DNA Markers Research Articles

Pheromonal variation and mating between two mitotypes of fall armyworm (Spodoptera frugiperda) in Africa

In the Americas, the fall armyworm (Spodoptera frugiperda) exists in two genetically distinct strains, the corn (C) and rice (R) strains. Despite their names, these strains are not associated with host plant preferences but have been shown to vary in pheromone composition and male responses. Recently, S. frugiperda was detected in Africa as an invasive species, but knowledge about variation in strain types, pheromone composition and inter-strain mating of populations of the pest in the continent has not been fully examined. Therefore, this study aimed to investigate variations, if any in the pheromone composition of female moths, male moth responses, and mating between C and R mitotypes of S. frugiperda populations in Kenya, as well as their geographic distribution. Strains (mitotypes) of S. frugiperda were identified using mitochondrial DNA (mtDNA) markers, and their pheromonal composition determined by coupled gas chromatography–mass spectrometric (GC–MS) analysis. Male moth responses to these compounds were evaluated using GC-electroantennographic detection (EAD), electroantennogram (EAG), and wind tunnel assays. Oviposition assays were used to determine whether R and C mitotype moths could mate and produce eggs. The results showed that both the R and C mitotypes were present, and there were no statistically significant differences in their distribution across all sampled locations. Five pheromone compounds including (Z)-7-dodecenyl acetate (Z7-12:OAc), (Z)-7-tetradecenyl acetate (Z7-14:OAc), (Z)-9-tetradecenyl acetate (Z9-14:OAc), (Z)-11-tetradecenyl acetate (Z11-14:OAc) and (Z)-11-hexadecenyl acetate (Z11-16:OAc), were detected in the pheromone glands of female moths of both mitotypes, with Z9-14:OAc being the most abundant. The relative percentage composition of Z9-14:OAc was similar in both mitotypes. However, the R mitotype had a 2.7 times higher relative percentage composition of Z7-12:OAc compared to the C mitotype moth, while the C mitotype moth had a 2.4 times higher relative percentage composition of Z11-16:OAc than the R mitotype moth. Male moths of both mitotypes exhibited similar responses to the pheromone compounds, showing the strongest responses to Z9-14:OAc and Z7-12:OAc in electrophysiological and behavioural assays. There was mating between R and C mitotypes with egg production comparable to mating within the same mitotype. Our results revealed that differences between the two S. frugiperda mitotypes are characterized by female moth pheromone composition rather than male moth responses to the pheromones, and that this does not prevent hybridisation between the mitotypes, which may have implications for their management.

Genetic and Epigenetic Aspects of Type 1 Diabetes Mellitus: Modern View on the Problem.

Omics technologies accumulated an enormous amount of data that advanced knowledge about the molecular pathogenesis of type 1 diabetes mellitus and identified a number of fundamental problems focused on the transition to personalized diabetology in the future. Among them, the most significant are the following: (1) clinical and genetic heterogeneity of type 1 diabetes mellitus; (2) the prognostic significance of DNA markers beyond the HLA genes; (3) assessment of the contribution of a large number of DNA markers to the polygenic risk of disease progress; (4) the existence of ethnic population differences in the distribution of frequencies of risk alleles and genotypes; (5) the infancy of epigenetic research into type 1 diabetes mellitus. Disclosure of these issues is one of the priorities of fundamental diabetology and practical healthcare. The purpose of this review is the systemization of the results of modern molecular genetic, transcriptomic, and epigenetic investigations of type 1 diabetes mellitus in general, as well as its individual forms. The paper summarizes data on the role of risk HLA haplotypes and a number of other candidate genes and loci, identified through genome-wide association studies, in the development of this disease and in alterations in T cell signaling. In addition, this review assesses the contribution of differential DNA methylation and the role of microRNAs in the formation of the molecular pathogenesis of type 1 diabetes mellitus, as well as discusses the most currently central trends in the context of early diagnosis of type 1 diabetes mellitus.

