Aluminium tolerance in lentil (Lens culinaris Medik.) with monogenic inheritance pattern

The objectives of this study were to determine genetics of Al tolerance and whether the Al tolerance observed is governed by the same gene. The lines ‘L‐7903’ and ‘L‐4602’ have been developed through breeding programme as Al‐tolerant lines. These lines showed maximum root regrowth and minimum accumulation of Al and callose as compared to sensitive genotypes (‘BM‐4’ and ‘L‐4147’). Al tolerance in the parents, F₁, F₂ and backcross generations was estimated using the regrowth of the primary root after staining and scoring of fluorescent signals. The F₁ hybrids responded similarly to the tolerant parents, indicating dominance of Al tolerance over sensitivity. The segregation ratios obtained for Al tolerance and sensitivity in the F₂ and backcross generations were 3 : 1 and 1 : 1, respectively. Test of allelism confirmed the same gene was conferring Al tolerance in both genotypes (‘L‐7903’ and ‘L‐4602’) as the F₁ was also tolerant and no segregation of tolerant : sensitive was recorded. These results indicated that Al tolerance is a monogenic dominant trait that can be easily transferred to agronomic bases through backcross breeding technique.     

Development,characterization and mapping of microsatellite markers for lentil (Lens culinaris Medik.)

Lentil is the sixth most important pulse crop terms of production in the world, but the number of available and mapped SSR markers are limited. To develop SSR markers in lentil, four genomic libraries for (CA)n, (GA)n, (AAC)n and (ATG)n repeats were constructed. A total of 360 SSR primers were designed and validated using 15 Turkish lentil cultivars and genotypes. The most polymorphic repeat motifs were GA and CT, with a mean number of alleles per locus of 7.80 and 6.55, respectively. Seventy‐eight SSR primers amplified a total of 400 polymorphic alleles, whereas 71 SSR primers produced markers within the expected size range. For 78 polymorphic SSR primers, the average number of alleles per locus was 5.1 and PIC value ranged from 0.07 to 0.89, with an average of 0.58. A linkage map was constructed using 92 individual F₂ plants derived from a cross between Karacada? × Silvan, with 47 SSR markers. The SSR markers developed in this study could be used for germplasm classification and identification and mapping of QTL in lentil.     

Breeding for boron tolerance in lentil (Lens culinaris Medik.) using a high‐throughput phenotypic assay and molecular markers

This study describes the identification of a quantitative trait locus (QTL) in the recombinant inbred line population of ILL2024 × ILL6788 and subsequent validation of associated molecular markers. A high‐quality genetic linkage map was constructed with 758 markers that cover 1,057 cM, with an average intermarker distance of 2 cM. QTL analysis revealed a single genomic region on Lc2 to be associated with B tolerance and accounted for up to 76% of phenotypic variation (Vp). The best markers for B tolerance were assessed for their utility in routine breeding applications using validation panels of diverse lentil germplasm and breeding material derived from ILL2024. A marker generated from the dense genetic map of this study was found to be the most accurate of all markers available for B tolerance in lentil, with a success rate of 93% within a large breeding pool derived from ILL2024. However, given the number of the unrelated lines for which the marker–trait association was not conserved, B tolerance screening is still required at later stages to confirm predicted phenotypes.     

Association of isozyme markers with quantitative trait loci in random single seed descent derived lines of lentil (Lens culinaris Medik.)

Summary Polymorphism at isozyme loci was used to locate factors responsible for variation in quantitative traits of lentil. Eight sets of random single seed descent (RSSD) derived lines were developed by advancing individual F₃ plants of interspecific (L. culinaris Medik. × L. orientalis Boiss.) hybrids to the F₆. The RSSD lines in each of the eight sets differed for alleles at 2–8 isozyme loci. In each set, association of isozyme loci with variation in seven quantitative traits (days to flower, days to mature, plant height, biomass, seed yield, harvest index, seed weight) was determined for each pairwise combination of a quantitative trait with a marker locus. Loci affecting variation in all seven quantitative traits were detected by their association with 14 isozyme markers (Aat-c, Aat-m, Aat-p, Adh-1, Fk, Gal-1, Gal-2, Lap-1, Lap-2, Pgd-p, Pgi, Pgm-c, Pgm-p, Skdh). The known position of 10 the 14 isozyme loci on the lentil genetic map was used to mark the genomic regions for possible location of associated quantitative trait loci (QTL). Detected QTL were found to be located in six of the seven linkage groups on lentil genetic map. Regions of the genome represented by linkage groups, 1, 5 and 7 appeared to affect a greater number of traits than other genomic regions represented by linkage groups 2, 3 and 4. Results indicated that the mean expression of quantitative traits at segregating marker locus classes can be used to locate the genetic factors in lentil which influence the behavior of economically important traits.     

