bluuebunny/crossref_metadata_2025 huggingface dataset:
Impact of Plant Breeding on the Genetic Diversity of Cultivated Strawberry as Revealed by Expressed Sequence Tag-derived Simple Sequence Repeat Markers
Authors: Gil-Ariza David Jesús , Amaya Iraida , López-Aranda José Manuel , Sánchez-Sevilla José Federico , Ángel Botella Miguel , Valpuesta Victoriano
Published: January 2019

Unlike other important crops analyzed so far for genetic diversity and population structure, the brief history and particularities of the genetics of the cultivated strawberry (Fragaria ×ananassa Duchesne) have limited its genetic characterization. The genomic composition and the pattern of inheritance have not been fully elucidated, although a number of studies have suggested a highly diploidized genome. In this study, the similarity relationships and structure of 92 selected strawberry cultivars with widely diverse origins have been established using simple sequence repeat (SSR) markers derived from expressed sequence tags (EST-SSR markers). Genetic analysis performed by the unweighted pair group method with arithmetic mean clustering revealed a distribution according to both date of cultivar release and breeding for a specific climatic adaptation. Additionally, a model-based clustering approach identified three populations among the strawberry cultivars with an overall FST value of 0.15 to 0.16. Both analyses support a limited differentiation of modern cultivars, most probably as a consequence of the methodology of strawberry breeding. Interestingly, the collection of strawberry cultivars here analyzed showed comparable genetic differentiation to that observed in natural populations of Fragaria chiloensis (L.) Mill., one of its wild ancestors. Our results suggest that breeding has produced a small but significant reduction on the genetic diversity of F. ×ananassa. The panel of 10 EST-SSRs described in this work provided an extremely low probability of confusion (less than 10−11), offering an efficient and accurate method for cultivar identification.
Polymorphism and Genetic Relationships among Tea Genotypes from Turkey Revealed by Amplified Fragment Length Polymorphism Markers
Authors: Kafkas Salih , Ercişli Sezai , Doğan Yıldız , Ertürk Yaşar , Haznedar Ayhan , Sekban Remzi
Published: January 2019

Individuals in most countries around the world drink tea (Camellia sinensis). Tea drinking has attained ceremonial status in many places as a social and medicinal beverage. Although tea is of great importance in Turkey's economy, little is known about the pattern of genetic variation among the various tea genotypes grown in Turkey. A total of 32 tea genotypes found at the Ataturk Tea and Horticulture Research Institute in the eastern Black Sea region of Turkey were sampled. Fluorescent dye amplified fragment length polymorphism (AFLP) markers and capillary electrophoresis were applied for molecular characterization. The AFLP analysis with six primer combinations generated 835 fragments of which 567 were polymorphic, corresponding to 69.8% polymorphism. Resolving powers of the AFLP primers ranged from 62.6 to 81.9, yielding a total of 437.8; the polymorphic information content (PIC) ranged from 0.76 to 0.83, with an average of 0.79. Genetic similarity values ranged from 0.68 to 0.92, with an average of 0.76. The dendrogram derived by unweighted pair group method with arithmetic mean algorithm (UPGMA) and principal coordinate analysis (PCoA) revealed that all tea genotypes could be clearly divided into four distinct clusters. The results of this study will provide valuable information to the tea cultivar breeding program for the purpose of parental selection.
Evaluation of Flowering Cherry Species, Hybrids, and Cultivars Using Simple Sequence Repeat Markers
Authors: Ma Hongmei , Olsen Richard , Pooler Margaret , Kramer Matthew
Published: January 2019

Flowering cherries belong to the genus Prunus, consisting primarily of species native to Asia. Despite the popularity of ornamental cherry trees in the landscape, most ornamental Prunus planted in the United States are derived from a limited genetic base of Japanese flowering cherry taxa. A diverse collection of ornamental Prunus germplasm is maintained at the U.S. National Arboretum as part of an ongoing flowering cherry improvement program, but the genetic backgrounds of many trees are unclear. We characterized this germplasm using five simple sequence repeat (SSR) primer pairs, including one chloroplast primer pair. These primers generated 140 unique alleles that were used to assess genetic relationships among species, hybrids, and cultivars in this collection. We found that these markers followed expected Mendelian inheritance from parents to progeny in controlled hybridizations. In general, species clustered according to published taxonomic groupings, including a distinct separation of the ornamental cherries (Prunus subgenus Cerasus section Pseudocerasus) from other subgenera. Individual accessions of several taxa did not cluster with other samples of the species, indicating possible misidentification or interspecific combinations. The resulting information will be useful in guiding decisions on breeding methodology and germplasm preservation.
Genetic Variation and Covariation of Yield and Phytochemical Traits in a Red Raspberry Factorial Study
Authors: Stephens M. Joseph , Scalzo Jessica , Alspach Peter A. , Beatson Ron A. , Connor Ann Marie
Published: January 2019

