DNA Molecular Markers Research Articles

The application of DNA molecular markers in the study of Codonopsis species genetic variation, a review

Codonopsis genus is comprised of species that are perennial plants primarily distributed across all east, southeast, and Central Asia. The most famous species of Codonopsis are C. tangshen, C. lanceolate, and C. pilosula. The records showed that they have a long story usage as traditional Chinese medicines, as they were alleged to be able to intensify the spleen and the lung as well as enriching blood and engendering liquid. Certain species have a culinary value in southern China and Southeast Asia, where they are considered as tea, wine, soup, plaster, and porridge. Codonopsis species were shown to be of great importance in medicine, due to their broad biological activity. Therefore, a clear understanding of their genetic diversity is needed. Adequate distinctions and descriptions of those species are necessary to preserve plant reservoir, investigations of genes associated with desirable traits, and understanding of evolutionary relationships. Subsequently, various molecular marker techniques such as Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP), Simple Sequence Repeats (SSR), and Inter Simple Sequence Repeat (ISSR), Single Nucleotide Polymorphism (SNP), internal transcribed spacer (ITS), and Sequence-Characterized Amplified Region (SCAR) have been improved to provide detailed informations about genomes, that historically were not possible to obtain based on only phenotypic methods. This review represents the usage of DNA molecular markers for molecular diversity analysis of medically important species belonging to the genus Codonopsis.

Production of allohexaploid Brassica hybrid between tuber mustard (Brassica juncea L. var. crassicaulis Chen &Yang) and Chinese kale (Brassica oleracea var. alboglabra Bailey)

Brassica is a key agricultural genus, containing oilseed and vegetable crops, which includes three major genomes (A, B and C), distributed in diploid (genomes: AA, BB and CC) and allotetraploid species (genomes: AABB, AACC and BBCC). Interspecific hybridization between Brassica species is an important way to combine valuable characters from the different species. In the present study, distant hybridization between inbred lines of tuber mustard (Brassica juncea var. crassicaulis Chen &Yang; AABB) and Chinese kale (Brassica oleracea var. alboglabra Bailey; CC) was conducted and allotriploid interspecific hybrids were identified. The hybrid seed set was improved by gibberellin treatment of the stigmas prior to pollination. Eight hybrid seedlings were obtained by bud pollination and ovary culture, and all seedlings were confirmed to be true hybrids by phenotypic analysis, cytological observation (27 chromosomes) and the use of parent-specific randomly amplified polymorphic DNA (RAPD) molecular markers. After tissue culture and colchicine-induced chromosome-doubling treatment, allohexaploid hybrids (AABBCC) were isolated and identified from the eight hybrids. The chromosomes of five plants were completely doubled, with a doubling rate of 5.68 %, and the number of chimeric plants was 24, at a rate of 27.27 %, when roots were immersed in 0.1 % colchicine solution. In comparison with the allotriploid hybrid, the synthetic hexaploid hybrids showed unique phenotypes with normal anthers, well-developed pods, larger flowers and petals, thicker stems than either parent and other phenotype types intermediate between the two parents. In the present study, synthetic tri-genomic Brassica vegetable germplasm, combining the A, B and C genomes, was created for use in vegetable breeding, as well as for a basis for further investigation of the mechanisms of Brassica evolution and adaptation. The current study is of great significance for the use of these allohexaploid plants to explore the heterotic effects of distant hybridization and for genetic improvement.

Genetic stability analysis using DNA barcoding and molecular markers and foliar micro-morphological analysis of in vitro regenerated and in vivo grown plants of Artemisia vulgaris L.

