A panel of microsatellites to individually identify leopards and its application to leopard monitoring in human dominated landscapes

<p>Abstract</p> <p>Background</p> <p>Leopards are the most widely distributed of the large cats, ranging from Africa to the Russian Far East. Because of habitat fragmentation, high human population densities and the inherent adaptability of this species, they now occupy landscapes close to human settlements. As a result, they are the most common species involved in human wildlife conflict in India, necessitating their monitoring. However, their elusive nature makes such monitoring difficult. Recent advances in DNA methods along with non-invasive sampling techniques can be used to monitor populations and individuals across large landscapes including human dominated ones. In this paper, we describe a DNA-based method for leopard individual identification where we used fecal DNA samples to obtain genetic material. Further, we apply our methods to non-invasive samples collected in a human-dominated landscape to estimate the minimum number of leopards in this human-leopard conflict area in Western India.</p> <p>Results</p> <p>In this study, 25 of the 29 tested cross-specific microsatellite markers showed positive amplification in 37 wild-caught leopards. These loci revealed varied levels of polymorphism (four-12 alleles) and heterozygosity (0.05-0.79). Combining data on amplification success (including non-invasive samples) and locus specific polymorphisms, we showed that eight loci provide a sibling probability of identity of 0.0005, suggesting that this panel can be used to discriminate individuals in the wild. When this microsatellite panel was applied to fecal samples collected from a human-dominated landscape, we identified 7 individuals, with a sibling probability of identity of 0.001. Amplification success of field collected scats was up to 72%, and genotype error ranged from 0-7.4%.</p> <p>Conclusion</p> <p>Our results demonstrated that the selected panel of eight microsatellite loci can conclusively identify leopards from various kinds of biological samples. Our methods can be used to monitor leopards over small and large landscapes to assess population trends, as well as could be tested for population assignment in forensic applications.</p

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Not AvailableElicitors released by Trichoderma upon colonizing the root system is known to confer induced systemic resistance (ISR) in plants through altered transcriptome and proteome of the host plants thus enabling them to fight against the diseases. Seedling blight disease, caused by Phytophthora parasitica var nicotiana, and wilt disease, caused by Fusarium oxysporum f.sp ricini (F. o. r), are among the major diseases of castor which cause yield reduction up to 80%. Efficacy of Trichoderma in inducing systemic resistance against these diseases was studied using different strains of Trichoderma. Colonization of castor roots by Trichoderma induced resistance against both the diseases. When compared with untreated seedlings seedling blight disease severity was reduced by 85.7% in Trichoderma (Th4d strain)-treated seedlings and similar results were obtained with respect to wilt disease. ISR by Trichoderma was confirmed by studying the expression pattern of a few signature genes known to be up-regulated during ISR. To further understand the differentially expressed genes during ISR, we have adopted RNA sequencing technique in a time series experiment and the results have shown global changes in the gene expression pattern. ISR is being further characterized by comparing proteome and metabolomes in Trichoderma treated and untreated castor seedlings.Not Availabl

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Not AvailableEarly events of root colonization in castor by the biocontrol agent TrichodermaNot Availabl

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Not AvailableTriacylglycerol (TAG) biosynthesis in plants is complex and involves several genes with specific roles in the Kennedy pathway. Analysis of the evolutionary pattern and diversity of these genes can help to improve understanding of TAG biosynthesis in oilseed crops. In this study, an attempt was made to explore the diversity of genes: DGAT1, DGAT2, GPAT9 and LPAT2 across 13 oilseed crops using the sequence information ofthe model species, Arabidopsis thaliana. A total of 213 protein sequences corresponding to these genes were retrieved from the NCBI database by BLAST, multiple sequence alignment was performed and a phylogenetic tree was constructed. DGAT1, DGAT2 and GPAT9 sequences produced distinct species-wise clusters with several distinct sub-clusters, indicatingmonophyletic and highlydivergent nature with specialized rolesin differentspecies. LPAT2 sequences did not produce species-wise distinct clusters, indicating their polyphyletic nature. Diverse candidate gene sequences and phylogenetic relationships presented in this study would help to study TAG biosynthesis through genome-wide analysis in oilseed crops.Not Availabl

