Document Type : Research Paper
Authors
- M Dehestani Ardakani 1
- M Kafi 1
- M Enayati Shariatpanahi 2
- M Jafarkhani-Kermani 3
- M.R Fattahi Moghadam 1
- M Oroojloo 3
1 Department of Horticultural Science & Landscape Architecture, Faculty of Agricultural Science & Engineering, College of Agriculture & Natural Resources, University of Tehran, Karaj, 31587, Iran
2 researcher and head of Tisue culture and Gene transformation Dep.
3 Department of Tissue Culture and Gene Transformation, Agricultural Biotechnology Research Institute of Iran, Mahdasht Road, P.O. Box 31535-1897 Karaj, Iran
Abstract
One of the most commonly used methods to produce doubled haploid plants is microspore culture. In this research work, the isolated microspore culture system in two rose cultivars i.e. Apollo and Amarosa was investigated. Important factors including isolation media (AB, B), AT3 induction medium with different carbohydrate sources (sucrose, maltose or glucose) and amino acid source (lactalbumin hydrolysate) were studied. Carbon starvation and temperature (heat and cold) treatments as two important stresses alone or in combination with each other for various periods were evaluated on the induction of symmetrical (sporophytic) divisions. A mixture of different developmental stages of microspores was used to initiate the cultures but the majority of them were at late uni-cellular stage. For eliminating bacterial or fungal contaminants, buds were surface-sterilized by immersion in 70% ethanol for 15, 30, 60 Sec. or 3.5% (w/v) sodium hypochlorite solution for 5, 10, 15 min prior to microspore isolation. The best sterilization procedure was observed when microspores were sterilized with sodium hypochlorite (%3.5) for 10 minutes. Two isolation media did not show a significant difference on the viability of microspores. Among induction media tested, in cv. Amarosa, the highest viability was observed in AT3 medium supplemented by glucose. Induction media in Apollo cultivar did not show a significant difference on viability of microspores. Combination of starvation (B medium) and cold (4°C) treatments for 3 days induced formation of pro-embryos in cv. Amarosa. Present investigation reports a protocol for induction of embryogenic developement in rose (Rosa hybrida) microspores.
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