Study of interspecific SSR polymorphism among 14 species from Triticum-Aegilops group

In the present study, using in-gel hybridization and PCR based approaches, interspecific SSR polymorphism was studied among 14 species of Triticum-Aegilops group. The material represented seven different genomes and three ploidy levels (2x,4x,6x). In-gel hybridization involved 13 probe-enzyme combinations (four SSR oligonucleotide probes in combination with 2-4 enzymes) and resolved 5 to 20 bands (0.40kb to > 23kb) in each of the 14 individual species. This suggested ubiquitous distribution and interspecific polymorphism of SSRs among different species of Triticum-Aegilops group. The available polymorphism also proved helpful in discriminating not only the species with different ploidy levels and possessing different genomes, but also those possessing similar or very closely related genomes. The amplification of SSR loci using 15 primer pairs derived from hexaploid wheat was also carried out in all the 14 species. The primer pairs, each amplified SSR loci not only in species containing A, B and D genomes, but also in 2 to 10 of the remaining species that contained other genomes. This suggested that wheat SSRS might have been derived from the corresponding SSRs in an ancestral genome and are conserved across a number of species in the Triticum-Aegilops group. Also, two pairs of SSRs (one consisting of WMC243 and WMC415 and the other consisting of WMC35 and WMC404) each discriminated all the 14 species examined during the present study. Therefore, one can infer from the present study that SSR primers can be used in studies on DNA polymorphism, genetic diversity, gene mapping and synteny conservation across different species of Triticum-Aegilops group

Development and characterization of large-scale simple sequence repeats in jute

Jute is an important crop of the Indian subcontinent and comprises tossa jute (Corchorus olitorius) and white jute (C. capsularis). The yield and fiber quality of this crop remained stagnant for many years and could not be improved through conventional plant breeding. Also, no effort has been made to develop molecular markers on a scale required for marker-assisted selection (MAS) to supplement conventional plant breeding. As a first step toward deploying MAS for jute improvement, 2469 simple sequence repeats (SSRs) were developed in tossa jute (JRO 524) using four SSR-enriched genomic libraries. A random subset of 100 SSRs (25 SSRs from each library) was used to detect polymorphism between the parental genotypes of each of the two recombinant inbred line (RIL) mapping populations. The RILs are being developed from JRO 524 × PPO4 (for fiber fineness) and JRC 321 × CMU 010 (for lignin content) crosses to prepare molecular maps and conduct quantitative trait loci (QTL) analyses. Both SSR length polymorphism and ± polymorphism (null alleles, i.e., presence and absence of specific SSR) were detected; 50 SSRs detected polymorphism between the two genotypes of tossa jute, whereas 45 SSRs detected polymorphism between the two genotypes of white jute. This SSR allelic polymorphism in jute is higher than that reported in other crops and is adequate for construction of genetic maps for QTL analysis. The large-scale SSRs will also prove useful in studying genetic diversity, population structure, and association mapping

Genetic dissection of grain weight in bread wheat through quantitative trait locus interval and association mapping

Genetic dissection of grain weight in bread wheat was undertaken through both genome-wide quantitative trait locus (QTL) interval mapping and association mapping. QTL interval mapping involved preparation of a framework linkage map consisting of 294 loci {194 simple sequence repeats (SSRs), 86 amplified fragment length polymorphisms (AFLPs) and 14 selective amplifications of microsatellite polymorphic loci (SAMPL)} using a bi-parental recombinant inbred line (RIL) mapping population derived from Rye Selection111 × Chinese Spring. Using the genotypic data and phenotypic data on grain weight (GW) of RILs collected over six environments, genome-wide single locus QTL analysis was conducted to identify main effect QTL. This led to identification of as many as ten QTL including four major QTL (three QTL were stable), each contributing >20% phenotypic variation (PV) for GW. The above study was supplemented with association mapping, which allowed identification of 11 new markers in the genomic regions that were not reported earlier to harbour any QTL for GW. It also allowed identification of closely linked markers for six known QTL, and validation of eight QTL reported earlier. The QTL identified through QTL interval mapping and association mapping may prove useful in marker-assisted selection (MAS) for the development of cultivars with high GW in bread whea

Development of SSR markers and construction of a linkage map in jute

Jute is an important natural fibre crop, which is only second to cotton in its importance at the global level. It is mostly grown in Indian subcontinent and has been recently used for the development of genomics resources. We recently initiated a programme to develop simple sequence repeat markers and reported a set of 2469 SSR that were developed using four SSR-enriched libraries (Mir et al. 2009). In this communication, we report an additional set of 607 novel SSR in 393 SSR containing sequences. However, primers could be designed for only 417 potentially useful SSR. Polymorphism survey was carried out for 374 primer pairs using two parental genotypes (JRO 524 and PPO4) of a mapping population developed for fibre fineness; only 66 SSR were polymorphic. Owing to a low level of polymorphism between the parental genotypes and a high degree of segregation distortion in recombinant inbred lines, genotypic data of only 53 polymorphic SSR on the mapping population consisting of 120 RIL could be used for the construction of a linkage map; 36 SSR loci were mapped on six linkage groups that covered a total genetic distance of 784.3 cM. Hopefully, this map will be enriched with more SSR loci in future and will prove useful for identification of quantitative trait loci/genes for molecular breeding involving improvement of fibre fineness and other related traits in jute

Transcriptional Upregulation of NLRC5 by Radiation Drives STING- and Interferon-Independent MHC-I Expression on Cancer Cells and T Cell Cytotoxicity.

Radiation therapy has been shown to enhance the efficacy of various T cell-targeted immunotherapies that improve antigen-specific T cell expansion, T regulatory cell depletion, or effector T cell function. Additionally, radiation therapy has been proposed as a means to recruit T cells to the treatment site and modulate cancer cells as effector T cell targets. The significance of these features remains unclear. We set out to determine, in checkpoint inhibitor resistant models, which components of radiation are primarily responsible for overcoming this resistance. In order to model the vaccination effect of radiation, we used a Listeria monocytogenes based vaccine to generate a large population of tumor antigen specific T cells but found that the presence of cells with cytotoxic capacity was unable to replicate the efficacy of radiation with combination checkpoint blockade. Instead, we demonstrated that a major role of radiation was to increase the susceptibility of surviving cancer cells to CD8+ T cell-mediated control through enhanced MHC-I expression. We observed a novel mechanism of genetic induction of MHC-I in cancer cells through upregulation of the MHC-I transactivator NLRC5. These data support the critical role of local modulation of tumors by radiation to improve tumor control with combination immunotherapy

T cell cytolytic capacity is independent of initial stimulation strength.

How cells respond to myriad stimuli with finite signaling machinery is central to immunology. In naive T cells, the inherent effect of ligand strength on activation pathways and endpoints has remained controversial, confounded by environmental fluctuations and intercellular variability within populations. Here we studied how ligand potency affected the activation of CD8+ T cells in vitro, through the use of genome-wide RNA, multi-dimensional protein and functional measurements in single cells. Our data revealed that strong ligands drove more efficient and uniform activation than did weak ligands, but all activated cells were fully cytolytic. Notably, activation followed the same transcriptional pathways regardless of ligand potency. Thus, stimulation strength did not intrinsically dictate the T cell-activation route or phenotype; instead, it controlled how rapidly and simultaneously the cells initiated activation, allowing limited machinery to elicit wide-ranging responses