Breeding for high yield is a major objective of most small fruit breeding programs worldwide. In recent years, research associated with health benefits of berry fruit has resulted in some breeding programs looking for material with improved health properties with the goal of incorporating these into genotypes with high yield and other favorable agronomic characters. In this study, we estimated variance components, heritabilities, and phenotypic and genotypic correlations for yield components and phytochemicals [total phenolics (TPH), antioxidant activity (FRAP), and total anthocyanins (TACY)] from 828 genotypes in a red raspberry (Rubus idaeus) factorial mating design based on 42 full-sib families derived from seven female and six male parents harvested in one season in New Zealand. Narrow sense heritability estimates varied from moderately low [0.23 for percent budburst (PCBB)] to high [0.82 for mean berry weight (BWT)]. Highest genotypic correlations with total yield (TYLD) were found for mean cane length (0.60) followed by moderate correlations for PCBB (0.38), BWT (0.34) and fruit number per unit lateral (0.31). For these four components, the correlation between the product of the empirical breeding values (eBV) and TYLD was as good (0.67) as the correlation between the product of the seven yield components measured (0.68). Moderately high negative genotypic correlations were found between TYLD and TPH (–0.67), FRAP (–0.68), and TACY (–0.64), suggesting that breeding for high-yielding genotypes may result in reduced phytochemical levels. A pigment-deficient R. parvifolius × R. idaeus hybrid derivative parent (R. parv deriv) had the most influence on berry weight as a yield component. Removal of the R. parv deriv progeny from the analysis increased the contribution of berry weight to total yield. Heritability estimates were generally lower when the R. parv deriv crosses were omitted, particularly for number of canes BWT, TYLD, and the phytochemicals. Implications of R. parv deriv crosses on these results are discussed. Of the 828 genotypes, only five had an eBV for total yield greater than that of the best yielding parent, whereas 49 had larger berries than the best parent. None of the 828 genotypes had an eBV for yield components or phytochemicals less than that of the lowest parent, whereas 13 had a higher eBV associated with phytochemicals than that of the highest parent.
Physiological and Morphological Changes Over the Past 50 Years in Yield Components in Tomato
Authors: Higashide Tadahisa , Heuvelink Ep
Published: January 2019

Greenhouse tomato (Solanum lycopersicum) yield in The Netherlands has increased tremendously over the past 50 years. The effects of breeding during this period were investigated. Eight Dutch cultivars and one typical current Japanese cultivar that were released over the past 50 years were compared in a short-term experiment conducted from summer to fall in The Netherlands. Fresh fruit yield of the Dutch cultivars significantly increased ≈0.9% per year with the year of release from 1950 to 2000. Dry weight fruit yield of the Dutch cultivars also increased with the year of release, whereas the fruit dry matter content was not correlated with the year of release. Total dry matter production of plants increased with the year of release, and the dry matter partitioning to fruit was not correlated with the year of release. An increase in dry matter production was caused not by an increase in fraction of intercepted light, but by light use efficiency based on correlations between each of them and the year of release. The light extinction coefficient in the plant canopy decreased, whereas leaf photosynthetic rate increased significantly with the year of release. Although fresh fruit yield of the Japanese cultivar was lower than that of the modern Dutch cultivars, fruit dry matter content of the Japanese cultivar was higher than that of the Dutch cultivars. An increase in yield over the past 50 years in Dutch tomato was caused by an increase in light use efficiency resulting from a decrease in light extinction coefficient (a morphological change) and an increase in leaf photosynthetic rate (a physiological change).
Volatiles Production and Lipoxygenase Gene Expression in Kiwifruit Peel and Flesh During Fruit Ripening
Authors: Zhang Bo , Yin Xue-Ren , Shen Ji-Yuan , Chen Kun-Song , Ferguson Ian B.
Published: January 2019

The relationship between lipoxygenase (LOX) pathway-derived volatiles and LOX gene expression was evaluated in kiwifruit [Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson var. deliciosa cv. Bruno] during postharvest ripening at 20 °C. The C6 aldehydes n-hexanal and (E)-2-hexenal were abundant in peel compared with flesh tissue and declined as kiwifruit ripened. Esters such as ethyl butanoate and methyl butanoate were lower in the peel than flesh and accumulated when the fruit underwent a climacteric rise in ethylene production. Total LOX activity was higher in the peel than in the flesh and increased as kiwifruit ripened. Expression of AdLox2, AdLox3, AdLox4 and AdLox6 was high in the peel, whereas AdLox1 and AdLox5 showed similar levels in the peel and flesh at the ethylene climacteric. AdLox1 and AdLox5 transcript levels increased and AdLox2, AdLox3, AdLox4 and AdLox6 levels decreased during postharvest fruit ripening. Principal component analysis showed that n-hexanal and (E)-2-hexenal were grouped with LOX genes that were downregulated as kiwifruit ripened. The possible roles of LOX genes in relation to kiwifruit volatile formation during fruit ripening are discussed.
Changes in the Phenolic Concentration during Flower Development of Rose ‘KORcrisett’
Authors: Schmitzer Valentina , Veberic Robert , Osterc Gregor , Stampar Franci
Published: January 2019

