Removal of nonnative fish results in population expansion of a declining amphibian (mountain yellow-legged frog, Rana muscosa)

The mountain yellow-legged frog (Rana muscosa) was once a common inhabitant of the Sierra Nevada (California, USA), but has declined precipitously during the past century due in part to the introduction of nonnative fish into naturally fishless habitats. The objectives of the current study were to describe (1) the effect of fish removal from three lakes (located in two watersheds) on the small, remnant R. muscosa populations inhabiting those lakes, and (2) the initial development of metapopulation structure in each watershed as R. muscosa from expanding populations in fish-removal lakes dispersed to adjacent habitats. At all three fish-removal lakes, R. muscosa population densities increased significantly following the removal of predatory fish. The magnitude of these increases was significantly greater than that observed over the same time period in R. muscosa populations inhabiting control lakes that remained in their natural fishless condition. Following these population increases, R. muscosa dispersed to adjacent suitable (but unoccupied) sites, moving between 200 and 900 m along streams or across dry land. Together, these results suggest that large-scale removal of introduced fish could result in at least partial reversal of the decline of R. muscosa. Continued monitoring of R. muscosa at the fish-removal sites will be necessary to determine whether the positive effects of fish eradication are sustained over the long-term, especially in light of the increasingly important role played by an emerging infectious disease (chytridiomycosis, caused by Batrachochytrium dendrobatidis) in influencing R. muscosa populations.     

Optimal release strategies for captive-bred animals in reintroduction programs: Experimental infections using the guppy as a model organism

Reintroduction of captive-bred individuals for supplementation or re-establishment of wild populations has become increasingly important to prevent the extinction of many endangered species. Despite the importance of reintroductions for conservation biology, few studies have empirically evaluated the potential impacts of reintroductions of naïve organisms on disease outbreaks in native wild populations. Here, we use a model organism, the guppy (Poecilia reticulata) and its ubiquitous parasite (Gyrodactylus turnbulli) to evaluate: (i) the most effective release procedure (i.e. en masse vs. gradual release) that minimises host mortality and parasite load, (ii) the effect of pre-exposure to the parasite on host susceptibility, and (iii) the potential advantage of removing the most susceptible individuals before release. We could not detect a statistically significant difference in host survival rates between release protocols and pre-exposure regime. However, the parasite population went extinct significantly more often when pre-exposed fish were released. Furthermore, the parasite load of individual guppies by the end of the reintroduction was significantly lower in treatments where fish had been pre-exposed to parasites than in treatments with naïve guppies. Additionally, pre-exposure of hosts provided important information about their level of resistance when reintroduced. These findings suggest that pre-exposure to native parasites could be beneficial for the survival rate of captive-bred animals when reintroduced in the wild. We discuss the practicalities of these potential release strategies.     

Molecular markers and a sequence deletion in intron 2 of the putative partial homologue of LEAFY reveal geographical structure to genetic diversity in the acutely threatened legume genus Clianthus

Clianthus is an acutely threatened, bird-pollinated genus endemic to New Zealand, represented in the wild by only one population of C. puniceus and 11 populations of C. maximus, each with very few individuals (typically <10 per population). A limited number of named Clianthus cultivars of indeterminate origin are commonly grown as ornamentals. Genomic DNA from individual Clianthus plants was extracted for genetic diversity analysis using a range of molecular markers, including amplified fragment length polymorphism (AFLP). Data were analysed by the unweighted pair-group method with arithmetic averaging (UPGMA), the generation of Neighbor-Joining trees, and analyses of molecular variance (AMOVA). Genetic distance between wild populations of C. maximus was highly correlated with geographical distance between populations. Sequencing of intron 2 of a putative partial homologue of the floral meristem identity gene LEAFY (CmLFY) revealed a 7 bp deletion that was exhibited homozygously in the more northern populations of C. maximus, and in all individuals tested from the sole population of C. puniceus. This deletion was not exhibited in more southern populations of C. maximus. Further, one geographically intermediate population contained some plants that were heterozygous for the deletion. Parallel analyses of cultivated Clianthus genotypes, more than half of which were also homozygous for the 7 bp deletion, showed that these were not representative of the broad, but threatened, diversity remaining in the wild. It is argued that wild populations of C. maximus are unlikely to have arisen from the escape of plants from cultivation. Conservation effort should focus on the protection and study of the extant plants in these wild populations, rather than on the introduction of disturbance regimes to uncover potential seed banks.     