Using genome-wide data to ascertain taxonomic status and assess population genetic structure for Houston toads (Bufo [= Anaxyrus] houstonensis)

The Houston toad (Bufo [= Anaxyrus] houstonensis) is an endangered amphibian with a small geographic range. Land-use changes have primarily driven decline in B. houstonensis with population supplementation predominant among efforts to reduce its current extinction risk. However, there has been historic uncertainty regarding the evolutionary and conservation significance of B. houstonensis. To this end, we used 1170 genome-wide nuclear DNA markers to examine phylogenetic relationships between our focal taxon, representatives of the Nearctic B. americanus group, and B. nebulifer, a sympatric Middle American species. Phylogenetic analyses indicate B. houstonensis is a taxon that is distinct from B. americanus. We corroborated such genetic distinctiveness with an admixture analysis that provided support for recent reproductive isolation between B. americanus and B. houstonensis. However, ABBA-BABA tests for ancient admixture indicated historic gene flow between Nearctic species while no signal of historic gene flow was detected between Nearctic and Middle-American species. We used an admixture analysis to recognize four Management Units (MU) based on observed genetic differentiation within B. houstonensis and recommend captive propagation, population supplementation, and habitat restoration efforts specific to each MU. Our results re-affirm the evolutionary novelty of an endangered relict.

Characterization of Eighty-Eight Single-Nucleotide Polymorphism Markers in the Manila Clam Ruditapes philippinarum Based on High-Resolution Melting (HRM) Analysis.

Single-nucleotide polymorphisms (SNPs) are the most commonly used DNA markers in population genetic studies. We used the Illumina HiSeq4000 platform to develop single-nucleotide polymorphism (SNP) markers for Manila clam Ruditapes philippinarum using restriction site-associated DNA sequencing (RAD-seq) genotyping. Eighty-eight SNP markers were successfully developed by using high-resolution melting (HRM) analysis, with a success rate of 44%. SNP markers were analyzed for genetic diversity in two clam populations. The observed heterozygosity per locus ranged from 0 to 0.9515, while the expected heterozygosity per locus ranged from 0.0629 to 0.4997. The value of FIS was estimated to be from -0.9643 to 1.0000. The global Fst value was 0.1248 (p < 0.001). After Bonferroni correction, 15 loci deviated significantly from the Hardy-Weinberg equilibrium (p < 0.0006). These SNP markers provide a valuable resource for population and conservation genetics studies in this commercially important species.

Unraveling phenotypic ambiguities of kagzi and ornamental lime accessions: a comprehensive exploration through morpho-biochemical and DNA marker profiling

Unraveling phenotypic ambiguities of kagzi and ornamental lime accessions: a comprehensive exploration through morpho-biochemical and DNA marker profiling

Sensitive and efficient surveillance of Japanese giant salamander (Andrias japonicus) distribution in western Japan using multi-copy nuclear DNA marker

Japanese giant salamander (Andrias japonicus) is one of the largest amphibian species in the world and an iconic species in Japan. However, as its distribution has recently declined across the country, rapid and extensive monitoring of the distribution is urgently needed for its efficient conservation. Here, we used environmental DNA (eDNA) analysis to assess the Japanese giant salamander’s distribution in western Japan and, for that purpose, we collected 410 water samples from 12 rivers. We then developed a new eDNA assay for multi-copy nuclear DNA (nuDNA) of the giant salamander and compared the eDNA detectability of the nuDNA marker with that of a previous mitochondrial DNA (mtDNA) marker. Throughout the survey, we detected target eDNA from 162 water samples using either of the markers, which generally corresponded to the known natural distribution of the species. Additionally, the use of the nuDNA marker allowed for higher detection rate of target eDNA than the mtDNA marker. Moreover, the detection rate of target eDNA decreased substantially in water samples with higher conductivity and also partly in those with higher pH, suggesting their negative impacts on the salamander’s ecology. Our results demonstrated that eDNA analysis with multi-copy nuDNA marker is highly useful for efficient and sensitive surveillance of Japanese giant salamander’s distribution. Our study provided the methodology for efficiently monitoring the Japanese giant salamander’s distribution via eDNA analysis and facilitating conservation activities for them.

