Taxonomic novelties in <Emphasis Type="Italic">Rhynchospora</Emphasis> (<Emphasis Type="Italic">Cyperaceae</Emphasis>) from South America

Summary  Two new species of Rhynchospora, R. acanthoma and R. rupestris, are described and illustrated. Their geographical distributions are discussed. The status of R. exilis and its synonymy is clarified.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.

     

Isolation and characterization of the <Emphasis Type="Italic">Larix gmelinii </Emphasis><Emphasis Type="Italic">ANGUSTIFOLIA</Emphasis> (<Emphasis Type="Italic">LgAN</Emphasis>) gene

ANGUSTIFOLIA (AN), a plant homolog of C-terminal binding protein, controls the polar elongation of leaf cells and the trichome-branching pattern in Arabidopsis thaliana. In the present study, degenerate PCR was used to isolate an ortholog of AN, referred to as LgAN, from larch (Larix gmelinii). The LgAN cDNA is predicted to encode a protein of 646 amino acids that shows striking sequence similarity to AN proteins from other plants. The predicted amino acid sequence has a conserved NAD-dependent 2-hydroxy acid dehydrogenase (D2-HDH) motif and a plant AN-specific LxCxE/D motif at its N-terminus, as well as a plant-specific long C-terminal region. The LgAN gene is a single-copy gene that is expressed in all larch tissues. Expression of the LgAN cDNA rescued the leaf width and trichome-branching pattern defects in the angustifolia-1 (an-1) mutant of Arabidopsis, showing that the LgAN gene has effects complementary to those of AN. These results suggest that the LgAN gene has the same function as the AN gene.     

Four new species of <Emphasis Type="Italic">Trattinnickia</Emphasis> from South America. Studies in Neotropical Burseraceae XXII

Four new species, Trattinnickia lorenziana, T. rhenocaulis, T. vacciniflora, and T. vulturina, are described and illustrated. This will bring the total number of accepted species in the genus to 16.     

<Emphasis Type="Italic">Oenothera speciosa</Emphasis> versus <Emphasis Type="Italic">Macroglossum stellatarum</Emphasis>: killing beauty

Hovering and dead individuals of the diurnal hawk-moth Macroglossum stellatarum (Linnaeus, 1758) (Lepidoptera: Sphingidae) were found with proboscides got stuck into flowers of the ornamental plant Oenothera speciosa Nutt (Onagraceae). The phenomenon was observed in several locations in Bulgaria where the plant has been introduced. Microscopic examination revealed that the reason for this unusual interaction is pubescence of thick-walled basiscopically oriented trichomes in the basal part of the hypanthium and style of the plant. When a foraging moth inserts its proboscis into this area, the tips of the trichomes are inserted into the transverse grooves of proboscis and hamper its back movement. As a result the moths are suspended for a long time, sometimes until death. Other trapped moth species were also observed but they always effected self-release. This plant–insect interaction is also a conservation issue as an estimation of its impact on wild insect populations is lacking.     

Three new species of <Emphasis Type="Italic">Stigmidium</Emphasis> and <Emphasis Type="Italic">Sphaerellothecium</Emphasis> (lichenicolous ascomycetes) on <Emphasis Type="Italic">Stereocaulon</Emphasis>

Sphaerellothecium stereocaulorum sp. nov., Stigmidium beringicum sp. nov., Stigmidium stereocaulorum sp. nov. and goniocysts are described on Stereocaulon species from the northern Holarctic. Endococcus nanellus is reported new to Alaska and Mongolia. Taxonomical novelties Sphaerellothecium stereocaulorum Zhurb. & Triebel, Stigmidium beringicum Zhurb. & Triebel, Stigmidium stereocaulorum Zhurb. & Triebel.     

Production of transgenic <Emphasis Type="Italic">Pinus armandii</Emphasis> plants harbouring <Emphasis Type="Italic">btCry</Emphasis>III(<Emphasis Type="Italic">A</Emphasis>) gene

A synthetic chimeric gene SbtCryIII(A) encoding the insecticidal protein btCryIII(A), was transformed into Pinus armandii embryos and embryogenic calli using Agrobacterium tumefaciens. Polymerase chain reaction and genomic DNA Southern blot analysis showed that the SbtCryIII(A) gene was integrated into the genome of transgenic Pinus armandii plants, and Northern blot analysis indicated that the SbtCryIII(A) gene was transcribed.     