Magnitude of the US trade in amphibians and presence of Batrachochytrium dendrobatidis and ranavirus infection in imported North American bullfrogs (Rana catesbeiana)

Amphibians are globally threatened by anthropogenic habitat loss, the wildlife trade and emerging diseases. Previous authors have hypothesized that the spread of the amphibian disease chytridiomycosis (Batrachochytrium dendrobatidis) and amphibian ranaviruses are associated with the international trade in live amphibians. The North American bullfrog (Rana catesbeiana) is thought to be a carrier of these pathogens, is globally traded as a live commodity, and is sold live in US markets. We obtained importation data for all live amphibians, and parts thereof, into three major US ports of entry (Los Angeles, San Francisco and New York) from 2000 to 2005. Importation of live amphibians into these ports totaled almost 28 million individuals over this 6-year period. We collected samples from freshly-imported market frogs and found infection with both pathogens in all three cities and all seasons, with an overall infection prevalence of 62% (306/493) and 8.5% (50/588) for B. dendrobatidis and ranaviruses, respectively, by PCR. This study definitively identifies these two important pathogens in recently imported live market frogs and suggests that the amphibian trade can contribute to introductions of these pathogens into new regions. It provides support for the recent listing of B. dendrobatidis and ranaviral diseases by the OIE, and provides evidence for measures to be taken to eradicate these pathogens from the trade.     

Recent, substantial, and unexplained declines of the common toad Bufo bufo in lowland England

A decline in numbers of a large common toad (Bufo bufo) population in south-east England during the 1990s, together with anecdotal reports of similar trends in other toad populations, prompted a nation-wide survey of this species. The survey also included the common frog (Rana temporaria) as a control for which there was no comparable evidence of recent decline. A questionnaire requesting information on the fate of toad and frog populations in the last 15 years of the 20th century was distributed to professional and amateur herpetologists during autumn 2001. Ninety-five respondents provided data on a total of 277 sites, including 232 frog and 202 toad populations in England, Scotland and Wales. More than 80% of the reported sites were rural for both species. Rural frog populations were generally doing well in most parts of Britain, with almost as many increasing as decreasing populations and a high proportion of stable populations. Rural toads also showed no overall trend across the country as a whole. However, regional analysis indicated that although toads were faring at least as well as frogs in the north and west of Britain, they were declining seriously in central and eastern/south-eastern areas. In these regions 50% or more of toad populations have experienced recent declines, whereas frogs have fared as well as they have elsewhere. Toads breeding alone have fared significantly worse than toads breeding at sites where frogs were also present. Reasons for apparently toad-specific declines in lowland England remain unknown.     

The recent decline of a New Zealand endemic: how and why did populations of Archey's frog Leiopelma archeyi crash over 1996-2001?

Dramatic changes have been documented in New Zealand's vertebrate faunas since human settlement, involving major declines and extinctions, but over recent years few species have declined in numbers so rapidly as the terrestrial Archey's frog Leiopelma archeyi (Anura: Leiopelmatidae). Long-term monitoring over more than 20 years revealed a major population reduction of the species over 1996-2001 and L. archeyi is now classified as Nationally Critical under the New Zealand threat classification system. The decline progressed northwards in the Coromandel ranges, and mostly larger (female) frogs survived. On a 100 m² study plot at Tapu Ridge, annual population estimates averaged 433 frogs (SE ±32) over 1984-1994, declining by 88% to average 53 frogs (SE ±8) over 1996-2002. A mean annual survival rate of 82% for most years declined to 33% over 1994-1997. There is mounting evidence to suggest that disease is the major agent of decline, supported by (1) the rapidity and severity of decline, (2) the progressive (south to north) nature of decline, and (3) finding frogs with chytriodiomycosis from Batrachochytrium dendrobatidis at the time of decline. Surprisingly, sympatric populations of the semi-aquatic Leiopelma hochstetteri have not declined dramatically, nor has a western population of L. archeyi at Whareorino, despite chytridiomycosis occurring in some frogs there. Sustaining and restoring populations of L. archeyi in New Zealand raises major challenges for conservation management.     

