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In efforts to understand the potential impacts of the Marcellus Shale extraction activities on stream health, we performed a baseline study on a focal pair of small streams in the Tenmile Creek watershed in Greene County, Pennsylvania. Bates Fork had intensive Marcellus Shale well drilling activity and several violations upstream from our study site. A tributary, Fonner Run, served as a control stream site with no drilling activity. Through two years of monitoring, we established baselines for water chemistry and biological communities of bacteria, fish and salamanders. In addition, we compared population genetic diversity of two darter species with microsatellite markers. Although no conclusive differences were found between the stream-pair, we established baseline parameters and gained insight into refining tools to detect the signature of Marcellus Shale extraction impacts on small streams in southwestern Pennsylvania. We conclude this chapter with lessons learned from this case study, future directions and suggestions for improved monitoring and detection of Marcellus Shale impacts on streams in southwestern Pennsylvania.
The widespread species Parmotrema crinitum (Ach.) M. Choisy and Parmotrema perlatum (Huds.) M. Choisy are mainly distinguished by their reproductive strategies. While P. crinitum propagates by isidia, P. perlatum produces soredia. In this study, we aim to evaluate the phylogenetic relationship between both species and to critically examine their species boundaries. To this purpose, 46 samples belonging to P. crinitum and P. perlatum were used in our analysis, including 22 for which we studied the morphology and chemistry, before extracting their DNA. We used 35 sequences of the internal transcribed spacer region of nuclear ribosomal DNA (ITS) of Parmotrema perlatum from Europe and Africa (20 of which were newly generated), and 11 of Parmotrema crinitum from Europe, North America and North Africa (two newly generated). Additionally, 28 sequences of several species from Parmotrema were included in the ITS dataset. The ITS data matrix was analyzed using different approaches, such as traditional phylogeny (maximum likelihood and Bayesian analyses), genetic distances, automatic barcode gap discovery (ABGD) and the coalescent-based method poisson tree processes (PTP), in order to test congruence among results. Our results indicate that all samples referred to P. crinitum and P. perlatum nested in a well-supported monophyletic clade, but phylogenetic relationships among them remain unresolved. Delimitations inferred from PTP, ABGD and genetic distance analyses were comparable and suggested that P. crinitum and P. perlatum belong to the same lineage. Interestingly, two samples of P. perlatum separate in a different monophyletic clade, which is supported as a different lineage by all the analyses.
HMGR, 3-hydroxy-3-methylglutaryl-CoA reductase, is a major rate-limiting enzyme in mevalonate (MVA) pathway for isoprenoids and subsequent tanshinone biosynthesis in the Chinese traditional bulk herbal medicine Danshen, Salvia miltiorrhiza, mainly for cardiovascular disorders. In this paper, the genomic SmHMGR genes of 38 cultivated populations of S. miltiorrhiza collected in China were for the first time sequenced to reveal the genetic diversity and phylogeny. The SmHMGR gene was shown to be intron-free, 1650~1659 bp in complete CDS with the majority being 1656 bp, and two unique populations (W-FJLY-V-1 and W-SCHY-W-1) being 1659 and 1650 bp respectively. A total of 103 SNP variation sites were detected with a variation rate of 6.22%, most of which occurred in S. miltiorrhiza f. alba population W-SCHY-W-1; a total of 25 amino acid variation sites were found, of which 19 was in W-SCHY-W-1. The same four populations, W-SCHY-W-1, V-HBAG-V-1, V-JLCC-V-1 and S-NM-V-1 could be discriminated from the remaining 34 by both the SNP fingerprints and the deduced amino acid variation sites. Other or composite DNA markers are needed for better identification. The SmHMGR gene of white flower S. miltiorrhiza f. alba population W-SCHY-W-1 is especially rich in variations and worthy of further studies. Phylogenetic trees based on both the gene and the deduced amino acid sequences showed a very similar two-clade topological structure. This research enriched the content and the genetic means for the molecular identification, genetic diversity and phylogenetic studies of the cultivated S. miltiorrhiza populations, and laid a solid foundation for further related and in-depth investigations.
