QTL mapping for morphological and physiological traits in RILs of spring wheat population of WH1021 × WH711

Aim: To identify the quantitative trait loci (QTLs) associated with various morphological and physiological traits. Methodology: A recombinant inbred line (RIL) population of wheat (Triticum aestivum L.) derived from cross of heat tolerant parent WH1021 with heat sensitive parent WH711 was used. For creation of heat stress delay in sowing (four weeks) was done in late sown from timely sown. Days to heading, days to maturity, plant height along with physiological traits viz. photosynthetic rate, transpiration rate, intrinsic water use efficiency (iWUE) and normalized difference vegetative index (NDVI) were recorded in both timely and late sown conditions over two years. Results: A linkage map consisting of 22 SSR was prepared covering eleven chromosome of wheat. Using the software WinQTL cartographer version 2.5, significant genomic regions associated with heat tolerance were observed on 2A, 2D, 4A and 5A chromosomes with composite interval mapping. QTLs for photosynthetic rate were found consistently for both years on chromosome 2D. In present study, significant variation was reported for days to heading and its QTLs were identified on chromosome 2A and 2D. Interpretation: The QTLs obtained for physiological and morphological traits in wheat can be used for marker-assisted selection for heat tolerance in wheat

Leveraging barrel medic genome sequence for the development and use of genomic resources for genetic analysis and breeding in legumes

BACKGROUND: A total of 62,591 cowpea expressed sequence tags (ESTs) were BLAST aligned to the whole-genome sequence of barrel medic (Medicago truncatula) to develop conserved intron scanning primers (CISPs). The efficacy of the primers was tested across 10 different legumes and on different varieties of cowpea, chickpea, and pigeon pea. Genetic diversity was assessed using the same primers on different cowpea genotypes. Singlenucleotide polymorphisms (SNPs) were detected, which were later converted to length polymorphism markers for easy genotyping. CISPs developed in this study were used in tagging resistance to bacterial leaf blight disease in cowpea. RESULTS: A total of 1262 CISPs were designed. The single-copy amplification success rates using these primers on 10 different legumes and on different varieties of cowpea, chickpea, and pigeon pea were approximately 60% in most of the legumes except soybean (47%) and peanut (37%). Genetic diversity analysis of 35 cowpea genotypes using 179 CISPs revealed 123 polymorphic markers with PIC values ranging from 0.05 to 0.59. Potential SNPs identified in cowpea, chickpea, and pigeon pea were converted to PCR primers of various sizes for easy genotyping. Using the markers developed in this study, a genetic linkage map was constructed with 11 linkage groups in cowpea. QTL mapping with 194 F3 progeny families derived from the cross C-152 × V-16 resulted in the identification of three QTLs for resistance to bacterial leaf blight disease. Conclusions: CISPs were proved to be efficient markers to identify various other marker classes like SNPs through comparative genomic studies in lesser studied crops and to aid in systematic sampling of the entire genome for well-distributed markers at low cost

Research progress in construction of genetic linkage maps of medicinal plants in recent five years / 中草药

Genetic linkage mapping is one of the key steps in genomic research, which is the cornerstone of a wide spectrum of biotechnology applications, including gene localization, map-assisted cloning, molecular marker-assisted selection in breeding, and quantitative trait loci. Furthermore, the prospering of the molecular marker technology laid the technological foundation of constructing high density genetic linkage map. This paper briefly reviewed the procedure of genetic mapping, types of molecular markers, mapping populations, and research progress in construction of the genetic linkage maps of medicinal plants in recent five years. In addition, the existing problems and the future direction have been discussed in this field.

Otimização do mapeamento genético vegetal de populações duplo-haplóide via simulação computacional / Optimization of vegetal genetic mapping of double-haploid populations via computational simulation

O mapeamento genético é um passo necessário para entender a organização genômica e a relação entre genes e o fenótipo. Um dos principais problemas está em encontrar a ordem, o espaçamento correto dos marcadores em um mapa genético, assim como o número de indivíduos a compor uma população. Deste modo, o objetivo deste estudo foi avaliar o nível de saturação do genoma e o tamanho ideal de populações simulada duplo-haplóide para a construção de mapas de ligação mais confiáveis por meio de simulação computacional. Foram simulados genomas parentais e populações duplo-haplóide considerando marcadores moleculares do tipo dominante, espaçados de forma equidistante a 5, 10 e 20 cM. Os tamanhos das populações geradas foram de 100, 200, 300, 500, 800 e 1000 indivíduos, com dez grupos de ligação cada e 100 repetições por amostra. Procedeu-se a análise de todas as populações geradas obtendo um genoma analisado o qual foi comparado com o genoma simulado inicialmente. Observou-se que o tamanho ideal de populações duplo-haplóide para mapeamento genético foi de no mínimo 200, 500 e 1000 indivíduos para genomas saturados, medianamente saturados e com baixa saturação. Populações de mesmo tamanho tendem a produzir mapas com maior acurácia em níveis de saturação do genoma mais elevados.