Assessing limitations on population growth in two critically endangered Acacia taxa

Flowering phenology, soil seedbank and the impact of fire, weeds and grazing were investigated in two rare Acacia taxa restricted to the agricultural district north of Perth Western Australia. Acacia aprica is known from six extant populations and one extinct population. Five of the extant populations are restricted to linear road reserves with the sixth population located in a small native vegetation remnant. Acacia cochlocarpa ssp. cochlocarpa is known from a single population also situated on a road verge. Size class structure, levels of canopy death and an absence of juveniles indicated that all populations are in decline. Flowering intensity and success varied between populations and years in A. aprica and between years in A. cochlocarpa ssp. cochlocarpa. Seed bank analysis indicated that seeds were patchily distributed while experimental fires demonstrated that such events could break seed dormancy and promote germination. Both taxa have similar germination physiologies and showed increased germination after seeds were exposed to heat. Following emergence, however, competition with annual weeds had a negative impact on seedling growth and survival in both taxa. While vertebrate grazing had some influence, weeds were the major inhibitory influence on recruitment. Reduced fire frequencies since fragmentation may be responsible for population decline in both taxa but other site specific factors such as weeds and grazing may affect the establishment of seedlings following fire.     

Does restocking with Japanese quail or hybrids affect native populations of common quail Coturnix coturnix?

Natural populations of the common quail Coturnix coturnix may hybridize in the wild with non-native individuals (Japanese quail Coturnix japonica or hybrids) as a result of restocking for hunting purposes. Several laboratory studies suggest that this could lead to a decline in the impulse to migrate in the common quail, and a drop in the frequency of phenotypes showing this tendency. This could lead to an increase in common quail populations in North Africa and a decrease in Europe. This paper provides new data on the proportion of hybrids in Catalonia (Northeast Spain) over 24 years (1983-2006) showing how restocking with Japanese quail or hybrids affects native populations of common quail. The first hybrids were detected in 1990 with an estimate of 4.65% of non-native individuals during the breeding season of wild common quail populations. No increase in non-native or hybrid numbers was detected during the study period, indicating that restocking poses no serious conservation problems at present. However, this may change in the near future, either with or without changes in the current scenario. A prudent policy with regard to restocking with non-native individuals is suggested. Moreover, further studies are needed to clarify the extent of this conservation problem.     

Conserving marginal populations of the food plant (Impatiens noli-tangere) of an endangered moth (Eustroma reticulatum) in a changing climate

Impatiens noli-tangere is scarce in the UK and probably only native to the Lake District and Wales. It is the sole food plant for the endangered moth Eustroma reticulatum. Significant annual fluctuations in the size of I. noli-tangere populations endanger the continued presence of E. reticulatum in the UK. In this study, variation in population size was monitored across native populations of I. noli-tangere in the English Lake District and Wales. In 1998, there was a crash in the population size of all metapopulations in the Lake District but not of those found in Wales. A molecular survey of the genetic affinities of samples in 1999 from both regions and a reference population from Switzerland was performed using AFLP and ISSR analyses. The consensus UPGMA dendrogram and a PCO scatter plot revealed clear differentiation between the populations of I. noli-tangere in Wales and those in the Lake District. Most of the genetic variation in the UK (H_T=0.064) was partitioned between (G_ST=0.455) rather than within (H_S=0.034) regions, inferring little gene flow occurs between regions. There was similar bias towards differentiation between metapopulations in Wales, again consistent with low levels of interpopulation gene flow. This contrasts with far lower levels of differentiation in the Lake District which suggests modest rates of gene flow may occur between populations. It is concluded that in the event of local extinction of sites or populations, reintroductions should be restricted to samples collected from the same region. We then surveyed climatic variables to identify those most likely to cause local extinctions. Climatic correlates of population size were sought from two Lake District metapopulations situated close to a meteorological station. A combination of three climatic variables common to both sites explained 81-84% of the variation in plant number between 1990 and 2001. Projected trends for these climatic variables were used in a Monte Carlo simulation which suggested an increased risk of I. noli-tangere population crashes by 2050 at Coniston Water, but not at Derwentwater. Implications of these findings for practical conservation strategies are explored.     

