A small effect of conservation agriculture on soil biodiversity that differs between biological kingdoms and geographic locations

The ITS2 barcode was used to accurately identify Albiziae Cortex, Albiziae Flos and their adulterants in this study. A total of46 samples from Albiziae Cortex, Albiziae Flos and their adulterants were collected. The ITS2 regions were amplified and sequenced. Sequences were assembled using the CodonCode Aligner. The genetic distances of ITS2 region were calculated using MEGA 5.0. BLAST1, nearest distance and phylogenetic tree (NJ-tree) methods were used to assess the identification efficiency of the ITS2 barcode. The results revealed that the intraspecific genetic distances of Albizia julibrissin were lower than the interspecific genetic distances between A. julibrissin and its adulterants. The identification efficiency of ITS2 barcode using BLAST1 was 100%. The NJ-tree showed that A. julibrissin and their adulterants can be easily differentiated according to their monophyly. The ITS2 barcode is suitable to be as a barcode to identify Albiziae Cortex, Albiziae Flos and their adulterants.

Aim of this study is to analyze the genetic structure of long-crowing Denizli chicken, a Turkish local fowl, using nucleotide sequences of the mitochondrial DNA 12S and D-loop regions. DNA isolation was carried out using feather samples of cocks of this local breed. D-loop and 12S regions were amplified using polymerase chain reaction technique and ca 1230 bp and 950 pb PCR products were obtained respectively. Native genotype Denizli fowl’s D-loop and 12S regions were sequenced and sequencing data were analyzed and compared with the related published data. Nucleotide content of the 12S region showed a 32% A, 19% T, 19% G and 30% C, whilst AT and GC ratios were found as 59.88% and 40.12% respectively for D-loop region. D-loop sequence data analysis clearly identified the Denizli fowl within the clade of long-crowing cocks. The results give the first informative data about the mitochondrial DNA D-loop and 12S regions of this local breed from Turkey.

Physalis species are used as an indigenous food and medicine in Kenya. However, species identification and an analysis of the health-promoting bioactive compounds and antioxidant properties are lacking. In this study, we report the molecular identification and mineral and phytochemical profiling of wild Physalis accessions. Leaf samples of 10 Physalis accessions were collected and used for species identification using nuclear ITS2 and plastid rbcL barcodes. Ripe fruits were collected from the same accessions and analyzed for mineral, total phenolic, tannin, and flavonoid contents, and antioxidant activities. The Physalis species were discriminated based on the ITS2 barcode and identified as Physalis purpurea. The genetic diversity, distance, and polymorphism of the ITS2 region of Physalis accessions were high due to the high rate of singleton and parsimony mutations. No genetic diversity, distance, or polymorphism was observed based on the rbcL barcode. The mineral content was significantly different (p < 0.05) for calcium, zinc, nickel, copper, and lithium among the Physalis accessions. No significant variation (p > 0.05) was found for phenolic acids or flavonoids, but the tannic acid content varied significantly (p < 0.05). DPPH free radical scavenging varied significantly (p < 0.05) among Physalis accessions. In conclusion, nuclear ITS2 was used to successfully identify the Physalis species of all the accessions as Physalis purpurea. The present study confirmed that Physalis purpurea has a significantly high mineral and phytochemical content and antioxidant activity. The findings from this study can be used to facilitate exploitation of Physalis purpurea in genetic breeding, their application in pharmaceutical, cosmetic, and nutritional value as well as conservation and sustainable use.

To identify Ephedrae Herba using the ITS2 barcode and to secure its quality and safety in medication. Total genomic DNA was isolated from Ephedrae Herba and its closely related species. Nuclear DNA ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. The Kimura 2-Parameter (K2P) distances were calculated using software MEGA 5.0. Identification analyses were performed using BLAST1, Nearest Distance and neighbor-joining (NJ) methods. The intra-specific genetic distances of Ephedrae Herba were ranged from 0 to 0.002. The inter-specific genetic distances between Ephedrae Herba and its closely related species were ranged from 0.004 to 0.034. All the three methods showed that ITS2 could discriminate Ephedrae Herba from its closely related species correctly. The ITS2 region is suitable to be used for authentication of Ephedrae Herba, and our study further confirmed the effectiveness of ITS2 to identify traditional Chinese medicinal materials.