The current study provides an enhanced and reproducible micropropagation system in Artemisia vulgaris L. using nodal explants of mature plants. The explants were cultured on basal Murashige and Skoog’s (MS) medium augmented with 6-benzylaminopurine (BAP) and Kinetin (Kin) concentrations individually. Direct regeneration of shoots (7.7 ± 0.36 shoots per explant) from nodal meristems was attained on MS medium combined with 1.5 mg L−1 BAP alone. Sub-culturing of these cultures on MS medium supplemented with 0.5 mg L−1 BAP and 0.5 mg L−1 inddole-3 acetic acid (IAA) helped in further multiplication of shoots (75.8 ± 1.00 shoots per culture vessel). Elongated shoots were transferred to half strength MS medium contained 1.0 mg L−1 IBA for roots formation. Rooting was also stimulated from the sliced ends of the shoots via ex vitro rooting method by pulse treating the shoots with 300 mg L−1 IBA for 5 min. Well rooted plants were shifted into small paper cups having manure: sand: soil in 1: 1: 1 proportion for hardening. The acclimatized plants were shifted to the natural field conditions for complete plant growth. Start codon targeted (SCoT), inter simple sequence repeats (ISSR) markers and universal DNA barcoding (rbcLa gene primers) were applied to analyze the genetic stability of micropropagated plants with the mother plant. The banding patterns of regenerants were found to be monomorphic in nature, representing the similarity with the mother plant and no somaclonal variations were observed. Similarly, foliar micro-morphological characters like stomata, trichome and vein pattern of leaves were studied from in vitro and acclimatized plants to focus on the developmental adaptation of micropropagated plantlets towards survival in field conditions. This study explores the highly reproducible procedure for A. vulgaris through nodal propagation without any somaclonal variations. It is the first report on foliar micro-morphological analysis at subsequent stages of in vitro propagation and genetic homogeneity of micropropagated plants with the mother plant. The current study paves the way for the development of genetically uniform A. vulgaris plants regenerated from nodal shoot segments and successful survival in field.

Detection of a 4 bp Mutation in the 3'UTR Region of Goat Sox9 Gene and Its Effect on the Growth Traits.

Simple SummaryThe sex determining region Y (SRY)-type high mobility group (HMG) box 9 (Sox9) gene is critically important in the formation and development of cartilage and is considered the “main regulator” of chondrogenesis. Additionally, a large number of studies have shown that mutations in a single allele of human Sox9 can lead to campomelic dysplasia syndrome. Therefore, the mutations of Sox9 have been the subject of increasing interest among researchers. However, no studies to date have examined the association between Sox9 gene variants and growth traits in goats. Here, we detected a 4 bp indel in the 3′Untranslated Regions (3′UTR) region of Sox9 in Shaanbei white cashmere (SBWC) goats (n = 1109) and studied the association between this indel and growth traits. The 4 bp indel of Sox9 was significantly associated with body length, heart girth, hip width, and all body measurement indexes (p < 0.05) in SBWC goats. Thus, this deletion could be used as an effective molecular marker for maximizing the growth traits of goats in breeding programs.The SRY-type HMG box 9 (Sox9) gene plays an important role in chondrocyte development as well as changes in hypertrophic chondrocytes, indicating that Sox9 can regulate growth in animals. However, no studies to date have examined the correlation between variations in Sox9 and growth traits in goats. Here, we found a 4 bp indel in the 3′UTR of Sox9 and verified its association with growth traits in Shaanbei white cashmere goats (n = 1109). The frequencies of two genotypes (ID and II) were 0.397 and 0.603, respectively, and polymorphic information content (PIC) values showed that the indel had a medium PIC (PIC > 0.25). The 4 bp indel was significantly correlated with body length (p = 0.006), heart girth (p = 0.001), and hip width (p = 4.37 × 10 −4). Notably, individuals with the ID genotype had significantly superior phenotypic traits compared with individuals bearing the II genotype. Hence, we speculated that the 4 bp indel is an important mutation affecting growth traits in goat, and may serve as an effective DNA molecular marker for marker-assisted selection in goat breeding programs.

Evolutionary Trends in the Mitochondrial Genome of Archaeplastida: How Does the GC Bias Affect the Transition from Water to Land?