Screening of bacterial isolates from various European soils for in vitro antagonistic activity towards Rhizoctonia solani and Fusarium oxysporum: Site-dependent composition and diversity revealed

A cultivation-based approach was used to determine the in vitro antagonistic potential of soil bacteria towards Rhizoctonia solani AG3 and Fusarium oxysporum f. sp. lini (Foln3). Four composite soil samples were collected from four agricultural sites with previous documentation of disease suppression, located in France (FR), the Netherlands (NL), Sweden (SE) and the United Kingdom (UK). Similarly, two sites from Germany (Berlin, G-BR; and Braunschweig, G-BS) without documentation of disease suppression were sampled. Total bacterial counts were determined by plating serial dilutions from the composite soil samples onto R2A, AGS and King's B media. A total of 1,788 isolates (approximately 100 isolates per medium and site) was screened for antifungal activity, and in vitro antagonists (327 isolates) were found amongst the dominant culturable bacteria isolated from all six soils. The overall proportion of antagonists and the number of isolates with inhibitory activity against F. oxysporum were highest in three of the suppressive soils (FR, NL and SE). Characterization of antagonistic bacteria revealed a high phenotypic and genotypic diversity. Siderophore and protease activity were the most prominent phenotypic traits amongst the antagonists. The composition and diversity of antagonists in each soil was site-specific. Nevertheless, none of the antimicrobial traits of bacteria potentially contributing to soil suppressiveness analyzed in this study could be regarded as specific to a given site.     

Spatial distribution of the nematode biocontrol agent Pasteuria penetrans as influenced by its soil habitat

Root-knot nematodes belonging to the Meloidogyne genus are ubiquitous plant-parasitic pests, especially on vegetables. The Pasteuria penetrans bacterium is an obligate parasite of nematodes, parasitizing most of the Meloidogyne species. Spatial distributions of Meloidogyne javanica populations infested or not by P. penetrans and of bacterial populations were studied in a vegetable plot naturally infested by these organisms. It was observed that distributions of M. javanica populations, of populations infested by P. penetrans, and of free bacteria populations were not overlapped. Soil factors involved were investigated. Soil texture and water flow in porosity are concerned, as they directly influence the level of the pool of bacteria and then the chances of both organisms to meet. The soil solution has a direct effect on the attachment of the bacterium on the nematode cuticle.     

An unseen foe in arthropod conservation efforts: The case of Wolbachia infections in the Karner blue butterfly

Infection by endosymbiotic bacteria is an underappreciated threat to endangered arthropods with serious implications for captive management programs. We examined the nature of Wolbachia infection in the North American endangered Karner blue butterfly, Lycaeides melissa samuelis, as a case study. Screening for Wolbachia across the range of the species confirmed widespread infection in the western half of the Karner blue’s range. Multilocus sequence typing using six genes confirmed that the infection in the western populations is attributable to a single strain of Wolbachia. This strain was also detected in the closely related Melissa blue butterfly, L. m. melissa, the presumed source of the infection. The infection in the Karner blue butterfly was perfectly correlated with the presence of a foreign mitochondrial DNA variant present in the Melissa blue butterfly, consistent with the hypothesis that the mitochondrial introgression was driven by the spread of Wolbachia. A single individual out of 71 screened from the eastern portion of the range of the Karner blue butterfly was also infected, however this infection was attributable to a different strain. Simulation models of the demographic effects of the spread of Wolbachia infection to uninfected populations and metapopulations suggest that such an infection might further reduce already small population sizes and substantially increase the probability of population extirpation. We discuss threats to other endangered arthropods in the light of this case study and make recommendations for minimizing the impact of endosymbiont infections in conservation plans, especially those including captive propagation and augmentation of endangered arthropod populations.     