The genus Jatropha belongs to the family Euphorbiaceae having significant economic importance. The present investigation was undertaken with an aim to understand phylogenetic relationships among seven species (J. curcas, J. glandulifera, J. gossypifolia, J. integerrima, J. multifida, J. podagrica, and J. tanjorensis.) which are widely distributed in India, using nuclear ribosomal DNA ITS sequence (nrDNA ITS) and to compare the results with multilocus marker analysis systems reported earlier for the same genus. The size variation obtained among sequenced nrDNA ITS regions was narrow and ranged from 647 to 654 bp. The overall mean genetic distance (GD) of genus Jatropha was found to be 0.385. Highest interspecific GD (0.419) was found between J. glandulifera and J. multifida. The least interspecific GD (0.085) was found between J. gossypifolia and J. tanjorensis. The highest intraspecific GD was observed in J. podagrica (0.011) and least in J. gossypifolia (0.002). The phylogram obtained using nrDNA ITS sequence showed congruence with the phylograms obtained using multilocus markers system reported earlier with minor variations. The present study also strongly supports high phylogenetic closeness of J. curcas and J. integerrima. The only exception found was J. podagrica which clustered with J. multifida in earlier based on multilocus marker analysis, was clustered with J. curcas in the present analysis. The sequence data generated in the present investigation will help for further studies in intraspecies population, and their phylogenetic analysis, biogeographical, molecular evolution studies and also pave way for future phylogenetic and/or evolution studies among the other groups belongs to the family Euphorbiaceae.

In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.

Cantaloupe (Cucumis melo L.) landraces have highly genetic diversity so that it is important to evaluate such distances for future breeding projects. Internal transcribed spacer regions (ITS1 and ITS4 regions) capability was used to evaluate genetic diversity of cantaloupe landraces and to identity genetic diversity among cantaloupe landraces. After DNA extraction from leaf tissue at the 4-true leaf stage and DNA quantification and qualification based on spectrophotometer and agarose gel electrophoresis, electrophoretic bands were sequenced. The sequence quality was assessed and compared with the ClustalW method. Phylogenetic relationship dendrograms were plotted based on genetic distance. Melon landraces could not be separated based on geographical location. The ITS1 region was more separable and diverse than the ITS4 region and was able to differentiate landraces related to the Hirmand region in a separate dendrogram. The numerical value (dN/dS) in ITS1 and ITS4 were 0.658 and 0.450, respectively, indicating pure selection had occurred in the gene and did not cause any important changes. The ITS region is not suitable to investigate intraspecific genetic diversity.

Stolotermes ruficeps is a widespread, primitive, lower termite occupying dead and decaying wood of many tree species in New Zealand's temperate forests. We identified core bacterial taxa involved in gut processes through combined DNA- and RNA (cDNA)-based pyrosequencing analysis of the 16S nucleotide sequence from five S. ruficeps colonies. Most family and many genus-level taxa were common to S. ruficeps colonies despite being sampled from different tree species. Major taxa identified were Spirochaetaceae, Elusimicrobiaceae and Porphyromonadaceae. Others less well known in termite guts were Synergistaceae, Desulfobacteraceae, Rhodocyclaceae, Lachnospiraceae and Ruminococcaceae. Synergistaceae, Lachnospiraceae and Spirochaetaceae were well represented in the RNA data set, indicating a high-protein synthesis potential. Using 130,800 sequences from nine S. ruficeps DNA and RNA data sets, we estimated a high level of bacterial richness (4024 phylotypes at 3% genetic distance). Very few abundant phylotypes were site-specific; almost all (95%) abundant phylotypes, representing 97% of data set sequences, were detected in at least two S. ruficeps colonies. This study of a little-researched phylogenetically basal termite identifies core bacteria taxa. These findings will extend inventories of termite gut microbiota and contribute to the understanding of the specificity of termite gut microbiota.

