Citrus kinokuni ‘Mukaku kishu’ PI539530 and its progeny were studied to identify random amplified polymorphic DNA (RAPD) primers associated with seedlessness. Ninety-one F1 [(Robinson op) × C. kinokuni] individuals showed a 1:1 segregation ratio between seedless and seeded phenotypes with seedless as a single dominant gene. Bulked segregant analysis was used to identify markers associated with the seedless locus. Eighteen RAPD primers were mapped into a partial linkage group (≈55.8 cM length) with four RAPD primers flanking the seedless locus: OPAI11-0.8 at 8.7 cM, OPAJ19-1.0 at 8.4 cM, OPM06r-0.85 at 4.3 cM, and OPAJ04r-0.6 at 6.4 cM. The identification of molecular markers linked to C. kinokuni Fs seedless locus constitutes an important and major tool for citrus breeding and selection.
Dario J. Chavez and José X. Chaparro
Dario J. Chavez and Paul M. Lyrene
In Vaccinium L., most tetraploid hybrids between tetraploid cultivated highbush blueberry (V. corymbosum L.) and diploid darrow's evergreen blueberry (V. darrowii Camp) have been produced by exploiting V. darrowii's tendency to produce 2n gametes and a strong triploid block, which greatly reduces the number of triploid hybrids produced. Colchicine-derived tetraploids offer an alternative method of producing V. darrowii plants that will easily cross with tetraploid Vaccinium species. V. darrowii (2n = 2x = 24) seeds were imbibed in 0.2% aqueous colchicine solution for 24 h. The seeds were germinated and seedlings whose morphology suggested colchicine effects were selected at various stages of development. No macromorphological changes were consistently associated with chromosome doubling. However, stomatal guard cells and pollen size increased substantially as a result of chromosome doubling. Several types of plants were identified after colchicine treatment: 1) plants with a doubled LI (epidermal tissues) and LII (internal tissues) plant layers; 2) periclinal chimeras with a doubled LI layer and a normal LII; and 3) periclinal chimeras with a doubled LII layer and a normal LI. Of ≈4000 seedlings that emerged from colchicine-treated seeds, 200 were selected for further examination based on leaf and stem morphology. Of the 200, five appeared to be tetraploid in LI, LII, or both layers based on stomatal guard cell size and pollen size. Crosses between colchicine-derived V. darrowii (4x) plants and southern highbush blueberry (V. corymbosum) cultivars (4x) were successful compared with 4x–2x and 2x–4x crosses using diploid V. darrowii and tetraploid southern highbush blueberry. Stomatal guard cells and pollen screening of the colchicine-treated plants were used as indicators of doubled V. darrowii plants and periclinal chimeras. The results from the crosses between colchicine-treated V. darrowii plants and tetraploid highbush blueberry cultivars confirmed the information obtained by stomata and pollen screening.
Dario J. Chavez and Paul M. Lyrene
Diploid Vaccinium darrowii Camp has been used in breeding tetraploid southern highbush blueberry (Vaccinium corymbosum L.) as a source of reduced chilling requirement, adaptation to hot, wet summers, and resistance to leaf diseases. V. darrowii in Florida is quite variable, but most crosses have involved only one V. darrowii clone, Fla. 4B. The use in breeding of a wider range of V. darrowii accessions would provide beneficial diversity in the blueberry cultivated gene pool. The purpose of this research was to determine the functional 2n gamete frequency of numerous V. darrowii genotypes when crossed with tetraploid V. corymbosum, and to study the pollen fertility and backcross ability of the interspecific (V. darrowii × V. corymbosum) hybrids to V. corymbosum. Crosses between diploid V. darrowii and tetraploid highbush blueberry cultivars had low fruit set compared with the V. darrowii × V. darrowii and highbush × highbush crosses. The unusually high number of hybrids per pollinated flower (HPF) in certain 4x-2x or 2x-4x crosses was attributed to high functional 2n gamete production in certain V. darrowii genotypes. Diploid Vaccinium fuscatum Aiton and diploid V. darrowii × V. fuscatum hybrids, when crossed with southern highbush blueberry cultivars, were equally productive of hybrids whether used as male or female parents. Variation in frequency of functional 2n gametes in V. darrowii, expressed as high HPF, was present within plants (megaspores vs. microspores) and among V. darrowii plants. Of the 114 interspecific (V. darrowii × V. corymbosum) hybrids studied, 106 had pollen stainability >50%. This indicated that most of these hybrids were tetraploid, because triploid blueberries, like most triploid plants, are highly sterile. Twenty-two V. darrowii × V. corymbosum hybrids were backcrossed to tetraploid highbush blueberry cultivars. Fruit set was variable, but large populations of vigorous hybrids were obtained. Lower fruit set was associated with hybrids that had lower pollen fertility. It should be possible to obtain plants of cultivar quality in a few generations of backcrosses.