Among the most intriguing mysteries in the evolutionary biology of photosynthetic organisms are the genesis and consequences of the dramatic increase in the mitochondrial and nuclear genome sizes, together with the concomitant evolution of the three genetic compartments, particularly during the transition from water to land. To clarify the evolutionary trends in the mitochondrial genome of Archaeplastida, we analyzed the sequences from 37 complete genomes. Therefore, we utilized mitochondrial, plastidial and nuclear ribosomal DNA molecular markers on 100 species of Streptophyta for each subunit. Hierarchical models of sequence evolution were fitted to test the heterogeneity in the base composition. The best resulting phylogenies were used for reconstructing the ancestral Guanine-Cytosine (GC) content and equilibrium GC frequency (GC*) using non-homogeneous and non-stationary models fitted with a maximum likelihood approach. The mitochondrial genome length was strongly related to repetitive sequences across Archaeplastida evolution; however, the length seemed not to be linked to the other studied variables, as different lineages showed diverse evolutionary patterns. In contrast, Streptophyta exhibited a powerful positive relationship between the GC content, non-coding DNA, and repetitive sequences, while the evolution of Chlorophyta reflected a strong positive linear relationship between the genome length and the number of genes.

Influence of different cytokinins on micropropagation of an important medicinal plant, Solanum erianthum D. Don, and assessment of the genetic fidelity of the regenerants

The present study describes the first successful report on an efficient in vitro protocol for direct and indirect organogenesis of an important medicinal and anticancerous plant, Solanum erianthum D. Don. An optimal response using leaf explants was achieved in half-strength Murashige and Skoog (MS)-medium supplemented with 5 mg L−1 naphthalene acetic acid for friable callus formation and 2 mg L−1 naphthaleneacetic acid and 0.5 mg L−1 6-benzylaminopurine amino purine for induction of compact callus. The maximum number of shoot buds (20.33 ± 11.55) was obtained from compact callus using indirect shoot organogenesis in MS medium supplemented with 0.5 mg L−1 thidiazuron among the four cytokinins tested, which included 6-benzyl amino purine, kinetin, 6-γ,γ-dimethylallylamino purine, and thidiazuron. Supplements of 3 mg L−1 6-benzyl amino purine played a crucial role in direct shoot bud formation from a leaf. The buds appeared as small protuberances, and finally developed into leafy shoots as the culture period progressed. Regenerated shoot buds were transferred to MS media that contained four cytokinins for shoot tip culture, in which 8 mg L−1 6-γ,γ-dimethylallylamino purine exhibited the best response (15 ± 2.52). Although efficient plant regeneration was achieved in the presence of all four tested cytokinins in the three organogenetic pathways, distinct differences exist in terms of shoot regeneration potential. Half-strength MS basal medium is recommended for root induction. An approximate 70% survival rate of regenerated plantlets was recorded following sequential transfer from culture conditions to the field. The genetic fidelity of the regenerated plants was evaluated using random amplified polymorphic DNA molecular markers, which showed a high number of uniform monomorphic bands.

A DNA toolbox for non-invasive genetic studies of sambar deer (Rusa unicolor)

Invasive sambar deer (Rusa unicolor) are having significant detrimental impacts on natural environments in south-eastern Australia. Little, however, is known about their ecology, limiting evidence-based management strategies directed at reducing deer impacts. Genetic data, generated from DNA isolated from deer scats, can be used to fill ecological knowledge gaps. This study outlines a non-invasive genetic sampling strategy by which good-quality DNA from a single deer scat can be used to determine (1) species of origin, (2) sex and (3) a unique DNA profile. DNA from deer tissue and sambar deer scat samples were used to develop and optimise molecular methods to collect reliable genetic information. A DNA toolbox is presented that describes how to find, collect and store scat samples, isolate DNA and use molecular markers to generate informative genetic data. Generating genetic data using this approach will support studies aimed at acquiring ecological knowledge about sambar deer. Such knowledge will be critical for developing evidence-based recommendations to improve on-ground management decisions for sambar deer.

