Molecular identification of house dust mites and storage mites

Mites are known causes of allergic diseases. Currently, identification of mites based on morphology is difficult if only one mite is isolated from a (dust) sample, or when only one gender is found, or when the specimen is not intact especially with the loss of the legs. The purpose of this study was to use polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) of the ITS2 gene, to complement the morphological data for the identification of mites to the species level. For this, six species were cultured: Dermatophagoides pteronyssinus, D. farinae, Blomia tropicalis, Tyrophagus putrescentiae, Aleuroglyphus ovatus and Glycycometus malaysiensis. Genomic DNA of the mites was extracted, quantified, amplified and digested individually with restriction enzymes. Hinf I and Ple I differentiated the restriction patterns of D. pteronyssinus and D. farinae. Bfa I and Alu I enzymes differentiated B. tropicalis and G. malaysiensis. Ple I enzyme was useful for the differentiation between T. putrescentiae and A. ovatus. Bfa I was useful for the differentiation of G. malaysiensis from the rest of the species. In conclusion, different species of mites can be differentiated using PCR–RFLP of ITS2 region. With the established PCR–RFLP method in this study, identification of these mites to the species level is possible even if complete and intact adult specimens of both sexes are not available. As no study to date has reported PCR–RFLP method for the identification of domestic mites, the established method should be validated for the identification of other species of mites that were not included in this study.     

Genetic characterisation of the intermediate hosts of schistosomiasis in its southern distribution limit in South America

ABSTRACT

Schistosomiasis is a tropical disease caused by the digenean parasite Schistosoma mansoni. In South America it is transmitted to humans via freshwater snails of the genus Biomphalaria. In a global warming scenario, disease range expansions to subtropical countries are possible. For the first time, the distributions and genetic identification of Biomphalaria specimens in border regions of Uruguay are reported. The inclusion of Uruguayan samples allows a better understanding of the relationships between and within taxa of Biomphalaria. Samples were collected between 2015 and 2016 using hand nets. Initially, they were classified morphologically. They were then classified genetically by analysing a fragment of the cytochrome c oxidase subunit I gene. Phylogenetic analysis revealed the presence of B. peregrina and B. occidentalis. Species belonging to the B. tenagophila complex were also recognised. Individuals initially identified from their external morphology as B. tenagophila tenagophila showed inconsistencies with the analysis of COI sequences that assigned them to B. occidentalis. Since the presence of schistosomiasis in Uruguay is likely to occur in the next few years, an exhaustive population survey of Biomphalaria taxa should be urgently developed to identify the presence of S. mansoni and the places most susceptible to be colonised by these snails.     

Research Note: Simple molecular discrimination of cultivated Porphyra species (Porphyra yezoensis and Porphyra tenera) and related wild species (Bangiales,Rhodophyta)

To discriminate between cultivated Porphyra species (Porphyra yezoensis and Porphyra tenera) and closely related wild Porphyra species, we developed a polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis of the rbcL gene using five restriction enzymes. Although our previous PCR‐RFLP analyses of internal transcribed spacer (ITS) rDNA and plastid RuBisCO spacer regions could not always discriminate wild P. yezoensis, wild P. tenera, and closely related wild species, the PCR‐RFLP profiles of the rbcL gene were useful in discriminating samples collected from natural habitats. Therefore, PCR‐RFLP analysis of the rbcL gene will help in the simple identification of a large number of samples, not only for the establishment of reliable cultures as breeding material, but also for the taxonomic investigations of species that are closely related to cultivated Porphyra.     

RESTRICTION ENZYME ANALYSIS OF DNA FROM SPECIES OF BULINUS (BASOMMATOPHORA: PLANORBIDAE) USING A CLONED RIBOSOMAL RNA GENE PROBE

A ribosomal RNA gene probe (pSM889) has been used to study restrictionenzyme digests of various species of Bulinus. In order to minimiseproblems of DNA shearing associated with snail tissues a methodof extracting nucleic acids from material embedded in agaroseblocks has been used. Restriction enzyme digests with Bgl IIand Bam HI hybridised to pSM889 showed clear differences betweenB. truncatus, B. wrighti, B. africanus and B. forskalii, representingthe four species groups of Bulinus. No differences were observedbetween samples of B. tropicus and B. truncatus digested withBam HI, Bgl II and Pst I. Intra-specific variation was observedbetween samples of B. forskalii from Säo Tomé andAngola digested with Bgl II and Hind III although restrictionprofiles for Bam HI, Pst I and Bst EII digests were similar.Intra-specific variation was also observed between two differentpopulation samples of B. wrighti from South Yemen using BamHI and Bgl II digested genomic DNA hybridised to pSM889. (Received 5 December 1989; accepted 19 April 1990)     