Marine Crabs of Guinea-Bissau, with Emphasis on the Deep Fauna, Supported by an Integrative Taxonomy

The updated checklist of the marine crabs of Guinea-Bissau presented in this work is the result of consolidating decades of research, ongoing systematic revisions of the regional carcinofauna, and the inclusion of new records. DNA markers and morphological analyses for accurate identifications are integrated. Sixty-one species are mentioned in this list after reviewing specimens from scientific collections and literature. Of these 61 species listed, the presence of 51 species in Guinea-Bissau is confirmed, and 21 are reported for the first time in the area. A total of 98 sequences were obtained from 41 species (51 of 16S for 38 species and 47 of COI for 35 species), and 48 sequences (24 of 16S and 24 of COI) were the first ones obtained for 29 species. The maximum or minimum bathymetric range is expanded by 16 species, and for other six species, a color description is provided for the first time. Merocryptus obsoletus is synonymized as Merocryptus boletifer. The present study will be a useful baseline for gathering further ecological information about globally important marine taxa, not only in Guinea-Bissau but about African brachyurans in general.

Polly: An R package for genotyping microsatellites and detecting highly polymorphic DNA markers from short-read data.

Highly polymorphic markers, such as microsatellites, are invaluable for the study of natural populations. However, contemporary methods for genotyping highly polymorphic variants have serious drawbacks that impede their efficiency. We created Polly, an R package with C++ source code that uses Illumina short-read data to genotype microsatellites, detect highly polymorphic variants and identify clusters of highly polymorphic SNPs, indels and microsatellites. We tested Polly on short-read data from Xiphophorus birchmanni (Teleostei: Poeciliidae) and Arabidopsis thaliana, finding it to be efficient and accurate both for microsatellite genotyping and polymorphic marker detection. This program can be applied to any diploid population for which there exists short-read data and at least one scaffolded reference genome.

Breeding approaches for improvement of temperate fruit crops and nuts - a review

Fruit breeding is an ancient technology with dynamic current technology and an exciting future (Janick and Moore, 1975, 1996).  In its broadest sense, fruit breeding refers to the purposeful genetic improvement of fruit crops through various techniques including selection, hybridization, mutation induction, and molecular techniques. Its origins trace to the domestication process in prehistory and antiquity, where useful species were chosen and cultivated, and improved by continuous selection (Janick, 2005, 2011). Much of the world fruit industry is still based on grower selection from chance seedlings as well as mutations (sports) and as a result many fruit species are characterized by a narrow germplasm base. Spontaneous hybridization between wild plants and cultivated clones was critical to the early domestication of fruits. The lengthy juvenile period and the large field space required to grow temperate fruits have imposed major limitations on conventional breeding and various molecular biological techniques have been employed to make fruit breeding easier. (Igarashi et al.2016). Marker-assisted selection (MAS) uses DNA markers to provide an early DNA-based evaluation of genetic performance potential of seedlings, with the aim of improving cost and/or genetic efficiency of seedling selection (Ru, et al., 2015). Transgenic technology has facilitated the development of fruit trees with resistance to fungal or bacterial diseases, improved fruit quality, or root stocks with better rooting or dwarfing ability. In recent years, new technologies for genetic improvement, such as transgrafting, and genome-editing, have emerged. (Mudge, et al. 2009; Jia and Wang 2014). Using these techniques, no foreign genes are present in the final product, and some of them show considerable promise for application to apple breeding. Furthermore, genome editing, by which only the target gene can be accurately modified, is emerging as a novel breeding technology. These new technologies will undoubtedly facilitate fruit breeding, and yield novel and attractive fruit cultivars.

DNA- and selectable-marker-free genome-editing system using zygotes from recalcitrant maize inbred B73.