Economic Evaluation of Early Sex Determination for Farmed Russian Sturgeon: The Case of Northern Israel

Sturgeon farming has high potential economic value, but the industry is characterized by late fish maturity, leading to a long payback period for sturgeon farms. Early sex determination (ESD) of fish is important for increasing the profitability of sturgeon farms due to early male separation, thus enabling growing more females and producing more caviar for the same farm area. The data for Russian sturgeons farmed in northern Israel and the results of molecular markers developed for ESD are used in this study. Two sturgeon sex determination technologies are examined and compared from an economic point of view: 1) using molecular DNA markers for ESD at the age of 0.5 years; and 2) using endoscopic methods at the age of 3.5 years, as is currently done in many sturgeon farms. For this purpose, a bio-economic model of a sturgeon farm was developed and an economic evaluation of a sturgeon farm was made to compare both technologies. The ESD technology results in a significant improvement in the economic performance of sturgeon farms. This improvement is greater when caviar prices are lower.

SENSITIVITY AND SPECIFICITY OF CONVENTIONAL CULTURE, LIGHT MICROSCOPY, SEROLOGICAL AND MOLECULAR METHODS FOR IDENTIFYING Mycobacterium tuberculosis COMPLEX

In this study, some different tools, i.e., conventional culture, light microscopy, serological and molecular methods were evaluated for their sensitivities to detect the Mycobacterium tuberculosis complex in forty specimens from animal and human sources. To achieve such goal 20 samples from each of raw milk, animal tissues, and human CSF were collected. Enzyme-linked immunosorbent assay (ELISA) was compared to conventional culture, light microscopy for its sensitivity and specificity to detect the presence of TB pathogen in these samples. Results showed that sensitivity and specificity of 10% &100% for IS6110-targed PCR, 30 and 90% for ELISA, 16.25% and 100% for conventional culturing and 17.5% and 87.0% for light microscopy were recorded. In other mean, the IS6110-targed PCR was relatively more useful in TB diagnosis followed by ELISA. Furthermore, the DNA fingerprinting of three standards TB strains using random amplified polymorphic of DNA-PCR (RAPD-PCR) was carried out in the hope of generating some DNA molecular marker for TB identification. RAPD-PCR finding showed some molecular DNA markers that could be very useful in the identification of tuberculosis strains.

ИЗУЧЕНИЕ ГЕНЕТИЧЕСКОГО РАЗНООБРАЗИЯ КОРМОВЫХ КУЛЬТУР С ПОМОЩЬЮ МОЛЕКУЛЯРНЫХ ДНК-МАРКЕРОВ

Genetic diversity is the precondition for any selection program. The collection and exploitation of natural variation from ecotypes and landraces has played a vital role in the improvement of forage crops. The review is devoted to the most important aspects of studying the genetic variation in populations, cultivars, samples and forms of wild and cultivated forage plants. The factors with negative impact on the biodiversity conservation have been determined. The main types of genetic markers that used for genetic recourses of perennial grasses evaluation were described. Particular attention was focused on the role of molecular DNA markers for the population genetics and phylogenetic studies. The main advantages of DNA markers application for the forage crops, due of its great variability of traits and properties, the complexity of the genetic system and a high degree of plasticity of this group of plants, have been discussed. The latest generation of genetic DNA markers allows conducting the objective and accurate assessment of genetic diversity, provides selection process intensification, increases the possibilities for identification and molecular-genetic certification of the selection achievements.

