The methods for isolation of cytoplasts in several crop plants

Autor:	Masami Watanabe, Hikoyuki Yamaguchi
Rok vydání:	1988
Předmět:	biology fungi food and beverages biochemical phenomena metabolism and nutrition Protoplast biology.organism_classification Cytoplast Phytotron Botany Centrifugation Hordeum vulgare Percoll Solanaceae Nicotiana
Zdroj:	Japanese Journal of Breeding. 38:43-52
ISSN:	2185-291X 0536-3683
DOI:	10.1270/jsbbs1951.38.43
Popis:	Leaf protoplasts were prepared from the plants grown in the phytotron with artificial illumination. Cytoplasts were obtained by ultracentrifugation of leaf protoplasts on the top of Percoll iso-osmotic discontlnuous density gradiants as described by LbRZ-et al. (1981). After centrifugation, the cytoplasts fraction was pipetted off from the top of the gradiant. According to microscopic observation, cytoplasts were highly vacuolated. chloroplast-poor cells and could be easily distinguished from the green leaf protoplasts. The number of cytoplasts and protoplasts in the cytoplast fractions was counted by Fuchus-Rosenthal haemocytometer. The optimal conditions for isolation of cytoplasts were determlned from a relationship of centrifugation speeds and running time. The yield of cytoplasts was decreased with increasing the diameter of leaf protoplasts. Cytoplasts isolated from Shiga's Cytoplasmic Male Sterile (CMS) rape (Brassica napus) were fused with leaf protoplasts of it's maintainer (B.napus cv. Bronowski) In the presence of polyethylene glycol. The Lusion process was observed with the passage of time by the color laser microscope. Cytoplasts and protoplasts were Lused immediately after their attachment.
Databáze:	OpenAIRE
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