Genetic mapping is a necessary step to understand the genomic organization and the relationship between genes and phenotypes. A major problem is to find the order, the correct spacing of the markers in a genetic map, and the number of individuals to compose a population. Thus, the objective of this study was to evaluate the saturation level of the genome and the optimal size of simulated double-haploid populations for the construction reliable linkage maps by means of computer simulation. Parental genomes and double-haploid populations were simulated considering dominant molecular markers, spaced equidistantly at 5, 10 and 20 cM. The sizes of the generated populations were 100, 200, 300, 500, 800 and 1000 individuals, with ten linking groups and 100 replicates per sample. It was proceeded the analysis of all generated population obtaining a genome which was compared with the first simulated genome. It was observed that the optimal size of double-haploid populations for genetic mapping has been at least 200, 500 and 1000 individuals for saturated genomes, medium unsaturated and low saturation. Populations of the same size tend to produce maps with greater accuracy in higher levels of genome saturation.

La biotecnología como herramienta para la propagación, conservación y el mejoramiento genético del guayabo / Biotechnology as a tool for propagation, con-servation and genetic breeding in guava

Las técnicas biotecnológicas contribuyen positiva y significativamente en los programas de propagación, conservación y mejoramiento de las especies vegetales. Dentro de éstas, el cultivo de tejidos, el desarrollo de mapas de ligamientos genéticos y de QTLs y la detección de genes de interés han demostrado ser de gran utilidad para los mencionados propósitos. En este sentido, se estandarizó una técnica para la multiplicación in vitro de la forma silvestre de guayabo en tres fases de cultivo: establecimiento, multiplicación de propágulos y enraizamiento. La misma constituye una vía de utilidad para la propagación, la conservación de germoplasma y el mejoramiento genético en la especie. Además, se estandarizó un método de conservación a corto-mediano plazo. Por otra parte, se construyó un mapa de ligamiento genético para la especie empleando marcadores AFLP y SSR. Los 11 grupos del mapa de ligamiento genético y los 50 QTLs relacionados con caracteres vegetativos y de calidad interna y externa del fruto, constituyen el punto de partida para el clonaje de genes de interés agrícola y la implementación futura de la selección asistida por marcadores en el guayabo. De igual forma, las 176 secuencias candidatas a genes de resistencia (RGL) y del desarrollo de la planta (MADS-box y HOMEO-box) detectadas pueden ser de gran utilidad en la saturación del mapa de ligamiento referido, el estudio de la variabilidad presente en el cultivo, así como en la solución de problemas relacionados con el rendimiento, la producción y la resistencia a estrés biótico y abiótico

Biotechnologies contribute positively and signifi¬cantly in the propagation, conservation and breeding programs of many plant species. From them, tissue culture, linkage maps and QTLs detection for interesting genes have been proved to be of great utility for these purposes. In this sense, a technique for in vitro multiplication of wild guava was standardized in three culture phases: establishment, multiplication and rooting. This technique constituted a useful way for propagation, germplasm conservation and genetic breeding in the specie. A method for short-medium term conservation was also standardized. On the other hand, a genetic linkage map was constructed for the specie using AFLP and SSR markers. The 11 groups of the genetic linkage map and the 50 QTLs related with vegetative and internal/external fruit characters constitute the starting point for genes cloning of agricultural interest and the future imple¬mentation of markers assisted selection in guava. Also, the 176 candidate sequences for resistance-gene-like (RGL) and plant development (MADS-box and HOMEO-box) genes detected can be of great utility in linkage map saturation, variability studies in this crop, as well as in the solution of problems rela¬ted with yielding and resistance to biotic and abiotic stresses.