Evaluation of RAPD markers for molecular identification of five bamboo genera from Indonesia

Abstract Conservation of bamboos for future exploitation as fuel, fibre and as an ingredient for cosmetics depends on knowledge of its natural genetic variation. The study of molecular genetic diversity in bamboos will provide important information for its conservation. This article reports on the genetic diversity in 25 species representing five genera of bamboos found in Indonesia using Random Amplified Polymorphic DNA (RAPD) molecular markers. Out of 40 primers, 24 primers produced 1107 total bands and 86.21% of polymorphic bands across the 25 species. Sixteen bands were uniquely found in one species only and their presence or absence helped to define nine bamboo species. RAPD band sizes ranged from 162 to 2247 base pairs. A dendrogram based on the similarity coefficient of Dice divided the bamboo species into three big clusters. In conclusion, RAPD can capture the diversity among five different bamboo genera and has a great potential to be used in the study of genetic diversity in Indonesian bamboos.

Transferability of Some Nuclear Microsatellite (SSRs) Markers from Related Species in Parkia biglobosa

Molecular DNA markers, particularly microsatellites (SSRs), are important tools for plant genetic resources characterization. The present study examined the transferability of nine polymorphic microsatellite markers developed for Parkia panurensis in Parkia biglobosa from Northern Benin, favorite area of Parkia biglobosa. Fourty (40) accessions have been considered. Five microsatellite loci (Parpan 3, Parpan 4, Parpan 9, Parpan 13 and Parpan 15) showed good amplifications in Parkia biglobosa. The size of the amplified markers, ranging from 100 to 200 bases pairs, was similar to those previously reported. Forest tree species are too numerous to benefit all a significant investment in molecular biology. Also, the present study has shown that the transfer of markers between species is possible. The transferability is possible for the species Parkia biglobosa, too. The microsatellites identified in this study are important to analyse the genetic structure and diversity of natural populations of Parkia biglobosa in Benin particularly and in the world generally. They are also important to analyse the evolution of species with phylogeny construction, to analyse the genome with its applications in genetic improvement and genetic identification of given material. For these different studies, the microsatellites of the present study could be completed to those defined for Parkia biglobosa.

A 21-bp indel within the LLGL1 gene is significantly associated with litter size in goat

The scribble cell polarity complex component (LLGL1) is part of the cytoskeletal network and is involved in maintaining cell polarity and epithelial integrity. Based on the whole-genome sequencing analysis in goat, LLGL1 gene is suggested as a putative important candidate gene affecting litter size in Shaanbei White Cashmere Goats (SBWC). Therefore, the objective of this study was to uncover the possible novel insertion/deletion (Indel) variant in goat LLGL1 gene and to evaluate its association with litter size of SBWC (n = 827). Using the PCR detection and DNA sequencing, the 21-bp indel in the upstream of LLGL1 was firstly founded and two genotypes were identified: II (insertion/insertion) and ID (insertion/deletion), respectively. Association analyses revealed that the 21-bp indel was significantly correlated with litter size (p = 0.017). Notably, the individuals with II genotype were significantly greater than that of the genotype ID, and the ‘I’ allele was dominant. Additionally, the remarkable influence of the indel on traits might be related to the change of DEAF-1-related (NUDR) binding site through bioinformatics analysis. Briefly, the 21-bp indel within the goat LLGL1 gene could be an effective DNA molecular marker and provide valuable theoretical basis for marker-assisted selection (MAS) in goat industry.