Resource competition between two rotifer species (Brachionus rubens and B.calyciflorus): an experimental test of a mechanistic model

A mechanistic model of competition on a single resource wastested experimentally with two freshwater rotifers, Brachionusrubens and B.calyciflorus, both grown on the alga Chlorellapyrenoidosa. Using open culture systems for each species wemeasured: (i) the resource-saturated exponential growth rate,µ_max, and (ii) the relationship between specific growthrate, µ, at steady-state and the residual algal concentrationover a range of system turnover rates, or dilution rates, D.The µ_max of B.calyciflorus was {small tilde}60% higherthan B. rubens. These results were then used to construct agraphical model for predicting the victor in interspecific competitionbetween the two rotifers. Since the two resource-dependent growthrates crossed, one species, B.calyciflorus, was predicted tobe the victor at a high D while B. rubens was predicted as thevictor at low D. Finally, the outcome of competition was determinedfor two turnover rates. As predicted by the graphical competitionmodels, B.calyciflorus was the dominant species at rapid D (0.029h^–1) and B.rubens was dominant at slow D (0.0044 h^–1).These studies support recent conclusions that mechanistic competitionmodels may be applied to predict dominant species from a prioriinformation on growth potential and resource levels, which isnot possible with traditional Lotka-Volterra models.     

Simultaneous identification of Grapholita molesta Busck and Grapholita dimorpha Komai by PCR‐RFLP

Although several molecular diagnostic techniques are available for the identification of the apple‐feeding pests Grapholita molesta Busck and Grapholita dimorpha Komai, these pests are severely affecting apple orchards in Korea. These two pests may be misidentified or the available molecular diagnostic techniques may not facilitate the simultaneous identification of the morphological features of both species. In this study, we developed a multiplex assay for these two species using the polymerase chain reaction – restriction fragment length polymorphism (PCR‐RFLP) method. Sixty‐two specimens were collected from apples presumed infested with moth larvae and from pheromone traps from 2013 to 2014. Both species were identified morphologically, and a partial region of the cytochrome b gene was sequenced to design primers for PCR‐RFLP. Digestion profiles of G. molesta and G. dimorpha, using the Sau3A1 restriction enzyme, were characterized using three DNA fragments each for G. molesta (363 bp, 91 bp and 31 bp) and G. dimorpha (220 bp, 234 bp and 31 bp). The RFLP assay developed for both species in this study was more efficient and accurate than other currently used diagnostic assays and would be helpful to identify field‐collected specimens for pest control research.     

Further studies on the molecular systematics of Biomphalaria snails from Brazil

The polymerase chain reaction and restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rRNA gene, using the enzyme DdeI were used for the molecular identification of ten species and one subspecies of Brazilian Biomphalaria. Emphasis is given to the analysis of B. oligoza, B. schrammi and B. amazonica. The RFLP profiles obtained using this enzyme were highly distinctive for the majority of the species and exhibited low levels of intraspecific polymorphism among specimens from different regions of Brazil. However, B. peregrina and B. oligoza presented very similar profiles that complicated their identification at the molecular level and suggested a very close genetic similarity between the two species. Others enzymes including HaeIII, HpaII, AluI and MnlI were tested for their ability to differentiate these species. For B. amazonica three variant profiles produced with DdeI were observed. The study demonstrated that the ITS contains useful genetic markers for the identification of these snails     