The concentration of major anthocyanins, quercetins, catechin, and phenolic acids during flower development of Rosa ×hybrida L. ‘KORcrisett’ was quantified using high-performance liquid chromatography/mass spectrometry. Additionally, the changes in petal color were monitored colorimetrically at four different stages of development (bud, partially open flowers, fully open flowers, senescent flowers) and correlation was calculated between the chromaticity parameters and major/total anthocyanins. Color parameters a*, b*, and h° decreased with the progression of flower development and a*/b* ratio and lightness (L*) increased. In rose petals, a negative trend in the content of major (pelargonidin-3,5-di-O-glucoside, cyanidin-3,5-di-O-glucoside) and minor (pelargonidin-3-O-glucoside, cyanidin-3-O-glucoside, peonidin-3-O-glucoside) anthocyanins was observed during flower development. Buds contained almost threefold higher concentrations of pelargonidin-3,5-di-O-glucoside and fourfold higher concentrations of cyanidin-3,5-di-O-glucoside than senescent flowers. Buds also contained significantly more quercetins (quercetin-3-O-rutinoside, quercetin-3-O-glucoside, and quercetin-3-O-rhamnoside), catechin, and phenolic acids (gallic acid, protocatechulic acid, chlorogenic acid, caffeic acid, p-coumaric acid) than flowers of subsequent developmental stages. The most significant differences were observed in the content of gallic acid; buds contained almost sixfold higher values than senescent flowers. Correlation analysis revealed a strong correlation between chromaticity parameters a*, b*, a*/b* ratio, h°, L*, and major/total anthocyanins with values ranging from 0.60 to –0.84.
Effects of Trinexapac-ethyl on Drought Responses in Creeping Bentgrass Associated with Water Use and Osmotic Adjustment
Authors: Bian Xiuju , Merewitz Emily , Huang Bingru
Published: January 2019

Understanding factors influencing drought resistance traits is important for improving turfgrass growth in water-limited environments. The objectives of this study were to examine effects of a plant growth regulator, trinexapac-ethyl (TE), on turf growth and water use for creeping bentgrass (Agrostis stolonifera L.) exposed to drought stress, and to determine changes in the accumulation of solutes involved in osmotic adjustment associated with TE application. Plant foliage of cultivar L-93 was sprayed with 1.95 mL·L−1 of TE at 0.113% a.i. 14 days before and at the initiation of drought stress. TE-treated and untreated plants were exposed to well-watered or drought stress conditions for 28 days in a growth chamber. TE-treated plants exhibited a reduced rate of water depletion from the soil as demonstrated by higher soil water content, lower evapotranspiration rates, and higher leaf relative water content during 28 days of drought stress compared with non-TE-treated plants. During the later phase of drought stress, TE-treated plants had a greater reduction in leaf ψS at full turgor or greater osmotic adjustment, which was associated with increased accumulation of soluble sugars and inorganic ions (Ca and K) in leaves of TE-treated plants. Proline content increased in response to drought stress, but was unaffected by TE application, suggesting that it may not contribute to the effects of TE on osmotic adjustment. TE-treated plants maintained significantly higher turf quality and leaf photochemical efficiency under drought stress. The results suggest that the promotive effects of TE application on turf growth during drought stress were associated with the reduction in water depletion or lower water use and increases in osmotic adjustment due to the accumulation of inorganic solutes and soluble sugars.
Profiling of Nondigestible Carbohydrate Products in a Complete Set of Alien Monosomic Addition Lines Explains Genetic Controls of Its Metabolisms in Allium cepa
Authors: Yaguchi Shigenori , Nakajima Tetsuya , Sumi Toshihisa , Yamauchi Naoki , Shigyo Masayoshi
Published: January 2019

Eight kinds of japanese bunching onion [Allium fistulosum L. (genomes FF)]-shallot [Allium cepa L. Aggregatum group (genomes AA)] monosomic addition line [MAL (FF+1A-FF+8A)] were used to study the effects of single alien chromosomes from A. cepa on the production of carbohydrates in the leaf tissues of A. fistulosum. Carbohydrate contents in green leaf blades of these MALs were measured during alternate months from May 2005 to Mar. 2006. The determination of soluble sugar content from leaf blades of each MAL and A. fistulosum revealed that nonreducing sugars (sucrose and fructan) accumulated in winter leaf blades. Reducing sugar (fructose) in the leaf blades of each MAL was lower than A. fistulosum in almost every time period. In the leaf blades of FF+4A, high fructan accumulation was observed from Nov. 2005 to Mar. 2006. A series of determinations on the pectin content showed that amounts of NaOH-soluble pectin and HCl-soluble pectin remained at low levels at all time periods. High pectin accumulations in FF+7A and FF+8A occurred in September and slightly decreased in November. A decrease in hexametaphosphoric acid-soluble pectin content was associated with the maturity of MALs from autumn to winter, whereas the water-soluble pectin content increased. The pectin methylesterase and polygalactronase genes of shallot were assigned to chromosome 7A and 4A, respectively. These results demonstrate that important genes related to pectin metabolism in shallot are located on chromosomes 4A, 7A, and 8A of shallot.
Pollen Tube Behavior in Different Mandarin Hybrids
Authors: Distefano Gaetano , Casas Giuseppina Las , La Malfa Stefano , Gentile Alessandra , Tribulato Eugenio , Herrero Maria
Published: January 2019

