Researchers developing new cultivars of red raspberry (Rubus idaeus subsp. idaeus L.) and black raspberry (R. occidentalis L.) observe progeny of breeding populations for several seasons to identify those that perform reliably. If a portion of any breeding population could be eliminated based on a qualitative character or molecular marker, resources used for that portion could be used for other progeny. Our objective is to identify such molecular markers for red raspberry and black raspberry. A black raspberry × red raspberry cross was made to develop a map of each parent, and an F2 population was generated to join the maps. Simple sequence repeat (SSR) markers derived from red raspberry and strawberry were used. The level of homozygosity for the red raspberry was 40%, and the level for the black raspberry was 80%. Severe segregation skewing was observed in the F2 generation and indicates problems with transmission. Our findings help quantify the relative levels of homozygosity previously reported for red raspberry and black raspberry. In addition, the severe skewing observed in the F2 generation provides a molecular perspective to the fertility problems previously reported for the black raspberry × red raspberry hybrids (purple raspberry). Since black raspberry is highly homozygous, purple raspberry has transmission and fertility problems, and black raspberry breeders have reported a frustratingly low level of diversity in this subgroup, development of a black raspberry map is expected to require twice the markers as for a red raspberry map, emphasizing the need for a black raspberry sequence from which to develop molecular markers.
Kim S. Lewers and Courtney A. Weber
Shiow Y. Wang and Kim S. Lewers
Fruit of the cultivated strawberry (Fragaria ×ananassa Duchesne ex Rozier) are a good source of natural antioxidants, which play an important role in protecting human health. Antioxidant levels vary considerably among strawberry genotypes. The cultivated strawberry is a hybrid of two very different wild progenitor species: F. virginiana Mill. and F. chiloensis (L.) Mill. The progenitor species are valued by strawberry breeders as sources of novel traits, but have not been evaluated for antioxidant capacity or levels of antioxidant compounds. The objectives of this study are 1) to evaluate the antioxidant contents and antioxidant activities in representatives of F. virginiana and F. chiloensis in comparison with representatives of the cultivated strawberry species (F. ×ananassa), 2) to determine which strawberry compounds are most closely correlated with antioxidant capacity among these three species, and 3) to identify wild strawberry genotypes with high antioxidant activity and bioactive compounds for use in cultivar development. Fruit of 19 accessions from each of the three species, cultivated strawberry plus the two progenitor species (F. ×ananassa, F. virginiana, and F. chiloensis), were evaluated for levels of antioxidant capacity (oxygen radical absorbance capacity), total phenolics, total anthocyanins, ellagic acid, quercetin 3-glucoside plus quercetin 3-glucuronide, kaempferol 3-glucoside, kaempferol 3-rutinoside, p-coumaryl–glucose, pelargonidin 3-glucoside, pelargonidin 3-glucoside–succinate, cyanidin 3-glucoside, and cyanidin 3-glucoside–succinate. Fruit of the 13 accessions tested from the wild progenitor species F. virginiana had significantly higher antioxidant capacity, total phenolics, and total anthocyanins than did the fruit of three accessions tested from the cultivated strawberry F. ×ananassa, or the three accessions tested from the other wild progenitor species (F. chiloensis), and will be valuable as a source of parent material for developing more nutritious strawberry cultivars. The high correlation with antioxidant capacity, relative efficiency, and lack of genotype-by-year interaction in this study suggests that the measurement of total phenolics may be the better assay to use for the later selection stages in a strawberry cultivar development program. Of the evaluated F. virginiana accessions, NC 95-19-1, JP 95-1-1, CFRA 0982, NC 96-5-3, and RH 30 fruit were highest in antioxidant capacity and should prove useful toward development of strawberry cultivars with high antioxidant capacities.
Eric T. Stafne, John R. Clark and Kim S. Lewers
A tetraploid blackberry population that segregates for two important morphological traits, thornlessness and primocane fruiting, was tested with molecular marker analysis. Both randomly amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers were used to screen a population of 98 genotypes within the population plus the two parents, `Arapaho' and `Prime-Jim' (APF-12). RAPD analysis averaged 3.4 markers per primer, whereas SSR analysis yielded 3.0 markers per primer pair. Similarity coefficient derived from the Dice index averaged over all individuals was 63% for RAPD markers, 73% for SSR markers, and 66% for RAPD and SSR markers together. The average similarity coefficients ranged from a high of 72% to a low of 38% for RAPD markers, 80% to 57% for SSR markers, and 73% to 55% for both. Comparison of the parents indicated a similarity of 67% for RAPD markers, 62% for SSR markers, and 67% for both. This is similar to a previous study that reported the similarity coefficient at 66%. Although inbreeding exists within the population, the level of heterozygosity is high. Also, evidence of tetrasomic inheritance was uncovered within the molecular marker analysis. This population will be used to identify potential markers linked to both morphological traits of interest. Further genetic linkage analysis and mapping is needed to identify any putative markers.