Spatial demographic and genetic consequences of harvesting within populations of the terrestrial orchid Cymbidium goeringii

Populations of many orchids, especially terrestrial species, have been rapidly decreasing due to mass collection by plant sellers and enthusiasts. Given the presumably negative demographic and genetic consequences, such anthropogenic activity should be taken into consideration for predicting ecological and evolutionary dynamics and for planning conservation strategies. To determine how recent human disturbance alters spatial demographic and population genetic processes, populations of the terrestrial orchid Cymbidium goeringii located in South Korea were examined to quantify the spatial distributions of individuals and genotypes with respect to three levels of disturbance: “disturbed” (four populations), “putatively disturbed” (two), and “undisturbed” (two). Undisturbed and putatively disturbed populations were found to possess significantly positive spatial clustering of individuals over a range of spatial scales. In contrast, disturbed populations exhibited little or no spatial aggregation, consistent with the selective removal of plants by collectors from higher density areas within these populations. Although overall genetic differentiation among populations was moderate and significant (F_ST = 0.082), levels of genetic diversity within populations were similar despite the different disturbance histories (mean H_e = 0.257-0.324). Spatial genetic autocorrelation analyses revealed that the undisturbed populations exhibited significant declines in kinship (F_ij) with distance, that mean kinship at interplant distances of ?4-6 m was significantly greater than zero and between plants ?0.5 m apart was in the range expected for first cousins to half-sibs. In contrast, only one putatively disturbed and one disturbed population exhibited significant declines in kinship with distance. These differences between disturbed versus putatively disturbed and undisturbed populations in the spatial distribution of individuals and genetic variation likely reflect the consequences of mass collections. Since these differences (and reduced population density) have important implications for future ecological and evolutionary trajectories, conservation managers of endangered terrestrial orchids may want to analyze the spatial distribution of individuals and their genotypes to infer whether a population with few individuals represents a natural state or the likely outcome of mass collection.     

Road crossing structures for amphibians and reptiles: Informing design through behavioral analysis

Road traffic causes significant amphibian and reptile mortality, which could be mitigated through the installation of road crossing structures that facilitate safe passage, but only if reptiles and amphibians are willing to use them. Through a series of behavioral choice experiments with frogs and turtles, we examined how aperture diameter, substrate type, length, and light permeability influenced individuals’ preferences for specific attributes of crossing structures, and how individuals responded to various heights of barrier fences. Snapping turtles (Chelydra serpentina), green frogs (Rana clamitans), and leopard frogs (Rana pipiens) preferred larger diameter tunnels (>0.5 m) whereas painted turtles (Chrysemys picta) preferred tunnels of intermediate (0.5-0.6 m) diameter. Green frogs preferred soil- and gravel-lined tunnels to concrete- and PVC-lined tunnels. Painted turtles showed non-random choice of different lengths of tunnel, possibly indicating some avoidance of the longest tunnel (9.1 m); although no species preferred to exit via the longest tunnels (9.1 m), members of all four species used such tunnels. Green frogs preferred tunnels with the greatest light permeability. Fences 0.6 m in height were effective barriers to green frogs, leopard frogs, and snapping turtles, whereas 0.3 m fences excluded painted turtles. We conclude that tunnels > 0.5 m in diameter lined with soil or gravel and accompanied by 0.6-0.9 m high guide fencing would best facilitate road crossing for these and likely other frog and turtle species.     

Pre-symbiotic and symbiotic interactions between Glomus intraradices and two Paenibacillus species isolated from AM propagules. In vitro and in vivo assays with soybean (AG043RG) as plant host

Two indole-producing Paenibacillus species, known to be associated with propagules of arbuscular mycorrhizal (AM) fungi, were examined for their mycorrhization helper bacteria activity at pre-symbiotic and symbiotic stages of the AM association. The effects were tested under in vitro and in vivo conditions using an axenically propagated strain of the AM fungus Glomus intraradices and Glycine max (soybean) as the plant host. The rates of spore germination and re-growth of intraradical mycelium were not affected by inoculation with Paenibacillus strains in spite of the variation of indole production measured in the bacterial supernatants. However, a significant promotion in pre-symbiotic mycelium development occurred after inoculation of both bacteria under in vitro conditions. The Paenibacillus rhizosphaerae strain TGX5E significantly increased the extraradical mycelium network, the rates of sporulation, and root colonization in the in vitro symbiotic association. These results were also observed in the rhizosphere of soybean plants grown under greenhouse conditions, when P. rhizosphaerae was co-inoculated with G. intraradices. However, soybean dry biomass production was not associated with the increased development and infectivity values of G. intraradices. Paenibacillus favisporus strain TG1R2 caused suppression of the parameters evaluated for G. intraradices during in vitro symbiotic stages, but not under in vivo conditions. The extraradical mycelium network produced and the colonization of soybean roots by G. intraradices were promoted compared to the control treatments. In addition, dual inoculation had a promoting effect on soybean biomass production. In summary, species of Paenibacillus associated with AM fungus structures in the soil, may have a promoting effect on short term pre-symbiotic mycelium development, and little impact on AM propagule germination. These findings could explain the associations found between some bacterial strains and AM fungus propagules.     

