Graduate Thesis Or Dissertation
 

Micropropagation and regeneration of hazelnut (Corylus species)

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  • A micropropagation system was developed for hazelnut cultivars and rootstocks. Culture establishment was affected by explant source, explant collecting season, and type of explant. The highest frequency of culture initiation and lowest tissue oxidation and contamination were obtained from internodes with single axillary buds from grafted greenhouse-grown plants early in the growing season (March). Nodal explants had a higher percentage of growth than shoot tips. For field collected materials, suckers collected in mid-season were the best option. Tissue contamination and oxidation were the main problems for the in vitro establishment of explants from mature field-grown trees. Attempts to decrease tissue oxidation by using forced outgrowth from field-grown trees and treatment of explants with antioxidants were not successful. There was a correlation between explant oxidation and phenolic content. Shoot multiplication was the best on a modified DKW medium (NCGR-COR medium) supplemented with N⁶-benzyladenine (BA) (1.5-3 mg*1⁻¹). Plants grown on medium with 3% glucose or fructose produced more and longer shoots than those on medium with sucrose. The general appearance and growth habit of shoots were better on medium with glucose than fructose. Shoot elongation varied with genotype. Changes in medium pH from 4.7 to 5.7 did not significantly affect the multiplication rate. Root formation was induced either in vitro by culturing shoots on NCGR-COR medium with half strength mineral salts and 1 mg*1⁻¹ indole-3-butyric acid (IBA) for four weeks or ex vitro by a brief dip in 0.2 or 1 g*1⁻¹ IBA and acclimatization. High transplant survival of plants was obtained. Internal bacterial contamination was present in micropropagated shoots. A combination of the antibiotics streptomycin and gentamicin was effective in eliminating contaminants from micropropagated shoots, but some phytotoxicity was noted. Adventitious shoots were induced at a low frequency from stem segments of 'Nonpareil', 'Willamette', and 'Tonda Gentile Romana' and leaf discs of 'Dundee' on medium containing glutamine, thidiazuron (TDZ), and naphthaleneacetic acid (NAA). Adventitious roots were produced from stem segments and leaf discs cultured on medium with NAA alone or high concentrations of a-NAA or IBA combined with low concentrations of BA.
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