High resolution mapping of a novel late blight resistance gene Rpi-avll, from the wild Bolivian species Solanum avilesii

Both Mexico and South America are rich in Solanum species that might be valuable sources of resistance (R) genes to late blight (Phytophthora infestans). Here, we focus on an R gene present in the diploid Bolivian species S. avilesii. The genotype carrying the R gene was resistant to eight out of 10 Phytophthora isolates of various provenances. The identification of a resistant phenotype and the generation of a segregating population allowed the mapping of a single dominant R gene, Rpi-avl1, which is located in an R gene cluster on chromosome 11. This R gene cluster is considered as an R gene “hot spot”, containing R genes to at least five different pathogens. High resolution mapping of the Rpi-avl1 gene revealed a marker co-segregating in 3890 F1 individuals, which may be used for marker assisted selection in breeding programs and for further cloning of Rpi-avl

Identifying Molecular Markers Suitable For Frl Selection in Tomato Breeding

Modern plant breeding heavily relies on the use of molecular markers. In recent years, next generation sequencing (NGS) emerged as a powerful technology to discover DNA sequence polymorphisms and generate molecular markers very rapidly and cost effectively, accelerating the plant breeding programmes. A single dominant locus, Frl, in tomato provides resistance to the fungal pathogen Fusarium oxysporum f. sp. radicis-lycopersici (FORL), causative agent of Fusarium crown and root rot. In this study, we describe the generation of molecular markers associated with the Frl locus. An F2 mapping population between an FORL resistant and a susceptible cultivar was generated. NGS technology was then used to sequence the genomes of a susceptible and a resistant parent as well the genomes of bulked resistant and susceptible F2 lines. We zoomed into the Frl locus and mapped the locus to a 900 kb interval on chromosome 9. Polymorphic single-nucleotide polymorphisms (SNPs) within the interval were identified and markers co-segregating with the resistant phenotype were generated. Some of these markers were tested successfully with commercial tomato varieties indicating that they can be used for marker-assisted selection in large-scale breeding programmes

Cloning and characterization of XiR1, a locus responsible for dagger nematode resistance in grape

The dagger nematode, Xiphinemaindex, feeds aggressively on grape roots and in the process, vectors grapevine fanleaf virus (GFLV) leading to the severe viral disease known as fanleaf degeneration. Resistance to X. index and GFLV has been the key objective of grape rootstock breeding programs. A previous study found that resistance to X. index derived from Vitis arizonica was largely controlled by a major quantitative trait locus, XiR1 (X. index Resistance 1), located on chromosome 19. The study presented here develops high-resolution genetic and physical maps in an effort to identify the XiR1 gene(s). The mapping was carried out with 1,375 genotypes in three populations derived from D8909-15, a resistant selection from a cross of V. rupestris A. de Serres (susceptible) × V. arizonica b42-26 (resistant). Resistance to X. index was evaluated on 99 informative recombinants that were identified by screening the three populations with two markers flanking the XiR1 locus. The high-resolution genetic map of XiR1 was primarily constructed with seven DNA markers developed in this study. Physical mapping of XiR1 was accomplished by screening three bacterial artificial chromosome (BAC) libraries constructed from D8909-15, V. vinifera Cabernet Sauvignon and V. arizonica b42-26. A total of 32 BAC clones were identified and the XiR1 locus was delineated within a 115 kb region. Sequence analysis of three BAC clones identified putative nucleotide binding/leucine-rich repeat (NB-LRR) genes. This is the first report of a closely linked major gene locus responsible for ectoparasitic nematode resistance. The markers developed from this study are being used to expedite the breeding of resistant grape rootstocks

A novel approach to locate Phytophthora infestans resistance genes on the potato genetic map

Mapping resistance genes is usually accomplished by phenotyping a segregating population for the resistance trait and genotyping it using a large number of markers. Most resistance genes are of the NBS-LRR type, of which an increasing number is sequenced. These genes and their analogs (RGAs) are often organized in clusters. Clusters tend to be rather homogenous, viz. containing genes that show high sequence similarity with each other. From many of these clusters the map position is known. In this study we present and test a novel method to quickly identify to which cluster a new resistance gene belongs and to produce markers that can be used for introgression breeding. We used NBS profiling to identify markers in bulked DNA samples prepared from resistant and susceptible genotypes of small segregating populations. Markers co-segregating with resistance can be tested on individual plants and directly used for breeding. To identify the resistance gene cluster a gene belongs to, the fragments were sequenced and the sequences analyzed using bioinformatics tools. Putative map positions arising from this analysis were validated using markers mapped in the segregating population. The versatility of the approach is demonstrated with a number of populations derived from wild Solanum species segregating for P. infestans resistance. Newly identified P. infestans resistance genes originating from S. verrucosum, S. schenckii, and S. capsicibaccatum could be mapped to potato chromosomes 6, 4, and 11, respectively