Herbicides, weeds and endangered species: management of bitou bush (Chrysanthemoides monilifera ssp. rotundata) with glyphosate and impacts on the endangered shrub, Pimelea spicata

Environmental weed invasion threatens the biodiversity of native species. Unfortunately, managing these weeds may also affect biodiversity adversely. A recent example occurred when glyphosate, a herbicide used to control the highly invasive weed, bitou bush (Chrysanthemoides monilifera ssp. rotundata), accidentally drifted over a small population of an endangered shrub, Pimelea spicata. Following concerns that the affected population would not recover and, thereby, cause the local extinction of P. spicata, we conducted a series of glasshouse and field experiments to explore the impacts of glyphosate on this endangered species. Seedlings and young plants of P. spicata, in which the tap root was undeveloped, were killed by a single application of glyphosate. Older plants with a well developed tap root also died back initially, but about 50% of individuals re-sprouted. This re-growth was associated with a significant decrease in tap root diameter, implying that further disturbance, including repeated treatment with glyphosate, would kill plants by impairing their potential for recovery. Unlike some sclerophyllous native shrubs, the tolerance of P. spicata to glyphosate was limited, even when its growth was slowed artificially by limiting water availability. Winter applications of glyphosate to manage infestations of bitou bush will impact adversely on populations of P. spicata and may also affect the other rare and endangered species whose survival is threatened by this species, even though some natives are unaffected by the herbicide. Protecting native biodiversity from bitou bush will involve sustainable weed management that minimises impacts on non-target native species.     

Population viability analysis of European bison populations in Polish and Belarusian parts of Bia?owie?a Forest with and without gene exchange

European bison (Bison bonasus) became extinct in the wild at the beginning of the 20th century. The contemporary Lowland line of bison was founded by seven individuals that survived in captivity. The largest population of Lowland bison live in the Bia?owie?a Forest, but the forest and the population are divided by a border fence between Poland and Belarus. This fence is a barrier to the movement of ungulates. In this study, we used population genetic models to predict the future viability of the Polish and Belarusian bison populations. We determined the founder contribution, founder equivalent, mean inbreeding coefficient, mean kinship, and the proportion of genetic diversity retained in the Belarusian bison population. Although the founding group of the Belarusian population was larger than that of the Polish population, the latter had more favourable genetic parameters. We assessed inbreeding depression for fecundity in free-ranging European bison compared to captive individuals. Using population viability analysis (PVA) we modelled both bison populations with or without gene exchange, and with or without incorporating the kinship of the founders. When founder kinship was included, in both populations the mean number of alleles and the gene diversity retained within extant populations decreased substantially compared to PVA models in which founders were not related. The worst genetic parameters were obtained for the Belarusian population under a scenario in which the founders are related and gene exchange is lacking, which is the closest to the real situation. Creation of passages for animals by partial removal of the border fence would have a favourable effect on the genetic variation and viability of both bison populations, especially the Belarusian.     