Genome-editing tools such as the clustered regularly interspaced short palindromic repeats/Cas9 (CRISPR/Cas9) system have become essential tools for increasing the efficiency and accuracy of plant breeding. Using such genome-editing tools on maize, one of the most important cereal crops of the world, will greatly benefit the agriculture and the mankind. Conventional genome-editing methods typically used for maize involves insertion of a Cas9-guide RNA expression cassette and a selectable marker in the genome DNA; however, with such methods, it is essential to eliminate the inserted DNA cassettes to avoid legislative concerns on gene-modified organisms. Another major hurdle for establishing an efficient and broadly applicable DNA-free genome-editing system for maize is presented by recalcitrant genotypes/cultivars, since cell/tissue culture and its subsequent regeneration into plantlets are crucial for producing transgenic and/or genome-edited maize. In this study, to establish a DNA-free genome-editing system for recalcitrant maize genotypes/cultivars, Cas9-gRNA ribonucleoproteins (RNPs) were directly delivered into zygotes isolated from the pollinated flowers of the maize-B73 cultivar. The zygotes were successfully developed and regenerated into genome-edited plantlets by co-culture with phytosulfokine, a peptide phytohormone. The method developed herein made it possible to obtain DNA- and selectable-marker-free genome-edited recalcitrant maize genotypes/cultivars with high efficiency. This method can advance the molecular breeding of maize and other important cereals, regardless of their recalcitrant characteristics.

Demographic dynamics and molecular evolution of the rare and endangered subsect. Gerardianae of Pinus: insights from chloroplast genomes and mitochondrial DNA markers.

The divergence of subsect. Gerardianae was likely triggered by the uplift of the Qinghai-Tibetan Plateau and adjacent mountains. Pinus bungeana might have probably experienced expansion since Last Interglacial period. Historical geological and climatic oscillations have profoundly affected patterns of nucleotide variability, evolutionary history, and species divergence in numerous plants of the Northern Hemisphere. However, how long-lived conifers responded to geological and climatic fluctuations in East Asia remain poorly understood. Here, based on paternally inherited chloroplast genomes and maternally inherited mitochondrial DNA markers, we investigated the population demographic history and molecular evolution of subsect. Gerardianae (only including three species, Pinus bungeana, P. gerardiana, and P. squamata) of Pinus. A low level of nucleotide diversity was found in P. bungeana (π was 0.00016 in chloroplast DNA sequences, and 0.00304 in mitochondrial DNAs). The haplotype-based phylogenetic topology and unimodal distributions of demographic analysis suggested that P. bungeana probably originated in the southern Qinling Mountains and experienced rapid population expansion since Last Interglacial period. Phylogenetic analysis revealed that P. gerardiana and P. squamata had closer genetic relationship. The species divergence of subsect. Gerardianae occurred about 27.18 million years ago (Mya) during the middle to late Oligocene, which was significantly associated with the uplift of the Qinghai-Tibetan Plateau and adjacent mountains from the Eocene to the mid-Pliocene. The molecular evolutionary analysis showed that two chloroplast genes (psaI and ycf1) were under positive selection, the genetic lineages of P. bungeana exhibited higher transition and nonsynonymous mutations, which were involved with the strongly environmental adaptation. These findings shed light on the population evolutionary history of white pine species and provide striking insights for comprehension of their species divergence and molecular evolution.

Ecological Speciation without Morphological Differentiation? A New Cryptic Species of Diodontus Curtis (Hymenoptera, Pemphredonidae) from the Centre of Europe.

Upon exploring the mitotype diversity of the aphid-hunting wasp, Diodontus tristis, we revealed specimens with highly divergent mitotypes from two localities in Lithuania and nesting in clayey substrate, while the specimens with typical mitotypes were found nesting in sandy sites. The comparison of inter- and intra-specific distances and application of delimitation algorithms supported the species status of the clay-nesting populations. Using a set of DNA markers that included complete or partial sequences of six mitochondrial genes, three markers of ribosomal operon, two homeobox genes, and four other nuclear genes, we clarified the phylogenetic relationships of the new cryptic species. The endosymbiotic bacteria infestation was checked, considering the option that the divergent populations may represent clades isolated by Wolbachia infection; however, it did not demonstrate any specificity. We found only subtle morphological differences in the new clay-nesting species, D. argillicola sp. nov.; the discriminant analysis of morphometric measurements did not reliably segregate it as well. Thus, we provide the molecular characters of the cryptic species, which allow confident identification, its phylogenetic position within the genus, and an updated identification key for the D. tristis species group.

Genetic diversity among maize (Zea mays L.) inbred lines adapted to Japanese climates.