Rice blast disease, caused by Magnaporthe oryzae, is one of the most damaging diseases of rice worldwide. Cultivation of rice varieties carrying resistance genes is the most economic and successful strategy to control the disease. In this study, 451 rice varieties from around the world including 363 Thai landrace rice varieties, 21 Thai improved rice varieties, 43 Japanese rice varieties and 24 worldwide rice varieties were screened by PCR technique using gene-specific markers for 10 rice blast resistance genes: Pi9, Piz-t, Pi50, Pigm(t), Pid2, Pid3, Pia, Pik, Pi54 and Pita. The results showed that 382 (99.48%) Thai rice varieties have at least one resistance gene and two rice varieties, ‘Hom’ and ‘Bak muay’, contained eight out of ten screened rice blast resistance genes. 320 rice varieties (83.33%) contained three or more rice blast resistance genes. The frequency of the rice blast resistance gene ranges from 87.76–9.64 per cent, of which the Pid3 gene has the highest frequency and the Pi54 gene has the lowest frequency. Two major resistance genes, found in Japanese rice varieties, are the Pik gene (76.74%) and the Pi9 gene (72.09%). While two major resistance genes, found in the international rice varieties are the Pi9 gene (66.67%) and the Pi54 gene (62.50%). The disease resistance gene profile of each rice variety obtained from this study will benefit the rice blast resistant breeding programme in the future.
A study was conducted to collect and characterize the indigenous Kala zeera (Bunium persicum Boiss. Fedtsch.) accessions from the hills of northwestern Himalayan sates of Jammu and Kashmir, Himachal Pradesh and Uttarakhand. Around 1000 accessions were collected during this exploration mission, out of which diversity of 252 accessions was established through morphological characterization. The morpho-agronomic characterization and the analysis of trait data revealed significant variability in number of branches per plant, number of umbelets per umbel, number of seeds per plant, seed yield per plant and 1000 seed weight. The collection and characterization of these diverse accessions can prove useful in future Kala zeera improvement programmes in the world as this is the first such comprehensive report of the crop from northwestern Himalayan region of India.
The fall armyworm (Spodoptera frugiperda), a destructive pest that originated in South and North America, spread to China in early 2019. Controlling this invasive pest requires an understanding of its population structure and migration patterns, yet the invasion genetics of Chinese S. frugiperda is not clear. Here, using the mitochondrial cytochrome oxidase subunit I (COI) gene, triose phosphate isomerase (Tpi) gene and eight microsatellite loci, we investigated genetic structure and genetic diversity of 16 S. frugiperda populations in China. The Tpi locus identified most S. frugiperda populations as the corn-strains, and a few were heterozygous strains. The microsatellite loci revealed that the genetic diversity of this pest in China was lower than that in South America. Furthermore, we found moderate differentiation among the populations, distinct genetic structures between adjacent populations and abundant genetic resources in the S. frugiperda populations from China sampled across 2 years. The survival rate of S. frugiperda was significantly higher when it was fed on corn leaves than on rice leaves, and the larval stage mortality rate was the highest under both treatments. Our results showed that S. frugiperda probably invaded China via multiple independent introductions and careful pesticide control, continuous monitoring and further studies will be needed to minimize its potential future outbreak.
Ascaridia galli (Nematoda: Ascaridiidae) is the most common intestinal roundworm of chickens and other birds with a worldwide distribution. Although A. galli has been extensively studied, knowledge of the genetic variation of this parasite in detail is still insufficient. The present study examined genetic variation in the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene among A. galli isolates (n = 26) from domestic chickens in Hunan Province, China. A portion of the cox1 (pcox1) gene was amplified by polymerase chain reaction separately from adult A. galli individuals and the amplicons were subjected to sequencing from both directions. The length of the sequences of pcox1 is 441 bp. Although the intra-specific sequence variation within A. galli is 0–7.7%, the inter-specific sequence differences among other members of the infraorder Ascaridomorpha were 11.4–18.9%. Phylogenetic analyses based on the maximum likelihood method using the sequences of pcox1 confirmed that all of the Ascaridia isolates were A. galli, and also resolved three distinct clades. Taken together, the findings suggest that A. galli may represent a complex of cryptic species. Our results provide an additional genetic marker for the management of A. galli in chickens and other birds.
