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Variegation is a common trait in plants that characteristically displays white or off-colored plant tissue. In grapevine, leaf variegation is expressed as white and pale green leaf tissue resulting in plants that are stunted in growth and hindered in development. In this study, several experiments were performed to investigate the impact of this mutation has on the anatomy of grape leaves and physiology of the plant. Histological staining of variegated and nonvariegated leaf tissue transections showed alterations to the leaf palisade mesophyll structure that affected leaf tissue width. An assay quantifying leaf pigments was performed to compare chlorophyll and carotenoid concentrations in leaves between variegated and wild-type seedlings, which showed that variegated leaf samples had reduced chlorophyll and carotenoid concentration. Through fluorescence imaging, we determined that photochemical efficiency of photosystem II (PSII) is reduced in variegated seedlings. By growing variegated and wild-type plants under high, medium, and low light intensities that variegated plants exposed to higher light intensity reduces the phenotypic expression of the variegation trait. Also, we found variegated plants to have significant reductions in growth traits such as plant height, leaf number, branch number, and dry weight compared with wild-type phenotype plants. Overall, our experiments revealed the variegation mutation altered normal leaf development causing significant effects to grapevine physiology.
Prairie junegrass [Koeleria macrantha (Ledeb.) Shultes] is a perennial, short-grass prairie species distributed throughout the Northern Hemisphere that is being evaluated for use as a low-input turf. In June 2007, 300 genotypes representing collection locations derived from Colorado, Nebraska, and Minnesota germplasm were grown and evaluated 3 years for turfgrass performance characteristics in a randomized complete block design with five clonal replications at two locations (St. Paul, MN, and Becker, MN). After establishment, plots received no supplemental irrigation or fertility and were mowed weekly to a height of 6.4 cm. Broad-sense heritability estimates were calculated on a clonal mean (Hc) and single-plant (Hsp) basis for turf quality (Hc = 0.62, Hsp = 0.13), crown density (Hc = 0.55, Hsp = 0.09), mowing quality (Hc = 0.59, Hsp = 0.09), and genetic color (Hc = 0.45, Hsp = 0.06). The heritability estimates indicate that selection for these traits should result in significant gains in germplasm improvement. Differences were observed in the means and variances among clones, collection locations, and/or collection regions for many of the traits evaluated including rust severity (Puccinia spp.), spring green-up, plant height, lateral spread, vertical regrowth, and flowering traits. The positive correlations among some of these traits and those with moderate heritability estimates should allow for multi-trait selection in cultivar development.
Grape (Vitis vinifera) cluster compactness is an important trait due to its effect on disease susceptibility, but visual evaluation of compactness relies on human judgement and an ordinal scale that is not appropriate for all populations. We developed an image analysis pipeline and used it to quantify cluster compactness traits in a segregating hybrid wine grape (Vitis sp.) population for 2 years. Images were collected from grape clusters immediately after harvest, segmented by color, and analyzed using a custom script. Both automated and conventional phenotyping methods were used, and comparisons were made between each method. A partial least squares (PLS) model was constructed to evaluate the prediction of physical cluster compactness using image-derived measurements. Quantitative trait loci (QTL) on chromosomes 4, 9, 12, 16, and 17 were associated with both image-derived and conventionally phenotyped traits within years, which demonstrated the ability of image-derived traits to identify loci related to cluster morphology and cluster compactness. QTL for 20-berry weight were observed between years on chromosomes 11 and 17. Additionally, the automated method of cluster length measurement was highly accurate, with a deviation of less than 10 mm (r = 0.95) compared with measurements obtained with a hand caliper. A remaining challenge is the utilization of color-based image segmentation in a population that segregates for fruit color, which leads to difficulty in differentiating the stem from the fruit when the two are similarly colored in non-noir fruit. Overall, this research demonstrates the validity of image-based phenotyping for quantifying cluster compactness and for identifying QTL for the advancement of grape breeding efforts.
Seedlessness is one of the most desirable traits for table and raisin grapes. Stenospermocarpic cultivars are desirable because they have large berries with superior quality. Embryo rescue techniques have been widely used to get progeny seedling populations from crosses using seedless mother plants. Selection of the female parent, sampling time, and the growth medium are the most crucial to the success of this technique. This study investigated the effect of best sampling time and media composition on embryo rescue efficiency in a cold-hardy hybrid grape breeding program. We sampled ovules 5 to 9 weeks after flowering, and we tested four media compositions. The greatest percentages of embryo germination and normal seedlings were obtained when ovules were harvest at 8 weeks after flowering, indicating that it is suitable to harvest ovules at veraison, when the extraction of ovules is easier as a result of softer berry flesh. For the media composition experiment, all ovules were harvested at 8 weeks after flowering. Nitsch & Nitsch culture medium had very low germination, and the resulting seedlings performed the lowest compared with the other treatments. Lloyd & McCown Woody Plant Basal Medium (WPM) increased the number of embryos germinated significantly, and a number of normal seedlings and plantlets developed. Although there was no significant difference among the other three media containing WPM supplemented with different doses of plant regulators, the WPM Plus medium [with cytokinin (6-benzlaminopurine), indole-3-butyric acid, gibberellin, and casein hydrolysate] promoted the greatest percentage of established plants (46.98%). Therefore, the 8-weeks-after-flowering harvest time and the WPM Plus medium were selected for use in the embryo rescue protocol at the University of Minnesota grape breeding program.
