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M.A. Dalbó, G.N. Ye, N.F. Weeden, W.F. Wilcox, and B.I. Reisch

The efficiency of marker-assisted selection for powdery mildew (Uncinula necator (Schw.) Burr) resistance in grapes (Vitis L. sp.) was studied using molecular markers associated with a major QTL (quantitative trait loci) for this trait. Initially, genetic maps were constructed from a segregating population of the cross `Horizon' × Illinois 547-1 (a hybrid between V. rupestris Scheele and V. cinerea Engelm.). A major QTL from Ill. 547-1, the resistant parent, explained 41% of the variation. One RAPD (randomly amplified polymorphic DNA) marker and one AFLP (amplified fragment length polymorphism) marker, obtained by bulked segregant analysis, showed the highest association with powdery mildew resistance in the mapping population. Segregation of the QTL was followed in different crosses by CAPS (cleaved amplified polymorphic sequence) markers developed from these two markers. An allele-specific amplified polymorphism that segregates as present/absent was also developed from the CS25b locus. Powdery mildew resistance was evaluated visually on a 1 to 5 scale in four different seedling populations. Two populations originated from crosses using Ill. 547-1 as the resistant parent. Two other populations were from crosses with NY88.0514.03, a resistant seedling from the original `Horizon' × Ill. 547-1 mapping population. Segregation ratio distortions were observed in some crosses. In these cases, the allele associated with the QTL for powdery mildew resistance was less frequent than the alternate allele. In all crosses, the markers were closely associated with resistance. If selection were based on markers, the percentage of susceptible individuals (classes 4 and 5) would decrease from 24% to 52% to 2% to 18%. Selection efficiency was greatest in crosses where segregation distortion was most intense.

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Philip L. Forsline, Warren F. Lamboy, James R. McFerson, and Cecil Stushnoff

The USDA–ARS germplasm collection of cold-hardy Vitis held at the Plant Genetic Resources Unit, Geneva, N.Y., has over 1300 clonal accessions maintained as field-grown vines. Security back-up using field-grown or potted vines at remote sites or via in vitro methods is costly. Cryopreservation offers a safe, cost-effective alternative. While we routinely employ cryogenic storage of dormant buds of Malus, dormant buds of Vitis generally do not appear to tolerate the desiccation levels required by our current cryopreservation protocol. Since tolerance to desiccation and cold appear to be correlated in Vitis, we tested desiccation tolerance of 60 germplasm accessions selected from the core subset to represent a range of cold hardiness. Budwood was collected in December 1995 in Geneva, stored at –4°C in sealed bags, and systematically desiccated to 30% and 20% moisture. In some treatments, additional desiccation was imposed by slow freezing to –25°C. Microscopic examination of rehydrated buds indicated 60% of accessions tolerated desiccation as low as 20% moisture. Freeze-desiccation at –25°C after desiccation at –4°C neither increased nor decreased viability in these accessions. Only slight modification so current protocols should be necessary for cryopreservation of this class. Of the remaining accessions, 25% tolerated desiccation to 30% moisture, but 15% were intolerant to any desiccation level tested. Techniques must be developed to successfully cryopreserve both these classes of accessions.

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S. Kaan Kurtural, Lydia F. Wessner, and Geoffrey Dervishian

photosynthesis and vine performance in Vitis vinifera L. cv. Barbera Aust. J. Grape Wine. Res. 17 351 357 California Department of Food and Agriculture 2012 Grape crush report. Final 2011. Sacramento. 1 Apr. 2012 < http

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Jiang Lu*, Hong Huang, Wayne Hunter, Phat Dang, and Stephen Leong

Vitis shuttleworthii is one of the most disease and pest resistant grape species originated and distributed in the southeast United States. It is highly resistant to Pierce's disease, anthracnose, black rot and downy mildew diseases, which are the limited factors to grow Vitis vinifera grapes in this region. A V. shuttleworthii cDNA library was constructed with mRNA isolated from leaves and flowers harvested during anthesis. 12,008 clean EST sequences were obtained and assembled. and generated 5776 unigenes 2106 contigs and 3670 singletons). ESTs distribution based on protein function using a modified MIPS MATDB Arabidopsis Scheme revealed that 7% of the V. shuttleworthii ESTs were related to disease/pest defense or stress tolerance genes. Over 300 contigs containing complete or 90% open coding region of known functional genes were obtained. The ESTs that were annotated as pathogenesis-related proteins, enzymes in salicylic acid, jasmonic acid, and ethylene signaling, were selected for further study in order to elucidate the role and interaction of them in the signal transduction cascade that leads to grape defense gene activation upon treatment of bacterial pathogen. We report the identification of novel disease resistant genes based on preliminary pathogenesis-relative pathways network analysis.