Distribution and Conservation Status of the Lao Leaf Monkey (<Emphasis Type="Italic">Trachypithecus</Emphasis> (<Emphasis Type="Italic">francoisi</Emphasis>) <Emphasis Type="Italic">laotum</Emphasis>)

Lao leaf monkeys (Trachypithecus (francoisi) laotum) are endemic to a small area of central and, marginally, north Lao. They are known from a few, mostly vague, historical records. We here present a detailed examination of the distribution of this little-known taxon and discuss its conservation status. Surveys since 1992 show its range to be centered upon the karst-dominated Phou Hin Poun National Protected Area (NPA), Nam Sanam Provincial Protected Area, and the southern part of Nam Kading NPA. The known range encompasses <2000 km2, within which occurrence is patchy, reflecting habitat availability. The taxonomic identity of leaf monkeys plausibly of this group reported to the north of this area is not known. In the south of Phou Hin Poun NPA, village reports that the monkeys have black heads are corroborated by the few sightings; their taxonomic relationship with typical Trachypithecus laotum is unknown. In the mid–late 1990s large populations remained and individuals were easily seen. There is no apparent large-scale threat to their habitat. There has been no significant reassessment of status since the late 1990s, nor is there any active conservation action in place. Although the monkeys are to a significant extent protected by the arduous terrain, this cannot be relied upon indefinitely: Trade-directed hunting, although apparently limited in the 1990s, is a potential threat that could cause rapid population declines. Local traditions offer significant starting points for conserving these monkeys.     

Role of Arabidopsis <Emphasis Type="Italic">ABF1</Emphasis>/<Emphasis Type="Italic">3</Emphasis>/<Emphasis Type="Italic">4</Emphasis> during <Emphasis Type="Italic">det1</Emphasis> germination in salt and osmotic stress conditions

Key message

Arabidopsis det1 mutants exhibit salt and osmotic stress resistant germination. This phenotype requires HY5, ABF1, ABF3, and ABF4.

Abstract

While DE-ETIOLATED 1 (DET1) is well known as a negative regulator of light development, here we describe how det1 mutants also exhibit altered responses to salt and osmotic stress, specifically salt and mannitol resistant germination. LONG HYPOCOTYL 5 (HY5) positively regulates both light and abscisic acid (ABA) signalling. We found that hy5 suppressed the det1 salt and mannitol resistant germination phenotype, thus, det1 stress resistant germination requires HY5. We then queried publically available microarray datasets to identify genes downstream of HY5 that were differentially expressed in det1 mutants. Our analysis revealed that ABA regulated genes, including ABA RESPONSIVE ELEMENT BINDING FACTOR 3 (ABF3), are downregulated in det1 seedlings. We found that ABF3 is induced by salt in wildtype seeds, while homologues ABF4 and ABF1 are repressed, and all three genes are underexpressed in det1 seeds. We then investigated the role of ABF3, ABF4, and ABF1 in det1 phenotypes. Double mutant analysis showed that abf3, abf4, and abf1 all suppress the det1 salt/osmotic stress resistant germination phenotype. In addition, abf1 suppressed det1 rapid water loss and open stomata phenotypes. Thus interactions between ABF genes contribute to det1 salt/osmotic stress response phenotypes.