Gyrodactylus sphinx Dmitrieva & Gerasev, 2000 is the only species of Gyrodactylus originally described from Aidablennius sphynx (Valenciennes) in the Black Sea. In the present study, monogeneans similar to G. sphinx are reported from the same host and from two other species of Blenniidae from the Black Sea, as well as from the Mediterranean Sea. This study aims to verify the taxonomic status of the specimens found in different hosts and localities, other than the type ones of G. sphinx. Twenty-two measurements of the haptoral structures of 169 gyrodactylids were used for the morphological study. Morphometric variability between different samples was analysed using analysis of variance, multivariate analysis of variance (MANOVA) and principal component analysis (PCA). Molecular studies were carried out using the nuclear internal transcribed spacer 2 and 5.8S ribosomal DNA regions. Network, Bayesian phylogenetic and species delimitation analyses were performed to infer the number of taxonomic units and the phylogeographic relationships occurring within and among them. MANOVA revealed a significant dependence of the morphometry of hamuli and marginal hooks on host species and regions, but a clear differentiation between samples was not confirmed by PCA. Moreover, the ranges of all dimensions overlapped between samples. However, molecular analyses suggested the occurrence of at least two taxonomic entities. The most common entity was present in individuals of the Black and Mediterranean seas, and is described here as Gyrodactylus gerasevi n. sp., whereas a second entity recognized as a G. sphinx was found only in individuals from two localities off Crimea. The monophyletic cluster grouping of these two species was placed within a large clade that also included a separate sister cluster with seven other species of the Gyrodactylus orecchiae cross-ocean species group.
Germplasm characterization is essential to estimate variation and identify desirable genotypes for crop improvement. The current study was conducted on eight qualitative and 14 quantitative traits related to growth and floral attributes for characterization and grouping of 54 chrysanthemum genotypes. Wide variability was observed among most of the traits and principal component analysis revealed that the first two principal components explained most of the existing variation (>98%) in germplasm. Genotypic coefficient of variation exhibited a wide range from 9.41% (ratio of leaf length/width) to 105.99% (corolla tube length of ray florets), while phenotypic coefficient of variation varied from 10.49% (ratio of leaf length/width) to 106.38% (corolla tube length of ray florets). Broad sense heritability estimates were higher (>96%) for most of the characters, except for traits such as ratio of leaf length/width and peduncle thickness. Forty-four pair-wise inter-trait combinations showed positive genotypic and phenotypic correlations for 91 possible combinations. D2 analysis revealed that genotypes of clusters I and IV to be highly distinct, and hybridization between them might have better chance to obtain desirable types. D2 analysis confirmed that the cultivars in cluster IV namely, Red D Spoon, Tokyo Soldier, Yellow Reflex and hierarchical clustering showed that the cultivar Tokyo Soldier to be highly diverse. It is recommended to use the above genotypes in crossing programme for obtaining better progenies.
Dichanthium annulatum is one of the dominant grasses of India, North Africa, Southeast Asia, China, Australia, Fiji, New Guinea, Cuba, Haiti and Puerto Rico. This drought-tolerant grass is an excellent fodder in mixed pastures. Developing varieties with improved quality and tolerance to various abiotic stresses is hampered due to its apomictic nature. Germplasm collection, characterization, genetic diversity analysis and core subset development followed by selection for desirable traits seems to be the most plausible breeding tool for developing new cultivars. In the present study, 498 genotypes collected from different agro-ecological zones in India were included. Genotypes were characterized for various metric and non-numeric traits; and the nutritional parameters. Agglomerative clustering analysis, using the Euclidean distance method, showed 14 distinct clusters. High variability was recorded for green forage yield, quantitative traits and nutritive quality parameters. A core subset of 50 accessions was identified, which captured most of the morphological and nutritional variability present in the total germplasm. Clustering of genotypes was observed to be related to the climatic conditions of the place of collection. High genetic variability observed for various morphological traits as well as forage yield indicated that these genotypes or subset of genotypes can be evaluated in different abiotic stress conditions such as salt, light and moisture stress for the identification of suitable varieties for the respective areas. Variability was attributed to inter-generic, inter-specific crossing together with the occasional presence of sexual plants in nature.