Herbal material is both a medicine and a commodity. Accurate identification of herbal materials is necessary to ensure the safety and effectiveness of medication. With this work, we initiated an identification method to investigate the species authenticity for herbal products of Celastrus orbiculatus and Tripterygum wilfordii utilizing DNA barcoding technology. An ITS2 (internal transcribed spacer two) barcode database including 59 sequences was successfully established to estimate the reliability of species-level identification for Celastrus and Tripterygium. Our findings showed that ITS2 can effectively and clearly distinguish C. orbiculatus, T. wilfordii and its congeners. Then, we investigated the proportions and varieties of adulterant species in the herbal markets. The data from ITS2 region indicated that 13 (62%) of the 21 samples labeled as “Nan-she-teng” and eight (31%) of the 26 samples labeled as “Lei-gong-teng” were authentic; the remaining were adulterants. Of the 47 herbal products, approximately 55% of the product identity were not in accordance with the label. In summary, we support the efficacy of the ITS2 barcode for the traceability of C. orbiculatus and T. wilfordii, and the present study provides one method and reference for the identification of the herbal materials and adulterants in the medicinal markets.

In this study, ITS2 barcode was used to identify Bupleurum chinense and B. longiradiatum. The ITS2 regions of 48 samples were amplified and sequenced. The sequences obtained above were aligned and the K2P distances were calculated. We used three methods, BLAST1, nearest distance and phylogenetic tree (NJ-tree), to test the identification ability. The results showed that the maximum intraspecific genetic distance of B. chinense was 0.013, and the minimum interspecific genetic distance between B. chinense and B. longiradiatum was 0.049. The NJ-tree can easily identify B. chinense and B. longiradiatum. Therefore, the ITS2 barcode is suitable to identify B. chinense and B. longiradiatum.

Solanaceae is an important angiosperm family having high economic and commercial importance being used as sources of food, spice, and medicine. The traditional method of plant identification using morphological characters often leads to inaccurate and unreliable results because of genetic and environmental factors. In Solanaceae family, highly similar morphological traits among the species also make the identification difficult. The present study evaluated the viability of employing DNA barcodes, ITS2 and rbcL regions to identify Solanaceae species. DNA barcoding is a rapid and reliable tool to identify and study the genetic relationship among plants and animals. The present study for the first time was conducted during 2017 to 2019 at the Department of Botany, Mizoram University to analyze the applicability of two DNA regions-rbcL (Ribulosebisphosphate Carboxylases) and ITS2 (Internal Transcribed Spacer gene), as the barcodes for the identification of Solanaceae species of Mizoram, India. Fifteen Solanaceae species were analyzed at ITS2 and rbcL regions. The success rates of PCR amplification and sequencing of ITS2 and rbcL were 100% and MEGA 7.0 was used to align the sequences and to compute genetic distances. The results from BLAST and nearest-distance methods revealed that ITS2 and rbcL can be used to identify all the studied species and high divergences at taxonomic levels were also found using K2P (Kimura 2-parameter) model. Hence, our study reveals that rbcL and ITS2 regions are suitable DNA barcodes for the identification of Solanaceae species and therefore can be successfully used to monitor the adulteration of plants.

Wild sunflower Helianthus annuus originates from North America and has naturalised in Argentina where it is considered invasive. The present study attempts to assess the genetic diversity using two different molecular marker systems to study the wild genetic patterns and to provide data applicable to conservation and breeding uses. Ten natural populations sampled throughout the wild range and six inbred lines were studied using inter-simple sequence repeat (ISSR) and simple sequence repeats (SSR) markers. A total of 64 ISSR bands and 29 SSR alleles were produced from 106 wild and cultivated plants. We found 9 ISSR private bands and 21 SSR private alleles in wild accessions, but no private bands/alleles were found in cultivated sunflowers. Molecular variability in wild populations was approximately 60% higher than in inbred lines. Local wild sunflowers kept considerable diversity levels in comparison with populations in the centre of origin (approximately 70%) and therefore they might possess a potential for adaptive evolutionary change. Analysis of molecular variance (AMOVA) indicated population structure with nearly 20% of genetic variability attributable to between-population differentiation. Principal coordinate analyses (PCO) grouped wild populations from different geographic locations, and a Mantel test showed low congruence between genetic distance (GD) and geographic distances (GGD); hence, molecular data could not rule out multiple wild introduction events. Low correlations were found between ISSR and SSR GD at individual and population levels; thus, divergent evolutionary groups were not evident in local wild sunflowers. Several genetic diversity criteria were utilised to assign conservation value and certain wild populations emerged as interesting sites for more extensive sampling.

