Graduate Thesis Or Dissertation
 

Improving initiation and mineral nutrition for hazelnut (Corylus avellana) micropropagation

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https://ir.library.oregonstate.edu/concern/graduate_thesis_or_dissertations/dv13zw55f

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  • Corylus avellana L. is a global commodity and a valuable crop for the U.S. Pacific Northwest. A tremendous amount of work has been put into the development of healthy disease free, high yielding and tasty hazelnuts for the in-shell and confectionary markets. Clonal propagation is required to provide nursery trees of these improved hazelnut cultivars. The rapid multiplication provided by micropropagation methods is an important part of meeting the demand. One challenge of hazelnuts micropropagation is the wide variation in growth response among the cultivars. The studies described in this thesis were involved in improving the mineral nutrients in the growth medium and to better determine culture initiation procedures. The first study was designed to determine what mineral nutrients are driving C. avellana in vitro shoot growth using a response surface design. Hazelnut genotypes 'Dorris,' 'Felix,' 'Jefferson', OSU 880.054, and 'Sacajawea' were used. Driver and Kuniyuki Walnut medium (DKW) mineral nutrients were separated into five factors: NH₄NO₃, Ca(NO₃)₂, mesos (MgSO₄ and KH₂PO₄), K₂SO₄, and minor nutrients (B, Cu, Mn, Mo, and Zn). The concentrations ranged from 0.5x to 2.0x the standard DKW concentrations with 33 treatments for use in modeling. Multifactor response surface analysis projected that optimum shoot proliferation was greatly influenced by the NH₄NO₃ to Ca(NO₃)₂ ratios, mesos, and minors. These factors were important to overall quality and shoot length for all genotypes. Improved shoot quality was observed with increased Ca(NO₃)₂, mesos, and minors for most of the cultivars tested. For 'Sacajawea' only the mesos components significantly improved shoot quality. Lower Ca(NO₃)₂ improved shoot multiplication while higher amounts increased shoot length for most cultivars. New media formulations will require higher Ca(NO₃)₂, mesos and minors as well as changes in the NH₄NO₃ to Ca(NO₃)₂ ratios. The second study determined the effects of individual minor-mineral nutrients including nickel on hazelnut shoot growth. Three hazelnut cultivars 'Dorris,' 'Jefferson,' and 'Sacajawea' were used. Six factors at 0.5x to 4.0x DKW concentrations, H₃BO₃, CuSO₄, MnSO₄, Na₂MoO₄, Zn(NO₃)₂, and NiSO₄, were tested in a response surface design with 39 treatment combinations. Ni, not present in DKW, ranged from 0 to 6 µM. Higher concentrations (4x) of B, Mo, and Zn increased overall shoot quality, length and multiplication. There were many significant interactions. High B concentrations significantly improved shoot quality for 'Jefferson'; shoot quality, length and number for 'Dorris'; and shoot length and number for 'Sacajawea'. Increased Mo improved some responses for each cultivar, and it also interacted with Cu and Zn. Interactions of Ni with other minor nutrients resulted in improved shoot quality and length in 'Sacajawea.' Ni interactions were significant for the other cultivars as well, altered the requirements for other nutrients, but did not necessarily improve the overall shoot response. Improved growth and shoot quality in 'Dorris' and 'Jefferson' required increased amounts of B, Mo, and Zn; 'Sacajawea' required increased B, Cu, Zn, and Ni. The diverse responses of these cultivars further confirmed that nutrient uptake or utilization was genotype dependent. Hazelnuts are generally difficult to initiate into culture due to internal microbial contaminants and a general lack of viability of the explants. The third study was designed to determine the effect of nodal position and collection techniques on the viability and contamination of shoot explants. Explants were collected from scion wood grafted onto seedling rootstocks and grown in the greenhouse. Single-node explants were collected from different locations on the scion wood. After surface sterilization, explants were first held in a liquid contaminant-detection medium for one week and the effect on explant viability was evaluated. Node position influenced the number of viable contaminant-free explants. Bacterial contamination increased with the distance from the shoot tip. The use of liquid detection medium as a part of the initiation procedure did not affect viability. Bacteria sampled from surface sterilized explants were identified as Brevundimonas vesicularis, Brevundimonas sp., and two Pseudomonas sp., by 16S ribosomal DNA sequences and API® 20CHB tests. The best procedure for collecting axenic, viable hazelnut explants was to collect from the first three nodes of actively-growing greenhouse plants and use detection medium to identify contaminant-free tissue. As a result of these studies several suggested growth media formulations are now available for use that may produce improved hazelnut shoot quality, multiplication and elongation for a wide range of C. avellana genotypes. Two base macro nutrient formulations, Hazelnut A and B, are likely to be suitable for many genotypes. Three micronutrient formulations provide options for growth of difficult genotypes with and without Ni. Continued optimization and testing of mesos and N components are needed for final medium formulations.
  • Keywords: Plant Mineral Nutrition, Plant tissue culture, Filberts, Micropropagation, Hazelnuts
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