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Philip L. Forsline, E.E Dickson, and A.D. Djangaliev

The USDA National Plant Germplasm System (NPGS) sponsored a 1993 collection of wild Malus in Kazakhstan and Kyrgyzstan which followed a collection in 1989 from sites in Tajikistan and Uzbekistan. There is strong evidence that the domestic apple originated in the foothills of the Tian Shari mountains where Malus sieversii (Ldb.) M. Roem remains as a primary forest species. The goal of the recent expedition was to obtain additional genetic diversity of apple from some of the remote sites in that area with the assistance of the Kazakh hosts. While there, isolated pockets of other fruit in the wild (especially Vitis) were discovered and collected. Seed collections from the expedition are stored with the NPGS and seedling populations are being evaluated for valuable traits.

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Hong Xu, Diane J. Wilson, S. Arulsekar, and Alan T. Bakalinsky

Randomly amplified polymorphic DNA (RAPD) markers were generated for identifying grape (Vitis) rootstocks. Seventy-seven primers (10 bases long) were screened using CsCl-purified leaf DNA derived from several field samples of nine rootstocks sampled in successive years. Nine RAPD markers were detected from six primers and, in combination, distinguished all nine rootstocks tested. Because inconsistencies were encountered in performing the RAPD assay, sequence-specific primers were derived from cloned RAPD bands for use under more stringent amplification conditions. Southern hybridization analysis of the RAPD gels with cloned RAPD bands as probes revealed deficiencies of scoring RAPD bands based solely on ethidium bromide staining. In some cases, bands of the same size generated by the same primer in different rootstocks-normally scored as the same marker-failed to cross-hybridize, implying lack of homology between the bands. More commonly, bands scored as absent based on ethidium bromide staining were detected by hybridization. Six of the nine cloned RAPD bands were partially sequenced, and sequence-specific primer pairs were synthesized. Two primer pairs amplified a product the same size as the original RAPD band in all rootstocks, resulting in loss of polymorphism. Two other pairs of sequence-specific primers derived from the same marker failed to amplify the expected band consistently. Three of the most useful primer pairs amplified apparent length variants in some accessions and will have value as polymerase chain-reaction markers for fingerprinting.

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Warren F. Lamboy and Christopher G. Alpha

Curators of plant genetic resources collections must preserve germplasm possessing known useful characteristics as well as material displaying general genetic diversity. In order to ensure that both types of germplasm are included in a collection, germplasm curators require three fundamental types of information about each accession: taxonomic identity, genetic identity, and genetic relationship. Because simple sequence repeat DNA fragments (SSRs) have been successfully used to determine the genetic identity of grape clones, we conducted a study to determine if SSRs would supply all three types of information for the accessions in the cold-hardy Vitis (grape) germplasm collection. SSR fragments were amplified at six different loci for 23 accessions of cold-hardy grape spanning the range of species diversity in the collection. The minimum number of different alleles found at a locus was 9; the maximum was 26. Heterozygosity values ranged between 0.565 and 0.783, while gene diversity values were in the range 0.785 to 0.944. Two hundred fifty-two pairs of plants out of a possible 253 could be distinguished by their SSR profiles. Nei's genetic identities were computed between all pairs of plants and used in a UPGMA cluster analysis. The relationships obtained did not correspond well to expected relationships based on geography and taxonomy. Four species of grapes were represented by two or more accessions in this study. No DNA fragments found at these six loci served to unambiguously distinguish one species from another. Thus, SSR fragments from the six loci studied were useful in determining genetic identity of accessions, but were not helpful in determining genetic relationships or taxonomic identities. We are searching for additional loci that are informative for these types of information. Meanwhile we highly recommend SSRs for determining genetic identity in germplasm resources collections.

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Stefano Poni, Alberto Palliotti, and Fabio Bernizzoni

This paper describes and evaluates the reliability of a model for prediction of daily carbon balance and dry matter (DM) accumulation in vertically shoot positioned grapevine (Vitis vinifera L.) canopies based on the user-friendly STELLA simulation software. Validation of the model was produced for potted `Cabernet Sauvignon' grapevines at both low canopy density [LD (≈10 shoots/m of row)] and high canopy density [HD (≈20 shoots/m of row)] by comparing, on a seasonal basis, the modelled daily CO2 balance with the diurnal net carbon exchange rate (NCER) measured using a whole-canopy enclosure method. Estimated daily total photosynthesis (Pn) was linearly correlated with measured NCER for LD (r 2 = 0.87) and HD (r 2 = 0.86), thereby indicating that despite its simplicity the model led to a fairly good degree of precision, although it tended to slightly underestimate (5% to 8% less) the measured rates and scattering increased at high values of CO2 fixations. Daily total respiration (R) for LD treatment was 29.0% of total daily Pn, with clusters, leaves and stems accounting for 11.8%, 46.7%, and 41.5%, respectively. Daily total R was 24.2% of total daily Pn in HD treatment and single organs contributed 22.3% (clusters), 41.6% (leaves), and 36.1% (stems). The model estimated that 1604 and 1893 g DM per vine accumulated at harvest for LD and HD treatment, respectively, whereas destructive sampling of leaves, stems and clusters yielded 1475 ± 64 g per vine for LD treatment and 1730 ± 96 g per vine for HD treatment, respectively, corresponding to the 91% and 92% of the DM estimated with STELLA, which in its present version does not take into account root respiration.

