Efficiency of Recurrent Selection by Marker and Phenotype for Multiple, Quantitative Yield Components in Four Cucumber Populations

Four cucumber (Cucumus sativus L.) inbred lines were intermated then bulked maternally to create four base populations denoted as cycle 0 (i.e., Pop.1 C0, Pop.2 C0, Pop.3 C0, Pop.4 C0). Each of these populations underwent phenotypic selection (PHE; open-field evaluations), selection by marker (MAS; genotyping at 20 marker loci), and random mating (RAN; no selection) for three cycles. The four traits under selection, multiple lateral branching (MLB), gynoecious sex expression (GYN), earliness (EAR), and fruit length to diameter ratio (L:D), are quantitatively inherited, controlled by relatively few (two to six) QTL per trait and are directly related to yield. Using the same C0 populations and selection scheme allowed a direct comparison of the effectiveness of MAS and PHE. Because each C0 population varied for any given trait, the response to MAS and PHE was not the same for each population. In general, C0 populations that were inferior for a trait either responded favorably to selection or remained constant, while those with superior trait values either did not change or decreased. Both MAS and PHE provided improvements in all traits under selection in at least one population, with the exception of MAS for EAR. MAS and PHE were equally effective at improving MLB and L:D, but PHE was generally more effective than MAS for GYN and EAR. When considering all traits, responses to PHE were superior in three of the four populations. The population for which MAS was superior, however, showed the only increase in yield (fruit per plant), which was not under direct selection. These results indicate that both MAS and PHE are useful for multi-trait improvement in cucumber, but their effectiveness depends on the traits and populations under selection.

(255) A Comparison of Phenotypic and Marker-assisted Selection in Cucumber

To increase yield in cucumber (Cucumussativus L.), we designed a recurrent selection program utilizing phenotypic (PHE) and marker-assisted (MAS) selection for the development of multiple lateral branching (MLB; branches per plant), gynoecious, early genotypes possessing high fruit length to diameter ratio (L:D). These yield components are under genetic control of few quantitative trait loci (QTL; 2-6 per trait), which have been placed on a moderately saturated molecular linkage map. Four inbred lines, complementary for the target traits, were intermated and the resulting population underwent MAS and PHE, as well as random mating (RAN), for three cycles. Selections by PHE were visually made for all four traits at the whole plant level. Selections based on MAS contained the highest number of desired marker genotypes from 20 marker loci (SSR, RAPD, SCAR, SNP). Using the same selection scheme and intensity allowed a direct comparison of MAS to PHE. Selection was equally effective for MLB and L:D by MAS (3.5 and 3.0) and PHE (3.6 and 3.0), which were both superior to RAN (2.8 and 2.8). For earliness (days to anthesis) and gynoecy (percent female), MAS (41.8 and 26.6) was less effective than PHE (40.5 and 81.8) and RAN (41.0 and 80.9), which were equal. For yield (fruit per plant), RAN (1.90) and MAS (1.88) were equal, but less than PHE (2.15). After three cycles of PHE, further selection by MAS identified superior genotypes, which were intermated. Superior hybrids were selected by MAS and underwent one backcross generation. In some backcrosses, gains were made in every trait compared to the PHE Cycle 3 mean, while in other backcrosses, gains were made only in some traits. Improvement by MAS was very effective during line extraction for these yield components.

Development and Characterization of PCR Markers in Cucumber

Highly polymorphic microsatellites or simple sequence repeat (SSR), along with sequence characterized amplified region (SCAR) and single nucleotide polymorphisms (SNP), markers are reliable, cost-effective, and amenable for large scale analyses. Molecular polymorhisms are relatively rare in cucumber (Cucumis sativus L.) (3% to 8%). Therefore, experiments were designed to develop SSR, SCAR and SNP markers, and optimize reaction conditions for PCR. A set of 110 SSR markers was constructed using a unique, strategically applied methodology that included the GeneTrapper (Life Technologies, Gaithersburg, Md.) kit to select plasmids harboring microsatellites. Of these markers, 58 (52%) contained dinucleotide repeats (CT, CA, TA), 21 (19%) possessed trinucleotide repeats (CTT, ATT, ACC, GCA), 3 (2.7%) contained tetranucleotide repeats (TGCG, TTAA, TAAA), 4 (3.6%) enclosed pentanucleotide repeat (ATTTT, GTTTT, GGGTC, AGCCC), 3 (2.7%) contained hexanucleotide repeats (CCCAAA, TAAAAA, GCTGGC) and 21 possessed composite repeats. Four SCARs (L18-3 SCAR, AT1-2 SCAR, N6-A SCAR, and N6-B SCAR) and two PCR markers based on SNPs (L18-2H19 A and B) that are tightly linked to multiple lateral branching (i.e., a yield component) were also developed. The SNP markers were developed from otherwise monomorphic SCAR markers, producing genetically variable amplicons. The markers L18-3 SCAR and AT1-2 SCAR were codominant. A three-primer strategy was devised to develop a codominant SCAR from a sequence containing a transposable element, and a new codominant SCAR product was detected by annealing temperature gradient (ATG) PCR. The use of a marker among laboratories can be enhanced by methodological optimization of the PCR. The utility of the primers developed was optimized by ATG-PCR to increase reliability and facilitate technology transfer. This array of markers substantially increases the pool of genetic markers available for genetic investigation in Cucumis.

Genetic Analysis of Yield Components in Cucumber at Low Plant Density

Plant architecture can be manipulated in cucumber (Cucumis sativus L.) to provide an array of phenotypes. Determinate, multiple-lateral plants are unique because they afford an opportunity to increase fruit yield per plant. Estimates of genetic variances, numbers of genetic factors, and genotypic and phenotypic correlations between traits were made in a population, segregating for sex expression, leaf size, and plant habit at low plant density (≈19,000 plants/ha). Replicated evaluation of 100 F3 families derived from an initial mating between a gynoecious, determinate, moderately branched line (G421) and a monoecious, indeterminate, multiple lateral branching line (`H-19') indicated that mainstem length and multiple lateral branching exhibit mostly additive genetic variance. For sex expression, additive and dominant genetic variances were important. The minimum number of genetic factors controlling sex expression, number of lateral branches, and mainstem length were estimated at five, four, and eight, respectively. Phenotypic and genotypic correlations between traits indicated that relative leaf size may influence fruit mass while having only limited influence on the number of fruit produced per plant. The amount and type of genetic variation suggests that the development of an array of determinate, multiple-lateral branching plant types with varying sex expression and plant stature is possible.