Potential of marker-assisted selection in hemp genetic improvement

Summary The development and applications of molecular markers to hemp breeding are recent, dating back only to the mid-1990s. The main achievements in this field are reviewed. The analysis of Cannabis germplasm by RAPD, AFLP and microsatellites is discussed, with its consequence for the still debated species concept in Cannabis. DNA-based markers have also been exploited in the field of forensic science, in an attempt to discriminate licit from illicit crop. The main applications of the molecular markers to the breeding, however, have been achieved with the development of markers closely linked to the male sex and to some of the most relevant chemotypes. Active research is carried out by several groups in the field of identification and characterization of the genes involved in fiber quality and quantity, and in the determination of monoecy, another very important target of hemp breeding. Besides, markers associated to new, potentially useful chemotypes are being developed, for the marker-assisted breeding of pharmaceutical Cannabis.     

QTL analysis of seed dormancy in rice (Oryza sativa L.)

Effective cumulative temperature (ECT) after heading would be a more reasonable parameter for seed sampling of pre-harvest sprouting/seed dormancy (SD) tests in segregating populations than the days after flowering. SD is an important agronomic trait associated with grain yielding, eating quality and seed quality. To identify genomic regions affecting SD at different grain-filling temperatures, and to further examine the association between SD and ECT during grain-filling, 127 double haploid (DH) lines derived from a cross between ZYQ8 (indica)/JX17 (japonica) by anther culture were analyzed. The quantitative trait loci (QTLs) and their digenic epistasis for SD were identified using a molecular linkage map of this population. A total of four putative QTLs for SD (qSD-3, qSD-5, 6 and 11) were detected on chromosomes 3, 5, 6 and 11, together explaining 41.4% of the phenotypic variation. Nine pairs of digenic epistatic loci were associated with SD on all but chromosome 9, and their contributions to phenotypic variation varied from 2.87%–8.73%. The SD QTL on chromosome 3 was identical to the QTLs found in other mapping populations with different genetic backgrounds, which could be a desirable candidate for gene cloning and marker-assisted selection in rice breeding.     

The association of culm anatomy with lodging susceptibility in modern spring wheat genotypes

Selection for lodging resistant cultivars in cereal breeding programs is difficult due to the challenge of screening for this trait under natural field conditions. The identification of easily measurable culm traits related to lodging resistance would simplify the selection process. The present study was conducted to determine if differences in culm anatomy exist among modern wheat genotypes differing in lodging susceptibility, and to determine the association between culm characters and lodging. From a 2-year field study conducted in Edmonton, Alberta, 13 spring wheat cultivars were chosen based on predetermined susceptibility to artificially induced lodging. Morphological and anatomical culm measurements were made visually and with an environmental scanning electron microscope. Genotypes differed (P < 0.05) for plant height, number of internodes per culm, basal internode length and diameter, culm wall thickness and the number of vascular bundles, but not for adventitious root frequency, lumen diameter or sclerenchyma ring thickness. Mean genotype field scores for artificially induced lodging were correlated (P < 0.05) with plant height (r= 0.51) and the length of the fourth basal internode (r= 0.51). Short, wide basal internodes and thick culm walls were characteristic of three lodging tolerant genotypes: Kohika, Sapphire and Olso. Nevertheless, despite such apparent genotype specific association between culm anatomy and field lodging, general applicable associations were not observed for most traits. The most practical and easily selectable trait for lodging resistance within a wheat breeding program remains plant height. This revised version was published online in July 2006 with corrections to the Cover Date.     

Segregation distortion in doubled haploid lines of barley (Hordeum vulgare L.) detected by simple sequence repeat (SSR) markers

A total of 147 simple sequence repeat (SSR) markers (including86 barley and 61 wheat microsatellite markers) were tested for their segregation in a doubled haploid (DH) and an F₂ population of barley. The DH population consisted of 71 doubled haploid lines, developed from F₁ plants of a cross between Tadmor and WI2291using isolated microspore culture technique. A genetic linkage map consisting of 43 microsatellite markers was constructed using the DH population. Particularly on chromosome 4H microsatellite markers showed distorted segregation ratios. Segregation of DH lines based on molecular markers were compared with segregation of 92 F₂ lines from the same cross. The proportion of loci deviating from the expected monogenic segregation ratios in the DH population was significantly higher (19/43loci, 44%) than in the F₂ population (7/43 loci, 16%). The deviation was biased towards the WI2291 parent alleles. In line with this observation, WI2291 was found to perform better than Tadmor in regenerating green plantlets with the isolated microspore-culture technique. This revised version was published online in July 2006 with corrections to the Cover Date.     

Homoeological relationships between the f chromosome of Brassica rapa and the e chromosome of Brassica oleracea

Eight plants of the putative double monosomic addition line (DMAL, 2n= 20) were developed by crossing a monosomic chromosome addition line of radish [f(A)‐type monosomic addition line (MAL) (2n= 19)] carrying the f chromosome of Brassica rapa (2n= 20, AA) with another [e(C)‐type MAL (2n= 19)] having the echromosome of Brassica oleracea (2n= 18, CC). The homoeological relationships between the two alien chromosomes were investigated by morphological, cytogenetic and random amplified polymorphic DNA (RAPD) analysis. Seventeen morphological traits that were not present in the radish cv. ‘Shogoin’ were observed in both MALs and these traits were substantially exhibited in DMAL plants. At the first metaphase of pollen mother cells (PMCs), the two parental MALs showed a chromosome configuration of 9II +1I, demonstrating impossibility of recombination between the R and the added chromosomes. The DMALs formed 10II in approximately 73% of PMCs, with one bivalent showing loose pairing between two chromosomes differing in size. In an attempt to identify the two MALs by RAPD‐specific markers using 26 selected random primers, 13 and 20 bands were specific for the f(A)‐type and the e(C)‐type MALs, respectively; 12 bands were common to both MALs (26.7%). In conclusion, the f chromosome of B. rapa is homoeologous to the e chromosome of B. oleracea. The genetic domain (genes) for 17 morphological traits are linked to each homoeologous chromosome bearing 27% of the corresponding RAPD markers.     

