Progress 04/19/14 to 09/30/18
Outputs
Target Audience:
Nothing Reported
Changes/Problems:Project Director is no longer at the University of Wyoming. What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?
Nothing Reported
What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
n/a
Publications
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:Presentations were made on grape production for updating grape growers and interested homeowners in research activities being carried out for expanding viticulture activities in Wyoming. Presentations and visits were carried out at the annual field days in Sheridan (280 people over 3 years) and Powell (200 people over 3 years). Information to several growers and homeowners was also provided via phone consultations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Community college and undergraduate students were involved in experiments involving vineyard management, data collection, and laboratory experiments including total RNA isolation and analyses. Two students were trained in RNA sequencing technology. How have the results been disseminated to communities of interest?The results have been presented at field days and undergraduate research days. What do you plan to do during the next reporting period to accomplish the goals?We will continue to screen additional genetic elements, genes and selectable markers for use in precision breeding of grapevines. Data will be collected on cold-hardiness, yield and fruit quality of grapevines growing in the cultivar evaluation trial.
Impacts
What was accomplished under these goals?
Data on bud break, number of inflorescences per vine, yield per vine, number of clusters, cluster weights, individual berry weights and TSS were recorded. Frontenac, Frontenac Gris and Osceola Muscat were the highest yielding cultivars. Most Vitis labrusca hybrids such as 'Elvira', 'Concord', 'Ives' and 'Fredonia' are extremely late maturing and ripening for the Sheridan area and not suitable for production due to the risk of crop being lost to an early Fall freeze. Efforts continued to advance precision breeding technology for genetic improvement of grapevine. Grapevine leaf samples were collected from field-grown grapevines exposed to low temperatures for RNA sequencing studies. Total RNA was isolated and will be sent for RNA sequencing to compare differential gene expression of grapevines exposed to abiotic stress. The Vitis rupestris-derived CBF1 gene is induced in response to multiple abiotic stress factors (cold/drought/salinity) and triggers the expression of functional genes involved in imparting stress tolerance. Preliminary results indicated that the constitutive expression of this transcription factor inhibited plant growth and development, which might necessitate expression of this regulatory gene under the control of stress inducible genetic elements. Additional grapevine-derived transporter and antiporter genes are also being tested to impart abiotic stress tolerance.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2017
Citation:
Baldwin, M. and Dhekney, S.A. 2017. The grape research program at the University of Wyoming. Proceedings of the 5th International Conference on Steep Slope and Mountain Viticulture, 55-60.
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Progress 10/01/15 to 09/30/16
Outputs
Target Audience:Research results were presented to grape growers statewide and at the world congress on in vitro biology held in San Diego in June 2016. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student completed his PhD dissertation as part of the project. How have the results been disseminated to communities of interest?Research results were presented at the 2016 World Congress on In Vitro Biology held in San Diego in June 2016. What do you plan to do during the next reporting period to accomplish the goals?Additional transcription factors will be isolated and placed under the control of tissue specific- and stress-inducible promoters for transformation in grapevine. Regenerated plants will be screened for abiotic stress tolerance.
Impacts
What was accomplished under these goals?
C-repeat binding factors (CBF) and ion-transporters were isolated from rootstocks of V. rupestris and gene sequences were analyzed for important regulatory elements conferring stress tolerance. The CBF1 gene was cloned into a binary vector and transferred into susceptible V. vinifera spp using genetic engineering. Somatic embryos at mid-cotyledonary stage of development were used to insert the gene construct. No significant difference was observed in transient and stable gene expression. Transgenic embryos was recorded to be 7.95 - 8.5% however a limited number of transgenic lines were recovered following embryo germination. Abnormality in embryo germination and plant regeneration was routinely observed in the form of malformation, stunted growth or tissue necrosis. It can be attributed to overexpression of CBF1 gene which activates several regulatory genes in a downstream pathway. Higher amount of regulatory proteins being produced could be the possible reason of negative phenotype appearance. Over expression of CBF1 may be due the same core promoter divergently placed upstream of the duplicated enhancer in a bi-directional dual promoter configuration in vector.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2017
Citation:
Kandel, R., Dutt, M., Gray, D.J., Li, Z.T., Bergey, D.R., Sitther, V., and Dhekney, S.A. 2017. Evaluation of a grapevine-derived reporter gene system for precision breeding of Vitis. Plant Cell Tissue and Organ Culture, 124: 599-609.
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Progress 10/01/14 to 09/30/15
Outputs
Target Audience:Grape growers, interested homeowners, master gardeners, and other scientists studying grape genetics. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?A graduate student completed his PhD program and several undergrduate students were involved in grape tissue culture experiments. How have the results been disseminated to communities of interest?Several scientific papers were published and presentations were made on grape production at the following locations for updating grape growers and interested homeowners in research activities being carried out for expanding viticulture activities in Wyoming. Talks were presented at the Master gardeners' conference in Sheridan, WY. A presentation was made to the Sheridan county master gardeners in April 2015. A grape production workshop was conducted in Powell in May 2015 to provide information on the use of the right training and pruning systems for cold-hardy grapevine cultivars. Presentations and visits were carried out at the annual field days in Sheridan and Powell. A number of prospective and current grape growers visited the vineyard and obtained information on suitable cultivars and vineyard establishment procedures. What do you plan to do during the next reporting period to accomplish the goals?Wewill continueto screengrapevine-derived screen abiotic-stress inducible genes, genetic elements and reporter genes for use in precision breeding.
