embolism. Calculated Ψ ₅₀ was highest (least negative) in N. alpina, indicating a relatively high susceptibility to cavitation. The xylem of the thermophilic N. obliqua and of N. dombeyi, which mainly occurs under oceanic climate, but can also survive at sporadically dry and warm sites, is not particularly adapted to periods of drought stress. Across all species, wood density was negatively correlated with the calculated hydraulic conductance. The xylem traits of N. alpina might contribute to its relatively high growth rate and facilitate its spread into forest gaps.     

<Emphasis Type="Italic">In vitro</Emphasis> propagation of endangered <Emphasis Type="Italic">Mammillaria</Emphasis> genus (Cactaceae) species and genetic stability assessment using SSR markers

In vitro propagation protocols were established for endangered species of cacti Mammillaria hernandezii, M. dixanthocentron, and M. lanata. In vitro-germinated seedlings were used as the explant source. Three explant types were evaluated as apical, basal, and lateral stem sections. Shoot multiplication was achieved using Murashige and Skoog (MS) medium supplemented with benzyladenine, kinetin, meta-topolin, and thidiazuron in equimolar concentrations (0.0, 0.4, 1.1, 2.2, 4.4, and 8.9 μM). Shoot regeneration was obtained primarily in the lateral stem section explants. In M. hernandezii, an average of 7.4 shoots was regenerated in MS medium with 2.2 μM meta-topolin. M. dixanthocentron and M. lanata averaged 16.7 and 17.9 shoots/explant, respectively, in MS medium supplemented with 1.1 μM meta-topolin. Rooting occurred in MS medium without growth regulators. Three in vitro culture cycles were performed to validate the propagation protocols and to verify genetic stability. Shoots were collected in each cycle and genomic DNA was extracted. Amplified microsatellites were used to compare each genotype with its respective donor plant. Polymorphic information content analysis showed low levels of intra-clonal polymorphisms—M. hernandezii 0.04 and M. dixanthocentron and M. lanata both 0.12. More than 95% of the plants were successfully acclimatized in the greenhouse. After 12 months, plants of M. hernandezii reached the flowering stage; M. dixanthocentron and M. lanata flowered at 24 mo.     

Some notes onCornaceae

Colmeiroa F. v.Muell. ist a later homonym and therefore a new name is proposed—Paracorokia. Its systematic position is discussed: it seems to belong toCornaceae rather than toEscalloniaceae. SubgenusDiscocrania Harms of the genusCornus is published in a generic rank.     

Comprehensive dataset of mangrove tree weights in Southeast Asia

We assembled a dataset tabulating the weights of Thai and Indonesian mangrove trees that we measured between 1982 and 2001. We selected four Thai study sites in Phang Nga, Ranong, Satun, and Trat Provinces and one site in eastern Indonesia on Halmahera Island in Maluku Province. The stands in Ranong Province and on Halmahera Island were in primary forests with data collected in the 1980s and the remaining stands were in secondary forests with data collected later. We collected 124 tree samples from ten species (Avicennia alba, Bruguiera cylindrica, B. gymnorrhiza, Ceriops tagal, Rhizophora apiculata, R. mucronata, Sonneratia alba, S. caseolaris, Xylocarpus granatum, and X. moluccensis) and measured the root weights of 32 individuals of nine species (A. alba, B. cylindrica, B. gymnorrhiza, C. tagal, R. apiculata, R. mucronata, S. alba, S. caseolaris, and X. granatum). All sampled trees were subjected to a standardized protocol to obtain aboveground weights. The trunks were divided into horizontal segments from which the leaves and branches were collected separately. Roots were collected by winching them out of the ground, by trench digging, or by complete excavation. Thus, we were able to compile the weights of the trunk, branches, leaves, and roots of each tree sampled. Aerial roots were included in root weight measurements, although they were collected above ground. We compiled separate lists of trunk diameters, trunk heights, heights of the lowest living branches, and the heights of aerial roots on the trunks of trees in different size categories. Our dataset includes a wide range of tree sizes (maximum trunk diameter 48.9 cm), geographical locations (1°10′N–12°24′N, 98°32′E–123°49′E) and organ weights (trunks, branches, leaves, and roots), and therefore should prove useful in future biomass studies of mangrove forests.     