LINKAGE MAPPING OF CANDIDATE GENES FOR INDUCE RESISTANCE AND GROWTH PROMOTION BY Trichoderma koningiopsis (Th003) IN TOMATO Solanum lycopersicum / Mapeo de genes candidatos relacionados con inducción de resistencia sistemica y promoción de crecimiento por Trichoderma koningiopsis (Th003) en tomate Solanum lycopersicum

Induced systemic resistance (ISR) is a mechanism by which plants enhance defenses against any stress condition. ISR and growth promotion are enhanced when tomato (Solanum lycopersicum) is inoculated with several strains of Trichoderma ssp. This study aims to genetically map tomato candidate genes involved in ISR and growth promotion induced by the Colombian native isolate Trichoderma koningiopsis Th003. Forty-nine candidate genes previously identified on tomato plants treated with Th003 and T. hamatum T382 strains were evaluated for polymorphisms and 16 of them were integrated on the highly saturated genetic linkage map named “TOMATO EXPEN 2000”. The location of six unigenes was similar to the location of resistance gene analogs (RGAs), defense related ESTs and resistance QTLs previously reported, suggesting new possible candidates for these quantitative trait loci (QTL) regions. The candidate gene-markers may be used for future ISR or growth promotion assisted selection in tomato.

La resistencia sistémica inducida (ISR) es un mecanismo mediante el cual las plantas aumentan sus defensas frente a cualquier condición de estrés. El objetivo de este trabajo fue localizar en el mapa genético de tomate, genes candidatos involucrados en ISR y promoción de crecimiento inducidos por la cepa colombiana nativa Th003 de Trichoderma koningiopsis. Se realizó una búsqueda de polimorfismos en cuarenta y nueve genes candidatos previamente identificados en plantas de tomate inoculadas con Th003 y la cepa T382 de T. hamatum. Diez y seis de estos genes candidatos fueron integrados en el mapa genético de tomate altamente saturado, llamado “TOMATO EXPEN 2000”. La ubicación de seis unigenes fue similar a la localización de genes análogos de resistencia (RGAs), ESTs relacionados con defensa y QTLs de resistencia previamente identificados, sugiriendo posibles nuevos candidatos para estas regiones de QTLs. Los genes candidatos o marcadores pueden ser usados en futuros programas de selección asistida relacionados con ISR o promoción de crecimiento en tomate.

Genetic mapping of EST-SSR, SSR and InDel to improve saturation of genomic regions in a previously developed sunflower map

In order to saturate a sunflower genetic map and facilitate marker-assisted selection (MAS) breeding for stress response, it is necessary to enhance map saturation with molecular markers localized in linkage groups associated to genomic regions involved in these traits. This work describes the identification and characterization of 1,134 simple sequence repeat (SSR) containing expressed sequence tags (EST) from unigenes available databases. Twelve of these functional markers as well as 41 public SSR markers were successfully localized in linkage groups, thus contributing to the saturation of specific regions on a reference genetic-linkage-map derived from recombinant inbred lines (RIL) mapping population from the cross between PAC2 x RHA266 lines. The enriched map includes 547 markers (231 SSR, 9 EST-SSR, 3 insertions/deletions (InDel) and 304 amplified fragment length polymorphisms (AFLP) distributed in 17 linkage groups (LG), spanning genetic size to 1,942.3 cM and improving its mean density to 3.6 cM per locus. As consequence, no gaps longer than 13.2 cM remain uncovered throughout the entire map, which increases the feasibility of detecting genes or traits of agronomic importance in sunflower.

Comparative linkage mapping of Oryza glumaepatula and Oryza sativa interspecific crosses based on microsatellite and expressed sequence tag markers

Molecular linkage maps representing the rice genome have been an important tool for breeding programs because they allow the elucidation of polygenic traits and are an efficient tool for monitoring wild introgressions in interspecific crosses. Common markers among rice genetic maps are important in defining the homology of chromosomes and the synteny between genomic target regions. We used 148 markers (expressed sequence tags, microsatellites and single nucleotide polymorphisms) to construct a molecular linkage map based on co-dominant markers for an interspecific backcross population using a wild rice (Oryza glumaepatula) from Brazil and performed a comparative analysis with other interspecific maps. The comparative analysis revealed a Spearman correlation index of 0.86 for marker order conservation to a previous map constructed for an interspecific cross using the same wild parent. Approximately 90 percent of markers common to other interspecific maps kept the same order. These results indicate that it will be possible to generate a unique genetic map using the wild donor and that it may be a helpful tool for breeding programs because plants derived from different interspecific populations can be rapidly scanned using markers associated with useful wild traits.