     

Tissue culture of <Emphasis Type="Italic">Sinningia speciosa</Emphasis> and analysis of the <Emphasis Type="Italic">in vitro</Emphasis>-generated <Emphasis Type="Italic">tricussate whorled phyllotaxis</Emphasis> (<Emphasis Type="Italic">twp</Emphasis>) variant

Two protocols were developed for the efficient regeneration of Sinningia speciosa from leaf explants via two developmental pathways. The first method involved formation of callus and buds, followed by subsequent root growth, in Murashige and Skoog medium (MS) containing 2.0 mg l⁻¹ 6-benzylaminopurine (BA) and 0.2 mg l⁻¹ α-naphthalene acetic acid (NAA), with a regeneration efficiency of 99.0%. The second method involved producing callus and roots, followed by subsequent formation of buds, in MS medium supplemented with 1.0–5.0 mg l⁻¹ NAA, and resulted in a regeneration efficiency of 90.4%. Our experiments indicate that the root-first pathway resulted in a lower plant regeneration efficiency. Through five continual generations using the buds-first method, a total of 215 regenerated plants were obtained in the last generation, and eight exhibited a phenotype we named tricussate whorled phyllotaxis (twp). Six of the regenerated twp variant plants maintained their tricussate whorled phyllotaxis phenotype, showing no other abnormalities, while one reverted to a wild type before flowering and another formed two rounds of sepals. Physiological analysis revealed that the twp plants responded differently than wild type to exogenous NAA and 2,3,5-triiodobenzoic acid (TIBA), while high-performance liquid chromatography (HPLC) analysis showed that the levels of endogenous indole-3-acetic acid (IAA) and gibberellin (GA) were lower in twp than wild-type plants. These results suggest that the formation of the twp mutant may be related to phytohormones and that the twp variant could be an important material for investigating the molecular mechanism of plant phyllotaxis patterning.     

Cryptic species of <Emphasis Type="Italic">Notophyllum</Emphasis> (Polychaeta: Phyllodocidae) in Scandinavian waters

The phyllodocid polychaete Notophyllum foliosum occurs in two colour morphs in Swedish and Norwegian waters, one palish yellow to grey form with black patches that is restricted to deeper waters and often associated with reefs of the deep-water coral Lophelia pertusa, and one usually yellow-orange form with black patches and white spots that is usually encountered on more shallow bottoms. We have sampled the two forms from sympatric occurrences in Norway, and the shallow form from the Swedish west coast. Phylogenetic and haplotype analyses based on the mitochondrial cytochrome c oxidase subunit I (COI) gene and the nuclear internal transcribed spacer region (ITS1-5.8SrDNA-ITS2) unequivocally indicate that the two forms represent different species. We apply the name N. foliosum (Sars, 1835) to the ‘shallow form’, and propose N. crypticum n. sp. for the ‘deep form’. A lectotype is fixed for N. foliosum.     

<Emphasis Type="Italic">Drosophila suzukii</Emphasis> in Southern Neotropical Region: Current Status and Future Perspectives

Non-native insect pests are often responsible for important damage to native and agricultural plant hosts. Since Drosophila suzukii Matsumura (Diptera: Drosophilidae) has become an important pest in North America and Europe (i.e., in 2008), the global production of soft thin-skinned fruits has faced severe production losses. In the southern Neotropical region, however, the first record of D. suzukii occurred in 2013 in the south of Brazil. It has also been recorded in Uruguay, Argentina, and Chile. Despite its recent occurrence in the southern Neotropical region, the fast dispersion of D. suzukii has inspired local research efforts in an attempt to mitigate the consequences of this insect pest invasion. In this forum, we explore the current status of D. suzukii in southern Neotropical regions, discussing its future perspectives. Additionally, we attempt to draft activities and a research agenda that may help to mitigate the losses caused by D. suzukii in native and commercial soft-skinned fruits produced in this region. Currently, D. suzukii appears to be well established in the south of Brazil, but considering the entire southern Neotropical region, the invasion panorama is still underinvestigated. The lack of studies and regulatory actions against D. suzukii has contributed to the invasion success of this species in this region. Considering several peculiarities of both the pest biology and the environmental of this region, the authors advocate for the need of intensive and integrative studies toward the development and implementation of area-wide integrated pest management programs against D. suzukii in the southern Neotropical region.     

