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Kromwijk A.,Wageningen UR Greenhouse Horticulture | Van Noort F.,Wageningen UR Greenhouse Horticulture | Verhoef N.,NSure | Balk P.,NSure | Van Wordragen M.,NSure
Acta Horticulturae | Year: 2013

During wintertime Viburnum opulus var. roseum (snowball) shrubs are forced in warm greenhouses to harvest early cut flowers. Early forcing is occasionally unsuccessful. This is probably due to a lack of hours with low temperatures that is needed to break bud dormancy. To gain more insight about the effect of temperature on breaking dormancy and early forcing results, shrubs were transferred into cold storage in week 41, 43 and 45 (2009) for 4, 6 or 8 weeks at a temperature of 2, 5 or 8°C. After storage, the shrubs were forced at a day-/night temperature of 28/23°C. Shrubs stored for at least 8 weeks from week 41, 6 weeks from week 43 and 4 weeks from week 45 gave good forcing results, regardless of storage temperature. Since growing conditions outside vary every year and differ between nurseries, it is difficult to determine when forcing can be started. To prevent poor forcing results, we aimed to develop a molecular diagnostic assay to determine the moment at which winter dormancy of snowball flower buds is sufficiently broken. In order to identify genes involved in bud breaking dormancy, an experiment was performed on a commercial nursery from October 2010 to January 2011. Flower buds of snowball shrubs outside were sampled weekly and in December and January batches of shrubs were placed every week in a greenhouse to observe forcing results. Genes involved in bud breaking dormancy were identified by performing next generation Illumina RNA sequencing on bud samples with poor and good forcing results. A set of candidate genes was validated during a second trial at two different commercial nurseries in 2011-2012. © ISHS 2013.

Stattin E.,Dalarna University | Verhoef N.,NSure | Balk P.,NSure | van Wordragen M.,NSure | Lindstrom A.,Dalarna University
New Forests | Year: 2012

In boreal forest regions, a great portion of forest tree seedlings are stored indoors in late autumn to prevent seedlings from outdoor winter damage. For seedlings to be able to survive in storage it is crucial that they store well and can cope with the dark and cold storage environment. The aim of this study was to search for genes that can determine the vitality status of Norway spruce (Picea abies (L.) Karst.) seedlings during frozen storage. Furthermore, the sensitivity of the ColdNSure™ test, a gene activity test that predicts storability was assessed. The storability of seedlings was tested biweekly by evaluating damage with the gene activity test and the electrolyte leakage test after freezing seedlings to -25°C (the SEL diff-25 method). In parallel, seedlings were frozen stored at -3°C. According to both methods, seedlings were considered storable from week 41. This also corresponded to the post storage results determined at the end of the storage period. In order to identify vitality indicators, Next Generation Sequencing (NGS) was performed on bud samples collected during storage. Comparing physiological post storage data to gene analysis data revealed numerous vitality related genes. To validate the results, a second trial was performed. In this trial, gene activity was better in predicting seedling storability than the conventional freezing test; this indicates a high sensitivity level of this molecular assay. For multiple indicators a clear switch between damaged and vital seedlings was observed. A collection of indicators will be used in the future development of a commercial vitality test. © 2012 The Author(s).

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