To facilitate population-genetic studies, we developed simple sequence repeat (SSR) markers and a molecular species identification assay for Peltigera membranacea (Ascomycota, Peltigerales), a common ground-dwelling lichen of forest and tundra ecosystems. Additional markers were developed for its Nostoc photobiont. Twenty-one fungal markers for P. membranacea were found to be polymorphic, with the number of alleles ranging from 3–21. Nei's unbiased gene diversity ranged from 0.588 to 0.640 in four significantly structured (FST = 0.059) mycobiont populations. For the Nostoc photobiont, 14 polymorphic SSR were developed, yielding 4–14 alleles each, with gene diversity ranging from 0.062 to 0.771 in four populations showing substantial population structure (FST = 0.278). The new markers developed are suitable for population genetic studies of Peltigera membranacea and of its cyanobiont, and at the same time allowed us to distinguish 98.5% of P. membranacea specimens from morphologically similar species of Peltigera.
The prevalence rates of trypanosomes, including those that require cyclical transmission by tsetse flies, are widely distributed in Africa. Trypanosoma brucei and Trypanosoma congolense are actively maintained in regions where there are no tsetse flies although at low frequencies. Whether this could be due to an independent evolutionary origin or multiple introduction of trypanosomes due to continuous movement of livestock between tsetse-free and -infested areas is not known. Thus, the aim of the study was to carry out microsatellite genotyping to explore intra-specific genetic diversity between T. (Trypanozoon), T. congolense and Trypanosoma vivax from the two regions: tsetse infested and tsetse free. Microsatellite genotyping showed geographical origin-based structuring among T. (Trypanozoon) isolates. There was a clear separation between isolates from the two regions signalling the potential of microsatellite markers as diagnostic markers for T. brucei and Trypanosoma evansi isolates. Trypanosoma vivax isolates also clustered largely based on the sampling location with a significant differentiation between the two locations. However, our results revealed that T. congolense isolates from Northern Kenya are not genetically separated from those from Coastal Kenya. Therefore, these isolates are likely introduced in the region through animal movement. Our results demonstrate the occurrence of both genetic connectivity as well as independent evolutionary origin, depending on the trypanosome species between the two ecologies.
The elephant seal nomenclature is arbitrary, based on fallacies, and is impractical. Elephant seals are true seals that originated in the North Atlantic 15 to 20 million years ago. Today, the northern species is found along the west coast of North America and the southern species inhabits sites around the Antarctic continent. The two species have been separated for a minimum of 5,000 years but the exact duration is unclear. The northern elephant seal population was abundant before the seals were hunted for their blubber oil in the first half of the nineteenth century. The slaughter was so extensive and relentless that by the late-nineteenth century, the species was considered extinct. Approximately 20 individuals survived, however, on a volcanic island in Mexico. The population experienced a severe population bottleneck and lost genetic diversity. The southern species was also hunted by sealers, but its population was not reduced as drastically.
The genus Echinochloa constitutes some of the most prominent weed species found in rice (Oryza sativa L.) production worldwide. The taxonomy of Echinochloa is complex due to its morphological variations. The morphophysiological diversity and taxonomic characteristics of Echinochloa ecotypes infesting rice fields in Texas are unknown. A total of 54 Echinochloa ecotypes collected during late-season field surveys in 2015 and 2016 were characterized in a common garden in 2017. Plants were characterized for 14 morphophysiological traits, including stem angle; stem color; plant height; leaf color; leaf texture; flag leaf length, width, and angle; days to flowering; panicle length; plant biomass; seed shattering; seed yield; and seed dormancy. Principal component analysis indicated that 4 (plant height, flag leaf length, seed shattering, and seed germination) of the 14 phenological traits characterized here had significantly contributed to the overall morphological diversity of Echinochloa spp. Results showed wide interpopulation diversity for the measured traits among the E. colona ecotypes, as well as diverse intrapopulation variability in all three Echinochloa species studied, including barnyardgrass [Echinochloa crus-galli (L.) P. Beauv.], junglerice [Echinochloa colona (L.) Link], and rough barnyardgrass [Echinochloa muricata (P. Beauv.) Fernald]. Taxonomical classification revealed that the collection consisted of three Echinochloa species, with E. colona being the most dominant (96%), followed by E. crus-galli (2%), and E. muricata (2%). Correlation analysis of morphophysiological traits and resistance status to commonly used preemergence (clomazone, quinclorac) and postemergence herbicides (propanil, quinclorac, imazethapyr, and fenoxaprop-ethyl) failed to show any significant association. Findings from this study provided novel insights into the morphophysiological characteristics of Echinochloa ecotypes in rice production in Texas. The morphological diversity currently present in Echinochloa ecotypes could contribute to their adaptation to selection pressure imposed by different management tools, emphasizing the need for a diversified management approach to effectively control this weed species.