Dario J. Chavez, Eileen A. Kabelka and José X. Chaparro
Phytophthora capsici causes seedling death, crown and root rot, fruit rot, and foliar blight on squash and pumpkins (Cucurbita spp. L.). A total of 119 C. moschata accessions, from 39 geographic locations throughout the world, and a highly susceptible butternut squash cultivar, Butterbush, were inoculated with a suspension of three highly virulent P. capsici isolates from Florida to identify resistance to crown rot. Mean disease rating (DR) of the C. moschata collection ranged from 1.4 to 5 (0 to 5 scale with 0 resistant and 5 susceptible). Potential resistant and tolerant individuals were identified in the C. moschata collection. A set of 18 PIs from the original screen were rescreened for crown rot resistance. This rescreen produced similar results as the original screen (r = 0.55, P = 0.01). The accessions PI 176531, PI 458740, PI 442266, PI 442262, and PI 634693 were identified with lowest rates of crown infection with a mean DR less than 1.0 and/or individuals with DR = 0. Further selections from these accessions could be made to develop Cucurbita breeding lines and cultivars with resistance to crown rot caused by P. capsici.
Dario J. Chavez, Thomas G. Beckman and José X. Chaparro
Prunus phylogeny has been extensively studied using chloroplast DNA (cpDNA) sequences. Chloroplast DNA has a slow rate of evolution, which is beneficial to determine species relationships at a deeper level. The chloroplast-based phylogenies have a limitation due to the transfer of this organelle by interspecific hybridization. This creates difficulties when studying species relationships. Interspecific hybrids in Prunus occur naturally and have been reported, which creates a problem when using cpDNA-based phylogenies to determine species relationships. The main goal of this project was to identify nuclear gene regions that could provide an improved phylogenetic signal at the species level in Prunus. A total of 11 species in Prunus and within section Prunocerasus were used. Two peach (Prunus persica) haploids were used to test the reliability of the molecular markers developed in this project to amplify single-copy genes. A total of 33 major genes associated with vernalization response, 16 with tree architecture, and 3 with isozymes, were tested. Similarly, 41 simple sequence repeat (SSR) markers, seven cpDNA regions, and the internal transcribed spacer (ITS) region, were used. Multiple gene regions were identified and provided the greatest number of characters, greatest variability, and improved phylogenetic signal at the species level in Prunus section Prunocerasus. Out of those, trnH-psbA, PGI, MAX4, AXR1, LFY, PHYE, and VRN1 are recommended for a phylogenetic analysis with a larger number of taxa. The use of potentially informative characters (PICS) as a measure of how informative a region will be for phylogenetic analyses has been previously reported beneficial in cpDNA regions and it clearly was important in this research. This will allow selecting the region(s), which can be used in phylogenetic studies with higher number of taxa.