FRESHWATER SNAILS OF THE BULINUS TRUNCATUS/TROPICUS COMPLEX IN KENYA: TETRAPLOID SPECIES

Studies in recent years on chromosome number in freshwater snailsbelonging to the Bulinus truncatus/tropicus complex demonstratedthe occurrence in Kenya of one diploid species, B. tropicus,and 2 tetraploid species, B. truncatus and B. permembranaceus.Further observations are now described on the morphologies ofthe tetraploid species, and their distributions. Observations were made on B. truncatus from 26 localities (11newly reported) and B. permembranaceus from 26 localities (7newly reported), in respect of chromosome number, egg proteins,enzymes (5 systems), shell (10 variables measured), male genitalsystem (normal or aphallic) and radula (1st lateral tooth, sizeand mesocone shape). B. permembranaceus differs most clearly from B. truncatus inegg proteins, enzyme-loci and in lacking aphallic individuals.Mesocone shape is generally less angular in B. permembranaceus.The shell of B. permembranaceus grows larger, has the columellamore commonly concave and its spire is proportionally higherand more acute. To distinguish these tetraploid species fromthe partly sympatric B. tropicus, observations on chromosomenumber and biochemical characters are necessary. The tetraploids are allopatric, B. truncatus being found rarelyas high as 1900 m altitude, whereas B. permembranaceus occupiesthe altitude range 1940–2760 m. B. truncatus may be expandingits distribution in Kenya in man-made waterbodies, while B.permembranaceus possibly is restricted by adaptation to coolconditions and by interaction with B. tropicus. Differencesbetween B. truncatus and B. permembranaceus, both morphologicaland biochemical, indicate their origins lie in independent episodesof tetraploidy. ^*Member of External Scientific staff, Medical Research Council (Received 31 October 1988; accepted 6 December 1988)     

Phytotoxic potential of Senna occidentalis (L.) Link extracts on seed germination and oxidative stress of Ipê seedlings

• Senna occidentalis is an invasive plant producing a series of allelochemicals that might inhibit the development of other plants. The objective of this study was to assess the phytotoxic effect of S. occidentalis extracts on the germination, development and antioxidant defence of the native species Tabebuia chrysotricha, T. pentaphylla, T. roseoalba and Handroanthus impetiginosus (Ipê species).
• We evaluated the effects of chemicals extracted from S. occidentalis on the germination rate, germination speed index (GSI) and biometric parameters of the test species under controlled conditions. The effect of the extracts on the pigment content, amount of H₂O₂ and malondialdehyde (MDA), and the activity of the antioxidant enzymes in roots and leaves were also tested.
• Alkaloids, coumarins, phenols, saponins, free steroids and condensed tannins were present in all extracts of S. occidentalis, while catechins were present only in leaf and stem extracts. Stem and root extracts caused a growth reduction in all Ipê species and total inhibition of seed germination in T. chrysotricha and T. roseoalba. All target species showed an increase in H₂O₂ and MDA in radicles and leaves. Oxidative stress contributed strongly to the morphological changes, such as seed blackening, thinning and darkening of radicle tips and reduction of biomass allocation in all Ipê species.
• Although there was activation of antioxidant defence mechanisms, such as an increase in activity of ascorbate peroxidase (APX) and peroxidase (POD) enzymes, the joint action of the allelochemicals caused phytotoxicity, leading to cell dysfunction in all Ipê species.

     

Diet selection by copepods in the presence of cyanobacteria

Boeckella, the dominant calanoid in many Southern Hemispherelakes, can survive, grow and reproduce to varying extents onmonocultures of cyanobacteria. In this study, we determinedthe effects of algal and cyanobacterial foods of different nutritionalvalue and concentration on food preferences of adult femaleBoeckella trianiculata and Boeckella hamata. Four species ofcyanobacteria (Anabaena flos-aquae, Nostoc sp. 2, OscillatoriatenuisandMicrocystis aeruginosa) were offered alone and mixedwith equal biomasses of Cryptomonas sp., Choricystis or a cyanobacterium.Food preferences were calculated as ratios of the rates at whichthe copepods removed each food at high and low food concentrations.In high-concentration mixtures with cyanobacteria, Cryptomonaswas consistently preferred by both Boeckella spp. In low-concentrationmixtures, both Boeckella spp. preferred Anabaena and Nostoc,which they removed at high rates(81–142 ml mg^–1h^–1), although Cryptomonas was selected in preferenceto Oscillatoria and Microcystis. When fed mixtures of filamentouscyanobacteria, both species of Boeckella showed invariant discriminationagainst Nostoc, andshifts in preference between Anabaena andOscillatoria that were related to food concentration. Microcystis,the least favouredfood, appeared to have a toxic effect on B.triarticulata. ¹Present address: Nursing and Midwifery Department, Otago Polytechnic,Forth Street, Dunedin, New Zealand     