Impacts
What was accomplished under these goals?
Field trials for year 2015 included collecting data on grapevine cultivar response and cold hardiness at two trials established in Sheridan and Powell. The unseasonal freeze that occurred in September 2014 caused significant delays in grapevine establishment at Sheridan. Additionally a field trial to estimate water requirement of mature vineyards and study the effect of landscape fabric on grape yield and quality was established at a grower's vineyard in Wheatland, WY. Experimental treatments were established and preliminary data was collected. Fruit obtained from the experimental plot was used to make wine. The wine was evaluated at a multistate meeting (NE1020) by comparing it for quality characteristics with wine made by other grape research programs in the United States. The laboratory component of my program focusses on studying the genetics of abiotic stress tolerance and utilizing this information for grapevine improvement. We continued to grapevine-derived screen abiotic-stress inducible genes, genetic elements and reporter genes for use in precision breeding. A Vitis-derived MybA1 transcription factor was optimized for use as a reporter gene.
Publications
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Dhekney, S.A., Kandel, R., Vardiman, J., Bergey, D.R., and Gray, D.J. 2015. Screening grapevine cultivars for adaptability to soil and climatic conditions in Wyoming. HortScience, S112.
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Dhekney, S.A., Kandel, R., Bergey, D.R., Shafer, H., Sitther, V., Li, Z.T. and Gray, D.J. 2015. Studying micropropagation response of cold-hardy grapevine cultivars. P-2038. In Vitro Cell. Dev. Biol. Plant, DOI 10.1007/s11626-015-9906-y
- Type:
Conference Papers and Presentations
Status:
Published
Year Published:
2015
Citation:
Kandel, R., Dutt, M., Gray, D.J., Li, Z.T., Bergey, D.R., Sitther, V., Quarterman, I., and Dhekney, S.A. 2015. Optimizing parameters for precision breeding of grapevine. HortScience, S112.
- Type:
Other
Status:
Published
Year Published:
2015
Citation:
Dhekney, S.A., Kandel, R., Bergey, D.R., Smith, D., and Vardiman, J. 2015. . Screening grapevine cultivars for adaptability to soil and climatic conditions in Wyoming.. UW AES Field Days Bulletin, 121.
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Progress 04/19/14 to 09/30/14
Outputs
Target Audience: Presentations were made on grape production to update grape growers and interested homeowners in research activities being carried out for expanding viticulture activities in Wyoming.Prospective grape growers were provided with information forvineyard establishment. Presentations on grape production and vineyard establishment were given at the Casper County master gardener program in Casper and Powell, WY, and at field days held at the Sheridan and Powell Research and Extension Centers. Additionally, information on grape production was also provided via email and phone conversations. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided? Two community college students (Sheridan College) and three UW undergraduate students carried out some internships in gene cloning and somatic embryogenesis. How have the results been disseminated to communities of interest? Presentations were made on grape production at the following locations for updating grape growers and interested homeowners in research activities being carried out for expanding viticulture activities in Wyoming. What do you plan to do during the next reporting period to accomplish the goals? Selected candidate gene sequences along with the assembled genetic elements for constitutive expressionwill be rapidly incorporated into binary vectors using the Gateway Clonase technology and transferred to Agrobacterium. Agrobacterium harboring the binary constructs will be used for cocultivation with embryogenic cultures and recovery of transgenic plant lines.
Impacts
What was accomplished under these goals?
Objective 1. The winter and spring of 2014 experienced record cold temperatures with minimum temperatures exceeding - 20F in December and February. Day/s of bud break in specific cultivars and at both locations was recorded. Following bud break, the total survival rates and individual survival rates in specific cultivars was recorded. All vines that exhibited bud break and some growth symptoms by June 25 were considered to have survived the 2013-14 winter and spring freeze-induced damage. Plant height was recorded by July 25, which is when most of the vines for all cultivars reached the top wire of the high wire cordon system (5 feet). We observed differences in survival rates between cultivars at the Sheridan and Powell location. While high survival rates (70% and higher) were observed in most cultivars in Sheridan, the overall survival rate in Powell was 40%. Objective 2. Vitis interspecific hybrid "Richter 110" was subjected to salt stress and total RNA was isolated from shoot tissues. Genes from the CBF, NAC, NHX, SOS and type 1 H+-phyrophosphatase families that are either induced during salt stress or implicated in salinity tolerance were recovered. The cDNA sequences were inserted in cloning vectors and sequenced. Sequences showing a 100% homology will be used for insertion in binary vectors and used in transformation studies. Objective 3. Embryogenic cultures were initiated and established in V. vinifera 'Thompson Seedless', 'Merlot' and Vitis interspecific hybrids 'Seyval Blanc', 'Bronx Seedless' and 'Richter 110'. Embryogenic cultures are currently being maintained on embryo development medium and will be used in gene insertion studies.
Publications
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Gray, D.J., Li, Z.T., and Dhekney, S.A. 2014. Precision breeding of grapevine for improved traits. Plant Science, 228: 3-10.
- Type:
Journal Articles
Status:
Published
Year Published:
2014
Citation:
Li, Z.T., Kim, K.H., Dhekney, S.A., Jasinski, J.R., Creech, M.R., and Gray, D.J. 2014. An optimized procedure for plant recovery from somatic embryos significantly facilitates the genetic improvement of Vitis. Horticulture Research, 1: 1-7 doi:10.1038/hortres.2014.27.
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