Brent L. Black, Stan C. Hokanson and Kim S. Lewers
In the perennial strawberry production system, removal of the harvested crop represents a loss of nitrogen (N) that may be influenced by cultivar. Eight strawberry (Fragaria ×ananassa Duch.) cultivars and eight numbered selections grown in advanced matted row culture were compared over three seasons for removal of N in the harvested crop. Replicated plots were established in 1999, 2000, and 2001 and fruited the following year. `Allstar', `Cavendish', `Earliglow', `Honeoye', `Jewel', `Northeaster', `Ovation', and `Latestar' and selections B37, B51, B244-89, B683, B753, B781, B793, and B817 were compared for yield and fruit N concentration. Harvest removal of N (HRN) was calculated from total season yield and fruit N concentration at peak harvest. There were significant differences in HRN among genotypes, ranging from 1.80 to 2.96 g N per meter of row for numbered selections B781 and B37, respectively. Among cultivars, HRN ranged from 2.01 to 3.56 g·m–1 for `Ovation' and `Jewel', respectively. The amount of HRN was largely determined by yield, however, there were also significant genotype differences in fruit N concentration, ranging from 0.608 to 0.938 mg N per gram fresh weight for B244-89 and `Jewel', respectively. These differences indicate that N losses in the harvested crop are genotype dependent.
Kim S. Lewers, John M. Enns and Patricia Castro
Shiow Y. Wang, Kim S. Lewers, Linda Bowman and Min Ding
Fruit extracts from 17 to 18 representatives of three strawberry species [Fragaria virginiana Mill., F. chiloensis (L.) Mill., and F. ×ananassa Duchesne ex Rozier] were tested for the ability to inhibit proliferation of A549 human lung epithelial cancer cells. The fruit extracts also were tested for activities against free radicals, (peroxyl radicals, hydroxyl radicals, singlet oxygen, and superoxide radicals), the activities of antioxidant enzymes [glutathione peroxidase (EC 188.8.131.52), superoxide dismutase (EC 184.108.40.206), guaiacol peroxidase (EC 220.127.116.11), ascorbate peroxidase (EC 18.104.22.168), monodehydroascorbate reductase (EC 22.214.171.124), dehydroascorbate reductase (EC 126.96.36.199), and glutathione reductase (EC 188.8.131.52)], and the activities of nonenzyme antioxidant components, ascorbic acid and glutathione. Correlations between the proliferation of cancer cells and these antioxidant activities were calculated. At the species level, F. virginiana fruit extract inhibited the proliferation of A549 human lung epithelial cancer cells to a significantly greater extent (34% inhibition) than the extracts from fruit of either F. chiloensis (26%) or F. ×ananassa (25%) (P < 0.0001). Extracts from fruit of F. virginiana also had significantly greater antioxidant activities and higher activities of antioxidant enzymes and nonenzyme components than did extracts from the other two species. Among individual genotypes, there was a high positive correlation between antiproliferation of A549 cancer cells, antioxidant activities against free radicals, activities of antioxidant enzymes, and activities of nonenzyme components. Although all fruit extracts from all the strawberry genotypes inhibited proliferation of A594 cancer cells, fruit extracts from seven F. virginiana genotypes showed significantly greater antiproliferative effects than any of the F. ×ananassa or F. chiloensis genotypes. These genotypes, CFRA 0982, JP 95-1-1, NC 95-19–1, RH 30, NC 96-48-1, JP 95-9-6, and LH 50-4, may be especially useful in developing cultivars with greater anticancer potential.
Julia M. Harshman, Wayne M. Jurick II, Kim S. Lewers, Shiow Y. Wang and Christopher S. Walsh
Raspberries are a delicate, high-value crop with an extremely short shelf life exacerbated by postharvest decay caused by Botrytis cinerea Pers. European red raspberry (Rubus idaeus L.) is the most widely grown variety. Yellow (R. idaeus L.), black (R. occidentalis L.), and purple raspberries (R. ×neglectus Peck. or R. occidentalis ×idaeus hybrids) are available mainly at local markets and U-pick farms. To compare the postharvest quality of the raspberry color groups, pesticide-free fruit from cultivars and breeding selections of red, yellow, purple, and black raspberries were examined for oxygen radical absorbance capacity (ORAC), phenolics, anthocyanins, soluble solids, titratable acids, pH, color, firmness, decay and juice leakage rates, ethylene evolution, and respiration. There were significant correlations between decay rate and physiochemical properties. Both decay and leakage rates were correlated with weather conditions before harvest, but each color group responded differently to different weather factors. There were no correlations among changes in color, firmness, decay, or juice leakage rates. All the other color groups were less acidic than the familiar red raspberry. Yellow raspberries had the worst decay rates but the best leakage rates. Black and purple raspberries, with the highest phenolics and anthocyanins and the lowest ethylene evolution rates, resisted decay the longest but bled soonest.