Molecular phylogeny of the Canary Island lacertids (Gallotia): mitochondrial DNA restriction fragment divergence in relation to sequence divergence and geological time

Restriction fragment length polymorphisms of 6 base pair recognising endonucleases are used to reconstruct the phylogeny of the endemic Canary Island lacertid, Gallotia. The division into conventional species is upheld by this molecular analysis and the western Canary Island lizard (G. galloti) and eastern Canary Island lizard (G. atlantica) are hypothesized to be sister species. A more comprehensive study of the intraspecific relationships of G. galloti, based on nineteen restriction enzymes, indicates that there are distinct southern and northern lineages within this species. The phylogenetic analysis does not uphold the conventional subspecies, but suggests an alternative arrangement with one northern (La Palma, Tenerife) and one southern (Gomera, Hierro) subspecies. The inferred timing of molecular divergence of populations of G. galloti, based on RFLP analysis, is compatible with the geological timing for island origin and fossil data. Mantel tests show that mitochondrial RFLP divergence is correlated with mitrochondrial 12s rRNA and cytochrome oxidase I sequence divergence and highly correlated with mitochondrial cytochrome b sequence divergence.     

A one‐step,single tube,duplex PCR to detect predation by native predators on invasive Bemisia tabaci MEAM1 and Frankliniella occidentalis

Bemisia tabaci (Gennadius) MEAM1 (Hemiptera: Aleyrodidae) and Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae), two important invasive species, are serious agricultural pests. In this study, a one‐step, single tube, duplex polymerase chain reaction (PCR) procedure was developed to allow rapid, specific, and sensitive identification of B. tabaci MEAM1 and F. occidentalis in predator guts. The system and conditions used for the duplex PCR were optimized. The species specificity of the duplex PCR determined by comparison against non‐targets that might interact with B. tabaci MEAM1 and F. occidentalis showed that oligonucleotide primers amplified nuclear gene target sequences present only in B. tabaci MEAM1 or F. occidentalis. The limits of detection were 9.53 ng μl^?1 for B. tabaci MEAM1 and 8.94 ng μl^?1 for F. occidentalis. Within a field cage study, in which predators Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae) and Orius sauteri (Poppius) (Hemiptera: Anthocoridae) were allowed to feed on B. tabaci MEAM1 and F. occidentalis for 10 h, the B. tabaci MEAM1 DNA was detectable in 100% of H. axyridis and O. sauteri, and F. occidentalis DNA was detectable in 80% of H. axyridis and 90% of O. sauteri; this implicated that B. tabaci MEAM1 and F. occidentalis remains could be detected in native predator guts simultaneously. The accuracy and reliability of the assay suggested strongly that the duplex PCR optimized for B. tabaci MEAM1 and F. occidentalis is sensitive and specific for both invasive insects and is therefore useful in early diagnosis and monitoring of B. tabaci MEAM1 and F. occidentalis infections, and can be used to identify domestic predator species and food web relationships.     

NITROGEN EXCRETION BY THE SANDY BEACH NASSARIID WHELKS BULLIA RHODOSTOMA REEVE AND B. DIGITALIS (DILLWYN)

Nitrogen excretion by two surf zone gastropods, Bullia rhodostomaReeve and Bullia digitalis (Dillwyn) was determined under laboratoryconditions. The forms of nitrogen excreted and the effects ofmass, temperature, short-term starvation and feeding on excretionrates were determined for each species. The excretion ratesof B. rhodostoma removed directly from the surf zone (within2 h of capture) were also determined at three temperatures.No significant differences in the forms of nitrogen excretedwere found between starved and fed whelks or between species.Ammonia was the major form of nitrogen excreted with amino acidsof secondary importance. Urea was not detected and a small percentageof total dissolved nitrogen (TDN) excreted was unaccounted forin both species. Mass significantly influenced the rate of ammoniaexcretion in both species. No significant difference in slopes(common b = 0.60) were found between starved B. rhodostoma andB. digitalis or between fed whelks of the two species (commonb = 0.54). Mass adjusted mean ammonia excretion rates of starvedB. rhodostoma and those removed directly from the surf zone(‘natural diet’) did not differ significantly. Adjustedmean ammonia excretion rates of fed whelks were significantlyhigher than ‘starved’ excretion rates in both species.Temperature did not significantly influence ammonia excretionrates of fed B. rhodostoma or starved and fed B. digitalis.Bullia recycle 14.45 to 23.98 g N per metre strip of surf zoneper year which constitutes less than 1% of total phytoplanktonrequirements in the surf zone. (Received 15 August 1989; accepted 16 October 1989)     

