Open AccessProtoplast isolation from leaf mesophyll of sugarbeet Beta vulgaris L. axenic seedlings
Author(s) -
Qutaiba Al-Nema,
Mozahim K. Al-Mallah
Publication year - 2013
Publication title -
maǧallaẗ markaz buḥūṯ al-taqniyyaẗ al-aḥyāʾiyyaẗ
Language(s) - English
Resource type - Journals
eISSN - 2708-1370
pISSN - 1815-1140
DOI - 10.24126/jobrc.2013.7.3.280
Subject(s) - protoplast , cellulase , axenic , biology , botany , somatic fusion , cell wall , horticulture , somatic cell , enzyme , gene , biochemistry , bacteria , genetics
Protoplasts were isolated from leaf mesophyll of sugarbeet (Beta vulgaris L.) axenic seedlings. Eight enzyme mixtures were tested for cell wall degrading ability. The efficient enzyme solution was mixture "II" that consist of 1.5% Cellulase RS, 2% Cellulase R10, 1% Macerozym R10 and 0.1% Pectolyase Y23. This mixture was efficient in releasing protoplasts and gave high yield of a density 7.3 × 104 protoplast / ml. These isolated protoplasts were viable 93%, their sizes ranged from 13 up to 52 µm, vacuolated and unvacuolated. This finding enable workers to focus on somatic hybridization through protoplast fusion to overcome some barriers facing gene transfer to improve plant species such as transfer of N2 fixation ability to non-legumes and producing resistant varieties to biotic stress and other traits.