Seedlessness is an important trait in the evaluation of commercial mandarin for fresh consumption. However, in the last decade, the presence of seeds in fruit of cultivars considered as seedless has become a problem in different citrus-growing areas because the commercial value is depreciated. Seeds have appeared concomitantly with the introduction of new cultivars that appear to be cross-compatible. To overcome this problem, different strategies have been explored, but a definitive answer is still elusive. The search for alternatives contrasts with how little is known about the basis of the problem: the pollen-pistil incompatibility reaction in mandarin, and the intercompatibility relationship between different cultivars. In this work, we characterized the pollen-pistil incompatibility in the two commonly grown mandarin cultivars Fortune and Nova, and evaluated the intercompatibility relationship between six cultivars with different genetic origins; these cultivars included Fortune (Citrus clementina Hort. ex Tan. × Citrus reticulata Blanco), Nova [(Citrus paradisi Macf. × C. reticulata) × C. clementina), Comune Clementine (C. clementina), Avana apireno (C. reticulata), Primosole (Citrus unshiu Marcov. × C. reticulata), and Simeto (C. unshiu × Citrus deliciosa Ten.). Following the controlled hand pollination in the field, we evaluated pollen tube performance after self- and cross-pollination, as well as intercultivar compatibility by observing pollen tube growth. The results show the self-incompatibility of these cultivars with the pollen tube having been arrested in the style, which explains their seedless condition when planted in solid blocks. The study of intercompatibility indicates a different pollen tube behavior depending on the genotype, as well as on the cross combinations. These results provide a basis to evaluate self- and intercompatibility in citrus, and the effect of close planting of some cultivars.
Genetic Analyses of Soluble Carbohydrate Concentrations in Onion Bulbs
Authors: Raines Steven , Henson Cynthia , Havey Michael J.
Published: January 2019

Fructans are the main soluble carbohydrate in onion (Allium cepa) bulbs and their concentrations show significant correlations with dry weights and pungency. In previous research, we identified regions on chromosomes 5 and 8 associated with higher amounts of soluble carbohydrates in onion bulbs. In this research, we estimated the genetic effects and interactions between these two chromosome regions using larger inbred families grown in field trials over 3 years. Bulbs were evaluated 30 and 90 days after harvest (DAH) for dry weights and soluble carbohydrates. Fructan concentrations decreased significantly between 30 and 90 DAH, consistent with loss of bulb dormancy over this period. Dry weights were negatively correlated with fructose and positively correlated with sucrose and fructans. Analyses of variance and interval mapping revealed that the region on chromosome 5 affected bulb dry weight, whereas the region on chromosome 8 significantly affected both dry weight and fructan concentrations. Regions on chromosomes 5 and 8 showed dominance for increased dry weights and/or soluble solids. Interactions between regions on chromosomes 5 and 8 were significant only for sucrose and the fructan neokestose, indicating that these regions independently contribute to higher amounts of soluble carbohydrates. These results demonstrate that onions with low concentrations of soluble carbohydrates were developed by selecting for relatively few recessively inherited chromosome regions.
Characterization of the Male-sterile Line Bcajh97-01A/B and Identification of Candidate Genes for Genic Male Sterility in Chinese Cabbage-pak-choi
Authors: Huang Li , Ye Wan-zhi , Liu Ting-ting , Cao Jia-shu
Published: January 2019

Cytological features of ‘Aijiaohuang’ chinese cabbage-pak-choi (Brassica campestris ssp. chinensis) Bcajh97-01A/B genic male-sterile AB line were examined to determine phenotypic reasons for male sterility. The sterile line Bcajh97-01A was found to undergo aberrant cytokinesis during male meiosis. Transcriptional profiling of the flower buds of both fertile and sterile plants was performed at the periods preceding meiosis, at the tetrad to uninucleate pollen period, and at the binucleate to mature pollen period. Transcript-derived fragments (TDFs) from corresponding genes that were expressed in flower buds at these three different stages could be divided into nine classes. We sequenced a total of 14 new TDFs that were differentially displayed at particular pollen developmental stages, including eight genes with unknown or hypothetical functions and six genes showing significant homology with known genes. This characterization of the Bcajh97-01A genic male-sterile line allowed the identification of candidate genes underlying genic male sterility.
Accumulation of End Products in Source Leaves Affects Photosynthetic Rate in Peach via Alteration of Stomatal Conductance and Photosynthetic Efficiency
Authors: Cheng Jieshan , Fan Peige , Liang Zhenchang , Wang Yanqiu , Niu Ning , Li Weidong , Li Shaohua
Published: January 2019

In ‘Beijing 24’ peach [Prunus persica (L.) Batch] trees, a series of source leaves with differing levels of end products were created by retaining fruit (“+fruit”), removing fruit (“−fruit”), or reducing the light period. To alter the light period, leaves were covered with a bag made of brown inner paper and outer silver paper, which was then removed at different times the next day. The highest level of end products were obtained by fruit removal, while reducing the light period resulted in a lower level than “+fruit.” Net photosynthetic rate (Pn) and stomatal conductance (gs) decreased, but leaf temperatures (Tleaf) increased, following an increase in end product levels in leaves. After the “−fruit” treatment, reduced Pn was correlated with lower gs, and Tleaf increase was concomitant with decreases in maximal quantum yield of photosystem II (Fv/Fm), actual photochemical efficiency of photosystem II (ΦPSII), and photochemical quenching, and with an increase in nonphotochemical quenching. However, there were no significant differences in chlorophyll fluorescence between “+fruit” and the two treatments reducing the light period. The ΦPSII decreased following an increase in foliar sorbitol level, and it linearly decreased as sucrose and starch increased. Although fruit removal resulted in a significant accumulation of sucrose, sorbitol, and starch in leaves throughout the day, the extractable activities of several important enzymes involved in carbohydrate leaf storage and translocation did not decrease. Therefore, instead of feedback regulation by the accumulation of end products in source leaves, a high Tleaf induced by decreased stomatal aperture may play a key role in regulation of photosynthesis by limiting the photochemical efficiency of the PSII reaction centers under high levels of the end products in peach leaves.
Applying Spermidine for Differential Responses of Antioxidant Enzymes in Cucumber Subjected to Short-term Salinity
Authors: Du Chang-Xia , Fan Huai-Fu , Guo Shi-Rong , Tezuka Takafumi
Published: January 2019