The cladistic classification of Phthiracaroidea (Niedbała 1986) and Euphthiracaroidea (Mahunka 1990) (Acari, Oribatida), based on morphological symplesio- and synapomorphic characters has been subjected to verification by molecular analysis of mitochondrial COI and nuclear ITS2, 18S and 28S DNA regions. Twenty-one selected species from representative genera of both superfamilies were analyzed. The phylogenetic trees generated by maximum likelihood method confirm the classification of ptyctimous mites resulting from the analysis of morphological characters. Mesoplophoridae, representing the supercohorts Enarthronota, were placed separately to the clade Mixonomata containing Phthiracaroidea and Euphthiracaroidea that are in a sister relationship. The percentage of genetic divergence between the main clades is high in three markers (COI, ITS2 and 28S), it ranges from 36.8% to 38.7% in the barcode marker COI from 26.0% to 35.4% in ITS2 and from 16.2% to 30.0% in 28S, while in 18S it is very low (1.1% - 3.3%). In the Phthiracaroidea, two sister clades Steganacaridae and Phthiracaridae are distinguished. Steganacaridae include three clades Atropacarus, Hoplophthiracarus and Steganacarus with Steganacarus and Tropacarus as the sister clades. Closely related S. (T.) carinatus and S. (T.) pulcherrimus are separate species. Within the Euphthiracaroidea, the four clades: Mesotritia, Microtritia, Acrotritia and Euphthiracarus are resolved. Mesotritia is the most distant, and Microtritia and Acrotritia show the closest relationship.

We present a multilocus phylogeny of the class Dacrymycetes, based on data from the 18S, ITS, 28S, RPB1, RPB2, TEF-1α, 12S, and ATP6 DNA regions, with c. 90 species including the types of most currently accepted genera. A variety of methodological approaches was used to infer phylogenetic relationships among the Dacrymycetes, from a supermatrix strategy using maximum likelihood and Bayesian inference on a concatenated dataset, to coalescence-based calculations, such as quartet-based summary methods of independent single-locus trees, and Bayesian integration of single-locus trees into a species tree under the multispecies coalescent. We evaluate for the first time the taxonomic usefulness of some cytological phenotypic characters, i.e., vacuolar contents (vacuolar bodies and lipid bodies), number of nuclei of recently discharged basidiospores, and pigments, with especial emphasis on carotenoids. These characters, along with several others traditionally used for the taxonomy of this group (basidium shape, presence and morphology of clamp connections, morphology of the terminal cells of cortical/marginal hyphae, presence and degree of ramification of the hyphidia), are mapped on the resulting phylogenies and their evolution through the class Dacrymycetes discussed. Our analyses reveal five lineages that putatively represent five different families, four of which are accepted and named. Three out of these four lineages correspond to previously circumscribed and published families (Cerinomycetaceae, Dacrymycetaceae, and Unilacrymaceae), and one is proposed as the new family Dacryonaemataceae. Provisionally, only a single order, Dacrymycetales, is accepted within the class. Furthermore, the systematics of the two smallest families, Dacryonaemataceae and Unilacrymaceae, are investigated to the species level, using coalescence-based species delimitation on multilocus DNA data, and a detailed morphological study including morphometric analyses of the basidiospores. Three species are accepted in Dacryonaema, the type, Da. rufum, the newly combined Da. macnabbii (basionym Dacrymyces macnabbii), and a new species named Da. macrosporum. Two species are accepted in Unilacryma, the new U. bispora, and the type, U. unispora, the latter treated in a broad sense pending improved sampling across the Holarctic.

The identity of the holotype specimen of G. campestre var. famatinum is investigated through both morphological and molecular approaches. Morphological data and molecular data of ITS, 28S, rpb1 and atp6 DNA regions confirm that G. campestre var. famatinum is a different species to G. campestre var. campestre, and that it should belong to G. sect. Hariotia instead of G. sect. Campestria. However, the analyses performed also show that the type specimen of G. campestre var. famatinum is better included within the intraspecific variation of a previously described species, G. parvisporum, so both names are considered as heterotypic synonyms.