Genetic diversity analysis of Moroccan lentil (Lens culinaris Medik.) landraces using Simple Sequence Repeat and Amplified Fragment Length Polymorphisms reveals functional adaptation towards agro‐environmental origins

In the absence of previous molecular characterization, we assessed genetic diversity of 53 Moroccan lentil landraces including two local cultivars using simple sequence repeat (SSR) and amplified fragment length polymorphisms (AFLP). Nineteen SSRs yielded 213 alleles, and seven AFLP primer combinations gave 766 fragments of which 422 were polymorphic. Moderate to high genetic variation was observed. Several small groups of landraces were differentiated. Interestingly, one of the smallest groups contained short‐cycle landraces with high early vegetative growth. Landraces in that group were from the dry land location of Abda, where they were likely selected for adaptation to drought and heat stress over centuries. Another group contained two landraces from highland areas that may have been selected for specific adaptation to cold stress. A third group contained one landrace from the Zear region known for its seed quality and has been proposed for the protected designation of origin (PDO) quality mark. Both techniques gave evidence of differentiation of the latter landrace supporting the idea of PDO attribution. Functional grouping according to agro‐environmental origins, cycle duration and early vegetative vigour was observed.     

Rapid generation cycling of an F2 population derived from a cross between Lens culinaris Medik. and Lens ervoides (Brign.) Grande after aphanomyces root rot selection

Cultivated lentil (Lens culinaris Medik.) is susceptible to aphanomyces root rot (ARR), whereas partial resistance is present in wild lentil including Lens ervoides (Brign.) Grande. Approximately six generations of selfing are required to fix a desired trait in a population, which usually requires 2 years in a breeding programme, so the primary objective was to develop a rapid generation cycling (RGC) technique that achieves this goal in 1 year. Rapid generation cycling was then tested on an F₂ population (LR‐59) derived from a L. culinaris × L. ervoides cross in combination with a reliable ARR screening technique, which generates a wide range of disease severities conducive to selection. Phenotyping of an F₂ population of more than 1,200 plants resulted in scores ranging from 2.4 to 4.0 on a scale from zero to five. Plants with scores lower than 4.0 were selected for advancement for five generations using a modified single‐seed descent method, optimum growing conditions, 20‐hr photoperiod and harvest of immature seeds. Seeds were germinated in a 100 μM gibberellin solution. Average generation length after phenotyping was 56 days resulting in five generations within approximately 300 days. Using a modified inoculation protocol, ARR phenotyping of the F₇ population resulted in scores ranging from 1.4 to 4.0. This inexpensive, nonsterile speed breeding protocol saves 1 year in the development of lentil varieties with improved ARR resistance.     

High‐throughput development of SSR marker candidates and their chromosomal assignment in rye (Secale cereale L.)

Shotgun survey sequences of flow‐sorted individual rye chromosomes were data mined for the presence of simple sequence repeats (SSRs). For 787,850 putative SSR loci, a total of 358,660 PCR primer pairs could be designed and 51,138 nonredundant SSR marker candidates were evaluated by in silico PCR. Of the 51,138 SSR primer candidates, 1,277 were associated with 1,125 rye gene models. A total of 2,112 of the potential SSR markers were randomly selected to represent about equal numbers for each of the rye chromosomes, and 856 SSRs were assigned to individual rye chromosomes experimentally. Potential transferability of rye SSRs to wheat and barley was of low efficiency with 4.3% (2,189) and 0.4% (223) of rye SSRs predicted to be amplified in wheat and barley, respectively. This data set of rye chromosome‐specific SSR markers will be useful for the specific detection of rye chromatin introgressed into wheat as well as for low‐cost genetic and physical mapping in rye without the need for high‐tech equipment.     