To examine whether spermidine (SPD) modifies plant antioxidant enzyme expression in response to short-term salt stress, cucumber (Cucumis sativus) seedlings were treated with NaCl in the presence or absence of SPD for 3 days. Compared with untreated control plants, free radical production and malondialdehyde content in leaves and roots increased significantly and plant growth was suppressed under 50 mm NaCl stress. Exogenous SPD sprayed on leaves at a concentration of 1 mm alleviated salinity-mediated growth reduction. Salt stress caused a consistent increase in soluble protein content, as well as peroxidase (POD) and superoxide dismutase (SOD) activities in cucumber seedlings. By native polyacrylamide gel electrophoresis, five POD isozymes were detected in cucumber seedling leaves, and seven in roots. We detected five SOD isozymes in leaves and four in roots, and two catalase (CAT) isozymes in leaves and two in roots. Our results indicate that salt stress induced the expression of POD and SOD isozymes in cucumber seedlings, but inhibited the expression of CAT isozymes in roots. Application of exogenous SPD further increased POD and SOD expression and activity, and led to the differential regulation of CAT in leaves and roots. These data show that antioxidant enzymes, especially POD and SOD, appear to protect cucumber seedlings against stress-related damage, and they appear to function as the molecular mechanisms underlying the response of cucumber seedlings to salinity. Moreover, SPD has potential to scavenge directly free radical and to alleviate growth inhibition and promote the activity and expression of antioxidant system enzymes in cucumber seedlings under short-term salt stress.
Salinity Tolerance and Leaf Water Use Efficiency in Citrus
Authors: Syvertsen James P. , Melgar Juan C. , García-Sánchez Francisco
Published: January 2019

In three separate experiments, the growth and water use of salinized citrus rootstock seedlings and grafted trees were modified using different growth substrates, elevated CO2, or 50% shade screen under field conditions. By reanalyzing previously published data, we tested the hypothesis that salinity tolerance in citrus can be characterized as the ability to maintain low levels of leaf Cl− accumulation through high plant growth and high water use efficiency (WUE) under saline conditions. Well-irrigated salinized seedlings of the relatively salt-sensitive Carrizo citrange [Carr (Citrus sinensis × Poncirus trifoliata)] were grown in sand, clay, or a peat-based soilless media. Salinity stress reduced plant growth and water use. Leaf Cl− concentration was negatively related to plant growth, but leaf Cl− increased with transpiration rate in low-saline treatments. In a second experiment using salinized seedlings of the relatively salt-tolerant Cleopatra mandarin [Cleo (Citrus reticulata)] grown along with Carr seedlings with or without elevated CO2, leaf Cl− was negatively related to growth and to shoot/root dry weight ratio, but was positively related to water use such that leaf Cl− was negatively related to leaf WUE. In a third experiment using salinized 2-year-old ‘Valencia’ orange (C. sinensis) trees grafted on Cleo or Carr rootstocks and grown with or without shadecloth, leaf Cl− was positively related to leaf transpiration as both were higher in the spring than in the fall, regardless of rootstock or shade treatment. Overall, leaf Cl− was positively related to water use and was negatively related to leaf WUE. High growth, low water use, and consequently, high WUE of salinized citrus were related to low leaf Cl−. Such relationships can be used as indicators of salinity tolerance.
Functional Identification of a C-repeat Binding Factor Transcriptional Activator from Blueberry Associated with Cold Acclimation and Freezing Tolerance
Authors: Polashock James J. , Arora Rajeev , Peng Yanhui , Naik Dhananjay , Rowland Lisa J.
Published: January 2019

Highbush blueberry (Vaccinium corymbosum) is susceptible to winter freezing injury and frost damage in the spring. As part of an ongoing project to understand the process of cold acclimation, we isolated a C-repeat binding factor (CBF) transcriptional activator gene-coding region from the highbush blueberry cultivar Bluecrop. Expression of the highbush blueberry CBF gene was compared in floral buds of the cold-tolerant northern highbush cultivar Bluecrop and the more cold-sensitive southern rabbiteye (V. virgatum) blueberry cultivar Tifblue. Relative gene expression was higher in ‘Bluecrop’ than in ‘Tifblue’. Expression in both cultivars was highest at the earliest time point in the fall (coincident with the first stage of cold acclimation), declined during the later fall and winter, and, in ‘Bluecrop’, increased again as buds deacclimated, when temperatures tend to fluctuate. To confirm the putative identity of the gene as a member of the CBF gene family, and to determine if expression in a heterologous system could enhance freezing tolerance, the blueberry gene coding sequence was overexpressed in transgenic Arabidopsis thaliana under the control of the cauliflower mosaic virus 35S promoter. Transgenic plants expressing the putative blueberry CBF gene exhibited induced expression of the A. thaliana cold-regulated (COR) genes COR78 and COR6.6, under non-inducing conditions (i.e., 23 °C); however, expression of two other COR genes was unaffected. Transgenic plants also exhibited enhanced freezing tolerance under non-acclimated conditions, but not to the level of acclimated control plants. Thus, the expression pattern in floral buds and the ability of the isolated gene to turn on a subset of COR genes and increase freezing tolerance in a heterologous system suggest it is a functional member of the CBF gene family in blueberry.
Identification and Mapping of Nucleotide Binding Site–Leucine-rich Repeat Resistance Gene Analogs in Bermudagrass
Authors: Harris Karen R. , Schwartz Brian M. , Paterson Andrew H. , Brady Jeff A.
Published: January 2019