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Not AvailableColonization of castor roots by Trichoderma induced resistance to seedling blight disease. When compared with untreated seedlings, Phytophthora disease severity was reduced by 93% in Trichoderma (Th4d strain)–treated ones. Induced systemic resistance (ISR) by Trichoderma was confirmed by the expression profiling of a few signature genes known to be up-regulated during ISR.NAS

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Not AvailableThe colonization of castor roots by the biocontrol agent Trichoderma has been studied in a time series experiment. The results showed penetration and colonization of castor roots by Trichoderma within 48 hours post-inoculation. Microscopy studies established that the colonization was restricted to cortical cells of the roots and during this interaction, there were architectural changes in the cell wall of the host cells.NAS

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Not AvailableThe main impediment in utilizing the de-oiled meal of castor as animal feed is the presence of two toxic proteins, ricin (also known as RCA II and RCA60) and RCA (Ricinus communis agglutinin, otherwise known as RCA I and RCA120) found in the endosperm of mature castor seeds. Their presence compromises with the high protein (around 25% with balanced amino acid composition) content in the left-over oilcake, which could otherwise find use as an excellent animal feed. This review is an attempt at providing insights into the biology, biogenesis of ricin and RCA and various approaches for reducing/eliminating ricin and RCA in castor.Not Availabl

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Not AvailableHost plant resistance is an effective means of managing aphid pests in safflower but its genetic basis is not known. In this study, F1 and F6- recombinant inbred lines (RILs) produced from the CO-1 × EC-523368-2 cross were phenotyped for tolerance to the aphid, Uroleucon compositae based on days-to-wilt (DW) after infestation. The F1 plants showed dominance effect and RILs exhibited quantitative variation. The RIL population was genotyped with 242 simple sequence repeat (SSR) markers, which included a set of 150 new SSRs designed through next generation sequencing and bioinformatics approaches. Two quantitative trait loci (QTLs), QUc-Ct3.1 and QUc-Ct5.1, putatively associated with DW after aphid infestation were detected. QUc-Ct3.1 was a major QTL located on linkage group (LG) 3 with the closest marker SafM-290 (LOD = 18.3), which explained 31.5% of the phenotypic variation. QUc-Ct5.1 was a minor QTL located on LG-5 with the closest marker CtDES-237 (LOD = 7.0), which explained 9.1% of the phenotypic variation. This is the first report and a significant lead towards discovering genes for tolerance to aphids in safflower.SERB, Government of India through National Post Doctoral Fellowship (PDF/2016/003679

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Not AvailableSafflower is grown primarily for edible oil rich in unsaturated fatty acids. Due to higher stability and longer shelf life, the oil with high oleic acid content is preferred for food and other industrial applications. Safflower cultivars with high oleic acid content (>70%) have been developed and commercially successful. Incorporation of the high oleic trait through conventional breeding techniques has been a slow process due to recessive inheritance and difficulties associated with phenotyping by biochemical methods. DNA based marker-assisted selection (MAS) for high oleic trait would help speed up the breeding efforts in safflower. In this study, a set of high oleic varieties were found to carry the same mutation in the fatty acid desaturase 2-1 gene, CtFAD2-1, which is presumed to be the ‘ol’ allele associated with high oleic acid content in safflower. Genotypic assays namely Kompetitive Allele Specific PCR (KASP®) and the Amplifluor™ SNPs Genotyping System (Amplifluor®) were designed for the prediction of high oleic trait based on the mutation in the CtFAD2-1. The assays were thoroughly validated in segregating populations derived from crosses between low and high oleic parents. Through marker-assisted backcrossing scheme, the high oleic allele, ‘ol’ from the exotic variety, Montola-2000 was incorporated into the background of popular Indian linoleic type variety, Bhima and a set of promising high oleic lines (75.2%– 81.8%) were developed. The MAS-derived lines showed consistent expression of high oleic acid content over seasons and comparable seed/oil yield performance with the local check varieties. The genotypic assays reported in this study were robust, non-destructive, co-dominant and accurately predicted high oleic trait in segregating populations; thus, recommended for fast-track breeding of high oleic cultivars in safflowerNot Availabl