The University of Minnesota Grape Breeding Program has developed cold-hardy wine grape cultivars that have facilitated the establishment of an economically important grape industry for the Midwest region. In recent years, the program has renewed efforts to breed cold-hardy table grapes. Table grapes might require postharvest storage if they are to be transported or stored for any period of time. Rachis dehydration, berry splitting, and decay can affect the postharvest quality of table grapes. In this study, we evaluated these postharvest traits in six released cultivars and nine advanced selections in the breeding program. For two growing seasons, we used industry standard packaging to assess postharvest traits (rachis dehydration, berry splitting, decay, and overall acceptability) at 2, 4, and 6 weeks of cold storage at 2.2 °C. The growing season had a significant effect on postharvest traits; therefore, the two were examined separately. There were significant differences in postharvest storage times for all traits, except berry splitting in 2020. Mean rachis dehydration reached unacceptable values (>3) after 4 weeks of postharvest storage in 2019 and after 6 weeks in 2020. All other trait means remained acceptable for many cultivars even after 6 weeks of postharvest storage. Advanced selections performed at and above the level of released cultivars, suggesting that selections will perform well in cold-hardy regions. The data collected regarding fruit quality and postharvest storage for two seasons will help to inform and improve breeding of cold-hardy grape cultivars.
Oakleaf hydrangea (Hydrangea quercifolia) is an understory shrub native to the southeastern United States. The species occupies a relatively small native range, and little is known about its demography, genetic diversity, or needs for conservation. Samples were collected from 188 plants in 73 locations throughout the species range and were genotyped using genotyping by sequencing. A structure analysis identified six genetic clusters that are geographically defined. Although these clusters are weakly differentiated, each has unique alleles. An environmental association analysis revealed that environmental variables explain 11.3% of genetic diversity, and population structure explains 13.5%. Further, 231 putative adaptive alleles were identified, most of which are correlated with precipitation-related variables, indicating that precipitation has an impact on genetic diversity in H. quercifolia. Many historically documented populations were found to be either extirpated or at risk of extirpation. The genetic clusters on the southern extent of the species range are relatively small and contain putative adaptive alleles at relatively high frequencies. These results highlight the importance of preserving representative germplasm from throughout the species range.
Oakleaf hydrangea (Hydrangea quercifolia Bartr.) is an understory shrub native to the southeastern United States. Hydrangeas are popular ornamental landscape plants; however, little is known about the diversity in horticulturally important traits for oakleaf hydrangea. Information regarding the variation in important traits could guide future breeding efforts for the species. Seed was collected from 55 populations throughout the range of the species for the purpose of conducting a horticultural characterization of the species compared with select cultivars. Plant architecture was characterized as plant height, number of nodes, internode length, number of branches, and plant width. Plant architecture was measured for container-grown and field-grown plants in two locations (Minnesota and Tennessee). Tolerance to leaf spot (Xanthomonas campestris L.) was characterized for wild-collected seedlings and cultivars by measuring disease severity under exposure to ambient inoculum. Cold hardiness was characterized during two winters with a controlled freezing experiment. During the first winter, seedlings were tested in January; during the second winter, seedlings and cultivars were tested monthly from October through April. Plant architecture varied by environment, with plants growing larger in Tennessee than in Minnesota. The heights of container-grown and field-grown plants were correlated with the collection site latitude (r = −0.66), with populations from the northeastern extent of the range of the species being the most compact, and populations from Florida being the tallest. Leaf spot severity varied significantly among populations and cultivars and was also correlated with latitude for the seedlings (r = 0.70). Two populations in Florida were identified as sources of high tolerance to leaf spot, whereas ‘Flemygea’ and ‘Alice’ were identified as having moderate tolerance to leaf spot. Cold hardiness varied among populations and cultivars and among months of the winter. The overall maximum cold hardiness was observed in February [mean lethal temperature (LT50) = −33.7 °C], and several populations maintained an extreme level of cold hardiness into late winter. Midwinter cold hardiness also varied by latitude (r = −0.65), with northern populations showing higher levels of cold hardiness. These results indicate that certain wild oakleaf hydrangea populations will be useful for introgressing novel variation into breeding programs.
Variegation in Vitis hybrids was investigated to confirm the inheritance as a single, recessive gene as previously proposed and commonly observed in breeding programs. Variegated leaves have ornamental appeal, but the phenotype is sublethal in some environments. Twenty-nine grape families were characterized for variegation including F1, S1, and S2 populations. The majority segregated 3 wild type (WT):1 variegated and were supported by chi-square tests. Four populations had segregation ratios supporting 15:1 or 1:1 models, and a unique flecking phenotype was identified in a Landot 4511 S1 population that suggested the interaction of two recessive loci. A variegated parent was selfed to produce progeny with no WT offspring and was segregated 0:1. Marker trait associations including bulk segregant analysis (BSA), genome-wide association mapping, and quantitative trait loci (QTL) mapping was used on three populations. On chromosome 14, Lvar1 was identified and mapped to 24.5 to 29.5 Mb and associated closely with rhAmpSeq marker 14_27607541. Lvar2 was associated with rhAmpSeq marker 11_18433819 on chromosome 11 at 12.2 to 18.4 Mb. The identification of two loci and the segregation data in some populations suggest that grape breeding germplasm segregates for two recessive loci. The pedigree records suggest that ‘Frontenac’ inherited one of these loci, and that Landot 4511, an ancestor of many populations tested in this experiment, may carry two loci. A total of 252 candidate genes were identified at these loci, including a key target—adenosine triphosphate (ATP)-dependent zinc metalloprotease FtsH6, involved in photosystem II and similar to the var2 mutant in Arabidopsis. This knowledge can help breeders select for ornamental grapevines or eliminate variegation from their breeding programs.