Kim S. Lewers*, Eric T. Stafne, John R. Clark, Courtney A. Weber and Julie Graham
Some raspberry and blackberry breeders are interested in using molecular markers to assist with selection. Simple Sequence Repeat markers (SSRs) have many advantages, and SSRs developed from one species can sometimes be used with related species. Six SSRs derived from the weed R. alceifolius, and 74 SSRs from R. idaeus red raspberry `Glen Moy' were tested on R. idaeus red raspberry selection NY322 from Cornell Univ., R. occidentalis `Jewel' black raspberry, Rubus spp. blackberry `Arapaho', and blackberry selection APF-12 from the Univ. of Arkansas. The two raspberry genotypes are parents of an interspecific mapping population segregating for primocane fruiting and other traits. The two blackberry genotypes are parents of a population segregating for primocane fruiting and thornlessness. Of the six R. alceifolius SSRs, two amplified a product from all genotypes. Of the 74 red raspberry SSRs, 56 (74%) amplified a product from NY322, 39 (53%) amplified a product from `Jewel', and 24 (32%) amplified a product from blackberry. Of the 56 SSRs that amplified a product from NY322, 17 failed to amplify a product from `Jewel' and, therefore, detected polymorphisms between the parents of this mapping population. Twice as many detected polymorphisms of this type between blackberry and red raspberry, since 33 SSRs amplified a product from NY322, but neither of the blackberry genotypes. Differences in PCR product sizes from these genotypes reveal additional polymorphisms. Rubus is among the most diverse genera in the plant kingdom, so it is not surprising that only 19 of the 74 raspberry-derived SSRs amplified a product from all four of the genotypes tested. These SSRs will be useful in interspecific mapping and cultivar development.
Eric T. Stafne, John R. Clark, Courtney A. Weber, Julie Graham and Kim S. Lewers
Interest in molecular markers and genetic maps is growing among researchers developing new cultivars of Rubus L. (raspberry and blackberry). Several traits of interest fail to express in seedlings or reliably in some environments and are candidates for marker-assisted selection. A growing number of simple sequence repeat (SSR) molecular markers derived from Rubus and Fragaria L. (strawberry) are available for use with Rubus mapping populations. The objectives of this study were to test 142 of these SSR markers to screen raspberry and blackberry parental genotypes for potential use in existing mapping populations that segregate for traits of interest, determine the extent of inter-species and inter-genera transferability with amplification, and determine the level of polymorphism among the parents. Up to 32 of the SSR primer pairs tested may be useful for genetic mapping in both the blackberry population and at least one of the raspberry populations. The maximum number of SSR primer pairs found useable for mapping was 60 for the raspberry population and 45 for the blackberry population. Acquisition of many more nucleotide sequences from red raspberry, black raspberry, and blackberry are required to develop useful molecular markers and genetic maps for these species. Rubus, family Rosaceae, is a highly diverse genus that contains hundreds of heterozygous species. The family is one of the most agronomically important plant families in temperate regions of the world, although they also occur in tropical and arctic regions as well. The most important commercial subgenus of Rubus is Idaeobatus Focke, the raspberries, which are primarily diploids. This subgenus contains the european red raspberry R. idaeus ssp. idaeus L., as well as the american black raspberry R. occidentalis L. and the american red raspberry R. idaeus ssp. strigosus Michx. Interspecific hybridization of these, and other raspberry species, has led to greater genetic diversity and allowed for the introgression of superior traits such as large fruit size, fruit firmness and quality, disease resistance, and winter hardiness.
Thomas M. Davis, Laura M. DiMeglio, Ronghui Yang, Sarah M.N. Styan and Kim S. Lewers
The cultivated strawberry, Fragaria ×ananassa Duchesne ex Rozier, originated via hybridization between octoploids F. chiloensis (L.) Mill. and F. virginiana Mill. These three octoploid species are thought to share a putative genome composition of AAA`A'BBB`B'. Diploid F. vesca L., is considered to have donated the A genome. Current attention to the development of a diploid model system for strawberry genomics warrants the assessment of simple sequence repeat (SSR) marker transferability between the octoploid and diploid species in Fragaria L. In the present study, 23 SSR primer pairs derived from F. ×ananassa `Earliglow' by genomic library screening were evaluated for their utility in six diploid Fragaria species, including eight representatives of F. vesca, four of F. viridis Weston, and one each of F. nubicola (Hook. f.) Lindl. ex Lacaita, F. mandshurica Staudt, F. iinumae Makino, and F. nilgerrensis Schltdl. ex J. Gay. SSR primer pair functionality, as measured by amplification success rate (= 100% - failure rate) in each species, was ranked (from highest to lowest) as follows: F. vesca (98.4%) > F. iinumae (93.8%) = F. nubicola (93.8%) > F. mandshurica (87.5%) > F. nilgerrensis (75%) > F. viridis (73.4%). The extent to which these octoploid-derived SSR primer pairs generated markers that could be added to the F. vesca linkage map also was assessed. Of the 13 F. ×ananassa SSR markers that segregated codominantly in the F. vesca mapping population, 11 were assigned to linkage groups based upon close linkages to previously mapped loci. These markers were distributed over six of the seven F. vesca linkage groups, and can serve as anchor loci defining these six groups for purposes of comparative mapping between F. vesca and F. ×ananassa.