Understanding the genetic diversity of inbred lines is vital for development of superior F1 varieties. The present study aimed to analyze Japanese maize parental inbred lines and determine their genetic diversity for future breeding. Genetic analyses were conducted using multiple methods. Principal component analysis (PCA), phylogenetic trees, and Bayesian clustering reflected borders between heterotic groups according to the derivation of each inbred line. A self-pollinated line derived from a classic F1 variety and another line from an open-pollinated population from the same derivation were classified as separate components by PCA and Bayesian clustering. The result suggests that open pollination could be essential in modern breeding. Of those classified as dent or flint based on their derivation, some had a combination of all components or clusters. Therefore, the classification of inbred lines should be based on their derivation and DNA markers. The findings will be valuable for breeding and genetic studies in Japan. Additionally, these techniques may be used to obtain a more significant number of SNPs and related phenotypic data.

Application of DNA barcode ITS2to identify herbal medicinal materials

Nowadays, the demand for using herb plants to treat and protect human health is increasing. Along with the growing herbal market, the adulteration of herbal medicinal materials is increasing in many countries, including Vietnam. It is difficult to determine the origin of herbal medicinal materials by morphological or chemical methods when they are substituted by herbs from closely related species or adulterated intentionally by materials from unrelated plants or processed materials. Therefore, it is necessary to use molecular methods based on DNA markers. ITS is a highly conservative nuclear ribosomal marker that has been widely used in the identification of plant species. In this study, the ITS2 marker was used to identify 8 herbal medicinal materials, including 2 samples of polyscias(DLTN, DLBG), 2 samples of morinda (BKTN, BKQN), 2 samples of black zingiber (GDTN, GDYB) and 2 samples of black curcuma (NDTN, NDYB). Gene analysis and sequencing results showed that the DLTN and DLBG were Polyscias fruticosa; BKTN and BKQN were different species, Gynochthodes officinalis (BKTN) and Morinda officinalis (BKQN); GDTN and GDYB were Kaempferia parviflora; NDTN and NDYB were Curcuma caesia. The research results show that the ITS2 indicator can be used to identify medicinal species.

New insights into the chromosomes of stoneflies: I. Karyotype, C-banding and localization of ribosomal and telomeric DNA markers in Skwalacompacta (McLachlan, 1872) (Polyneoptera, Plecoptera, Perlodidae) from Siberia.

This study provides data on chromosome number (2n♂♀=26), sex determination mechanism (XY♂/XX♀), C-banding pattern, distribution of clusters of telomeric TTAGG repeats and 18S ribosomal DNA in the karyotype of the stonefly Skwalacompacta (McLachlan, 1872). For the first time in the history of stoneflies cytogenetics, we provide photos of the chromosomes of the Plecoptera insects. The karyotype of males and females of S.compacta consists of 12 pairs of autosomes. Three pairs of large autosomes and four pairs of medium-sized autosomes are subacrocentric. The remaining pairs of autosomes are small, with unclear morphology. Pericentromeric C-bands were revealed in all autosomes. The sex chromosomes are also subacrocentric. The short arms of X and Y chromosomes are entirely heterochromatic and are rich in ribosomal DNA sequences. In the X chromosome this arm is larger than in the Y chromosome. It is likely that this arm associated with the nucleolar organizer (NOR). Telomeric DNA (TTAGG)n repeats were detected in the terminal regions of all chromosomes.

A microfluidic card-based electrochemical assay for the detection of sulfonamide resistance genes