Thirty-one partial bermudagrass (Cynodon spp.) disease-resistance gene analogs (BRGA) were cloned and sequenced from diploid, triploid, tetraploid, and hexaploid bermudagrass using degenerate primers to target the nucleotide binding site (NBS) of the NBS–leucine-rich repeat (LRR) resistance gene family. Alignment of deduced amino acid sequences revealed that the conserved motifs of the NBS are present and all sequences have non-Drosophila melanogaster Toll and mammalian interleukin-1 receptor (TIR) motifs. Using a neighbor-joining algorithm, a dendrogram was created and nine groups of deduced amino acid sequences from bermudagrass could be identified from those sequences that span the NBS. Four BRGA markers and 15 bermudagrass expressed sequence tags (ESTs) with similarity to resistance genes or resistance gene analogs were placed on a bermudagrass genetic map. Multiple BRGA and EST markers mapped on T89 linkage groups 1a and 5a and clusters were seen on T89 19 and two linkage groups previously unidentified. In addition, three primers made from BRGA groups and ESTs with similarity to NBS-LRR resistance genes amplify NBS-LRR analogs in zoysiagrass (Zoysia japonica or Z. matrella) or seashore paspalum (Paspalum vaginatum). This gives evidence of conservation of NBS-LRR analogs among the subfamilies Chloridoideae and Panicoideae. Once disease resistance genes are identified, these BRGA and EST markers may be useful in marker-assisted selection for the improvement of disease resistance in bermudagrass.
Changes in Cell/Tissue Organization and Peroxidase Activity as Markers for Early Detection of Graft Incompatibility in Peach/Plum Combinations
Authors: Zarrouk Olfa , Testillano Pilar S. , Risueño María Carmen , Moreno María Ángeles , Gogorcena Yolanda
Published: January 2019

Changes in cell and tissue organization and in peroxidase activity have been analyzed to find early markers to predict graft incompatibility occurrence in peach/plum combinations (Prunus persica/Prunus spp.) at 5 months after grafting in the dormancy period. Different compatible and incompatible peach/plum grafts were grown for 5 months in a nursery. The cellular study of the graft interface revealed structural changes associated with graft incompatibility symptoms. The main structural features were cambium cell disorganization, less differentiation of vascular tissues, degeneration of phloem and xylem cells, and accumulation of phenols at the graft interface after 5 months of graft development. The peroxidase study was performed during dormancy and the vegetative growth period, and revealed a significant increase in peroxidase activity in the incompatible unions, with significant differences between compatible and incompatible grafts. Analysis of gel profiles of nonbudded rootstocks and scions revealed an anodal isoperoxidase band [relative front (Rf) = 0.48] present in scions and compatible rootstocks, and another isoperoxidase band (Rf = 0.53) only present in the incompatible rootstocks. Our results show that the analysis of cell organization to detect early structural events and the evaluation of peroxidase activity at graft unions constituted feasible and convenient methods for early diagnosis of graft incompatibility. Also, it was suggested that the presence of band Rf = 0.48 in plum rootstocks and peach cultivars could be used as a marker to predict graft compatibility for peach scions and plum rootstocks.
Growth and Physiological Traits Associated with Drought Survival and Post-drought Recovery in Perennial Turfgrass Species
Authors: Chai Qi , Jin Fang , Merewitz Emily , Huang Bingru
Published: January 2019

The objective of this study was to determine physiological traits for drought survival and post-drought recovery upon re-watering in two C3 perennial grass species, kentucky bluegrass [KBG (Poa pratensis)] and perennial ryegrass [PRG (Lolium perenne)]. Plants were maintained well watered or exposed to drought stress by withholding irrigation and were then re-watered in a growth chamber. KBG had significantly higher grass quality and leaf photochemical efficiency, and lower electrolyte leakage than PRG during 20 days of drought. After 7 days of re-watering, drought-damaged leaves were rehydrated to the control level in KBG, but could not fully recover in PRG. KBG produced a greater number of new roots, while PRG had more rapid elongation of new roots after 16 days of re-watering. Superior drought tolerance in KBG was associated with osmotic adjustment, higher cell wall elasticity, and lower relative water content at zero turgor. Osmotic adjustment, cell wall elasticity, and cell membrane stability could play important roles in leaf desiccation tolerance and drought survival in perennial grass species. In addition, post-drought recovery of leaf hydration level and physiological activity could be associated with the accumulation of carbohydrates in leaves and rhizomes during drought stress and new root production after re-watering.
Effect of Receiver pH on Infinite Dose Diffusion of 55FeCl3 across the Sweet Cherry Fruit Exocarp
Authors: Weichert Holger , Peschel Stefanie , Knoche Moritz , Neumann Dieter
Published: January 2019