Review on medicinal importance of Vigna genus

Globally, there are more than 500,000 plant species (green scum, duckweed, lichens, liverworts, fungi, ferns, conifers, mosses and flowering plants etc.) that maintain earth’s environmental equilibrium, ecosystem stability and also possess vast endemic, aesthetic and cultural importance, provide medicine, food, fuel, shelter and clothing. Plants are used as therapeutic agents to improve health by a large part of population. Several clinical facts suggest that plant derived foods hold various potential health benefits and well known as neutraceuticals. These are the products that are used as food or as a part of food (foodstuffs), able to cure and prevent diseases in addition to their basic nutritional value. More than 200,000 chemical compounds are synthesized by plants and no doubt, also possess medicinal importance. Worldwide, about 70% plant based preparations are used as traditional medicines and also facilitate the base for 50 percent of prescription and/or over the counter drugs used in the Western-type practice of medicine. For underdeveloped and developing countries, it is a need to provide safe, efficient and cheap medications. In various part of India medicinal plants are widely distributed and always have increasing demand due to their medicinal properties. The present review is focused on the genus Vigna which is widely cultivated and used as neutraceuticals. They grow in varied climatic zones, in high temperatures, low rainfall and poor soils with low input in form of fertiliser and irrigation that make them valuable crop plants. As Vigna is an important genus that fulfils the food demand, useful in cosmetics and medicines, there is scope to enhance its productivity via resource conservation, optimum use of rainwater, bridging the yield gaps and innovations in technology transfer and up scaling. One of the important steps to find out a way to increase the production is the detection and analysis of naturally occurring DNA sequence variation by using DNA markers or molecular markers as these markers are indispensable tool that construct maps of genetic linkage and mark the agronomically important traits.

The influential role of polyamines on the in vitro regeneration of pea (Pisum sativum L.) and genetic fidelity assessment by SCoT and RAPD markers

A proficient and reliable in vitro plant regeneration protocol was established for pea by utilizing cotyledonary node as an explants, which excised from 3-days old imbibed seeds. The present research work explains the positive role of polyamines (PAs) accompanied by plant growth regulators (PGRs) on the multiple shoot induction and rooting in pea plant regeneration. The highest multiple shoots (65.1 shoots/explant) was attained from the cotyledonary node explant on Murashige and Skoog (MS) medium accompanied with 20 mg l−1 spermidine (SPD) along with 1.5 mg l−1 6-benzyladenine (BA). Moreover, the highest elongation of multiplied shoots (10 cm length/shoot) was noted on MS medium enriched with 1 mg l−1 of gibberellic acid (GA3). The elongated shoots produced the highest number of roots (33.66 roots/shoot) on MS medium augmented with 30 mg l−1 putrescine (PUT) along with 0.6 mg l−1 1-naphthaleneacetic acid (NAA). Rooted plants were hardened and acclimatized in the greenhouse with an existence rate of 98%. The standardized procedure with the use of PAs has enhanced the multiple shoot induction to threefold higher than the plants raised from PGRs treatments. The regenerated pea plants revealed the fivefold enhanced photosynthetic pigment, twofold enhanced antioxidant profile and the substantial growth in chloroplast count have been achieved in optimized SPD and BA supplemented MS medium compared to control plant. Start Codon Targeted polymorphism and Random Amplified Polymorphic DNA molecular markers recognized the genetic purity of the in vitro regenerated pea plants for their true type of nature. An enhanced and consistence in vitro regeneration protocol was established for pea, by the application of polyamines along with PGRs, which significantly improved the multiple shooting, shoot elongation, rooting, total chlorophyll and antioxidant profile of regenerated plants, further not hampering their genetic fidelity.

Application of DNA Molecular Markers in Plant Breeding (Review article)

Application of DNA Molecular Markers in Plant Breeding (Review article)

Identification of Corydalis yanhusuo,C. turtschaninovii,C. decumbens by allele-specific PCR

To establish a DNA molecular markers method for identification of Corydalis yanhusuo,C. turtschaninovii and C. decumbens,the mat K,trn G and psb A-trn H sequences of 56 samples from 14 species of C. yanhusuo,C. turtschaninovii,C. decumbens and their related species were obtained by sequencing. The SNP loci were obtained by Bio Edit 7. 2. 2 software. The primers for AS-PCR identification were designed based on the mutation sites,and the conditions of PCR were optimized to identify C. yanhusuo,C. turtschaninovii,and C. decumbens according to the specific bands. The results showed that the amount of template( 0. 6-1 200 ng)and annealing temperature( 42-60 ℃) had little influence on the amplification results,and the number of cycles had much influence on the amplification results. When the number of cycles was 20,the specific bands of 297 bp( mat K),353 bp( trn G) and 544 bp( mat K) were amplified from C. yanhusuo,C. turtschaninovii and C. decumbens,respectively. The method established in this study had a minimum detection limit of 6 ng for C. yanhusuo,60 ng for C. decumbens and less than 0. 6 ng for C. turtschaninovii. Thus,the allelespecific PCR method established in the research can specifically identify C. yanhusuo,C. turtschaninovii,and C. decumbens.