Jun Liu, Orville M. Lindstrom and Dario J. Chavez
Differential thermal analysis (DTA) has great potential as a quick and convenient cold hardiness determination method in plants. It measures freezing events inside of plant samples by detecting exotherm(s) produced when water changes from liquid to solid phase. DTA is highly sensitive to the experimental conditions and it has been reported to be ineffective among different fruit crops after acclimation of floral buds has occurred. The objective of this project was to establish DTA as a rapid and accurate method to predict peach floral bud cold hardiness from acclimation to deacclimation as compared with the traditional standard artificial freezing test. Floral buds of ‘Elberta’ and ‘Flavorich’ peach cultivars were subjected to DTA and artificial freezing tests throughout the winters of 2015–16 and 2016–17. Before deacclimation, two distinct exotherms, low-temperature exotherms (LTE) and high-temperature exotherms (HTE), were normally detected from floral bud DTA analyses. After deacclimation, DTA tests yielded only a few LTEs. However, incubation of floral buds at −2 °C overnight before the cooling process of DTA tests yielded an increased number of LTEs for both seasons in comparison with samples directly run using DTA without incubation. Similarly, after deacclimation started, the temperature in which LTE occurred was correlated (r = 0.59–0.86) with LT50 (lethal temperature that damaged 50% of floral buds) when DTA samples were treated overnight at −2 °C. In our study, pretreatment of floral buds at −2 °C overcame the inability of DTA to detect LTEs after deacclimation, which improved the ability and reliability of DTA to detect LTEs for more than 50% of the buds used per date per cultivar. DTA is a promising method to predict cold hardiness of peach plants.
Catherine Belisle, Uyen T.X. Phan, Koushik Adhikari and Dario J. Chavez
Peach (Prunus persica) production in the southeastern United States extends from mid-May to mid-September. There are ≈60 peach cultivars commercially grown. Each cultivar has unique fruit quality characteristics, which could influence consumer perception and likability. The present study is a survey of chemical and physical characteristics of mature, commercially grown, fresh peaches in Georgia. A collection of 30 cultivars was evaluated in 2015 and 2016 for soluble solids concentration (SSC), total titratable acidity (TTA), SSC/TTA ratio, texture (compression, puncture, and Kramer shear), and skin and flesh color (CIE L*, chroma, and hue color space values). There was significant variation between seasons for all variables (P < 0.05) except for TTA (P = 0.12), and flesh hue values (P = 0.38). Statistical differences among cultivars within each year were reported for all variables (P < 0.0001). SSC showed variation seasonally and among cultivars, whereas TTA variation was mainly attributed to cultivar differences. Similarly, cultivar-to-cultivar differences were found when comparing the different texture tests evaluated with cultivars such as Goldprince, Early August Prince, Flameprince, Majestic, and Red Globe having the most variation between seasons. Other cultivars analyzed had little variation between seasons. Moreover, firmness differences observed across the three texture tests were inconsistent. In peach skin color, significant variation was observed for L*, chroma, and hue among cultivars. Skin hue and chroma were highly correlated within each season (r = 0.77 for 2015, r = 0.72 for 2016). The results of this survey demonstrate the variation of quality characteristics for a large selection of peach cultivars grown in Georgia. The information reported in this paper will be used as a baseline for further examining and understanding peach fruit quality.
Thomas G. Beckman, Philip A. Rollins, James Pitts, Dario J. Chavez and Jose X. Chaparro
The primary focus of the stone fruit rootstock program at Byron, GA, has been the development of disease-resistant rootstocks for peach (Prunus persica L. Batsch). Historically peach tree short life (PTSL), aka bacterial canker complex, and Armillaria root rot (ARR) have been the two most important causes of premature mortality of commercial peach trees in the southeastern United States. Guardian®, a seedling peach rootstock, was cooperatively released in 1993 by the U.S. Department of Agriculture (USDA)-Agricultural Research Service (ARS) and Clemson University. It has since been widely adopted by the southeastern peach industry. As a result, trees losses to PTSL have declined sharply. However, Guardian, like most other peach seedling rootstocks, is susceptible to ARR. ARR has now moved to the forefront as the primary cause of premature peach tree death in the Southeast. In response to this threat, the USDA-ARS in cooperation with the University of Florida, released ‘Sharpe’, a plum hybrid rootstock in 2007. Despite its broad disease resistance, ‘Sharpe’ proved unsuited for widespread commercial utilization due to its relatively poor cropping performance. In 2011, ‘MP-29’, a semidwarf, clonal, plum × peach hybrid, was released for commercial trial. ‘MP-29’s broad disease and nematode resistance, in combination with its dwarfing ability and excellent productivity, offered great promise for use in this production area and in others suffering from similar issues. Since its release, testing of ‘MP-29’ has continued both in researcher and grower trials. To date, performance has exceeded all expectations.