Most electroanalytical detection schemes for DNA markers require considerable time and effort from expert personnel to thoroughly follow the analysis and obtain reliable outcomes. This work aims to present an electrochemical assay performed inside a small card-based platform powered by microfluidic manipulation, requiring minimal human intervention and consumables. The assay couples a sample/signal dual amplification and DNA-modified magnetic particles for the detection of DNA amplification products. Particularly, the sul1 and sul4 genes involved in the resistance against sulfonamide antibiotics were analyzed. As recognized by the World Health Organization, antimicrobial resistance threatens global public health by hampering medication efficacy against infections. Consequently, analytical methods for the determination of such genes in environmental and clinical matrices are imperative. Herein, the resistance genes were extracted from E. coli cells and amplified using an enzyme-assisted isothermal amplification at 37 °C. The amplification products were analyzed in an easily-produced, low-cost, card-based set-up implementing a microfluidic system, demanding limited manual work and small sample volumes. The target amplicon was thus captured and isolated using versatile DNA-modified magnetic beads injected into the microchannel and exposed to the various reagents in a continuously controlled microfluidic flow. After the optimization of the efficiency of each phase of the assay, the platform achieved limits of detections of 44.2 pmol L−1 for sul1 and 48.5 pmol L−1 for sul4, and was able to detect down to ≥500-fold diluted amplification products of sul1 extracted from E. coli living cells in around 1 h, thus enabling numerous end-point analyses with a single amplification reaction.

Effect of dietary tall oil fatty acids and hydrolysed yeast in SNP2-positive and SNP2-negative piglets challenged with F4 enterotoxigenic Escherichia coli

Reduction of post-weaning diarrhoea caused by ETEC is a principal objective in pig farming in terms of welfare benefits. This study determined the effects of genetic susceptibility and dietary strategies targeting inflammation and fimbriae adherence on F4-ETEC shedding and diarrhoea in weaned piglets in an experimental challenge model. A DNA marker test targeting single nucleotide polymorphism 2 (SNP2) identified piglets as heterozygous (SNP2+, susceptible) or homozygous (SNP2-, resistant) to developing F4ac-ETEC diarrhoea. A total of 50 piglets, 25 SNP2+ and 25 SNP2-, were weaned at 30 days of age and equally distributed to different treatments (n = 10): Positive control (PC): piglets fed with a negative control diet and provided with colistin via drinking water; Negative control (NC): piglets fed with a negative control diet; Tall oil fatty acids (TOFA): piglets fed with a negative control diet + 1.0 g TOFA/kg feed; Yeast hydrolysate (YH): piglets fed with a negative control diet + 1.5 g YH/kg feed derived from Saccharomyces cerevisiae; and Combination (COM): piglets fed with a negative control diet + 1.0 g TOFA and 1.5 g YH/kg feed. On day 10 post-weaning, all piglets were infected with F4-ETEC by oral administration. Piglets fed with PC, TOFA, YH or COM had a lower faecal shedding of F4-ETEC than NC piglets (P < 0.001), which was also shorter in duration for PC and TOFA piglets than for NC piglets (P < 0.001). Piglets in PC, TOFA, YH and COM had a shorter diarrhoea duration versus NC when classified as SNP2+ (P = 0.02). Furthermore, PC, TOFA and YH piglets grew more than NC and COM piglets in the initial post-inoculation period (P < 0.001). In addition, the level of faecal F4-ETEC shedding and the percentage of pigs that developed F4-ETEC diarrhoea (72 vs. 32%, P < 0.01) following infection were higher, and the duration of F4-ETEC diarrhoea longer (2.6 vs. 0.6 days, P < 0.001), in SNP2+ piglets than in SNP2- piglets, and led to reduced growth performance (P = 0.03). In conclusion, piglets fed with TOFA, YH or their combination, irrespective of their SNP2 status, are more resilient to F4-ETEC infection. Moreover, SNP2+ piglets show a higher level of F4-ETEC shedding and diarrhoea prevalence than SNP2- piglets, confirming an association between SNP2 and F4ac-ETEC susceptibility.

Rapid In Vitro Regeneration and Genetic Fidelity Assessment of Regenerated Plants in Ayapana Triplinervis (Vahl) R.M. King &amp; H. Robinson: An Ethnomedicinal and Ornamental Herb