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Anne Fennell and Michael J. Line

Physiological and biophysical changes were monitored during shoot maturation and bud endodormancy induction in grape (Vitis riparia Michx.) under controlled environments. Growth, dry weight (DW), periderm development, bud endodormancy, and nuclear magnetic resonance imaging (MRI) T2 relaxation times were monitored at 2, 4, or 6 weeks of long-photoperiod [long day (LD), 15 h, endodormancy inhibition] or short-photoperiod [short day (SD), 8 h, endodormancy induction] treatments at 15/9 h day/night thermoperiod of 25/20 ± 3 °C. Shoots on LD plants grew throughout the entire study period, although the rate of growth decreased slightly during the 6th week. Shoot growth slowed significantly after 2 weeks of SD, was minimal by the 4th week of SD and most of the shoot tip meristems had abscised after 6 weeks of SD. Endodormancy was induced after 4 weeks of SD. DW of the stem and buds increased with increasing duration of LD and SD. While bud DW increased more under SD than LD, stem DW increased more under LD than SD. T2 relaxation times were calculated from images of transverse sections of the grape node. There was a slight decrease in the T2 times in the node tissues with increased duration of LD treatment, whereas SD induced a significant decrease in T2 times during endodormancy induction. T2 values for the node decreased after 4 weeks of SD, coinciding with endodormancy induction. Separation of node tissues into bud, leaf gap, and the remainder of the stem and analysis of the proportion of short and long T2 times within those tissues indicated differential tissue response. A greater proportion of short T2 times were observed in the 2-week SD leaf gap tissue than in the LD and the proportion of short T2 times continued to increase with subsequent SD treatment. Bud and all other stem tissues had a greater proportion of short T2 times after 4 weeks of SD, coinciding with bud endodormancy induction. The proportion of short and long T2 times in a tissue was a better indicator of endodormancy than the averaged T2 time for the tissue. Thus, MRI allows nondestructive identification of differential tissue response to photoperiod treatments and makes it possible to separate normal vegetative maturation responses from endodormancy induction.

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Li-Song Chen and Lailiang Cheng

One-year-old grapevines (Vitis labrusca L. `Concord') were supplied twice weekly for 5 weeks with 0, 5, 10, 15, or 20 mm nitrogen (N) in a modified Hoagland's solution to generate a wide range of leaf N status. Both light-saturated CO2 assimilation at ambient CO2 and at saturating CO2 increased curvilinearly as leaf N increased. Although stomatal conductance showed a similar response to leaf N as CO2 assimilation, calculated intercellular CO2 concentrations decreased. On a leaf area basis, activities of key enzymes in the Calvin cycle, ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), and key enzymes in sucrose and starch synthesis, fructose-1,6-bisphosphatase (FBPase), sucrose phosphate synthase (SPS), and ADP-glucose pyrophosphorylase (AGPase), increased linearly with increasing leaf N content. When expressed on a leaf N basis, activities of the Calvin cycle enzymes increased with increasing leaf N, whereas activities of FBPase, SPS, and AGPase did not show significant change. As leaf N increased, concentrations of glucose-6-phosphate (G6P), fructose-6-phosphate (F6P), and 3-phosphoglycerate (PGA) increased curvilinearly. The ratio of G6P/F6P remained unchanged over the leaf N range except for a significant drop at the lowest leaf N. Concentrations of glucose, fructose, and sucrose at dusk increased linearly with increasing leaf N, and there was no difference between predawn and dusk measurements. As leaf N increased, starch concentration increased linearly at dusk, but decreased linearly at predawn. The calculated carbon export from starch degradation during the night increased with increasing leaf N. These results showed that 1) grapes leaves accumulated less soluble carbohydrates under N-limitation; 2) the elevated starch level in low N leaves at predawn was the result of the reduced carbon export from starch degradation during the night; and 3) the reduced capacity of CO2 assimilation in low N leaves was caused by the coordinated decreases in the activities of key enzymes involved in CO2 assimilation as a result of direct N limitation, not by the indirect feedback repression of CO2 assimilation via sugar accumulation.