Addenda to the Revisions of the Genera <Emphasis Type="Italic">Gergithus</Emphasis> Stål and <Emphasis Type="Italic">Hemisphaerius</Emphasis> Schaum (Hemiptera,Auchenorrhyncha, Fulgoroidea: Issidae)

The genus Gnezdilovius Meng, Webb et Wang, 2017 is revised. Maculergithus Constant et Pham, 2016, which was described as a subgenus of Gergithus Stål, 1870, is upgraded to a genus. Ishiharanus Hori, 1969 is reinstalled from synonymy with Gergithus and considered a valid name. Two new genera are erected, Ceratogergithus Gnezdilov, gen. n. (type species: Gergithus spinosus Che, Zhang et Wang, 2007) and Ophthalmosphaerius Gnezdilov, gen. n. (type species: Hernisphaerius trilobulus Che, Zhang et Wang, 2006). Hernisphaerius bistriatus Schumacher, 1915, Gergithus carbonarius Melichar, 1906, G. rosticus Chan et Yang, 1994, G. nummarius Chan et Yang, 1994, and G. rotundus Chan et Yang, 1994 are transferred to the genus Epyhemisphaerius Chan et Yang, 1994, Gergithus quinquemaculatus Che, Zhang, Wang, 2007—to the genus Maculergithus, Gergithus chelatus Che, Zhang, Wang, 2007 and G. pseudotessellatus Che, Zhang, Wang, 2007—to the genus Ceratogergithus, Hernisphaerius binocularis Chen, Zhang, Chang, 2014—to the genus Ophthalmosphaerius, and Gergithus robustus hoozanensis Schumacher, 1915—to the genus Gnezdilovius. The male genitalia of Gergithus herbaceus (Kirby, 1891) and Hernisphaerius interclusus Noualhier, 1896 are illustrated for the first time.     

Isolation and characterization of the 4-coumarate:coenzyme A ligase (4CL1) promoter from <Emphasis Type="Italic">Eucalyptus camaldulensis</Emphasis>

The most important enzyme of the phenylpropanoid pathway, 4-coumarate:coenzyme A ligase (4CL), is encoded by several homologous genes including 4CL1. The 4CL1 promoter is a tissue-specific gene expression element, particularly active in the secondary xylem or older stems. In this study, the 1127 bp 5′- upstream region of the 4CL1 coding sequence from Eucalyptus camaldulensis, Euc4CL1, was isolated and characterized. Essential putative cis-elements in the Euc4CL1 promoter included: a TATA-box at ?22/?28 position, two CCAAT-boxes at ?256/?260 and ?277/?281 positions, respectively, an AC-element at ?328/?336 and A-boxes at ?115/?120 and ?990/?995 positions. To investigate the effect of the Euc4CL1 promoter on gene expression, a plant transformation vector, pEuc4CL1p, containing the reporter gene for β-glucuronidase (GUS) under the control of Euc4CL1 promoter was constructed based on the pBI101 backbone and introduced in tobacco plants. Stable expression of the GUS gene in transgenic lines was analysed by a histochemical GUS assay. The results indicated the specific expression of the GUS gene in the stem xylem cells of transgenic tobacco lines was controlled by the Euc4CL1 promoter. The observations suggest the isolated Euc4CL1 promoter is a potential candidate for driving the expression of a foreign gene in plant xylem tissues.     