Background: Ayapana triplinervis is a popular ethnomedicinal and ornamental plant species. Due to its high medicinal importance, A. triplinervis was recently documented in the French Pharmacopeia. Objective: Rapid and efficient tissue culture protocol development is crucial for the high production and biotechnological applications of this plant. Methods: In this study, an efficient tissue culture protocol was developed for plant regeneration using nodal explants of A. triplinervis. The nodal explants were treated in Murashige and Skoog’s (MS) medium supplemented with various individual concentrations of cytokinins (BAP and KIN) and auxins (IAA and IBA). The nodal explant was regenerated in three different steps: (1) initial shoot induction, (2) shoot multiplication and elongation, and (3) rooting. Results: The results revealed that all individual concentrations (10, 20, 30, or 40 mg/L) of BAP or KIN responded to induce shoot initiation. The highest shoot multiplication and elongation were achieved in the MS medium with 20 mg/L BAP and 20 mg/L KIN. The regenerated plantlets produced better roots on MS medium containing 1.0 mg/L of each IAA or IBA. The well-established rooted plantlets were maintained in the culture room and greenhouse for better acclimatization and achieved a 100% survival rate. We analyzed the genetic fidelity of in vitro regenerated plants using random amplified polymorphic DNA (RAPD) markers. No genetic polymorphisms were observed in vitro plants compared to the mother plants. Conclusion: This efficient protocol could benefit future biotechnological applications like mass multiplication, genetic transformation and gene editing for improving the bioactive molecules in A. triplinervis.

Genome-Wide Association Study of Milk Composition in Karachai Goats.

This study is first to perform a genome-wide association study (GWAS) to investigate the milk quality traits in Karachai goats. The objective of the study was to identify candidate genes associated with milk composition traits based on the identification and subsequent analysis of all possible SNPs, both genome-wide (high-confidence) and suggestive (subthreshold significance). To estimate the milk components, 22 traits were determined, including several types of fatty acids. DNA was extracted from ear tissue or blood samples. A total of 167 Karachai goats were genotyped using an Illumina GoatSNP53K BeadChip panel (Illumina Inc., San Diego, CA, USA). Overall, we identified 167 highly significant and subthreshold SNPs associated with the milk components of Karachai goats. A total of 10 SNPs were located within protein-coding genes and 33 SNPs in close proximity to them (±0.2 Mb). The largest number of genome-wide significant SNPs was found on chromosomes 2 and 8 and some of them were associated with several traits. The greatest number of genome-wide significant SNPs was identified for crude protein and lactose (6), and the smallest number-only 1 SNP-for freezing point depression. No SNPs were identified for monounsaturated and polyunsaturated fatty acids. Functional annotation of all 43 SNPs allowed us to identify 66 significant candidate genes on chromosomes 1, 2, 3, 4, 5, 8, 10, 13, 16, 18, 21, 23, 25, 26, and 27. We considered these genes potential DNA markers of the fatty acid composition of Karachai goat milk. Also, we found 12 genes that had a polygenic effect: most of them were simultaneously associated with the dry matter content and fatty acids (METTL, SLC1A 8, PHACTR1, FMO2, ECI1, PGP, ABCA3, AMDHD2). Our results suggest that the genes identified in our study affecting the milk components in Karachai goats differed from those identified in other breeds of dairy goats.

Molecular characterization of red ginger varieties (Zingiber officinale Roxb. var. rubrum) by DNA markers

Abstract. Yunita O, Fadhilah NA, Pramadiyanti S, Jonatan S. 2023. Molecular characterization of red ginger varieties (Zingiber officinale Roxb. var. rubrum) by DNA markers. Biodiversitas 24: 6905-6913. Red ginger (Zingiber officinale Roxb. var. rubrum) rhizomes have been traditionally used in Indonesia to increase human health. Therefore, the assurance of raw material quality holds significant importance in the industrial application of red ginger. The primary step in ensuring the quality, safety, and effectiveness of the final product in the industrial environment is the identification of plants used as raw materials for supplements and herbal medications. DNA-based molecular characterization approaches have been specifically developed to aid in the identification of herbal raw materials prior to their application in production processes. Based on the findings derived from the conducted research, the Random Amplified Polymorphic DNA (RAPD) technique can be employed to amplify the DNA isolates of red ginger rhizome samples. The results obtained from the PCR-RAPD amplification of DNA samples extracted from red ginger plants indicated that 90.48% of the total 147 bands seen exhibited polymorphism. The findings suggest a tendency for DNA banding patterns derived from red ginger samples planted in the same geographic area to display clustering tendencies.