Recent studies established that some ferric salts, including FeCl3, decrease water permeability of the sweet cherry (Prunus avium L.) fruit exocarp and fruit cracking, presumably by a pH-dependent precipitation reaction that blocks high-flux pathways across the fruit surface. The objectives of our study were the following: to establish the effect of receiver pH on penetration of 55FeCl3 through excised exocarp segments (ES) and isolated cuticular membranes (CM) and to localize any Fe precipitates in the epidermal system of mature sweet cherry fruit. Penetration was studied using an infinite dose diffusion system where 55Fe penetrated from donor solutions of ferric salts (10 mm, pH 2.2–2.6) or EDTA-Na-Fe(III) (10 mm, pH 5.0) across an interfacing ES or CM into aqueous receiver solutions of pH values ranging from 2.0 to 6.0. For receiver pH 2.0, 55Fe penetration of the ES from a 10 mm FeCl3 donor (pH 2.6) was linear with time, but for receiver pH ≥ 3.0, penetration was low and insignificant. Increasing the pH of the water receiver from 2.0 to 6.0 in the course of an experiment resulted in an immediate halt of penetration regardless of whether 55Fe penetration occurred from FeCl3 (pH 2.6), Fe(NO3)3 (pH 2.6), or Fe2(SO4)3 (pH 2.4) as donor solutions (all at 10 mm). Only from EDTA-Na-Fe(III) (pH 5.0) 55Fe penetration continued to occur albeit at a decreased rate (−30%). At receiver pH 2.0, the 55FeCl3 flux through stomatous ‘Sam’ ES averaged 10.4 ± 2.3 pmol·m−2·s−1 and was positively correlated to stomatal density. Conventional and analytical electron microscopy (energy dispersive X-ray analysis, electron spectroscopic imaging, and electron energy loss spectroscopy) identified ferric precipitates in periclinal and anticlinal cell walls of epidermal cells underlying the cuticle, but not within the cuticle. These data indicate that the lack of 55Fe penetration from donor solutions of ferric salts through the ES into a receiver solution at pH ≥ 3 and the previously reported decrease in water uptake and cracking as a response to immersing fruit in solutions of ferric salts are the result of a precipitation reaction at the cuticle/cell wall interface in the sweet cherry exocarp. Although spray application of ferric salts is prohibitive for ecotoxicological reasons, understanding their mechanism in decreasing water uptake and fruit cracking may be helpful in the search for alternate compounds that are effective and ecotoxicologically acceptable.
Regulated Water Deficits Improve Phytochemical Concentration in Lettuce
Authors: Oh Myung-Min , Carey Edward E. , Rajashekar C.B.
Published: January 2019

In a growth chamber study, lettuce (Lactuca sativa) plants were used to evaluate the effects of water deficits on health-promoting phytochemicals with antioxidant properties. Lettuce plants were treated with water stress by withholding water once at 6 weeks after sowing for 2 days or multiple times at 4 weeks for 4 days, at 5 weeks for 3 days, and at 6 weeks for 2 days. Water stress increased the total phenolic concentration and antioxidant capacity in lettuce. Young seedlings, 7 days after germination, had the highest total phenolic concentration and antioxidant capacity, and also, younger plants were typically more responsive to water stress treatments in accumulating the antioxidants than older plants. Phenylalanine ammonia lyase and γ-tocopherol methyltransferase genes, involved in the biosynthesis of phenolic compounds and vitamin E, respectively, were activated in response to water stress, although no activation of L-galactose dehydrogenase was detected. Lettuce plants subjected to multiple water stress treatments accumulated significant amounts of chicoric acid compared with the control plants. Although the increase in antioxidant activity in water stress-treated plants at harvest was not as great as in young seedlings, it was significantly higher than the control. One-time water stress treatment of lettuce at the time of harvest did not result in any adverse effect on plant growth. Thus, these results show that mild water stress in lettuce applied just before harvest can enhance its crop quality with regard to its phytochemical concentration without any significant adverse effect on its growth or yield.
Transcriptional Profiling of Rapidly Growing Cucumber Fruit by 454-Pyrosequencing Analysis
Authors: Ando Kaori , Grumet Rebecca
Published: January 2019

Fruit development proceeds from cell division to expansion, maturation, and ripening. Expansion is critical for size, yield, and quality; however, this period of development has received little attention. We used 454-pyrosequencing to develop a cucumber (Cucumis sativus) fruit transcriptome, identify highly expressed transcripts, and characterize key functions during exponential fruit growth. The resulting 187,406 expressed sequence tags (ESTs) were assembled into 13,878 contigs. Quantitative real-time polymerase chain reaction (qRT-PCR) verification of differentially expressed genes from fruit of different ages, and high correlation in transcript frequency between replicates, indicated that number of reads/contig reflects transcript abundance. Putative homologs were identified in Arabidopsis thaliana for 89% of the contigs represented by at least 10 ESTs; another 4% had homologs in other species. The remainder had homologs only in cucurbit species. The most highly expressed contigs were strongly enriched for growth (aquaporins, vacuolar ATPase, phloem proteins, tubulins, actins, cell wall-associated, and hormone-related), lipid, latex, and defense-related homologs. These results provide a resource for gene expression analysis in cucumber, profile gene expression in rapidly growing fruit, and shed insight into an important, but poorly characterized, developmental stage influencing fruit yield and quality.
Insertion and Characterization of the cry1Ia1 Gene in the Potato Cultivar Spunta for Resistance to Potato Tuber Moth
Authors: Zarka Kelly A. , Greyling Ria , Gazendam Inge , Olefse Dean , Felcher Kimberly , Bothma Gurling , Brink Johan , Quemada Hector , Douches David S.
Published: January 2019