Apricot is considered an ecologically and economically important tree species of the stone-fruit crops that is widely grown in temperate regions of the world. Very few studies on apricot genetic diversity assessment have been carried out from the regions of Kashmir and Ladakh. In this backdrop, the present study was carried out to analyse the genetic diversity and population structure of 120 apricot genotypes collected from both the regions using 21 SSR markers. A total of 52 alleles were amplified with average values of marker index (MI) = 0.7084, resolving power (RP) = 2.8690, polymorphism information content (PIC) = 0.3132, Na = 2.317, Ne = 1.720, I = 0.572, Ho = 0.284, He = 0.360 and an average polymorphism of 91.2% per assay indicating high level of genetic diversity. The neighbour-joining (NJ) dendrogram generated three main clusters among selected apricot genotypes independent of their geographical locations. Interestingly, the result of the dendrogram coincides with the results of structure analysis which showed that the 120 apricot genotypes could be assigned to three (K = 3) sub-populations and the grouping of genotypes did not follow their geographical location suggesting that they share the same genetic pool. Moreover, analysis of molecular variance showed that 73% of the variation was attributed to differences within the individuals, 25% among individuals while only 2% of the variation was observed among the populations. The present study represents the most comprehensive analysis of the genetic diversity and population structure of apricot genotypes in Kashmir and Ladakh regions of India.
There was little information regarding the occurrence of canine vector-borne disease (CVBDs) in shelter dogs in Thailand. This work is the first report regarding a molecular method used to determine the occurrence and genetic diversity of three canine tick-borne pathogens (TBPs) (Hepatozoon canis, Anaplasma platys and Ehrlichia canis) in blood samples from 275 shelter dogs in the north and central areas of Thailand. The PCR results based on the 18S rRNA and 16S rRNA genes showed that 71 (25.82%) dogs were positive for at least a TBP. The overall occurrence rates of H. canis, A. platys and E. canis infections were 1.81, 16.36 and 7.64%, respectively. For the phylogenetic analysis, A. platys 16S rRNA gene was genetically diverse, while H. canis 18S rRNA and E. canis 16S rRNA genes were conserved. The haplotype diversity exhibited 12 and 2 haplotypes as well as 78 and 178 polymorphic sites of A. platys and E. canis 16S rRNA genes, respectively. Our findings could be used to improve the understanding of phylogeny and genetic diversity of TBP rRNA genes and used to ameliorate the diagnosis and control programmes for the diseases in Thailand.
Tall fleabane [Conyza sumatrensis (Retz.) E. Walker] is commonly invasive in agricultural fields, reducing yield in various infested crops. The current study investigates the genetic diversity within and between a significant number of invasive C. sumatrensis biotypes in soybean fields in southern, southeastern, and midwestern Brazil, using microsatellites as molecular markers. High and low observed and expected heterozygosity estimated in microsatellite loci supported our hypothesis that different levels of genetic diversity may be detected within biotypes from different invaded fields. Analysis of a significant number of biotypes in several fields showed high and low genetic diversity not associated with geographic distribution, bottleneck effect, or susceptibility to glyphosate. A deficit of heterozygous plants, high genetic divergence, and moderate allelic transference were also observed. Allelic fixation was different in the different biotypes. The bottleneck effect was seen in biotypes with reduced genetic diversity and in biotypes with the highest genetic diversity. Data on genetic diversity, bottleneck effect, and glyphosate resistance showed contrasts in biotypes from nearby invaded fields. Our study showed different genetic diversity levels in biotypes from invaded areas under the same climatic conditions.