The <Emphasis Type="Italic">Campylobacter jejuni</Emphasis>/<Emphasis Type="Italic">coli</Emphasis><Emphasis Type="Italic"> cjaA</Emphasis> (<Emphasis Type="Italic">cj0982c</Emphasis>) Gene Encodes an N-Glycosylated Lipoprotein Localized in the Inner Membrane

The Campylobacter coli 72Dz/92 cjaA gene (orthologue of cj0982c of C. jejuni NCTC 11168) product is a highly immunogenic, amino acid-binding protein. CjaA was palmitic acid-modified when processed in E. coli. In addition, site-directed mutagenesis of the Cys residue of the LAAC motif of its signal sequence confirmed that CjaA is a lipoprotein when processed in Campylobacter. Localization of the protein appeared to be host dependent. In Campylobacter, CjaA was recovered mainly as an inner-membrane protein, whereas in E. coli most of the protein was present in the periplasmic space. Interestingly, antiserum raised against Campylobacter glycine-extracted material also recognized CjaA produced by Campylobacter and Escherichia coli, indicating that at least part of the protein may be surface exposed. Site-directed mutagenesis of the Asn residues of two putative N-linked glycosylation sites (NIS and NFT) showed that CjaA is glycosylated and that only the first N-X-S/T sequeon serves as a glycan acceptor.     

Genomewide analysis of <Emphasis Type="Italic">ABCB</Emphasis>s with a focus on <Emphasis Type="Italic">ABCB1</Emphasis> and <Emphasis Type="Italic">ABCB19</Emphasis> in <Emphasis Type="Italic">Malus domestica</Emphasis>

The B subfamily of ATP-binding cassette (ABC) proteins (ABCB) plays a vital role in auxin efflux. However, no systematic study has been done in apple. In this study, we performed genomewide identification and expression analyses of the ABCB family in Malus domestica for the first time. We identified a total of 25 apple ABCBs that were divided into three clusters based on the phylogenetic analysis. Most ABCBs within the same cluster demonstrated a similar exon–intron organization. Additionally, the digital expression profiles of ABCB genes shed light on their functional divergence. ABCB1 and ABCB19 are two well-studied auxin efflux carrier genes, and we found that their expression levels are higher in young shoots of M106 than in young shoots of M9. Since young shoots are the main source of auxin synthesis and auxin efflux involves in tree height control. This suggests that ABCB1 and ABCB19 may also take a part in the auxin efflux and tree height control in apple.     

<Emphasis Type="Italic">Caragana aliensis</Emphasis>, a new species of <Emphasis Type="Italic">Caragana</Emphasis> (<Emphasis Type="Italic">Leguminosae</Emphasis>) from Tibet,China

Summary   Caragana aliensis a new species from Tibet, China, is described and illustrated.     

<Emphasis Type="Italic">Rhizobium</Emphasis> strains in the biological control of the phytopathogenic fungi <Emphasis Type="Italic">Sclerotium</Emphasis> (<Emphasis Type="Italic">Athelia</Emphasis>) <Emphasis Type="Italic">rolfsii</Emphasis> on the common bean

Aims

To identify Rhizobium strains’ ability to biocontrol Sclerotium rolfsii, a fungus that causes serious damage to the common bean and other important crops, 78 previously isolated rhizobia from common bean were assessed.

Methods

Dual cultures, volatiles, indole-acetic acid (IAA), siderophore production and 16S rRNA sequencing were employed to select strains for pot and field experiments.

Results

Thirty-three antagonistic strains were detected in dual cultures, 16 of which were able to inhibit ≥84% fungus mycelial growth. Antagonistic strains produced up to 36.5 μg mL^?1 of IAA, and a direct correlation was verified between IAA production and mycelium inhibition. SEMIA 460 inhibited 45% of mycelial growth through volatile compounds. 16S rRNA sequences confirmed strains as Rhizobium species. In pot condition, common bean plants grown on S. rolfsii-infested soil and inoculated with SEMIA 4032, 4077, 4088, 4080, 4085, or 439 presented less or no disease symptoms. The most efficient strains under field conditions, SEMIA 439 and 4088, decreased disease incidence by 18.3 and 14.5% of the S. rolfsii-infested control.

Conclusions

Rhizobium strains could be strong antagonists towards S. rolfsii growth. SEMIA 4032, 4077, 4088, 4080, 4085, and 439 are effective in the biological control of the collar rot of the common bean.