Potato tuber moth (Phthorimaea operculella) is a serious pest of potatoes in tropical and subtropical regions of the world, including South Africa. The cry1Ia1 gene (from Bacillus thuringiensis) under the control of the 35S cauliflower mosaic virus promoter was transformed into the potato (Solanum tuberosum) cultivar Spunta to develop a cultivar with resistance to potato tuber moth for release in South Africa. Two transformation events, ‘SpuntaG2’ and ‘SpuntaG3’, were selected and subjected to extensive molecular analyses as required by the regulatory agencies of South Africa. Southern hybridization experiments indicated that ‘SpuntaG2’ and ‘SpuntaG3’ had one and three copies of the cry1Ia1 gene, respectively, and that the gene insertion was stable through multiple clonal generations. Furthermore, the sequence of the cry1Ia1 gene in ‘SpuntaG2’ was compared with the known sequence of the cry1Ia1 gene and found to be identical. Polymerase chain reaction (PCR) amplification using primers for plasmid “backbone” genes demonstrated that ‘SpuntaG2’ contained no backbone plasmid genes, whereas ‘SpuntaG3’ contained several backbone plasmid genes. Therefore, further analyses were limited to ‘SpuntaG2’, and event-specific primers were developed for this cultivar. Analysis of the left and right border regions in ‘SpuntaG2’ demonstrated that the insertion of the cry1Ia1 gene did not disrupt any functional genes nor did it create new open reading frames that encoded proteins with a significant match to the non-redundant sequence database queried by the BLASTP program. Enzyme-linked immunoabsorbent assays (ELISA) tests indicate that the cry1Ia1 gene was expressed at a mean concentration of 2.24 μg·g−1 fresh weight in leaf tissue and 0.12 μg·g−1 fresh weight in tubers. This study demonstrates the extensive molecular characterization that is necessary to apply for deregulation of a genetically modified crop and these data have been used in a regulatory package for the general release of ‘SpuntaG2’.
Safety Evaluations of the Cry1Ia1 Protein Found in the Transgenic Potato ‘SpuntaG2’
Authors: Quemada Hector , Zarka Kelly , Pett Walter , Bothma Gurling , Felcher Kimberly , Mirendil Hope , Koch Muffy , Brink Johan , Douches David
Published: January 2019

The transgenic potato ‘SpuntaG2’ (Solanum tuberosum), which is resistant to potato tuber moth (Phthorimaea operculella), was subjected to protein safety evaluations including protein equivalency tests for the Cry1Ia1 protein from ‘SpuntaG2’ and bacterially produced Cry1Ia1, toxicity and allergenicity evaluations of Cry1Ia1 protein, and compositional equivalency of ‘SpuntaG2’ compared with non-transgenic ‘Spunta’. Western blot analysis and biological activity assays showed molecular and functional equivalency between ‘SpuntaG2’-derived Cry1Ia1 protein and bacteria-derived Cry1Ia1 protein. Comparison of the Cry1Ia1 amino acid sequence to known amino acid sequences revealed no significant homology to known toxins or known allergens. Acute toxicity studies using rodents were used to calculate an acceptable daily intake (ADI) value of 20 mg·kg−1 body weight per day. The ADI value was then used to calculate a margin of exposure (MOE) of 2,222,222, which is more than 22,000 times greater than the commonly used target MOE of 100. Digestibility and thermostability assays determined that Cry1Ia1 was fully digested within 30 s of exposure to pepsin and inactive after 3 to 4 minutes at 100 °C, indicating that it would not be a potential allergen. Compositional analyses revealed no difference between ‘SpuntaG2’ and non-transgenic ‘Spunta’. These results strongly indicate that the Cry1Ia protein and the transgenic potato ‘SpuntaG2’ is not a human health risk.
Field and Storage Evaluations of ‘SpuntaG2’ for Resistance to Potato Tuber Moth and Agronomic Performance
Authors: Douches David , Pett Walter , Visser Diedrich , Coombs Joseph , Zarka Kelly , Felcher Kimberly , Bothma Gurling , Brink Johan , Koch Muffy , Quemada Hector
Published: January 2019

‘SpuntaG2’ is a transgenic potato (Solanum tuberosum) cultivar that contains the cry1Ia1 gene for resistance to potato tuber moth (Phthorimaea operculella), which is a serious pest of potato in many parts of the world. Previous studies have characterized ‘SpuntaG2’ at the molecular level and evaluated it for safety as a human food source. The objective here was to determine the efficacy of ‘SpuntaG2’ against the potato tuber moth in the field and in storage and to evaluate its agronomic performance. Efficacy trials at seven South African locations over 5 years indicated that ‘SpuntaG2’ gave complete control of potato tuber moth in the field and storage. The agronomic performance (tuber size and yield) of ‘SpuntaG2’ was not statistically different from ‘Spunta’ or was better than ‘Spunta’ at all locations/years with two exceptions. Three years of trials in Michigan further support these conclusions. Evaluations done during the 2006–07 South African season showed that ‘SpuntaG2’ did not differ from non-transgenic ‘Spunta’ for the following traits: maturity, growth habit, eye morphology, tuber size distribution, tuber shape, skin and flesh color, growth cracks, internal defects, specific gravity, chip color, cooking quality, disease resistance, and pollen fertility. Based on the results of these trials, it was concluded that ‘SpuntaG2’ provides the agronomic performance of ‘Spunta’ with the added benefit of resistance to potato tuber moth.