Low-dose oral immunization with lyophilized tissue of herbicide-resistant lettuce expressing hepatitis B surface antigen for prototype plant-derived vaccine tablet formulation

Efficient immunization against hepatitis B virus (HBV) and other pathogens with plant-based oral vaccines requires appropriate plant expressors and the optimization of vaccine compositions and administration protocols. Previous immunization studies were mainly based on a combination of the injection of a small surface antigen of HBV (S-HBsAg) and the feeding with raw tissue containing the antigen, supplemented with an adjuvant, and coming from plants conferring resistance to kanamycin. The objective of this study was to develop a prototype oral vaccine formula suitable for human immunization. Herbicide-resistant lettuce was engineered, stably expressing through progeny generation micrograms of S-HBsAg per g of fresh weight and formed into virus-like particles (VLPs). Lyophilized tissue containing a relatively low, 100-ng VLP-assembled antigen dose, administered only orally to mice with a long, 60-day interval between prime and boost immunizations and without exogenous adjuvant, elicited mucosal and systemic humoral anti-HBs responses at the nominally protective level. Lyophilized tissue was converted into tablets, which preserved S-HBsAg content for at least one year of room temperature storage. The results of the study provide indications on immunization methodology using a durable, efficacious, and convenient plant-derived prototype oral vaccine against hepatitis B

Genetics, fertility behaviour and molecular marker analysis of a new TGMS line, TS6, in rice

The thermosensitive genic male sterility (TGMS) system has great potential for revolutionizing hybrid rice production through simple, less expensive and more efficient seed production technology. For the successful utilization of this novel male sterility system, knowledge of the breeding and fertility behaviour of a TGMS line is essential. In this study, the fertility transformation behaviour, the critical fertility and sterility temperatures and the mode of inheritance of male sterility were studied for a new TGMS line, TS6, identified at Tamil Nadu Agricultural University, Coimbatore, India. The pollen and spikelet fertilities recorded on plants raised at fortnightly intervals revealed that this line was completely sterile for 78 consecutive days (35/22 to 32/23�C, maximum/minimum temperatures) and reverted to fertile when the temperature was 30/18�C. It remained fertile continuously for 69 days and the maximum pollen and spikelet fertilities recorded were 75 and 70%, respectively. The fertility was highly influenced by daily maximum temperature followed by average and minimum temperatures. It was not influenced by relative humidity, sunshine hours or photoperiod. The critical temperature inducing sterility and fertility was 26.7 and 25.5�C, respectively. The male sterility in TS6 was inherited as a monogenic recessive in the F2 and BC1 populations of TS6 · MRST9 as well as TS6 · IR68281B. Using bulked segregant analysis on an F2 population of TS6 · MRST9, an RAPD marker, OPC052962, was identified to be associated with TGMS in TS6

The ‘mosaic habitat’ concept in human evolution: past and present

The habitats preferred by hominins and other species are an important theme in palaeoanthropology, and the ‘mosaic habitat’ (also referred to as habitat heterogeneity) has been a central concept in this regard for the last four decades. Here we explore the development of this concept – loosely defined as a range of different habitat types, such as woodlands, riverine forest and savannah within a limited spatial area– in studies of human evolution in the last sixty years or so. We outline the key developments that took place before and around the time when the term ‘mosaic’ came to wider palaeoanthropological attention. To achieve this we used an analysis of the published literature, a study of illustrations of hominin evolution from 1925 onwards and an email survey of senior researchers in palaeoanthropology and related fields. We found that the term mosaic starts to be applied in palaeoanthropological thinking during the 1970’s due to the work of a number of researchers, including Karl Butzer and Glynn Isaac , with the earliest usage we have found of ‘mosaic’ in specific reference to hominin habitats being by Adriaan Kortlandt (1972). While we observe a steady increase in the numbers of publications reporting mosaic palaeohabitats, in keeping with the growing interest and specialisation in various methods of palaeoenvironmental reconstruction, we also note that there is a lack of critical studies that define this habitat, or examine the temporal and spatial scales associated with it. The general consensus within the field is that the concept now requires more detailed definition and study to evaluate its role in human evolution