Molecular phylogenetics and anti-<Emphasis Type="Italic">Pythium</Emphasis> activity of endophytes from rhizomes of wild ginger congener, <Emphasis Type="Italic">Zingiber zerumbet</Emphasis> Smith

Zingiber zerumbet, a perennial rhizomatous herb exhibits remarkable disease resistance as well as a wide range of pharmacological activities. Towards characterizing the endophytic population of Z. zerumbet rhizomes, experiments were carried out during two different growing seasons viz., early-June of 2013 and late-July of 2014. A total of 34 endophytes were isolated and categorized into 11 morphologically distinct groups. Fungi were observed to predominate bacterial species with colonization frequency values ranging from 12.5 to 50 %. Among the 11 endophyte groups isolated, molecular analyses based on ITS/16S rRNA gene sequences identified seven isolate groups as Fusarium solani, two as F. oxysporum and one as the bacterium Rhizobium spp. Phylogenetic tree clustered the ITS sequences from Z. zerumbet endophytes into distinct clades consistent with morphological and sequence analysis. Dual culture assays were carried out to determine antagonistic activity of the isolated endophytes against Pythium myriotylum, an economically significant soil-borne phytopathogen of cultivated ginger. Experiments revealed significant P. myriotylum growth inhibition by F. solani and F. oxysporum isolates with percentage of inhibition (PoI) ranging from 45.17 ± 0.29 to 62.2 ± 2.58 with F. oxysporum exhibiting higher PoI values against P. myriotylum. Using ZzEF8 metabolite extract, concentration-dependent P. myriotylum hyphal growth inhibition was observed following radial diffusion assays. These observations were confirmed by scanning electron microscopy analysis wherein exposure to ZzEF8 metabolite extract induced hyphal deformities. Results indicate Z. zerumbet endophytes as promising resources for biologically active compounds and as biocontrol agents for soft rot disease management caused by Pythium spp.     

Steps to a Semiotics of Being

The following points, which represent a path to a semiotics of being, are pertinent to various sub-fields at the conjunction of semiotics of nature (biosemiotics, ecosemiotics, zoosemiotics) and semiotics of culture—semioethics and existential semiotics included. 1) Semiotics of being entails inquiry at all levels of biological organization, albeit, wherever there are individuals, with emphasis on the living qua individuals (integrated biological individualism). 2) An Umwelt is the public aspect (cf. the Innenwelt, the private aspect) of a phenomenal/experienced world that is organism-specific (rather than species-specific) and ultimately refers to an existential realm. 3) Existential universals at work on Earth include seeking out the edible, dwelling in a medium, holding a phenomenal world (possibly an Umwelt) and being endowed with life, and consequently being mortal. 4) Human Umwelten include speechless Umwelten, spoken Umwelten and alphabetic Umwelten. 5) An Uexküllian phenomenology—stating that semiotic states represent the general class to which all mental/cognitive states belong—can draw on the works of the phenomenologists David Abram and Ted Toadvine (The notion of semiotic states is treated in Tønnessen 2009a: 62–63. For an introduction to eco-phenomenology, see Brown and Toadvine (eds.) 2003). 6) A task for such a phenomenology is to portray the natural history of the phenomenal world. 7) An imperative task in our contemporary world of faltering biological diversity is that of Umwelt mapping, i.e. a mapping of ontological niches. 8) The ecological crisis is an ontological crisis with historical roots in humankind’s domestication of animals and plants, which can be taken as archetypical for our attempted planet-scale taming of the wild. 9) The process of globalization is expressed by correlated trends of depletion of semiotic diversity and semiotic diversification. 10) Semiotic economy is a field which task it is to map the human ontological niche insofar as its semiotic relations are of an economic nature. All ten points will be commented (explicitly or implicitly) in due time.     

<Emphasis Type="Italic">Athyrium sessilipinnum</Emphasis>: A new lady fern (Athyriaceae) from southern China

A new species of Athyrium from southern China is described and illustrated as Athyrium sessilipinnum. Morphological comparisons among this new species and similar ones (e.g., A. arisanense, A. gedeanum, A. epirachis, and A. otophorum) are provided. Molecular evidence reveals a sister relationship between the new species and A. arisanense. The most distinctive characters of A. sessilipinnum are the sessile, non-pinnatifid and non-shortened pinnae.     