Lotononis angolensis forms nitrogen fixing, lupinoid nodules with phylogenetically unique, fast-growing, pink-pigmented bacteria, which do not nodulate L. bainesii or L. listii

Root-nodule bacteria that nodulate the legume genus Lotononis are being investigated to develop new forage species for agriculture. Bacteria isolated from nodules of Lotononis angolensis were fast-growing, highly mucoid and pink-pigmented, and on the basis of 16S rRNA phylogeny <94% related to other genera in the Alphaproteobacteria. Root-nodule bacteria isolated from other Lotononis species (L. bainesii, L. solitudinis and L. listii) resembled the more common dry, slow-growing, pink-pigmented rhizobia previously described for L. bainesii. These isolates could be attributed to the Methylobacterium genus, although not to the type species Methylobacterium nodulans. Further differences were uncovered with nodulation studies revealing that nodule isolates from L. angolensis were effective at nitrogen fixation on their host plant, but could nodulate neither L. bainesii nor L. listii. Reciprocal tests showed isolates from L. bainesii, L. listii and L. solitudinis were incapable of nodulating L. angolensis effectively. Nodule morphology for L. bainesii, L. angolensis and L. listii was characteristically lupinoid, with little structural divergence between the species, and with nodules eventually enclosing the entire root.     

Genetic variation in captive population of chinese alligator, Alligator sinensis, revealed by random amplified polymorphic DNA (RAPD)

Chinese alligator (Alligator sinensis) is an endemic species in China. The likely extinction of it in the wild has been recognised. To prevent this species becoming extinct, the Anhui Research Centre of Chinese Alligator Reproduction (ARCCAR) was established in Xuanzhou, Anhui Province in 1979, where has been established the largest captive population of Chinese alligator (XZSP) in the world. Another farm (CXSP) was established by villagers in Changxin, Zhejiang Province. The results of an investigation of the two captive subpopulation structures by genetic analysis are presented in this paper. We examined the genetic variation in the two captive subpopulations using RAPDs. Thirty-one random primers were selected among 199 random primers screened. A total of 193 reproducible RAPD fragments were scored among 43 individuals, of which 21 (10.88%) were polymorphic. The genetic distances between 43 individuals ranged from 0 to 0.0376 with average of 0.0104±0.0055 S.E. The genetic similarity in CXSP (0.9948±0.0029 S.E.) was higher than that in XZSP (0.9894±0.0055 S.E.). The founder effect is a possible explanation for very low genetic variation in CXSP. Analysis of the RAPD data showed that the mean phenotypic band frequencies of each polymorphic loci was 0.6656±0.3730 S.E. The lowest phenotypic band frequency (0.0233) was found in four of those polymorphic loci. There was no genetic difference between the two subpopulations (D_ij=0.0009). According to the dendrogram and the distribution of polymorphic fragments in two subpopulations, CXSP originated genetically from XZSP. This paper summarises a preliminary research on genetic structure in populations of Chinese alligator. Although there is higher genetic similary (0.9896±0.0055 S.E.) in captive population of A. sinensis, we did not determine whether or not loss of genetic variation had occurred in relation to a wild control population. The data of malformed offspring will be collected carefully, and wild samples be added to set up a control population in future study.     

Brassica napus seed meal soil amendment modifies microbial community structure, nitric oxide production and incidence of Rhizoctonia root rot

A low glucosinolate content (21.8 μmol g⁻¹) Brassica napus seed meal (RSM) applied to orchard soils altered communities of both pathogenic and saprophytic soil micro-organisms. RSM amendment reduced infection by native and introduced isolates of Rhizoctonia spp. and recovery of Pratylenchus spp. from apple roots. Root infection by Rhizoctonia solani AG-5 was also suppressed in split-root assays where a portion of the root system was cultivated in RSM-amended soils and the remainder grown in the presence of the pathogen but lacking RSM. R. solani hyphal growth was not inhibited by RSM amendment. Suppression of Pratylenchus was attained to an equivalent extent by amending soils with either RSM or soybean meal (SM) when applied to provide a similar N content. Thus, glucosinolate hydrolysis products did not appear to have a significant role in the suppression of Rhizoctonia spp. or Pratylenchus spp. obtained via RSM amendment. RSM amendment elevated populations of Pythium spp. and of ammonia-oxidizing bacteria that release nitric oxide but suppressed fluorescent pseudomonad numbers. Streptomyces spp. soil populations increased significantly in response to RSM but not SM amendment. The vast majority of Streptomyces spp. recovered from the apple rhizosphere produced nitric oxide and possessed a nitric oxide synthase homolog. We propose that transformations in the bacterial community structure are associated with the observed control of Rhizoctonia root rot, with NO production by soil bacteria potentially having a role in the induction of plant systemic resistance.