The study of inter-specific relationships of Bromus genus based on SCoT and ISSR molecular markers.

The genus of Bromus is one of the most important collection of rangeland plants, which are distributed in a wide range of natural areas of Iran. Interspecific relationships were evaluated in 90 accessions of 18 Bromus species based on 15 ISSR and 15 SCoT primers. SCoT markers separated the accessions better than ISSR marker. In addition, there was a high interspecific diversity between surveying germplasm. The sections of Bromus genus completely separated based on DNA molecular markers. SCoT markers could separate the accessions in each species. The primers of SC5 and SC35 from SCoT marker and UBC861, UBC857 and UBC844 primers from ISSR marker were identified as the best primers in revealing of genetic diversity between accessions. The sections of Ceratochloa, Genea, Pnigma and Bromus were monophyletic and were placed in one cluster. The section Bromus had a direct relationship with section Genea. In other words, section Ceratochloa has a direct relationship with Pnigma. B. tectorum and B. sericeus. B. sterilis had the most distance with other species in section Genea. B. squarrosus and B. japonicus had the most similarity and B. briziformis with B. danthoniae and B. scoparius with B. rechingeri had a moderate relationship in section Bromus. B. tomentosus and B. persicus had the highest similarity and B. riparius with B. biebersteinii and B. tomentellus with B. inermis had a moderate similarity in section Pnigma.

Molecular identification of water molds (oomycetes) associated with chum salmon eggs from hatcheries in Japan and possible sources of their infection

Oomycete infection of various freshwater animals, including salmonid eggs, causes significant economic damage to aquaculture worldwide. In this study, we detected oomycetes in infected chum salmon Oncorhynchus keta eggs at two hatcheries in northern Japan, in the source water used for egg incubation, and in the air at the hatcheries to clarify the source(s) of oomycete transmission using a DNA molecular marker. Seven oomycete taxa, belonging to Saprolegniaceae and Pythiaceae, were detected from the infected eggs. From the source water used for egg incubation and the air at the hatcheries, nine oomycete taxa, including those found on infected eggs, were detected, suggesting that both water and air are potential sources of oomycete transmission. There is no report of airborne transmission of these oomycetes detected in this study so far. Regarding protection and sterilization against oomycete infection in aquaculture hatcheries, not only water used at hatcheries but also the air in hatcheries may need to be considered hereafter.

A DNA toolbox for non-invasive genetic studies of sambar deer (Rusa unicolor)

Invasive sambar deer (Rusa unicolor) are having significant detrimental impacts on natural environments in south-eastern Australia. Little, however, is known about their ecology, limiting evidence-based management strategies directed at reducing deer impacts. Genetic data, generated from DNA isolated from deer scats, can be used to fill ecological knowledge gaps. This study outlines a non-invasive genetic sampling strategy by which good-quality DNA from a single deer scat can be used to determine (1) species of origin, (2) sex and (3) a unique DNA profile. DNA from deer tissue and sambar deer scat samples were used to develop and optimise molecular methods to collect reliable genetic information. A DNA toolbox is presented that describes how to find, collect and store scat samples, isolate DNA and use molecular markers to generate informative genetic data. Generating genetic data using this approach will support studies aimed at acquiring ecological knowledge about sambar deer. Such knowledge will be critical for developing evidence-based recommendations to improve on-ground management decisions for sambar deer.