The Sikkim-Himalaya represents one of the unique reservoirs of rice genetic resources in India owing to the presence of a large number of landraces adapted to extreme climatic and edaphic conditions. This valuable gene pool is now under threat due to the introduction of high-yielding varieties, wanting suitable genetic intervention for enhancing their productivity, and thus economic viability. Development of lodging resistant, high-yielding varieties tolerant to soil acidity through association mapping aided marker-assisted breeding programmes can help achieve this in a fast and efficient manner. But this requires information on genetic diversity, population structure, etc., of the germplasm collection, which is strikingly lacking. We, therefore, characterized a set of 53 rice landraces from Sikkim to address the above issues. The results revealed moderate diversity, poor divergence and high gene flow in our germplasm collection attesting its utility as an association mapping panel. Further, a total of 115 putative marker-trait associations (P < 0.05, R2 ≥ 10%) were obtained using the general linear model and mixed linear models of which 25 were identical in both. Some of the associated markers were positioned in those regions where Qualitative Trait Loci have been previously identified in rice, providing credence to our results. The resources generated from this study will benefit the rice breeders from this region and elsewhere for targeting the yield and related traits, in addition to conservation efforts by the interested researchers.
The taxonomy of ruminant Trypanosoma theileri and its relatives (Kinetoplastida: Trypanosomatidae) is controversial, with recent phylogenetic studies segregating T. theileri in cattle and other ruminants worldwide into two major genetic lineages (the TthI and TthII clades) based on genetic markers. In the present study, T. theileri-like trypanosomes isolated from Honshu sika deer (Cervus nippon) in the western Japan (YMG isolate) were genetically characterized using a number of genetic markers. Sika deer trypanosomes of the YMG isolate were genetically different from the Trypanosoma sp. TSD1 isolate previously recorded from Hokkaido sika deer in northern Japan, with the former trypanosome isolate being genetically closer to European cervid trypanosomes and the bovine T. theileri TthII lineage. In contrast, the latter isolate exhibited greater relatedness to North American cervid trypanosomes and the bovine T. theileri TthI lineage, although a clear genetic distinction between these was apparent. Furthermore, trypanosomes in Honshu sika deer from the central part of Japan harboured additional genetic diversity and were closer to either TSD1 or YMG isolates, while distinct from known T. theileri-related genotypes. Importantly, cervids and wild ruminants worldwide might harbour divergent descendants of a T. theileri ancestor, which exhibit rigid host specificity to either bovines or cervid species.
Participatory plant breeding (PPB), commonly applied in the Global South to address the needs of underserved farmers, refers to the active collaboration between researchers, farmers and other actors throughout the breeding process. In spite of significant public and private investments in crop variety improvement in the Global North, PPB is increasingly utilized as an approach to address cropping system needs. The current study conducted a state-of-the-art review, including a comprehensive inventory of projects and five case studies, to explore the emergence of PPB in the Global North and inform future PPB efforts. Case studies included maize (Zea mays), tomato (Solanum lycopersicum), Brassica crops (Brassica oleracea), wheat (Triticum aestivum) and potato (Solanum tuberosum). The review identified 47 projects across the United States, Canada and Europe including 22 crop species representing diverse crop biology. Improved adaptation to organic farming systems and addressing principles and values of organic agriculture emerged as consistent themes. While projects presented evidence that PPB has expanded crop diversity and farmer's access to improved varieties, obstacles to PPB also emerged including challenges in sustained funding as well as addressing regulatory barriers to the commercial distribution of PPB varieties. Agronomic improvements were only one lens motivating PPB, with many projects identifying goals of conservation of crop genetic diversity, farmers' seed sovereignty and avoidance of certain breeding techniques. The authors conclude that a multidisciplinary approach is needed to fully understand the social, political and agroecological influences driving the emergence of projects in the Global North and factors impacting success.