Cross‐genera amplification of informative microsatellite markers from common bean and scarlet runner bean for assessment of genetic diversity in mungbean (Vigna radiata)

Mungbean (V. radiata) is an important Asiatic legume supplying inexpensive protein to a vast majority of vegetarian masses. To increase markers repertoire in mungbean, a study was conducted to analyse 384 microsatellite markers derived from common bean, scarlet runner bean and adzuki bean for their transferability and polymorphism. The results showed that 87 (24.71%) primer pairs could amplify DNA loci of 20 mungbean genotypes including one accession of V. trilobata, while 52 showed reliable banding and polymorphism. These showed different degrees of variability at each locus producing 250 alleles with the number of alleles varying from 2 to 9. The major allele frequency varied from 0.17 to 0.95, while the polymorphic information content of SSRs ranged between 0.09 and 0.86 with an average of 0.60 ± 0.16. UPGMA revealed three major clusters accommodating ~95% of the accessions while one accession of V. trilobata (‘NSB‐007’) did not group with any other genotype describing the discriminating power of informative microsatellites. This study identified a set of useful microsatellite markers to accelerate the genetic studies and breeding programme of mungbean.     

Phenotypic and marker‐assisted pyramiding of genes for resistance to zucchini yellow mosaic virus in oilseed pumpkin (Cucurbita pepo)

We report on the identification of phenotypic and molecular markers for genes introgressed into oilseed pumpkin Cucurbita pepo from C. moschata germplasm originating in Nigeria, Portugal and Puerto Rico, which provide resistance against zucchini yellow mosaic virus (ZYMV) and on pyramiding these genes for improved and long‐lasting field protection of oilseed pumpkins. One SCAR and two SSR markers have been found for three dominant resistance genes, Zym‐0, Zym‐1 and Zym‐2. Characteristic reactions to ZYMV inoculation of plants carrying the recessive genes for resistance zym‐4* and zym‐6 have been defined. Described are procedures and results of pyramiding various combinations of these genes in oilseed pumpkin using the three markers and the specific phenotypic reactions to infection of some of these genes. The putative combination of all six resistance genes in one genotype resulted in a resistance that appeared to be at least as strong as or even stronger than that of the resistance source germplasm in C. moschata.     

Inter simple sequence repeat (ISSR) markers as a tool for the assessment of both genetic diversity and gene pool origin in common bean (Phaseolus vulgaris L.)

In this study, we report the use of ISSR to assess genetic diversity and to determine the relationships among ten cultivars of common bean developed in Argentina and three materials from France. ISSR markers resolved two major groups corresponding to the Andean and Mesoamerican gene pools of common bean. We compared the results of previous analysis, performed with RAPD markers (Galván et al., 2001), with the results generated by means of ISSR. It appears that ISSR are better tools than RAPDs to identify beans by gene pool of origin though they did not revealed as many differences between individuals as RAPDs. This revised version was published online in July 2006 with corrections to the Cover Date.     

Validation of a set of informative simple sequence repeats markers for variety identification in Pak‐choi (Brassica rapa L. ssp. chinensis var. communis)

A core set of 21 simple sequence repeats (SSR) markers was developed for Pak‐choi (Brassica rapa ssp. chinensis var. communis) variety identification. We initially selected 74 SSR markers which exhibited high polymorphism and reproducibility in SSR detection from 2129 SSRs. Using the 74 SSR‐based dendrogram for 45 inbred lines as calibration, 21 core SSRs were selected out. The utility of this core set SSRs was firstly tested in 45 inbred lines and finally verified in 102 commercial varieties. We also constructed a molecular ladder for each core SSR as a reference standard. Diversity analysis of this core SSR panel in 102 varieties demonstrated that each marker generates 2–3 alleles (averaged 2.33), with polymorphism information content values ranging from 0.01 to 0.56 (averaged 0.31). The averaged values of Shannon information index, observed heterozygosity, expected heterozygosity and Wright's fixation index were 0.59, 0.43, 0.38 and −0.09, respectively. Furthermore, the 21 SSR‐based classifications for 102 varieties were consistent with traditional classification based on morphology. This core SSR panel represents an effective tool for genetic variation analysis in Pak‐choi.