Morphometric and molecular identification of the female castes of Bombus ignitus and B. ardens (Apidae: Hymenoptera)

Among Korean bumblebees, Bombus ignitus and B. ardens are relatively abundant and important for pollination of wildflowers and agricultural crops. Although the males are easily distinguishable phenotypically, the female castes are difficult to identify from each other. Here we evaluated the value of some morphometric characters in species identification. Also, we developed a polymerase chain reaction followed by restriction fragment length polymorphism (PCR-RFLP) to discriminate these similar species. In spite of statistically significant differences of some morphological characters between two species, overlapping quantitative traits hindered accurate identification of the species. However, using 435 bp of COI gene and AluI, BspHI and Earl restriction enzymes allowed molecular identifications of these two species with unique profiles from the digestion by these restriction enzymes. This method can also be applied for older specimens with some morphological characters damaged. We also developed species-specific primers for fast and cost-effective identification of these species.     

Biochemical characterization of the putative isoforms of myoglobins from mollusks of the Biomphalaria genus

Although gastropod myoglobin has been extensively studied, there are knowledge gaps in its biological characteristics. We describe for the first time the presence of a myoglobin in the triturative stomach of Biomphalaria gastropods. We compared biochemical parameters of myoglobins of stomach and radular muscles of Biomphalaria glabrata and Biomphalaria tenagophila. Apomyoglobin and holomyoglobin were obtained. Myoglobin was the most abundant protein in the stomach (85.0%) and radular muscles (80.0%) of the two Biomphalaria species evaluated. The Molecular mass and isoeletric point of stomach myoglobins were 16,124.93 Da and 7.98 and 16.095.28 Da and 7.77 for B. glabrata and B. tenagophila, respectively. Stomach myoglobins of B. glabrata and B. tenagophila rate autoxidation were equal to 8.0 × 10⁻⁴ h⁻¹ ± 0.0002 and 1.0 × 10⁻⁴ h⁻¹ ± 0.00142, respectively. Analysis of N-terminal amino acid sequencing revealed that stomach myoglobins are blocked in this region for a chemical group. Concluding, the differences we observed in the biochemical properties of stomach and radular myoglobins of B. glabrata and B. tenagophila suggest they may be isoform representing an evolutionary event related to the adaptation of these proteins.     

Systematic revision of the living species of Bullidae (Mollusca: Gastropoda: Cephalaspidea), with a molecular phylogenetic analysis

Bullidae are a worldwide family of marine shelled cephalaspidean gastropods with a mainly tropical distribution, but also with some representatives in temperate waters. The taxonomy of the group has in the past been based only on shell characters, and the few anatomical accounts available have not addressed more than one to three species, so there has been no agreement about the number of valid species. Seventy‐two specific names and 16 varietal names have been proposed worldwide. The systematics of the family Bullidae are revised, based not only on shells but also on anatomy of all extant species and on DNA sequence data. Twelve species are recognized worldwide, including one new species here described, and all are assigned to the genus Bulla. Two species occur in the eastern Atlantic, B. striata and B. mabillei; two in the western Atlantic, B. occidentalis and B. solida; two in the eastern Pacific, B. gouldiana and B. punctulata; and six in the Indo‐West Pacific, B. ampulla, B. arabica sp. nov. , B. orientalis, B. peasiana, B. quoyii and B. vernicosa. Full synonymies and taxonomic histories are provided for each species. In order to promote taxonomic stability, neotypes are designated for B. striata, B. solida, B. nebulosa (valid name B. gouldiana) and B. vernicosa, and lectotypes for B. occidentalis, B. mabillei, B. punctulata, B. ampulla and B. quoyii. The type locality of B. ampulla is restricted to Mauritius. Bullidae show a general morphological stasis, with anatomy being very similar between species. However, there are high levels of intraspecific variability in the shell, radula and male genital system. In some cases species could only be separated based on molecular data . After defining the characters and geographical range of each species it became clear that sympatric species (a maximum of three) show distinctive shells and reproductive structures, which makes identification straightforward. This study employs an integrative approach, combining information on shells, anatomy, DNA and geographical distribution, in order to resolve the systematics of a difficult taxonomic group. © 2008 The Linnean Society of London, Zoological Journal of the Linnean Society, 2008, 153 , 453–543.     