Pyramiding genes for resistance to bean common mosaic virus

Summary Gene pyramiding in Phaseolus vulgaris is being utilized to develop more effective resistance to the temperature-insensitive-necrosis-inducing (TINI) strains of Bean Common Mosaic Virus (BCMV) present in the USA. Our data indicate that contrary to previous work, the bc-3 gene is effective against these strains in the absence of the strain unspecific bc-u gene in genotypes possessing the I gene. The epistatic bc-3 gene interferes with traditional efforts to pyramid the other recessive resistance genes by masking their activity. Indirect selection based on markers linked to the other recessive resistance genes would likewise be ineffective without the ability to also select for the bc-u gene which is required for expression of the bc-2 ² gene in germplasm carrying the I gene. Because the most resistant genotype (I, bc-u, bc-I ², bc-2², bc-3) can only be introduced into a wide range of germplasm through the use of molecular markers linked to the different resistance genes, the search for a marker linked to the strain unspecific bc-u gene should also be given priority.     

Comparison of pedigree selection and single seed descent for oil yield in linseed (Linum usitatissimum L.)

Summary Four populations of linseed derived from five parents were advanced from the F₂ to the F₆ generation by SSD and to the F₅ by pedigree selection. These populations were used to compare the efficiency of the two methods in order to provide superior genotypes with respect to grain yield and oil content, i.e. oil yield. The results showed minimum differences between pedigree and SSD lines for grain yield, where in only one cross the SSD lines were significantly superior to the pedigree lines. Since pedigree selection was carried out for both, seed yield and oil content, a positive response to selection was expected. However, early selection for yield, a character with low heritability, was not successful. On the contrary, significant differences for oil content were detected between the two groups of lines in three of the four crosses studied. In these cases the pedigree lines were superior to the SSD lines. These results demonstrate that strict selection in early generations for oil content, a character with comparatively high heritability, is feasible and successful in linseed. However, selection for seed yield should be postponed to later inbred generations. Consequently, in breeding for maximum oil yield of linseed a two-step selection procedure is recommended.Abbreviations PS pedigree selection - SSD single seed descent - TGW thousand grain weight     

Oligonucleotide fingerprinting of tomato DNA

Hinfl -digested DNA samples of wild and cultivated species of tomato were hybridized in gel to different oligonucleotide probes complementary to simple-sequence repeats. All motifs tested i.e. (GATA)₄, (GGAT)₄, (CA)₈. (CAG)₅, (GTG)₅, (GAAGTGGG)₂ and (CCTA)₄, produced multilocus fingerprints in all tomato species and cultivars. The extent of inter- and intraspecific polymorphism of the banding patterns varied considerably between different probes, with (GGAT)₄, (CCTA)₄, (GATA)₄ and (CA)₈ showing a much higher discriminatory potential than the other probes. Cluster analysis of band-sharing data yielded dendrograms which were consistent with the known history of tomato cultivation. (GATA)₄ was also successfully used for purity testing of breeding lines, as well as for analysing an F₁ progeny of a cross between two lines.     

Path analysis of genotype-environment interactions of potatoes exposed to increasing warm-climate constraints

Summary Twenty potato cultivars were tested in replicated trials in three seasons. The weather gradually changed from relatively high humidity and cold air temperatures in February to dry and hot in July. Fresh and dry weights, sucrose, reducing sugar and starch contents were recorded on tubers sequentially harvested over four stages during the growing period of each of the three trials. The data were analyzed for genotype-environment interactions by a path regression model which is composed of a main effect and four multiplicative effects. The model fitted well to the observed data for all traits but reducing sugar. Results of dry tuber weight were used to calculate a heat susceptibility index for each of the cultivars. Four components of the index were identified based on the path model. Comparison of sizes of the components provided information on the degree of response to the environmental stress of each of the cultivars during successive stages of growth. High yielding and heat tolerant cultivars were identified based on the experimental results.     

Progress in mapping agronomic genes in apple (The European Apple Genome Mapping Project)

Summary The progress of the European Apple Genome Mapping Project is described. Populations segregating for a range of agronomic genes have been established in six European countries. The need for robust methods of analysis has been identified, especially with regard to the development of molecular markers. Isozyme systems, RAPDs, RFLPs and amplified genes are being used to construct a reference genetic linkage map. Standardisation and precise definition of both genotypic and phenotypic measurements has been recognised as being essential for future exploitation of genetic markers in apple breeding. Phenotypic measurements are being replicated in different geographical locations over several years. Statistical and genetic analyses are aimed at defining components of genetic variation which account for ‘genes’, as defined by apple breeders. A relational database is being constructed which will combine disparate sources of data relating to the genetics of apple. Comparative mapping has been identified as an efficient means of expanding genetic knowledge within and between Rosaceae genomes.