Improving biocontrol of black vine weevil (<Emphasis Type="Italic">Otiorhynchus sulcatus</Emphasis>) with entomopathogenic fungi in growing media by incorporating spent mushroom compost

Amending a peat-based growing medium with 10% v/v spent mushroom compost, a source of fungal chitin and other nutrients, prolonged the persistence of entomopathogenic fungi (Metarhizium brunneum Petsch and Beauveria bassiana (Balsamo) Vuillemin; Hypocreales: Clavicipitaceae). This resulted in improved efficacy of M. brunneum against black vine weevil, Otiorhynchus sulcatus F. (Coleoptera: Curculionidae) larvae compared with using inoculum without spent mushroom compost. B. bassiana only controlled larvae when used in combination with spent mushroom compost (75?±?7% reduction in live larvae). Mixing entomopathogenic fungal inoculum with spent mushroom compost and growing medium was as effective in controlling black vine weevil larvae as using spent mushroom compost colonised with M. brunneum or B. bassiana in the growing medium (80?±?12% reduction in live larvae). The former method is preferable since it does not require production and storage of colonised spent mushroom compost, or registration of new substrate formulations of M. brunneum or B. bassiana.     

Trematodes (Plathelminthes) of <Emphasis Type="Italic">Clarias gariepinus</Emphasis> (Pisces: Clariidae) in Lake Tana,Ethiopia

All of the 166 Clarias gariepinus catfishes in Lake Tana, Ethiopia, were examined for trematode infestation in 2006—2009. Seven trematode species—Eumasenia bangweulensis, Astiotrema reniferum, Orientocreadium batrachoides, Paralecithodendrium chilostomum, Phyllodistomum bavuri, P. tana, and Cladorchiidae gen. sp.—as adult were found. The common catfish parasites were Eumasenia bangweulensis (20% prevalence and 1—62 intensity of invasion), Orientocreadium batrachoides (30% prevalence and 1–31 intensity of invasion), Phyllodistomum bavuri (24.8% prevalence and 1–8 intensity of invasion), Ph. tana (17.6% prevalence and 1–23 intensity of invasion), and Ph. bavuri. Astiotrema reniferum (three specimens were only found) was a rare species; Paralecithodendrium chilostomum was an accidental parasite of catfish. All these trematodes were first recorded in Ethiopia and Eastern Africa.     

<Emphasis Type="Italic">Melampsora pakistanica</Emphasis> sp. nov., a new rust fungus on <Emphasis Type="Italic">Euphorbia helioscopia</Emphasis> (Sun spurge) from Pakistan

A rust fungus was found on the leaves of Euphorbia helioscopia during a field study in Pakistan. Previously, Melampsora euphorbiae, M. euphorbiae-gerardianae and M. helioscopiae have been reported on E. helioscopia, the first two of which are also known from Pakistan. Morphological observations of the newly collected rust samples detected some differences from the previously described Melampsora species on E. helioscopia. The molecular analysis of the ITS and LSU sequences also detected that the rust is different from the previously reported rusts described from E. helioscopia. Based on both morphological comparisons and sequence analysis, the rust is described here as M. pakistanica sp. nov. This species could have potential as a bio-control agent against its host plant—E. helioscopia—which is a weed of wheat fields in Pakistan and elsewhere in the world.     

Distribution and ecological relations among the alien crab, <Emphasis Type="Italic">Percnon gibbesi</Emphasis> (H. Milne-Edwards 1853) and autochthonous species,in and out of an SW Mediterranean MPA

The distribution of the allochthonous crab Percnon gibbesi and its relationships with other benthic invertebrate species was assessed inside the marine reserve of Cabo de Palos—Islas Hormigas (Mediterranean sea, Spain) and neighbouring non-protected sites. Although a significant spatial variability was detected at finer spatial scale, there was no influence of protection measures or insularity on the abundance of P. gibbesi. The presence of small holes, encrusting algae and low slope favour the colonization success of this crustacean, and the spatial distribution of these habitat features could explain the observed pattern. The abundance of P. gibbesi was similar to that of native crab species; however, a non-significant negative relationship between the abundance of P. gibbesi and native crabs (Pachygrapsus marmoratus and Eriphia verrucosa), urchins (Arbacia lixula and Paracentrotus lividus) and a snail (Phorcus turbinatus) was observed. This work highlights the importance of monitoring alien crab population densities taking structural habitat and other potentially influential factors into account and the likely effect of this alien species on the native ones.