Effect of Graft and Sexual Hybridization on the Nodulation of Trifolium ambiguum M.B.

Trifolium ambiguum M.B. is a potentially important clover species,but it has not been used widely as a forage legume because itnodulates sparsely or not at all and the nodules formed areusually ineffective. In some compatible grafts of T. hybridumon T. ambiguum effective nodules developed on the roots of thestock. In addition, effective nodulation was obtained on thesexual hybrid between these two species. The investigation dealswith the effectiveness of nodulation of the graft and sexualhybrids and also of the two host species when inoculated withisolates from the graft hybrid, the sexual hybrid, T. hybridum,and from effectively nodulating T. ambiguum.     

Mapping of RFLP and qualitative trait loci in Brassica rapa and comparison to the linkage maps of B. napus,B. oleracea,and Arabidopsis thaliana

A linkage map of restriction fragment length polymorphisms (RFLPs) was constructed for oilseed, Brassica rapa, using anonymous genomic DNA and cDNA clones from Brassica and cloned genes from the crucifer Arabidopsis thaliana. We also mapped genes controlling the simply inherited traits, yellow seeds, low seed erucic acid, and pubescence. The map included 139 RFLP loci organized into ten linkage groups (LGs) and one small group covering 1785 cM. Each of the three traits mapped to a single locus on three different LGs. Many of the RFLP loci were detected with the same set of probes used to construct maps in the diploid B. oleracea and the amphidiploid B. napus. Comparisons of the linkage arrangements between the diploid species B. rapa and B. oleracea revealed six LGs with at least two loci in common. Nine of the B. rapa LGs had conserved linkage arrangements with B. napus LGs. The majority of loci in common were in the same order among the three species, although the distances between loci were largest on the B. rapa map. We also compared the genome organization between B. rapa and A. thaliana using RFLP loci detected with 12 cloned genes in the two species and found some evidence for a conservation of the linkage arrangements. This B. rapa map will be used to test for associations between segregation of RFLPs, detected by cloned genes of known function, and traits of interest.     

Preliminary results on the genetic variability of mitochondrial DNA in the signal crayfish,Pacifastacus leniusculus Dana

This study presents a revised method for the extraction of mitochondrial genome (mtDNA) from crayfish species of aquacultural interest, Pacifastacus leniusculus. The mean size of the mitochondrial genome is approximately 18 000 bp. Restriction fragment length polymorphism (RFLP) analysis performed on the mtDNA has revealed extensive genetic variability within this species. The estimated percentage of nucleotide sequence divergence among the four haplotypes found for P. leniusculus ranges from 1.16 to 3.99%. These data suggest that RFLP analysis of mtDNA may provide greater resolution than protein electrophoresis for population identification among signal crayfish populations. This marker offers new avenues for aquaculture and for understanding the genetic structure and evolutionary history of crayfish populations.     

Effect of Dormancy Relieving Compounds on the Seed Germination of Non-dormantAllenrolfea occidentalisunder Salinity Stress

Allenrolfea occidentalis(Chenopodiaceae) is a highly salt tolerantplant species that is widely distributed in inland salt marshesand salt playas of the western United States. We investigatedthe influence of dormancy-relieving compounds (fusicoccin, ethephon,nitrate and thiourea) in alleviating salinity stress on theseed germination ofA. occidentalis. Seed germination decreasedwith an increase in salinity and no seed germinated at 800 mMNaCl.Fusicoccin (5 µM), ethephon (10 mM) and nitrogenous compounds(20 mMnitrate and 10 mMthiourea) were able to counteract theinhibition produced by salinity treatments. All dormancy relievingcompounds significantly (P<0.0001) promoted germination atall salinity concentations. Fusicoccin completely reversed theinhibitory effects of salinity on seed germination ofA. occidentalis.Ethephon application significantly promoted germination at allsalinities. Nitrate and thiourea were relatively less effectivein alleviating the effects of high salinity on germination.Copyright1998 Annals of Botany Company Allenrolfea occidentalis, ethephon, fusicoccin, halophyte, dormancy, nitrate, salinity, seed germination, thiourea.