Pre-isolation, isolation and regeneration protoplasts from leaf mesophyll of in vivo Malus domestica 'Anna' cv.

文献类型: 外文期刊

第一作者: El-Gioushy, Sherif Fathy Eid El-Sayed

作者: El-Gioushy, Sherif Fathy Eid El-Sayed;Kareem, Abdul;Baiea, Mohamed Hemdan Mohamed

作者机构:

关键词: Apple; Hybridization; Cellulose; Pectianase and Macrozyme

期刊名称:REVISTA BRASILEIRA DE FRUTICULTURA ( 影响因子:0.912; 五年影响因子:0.968 )

ISSN: 0100-2945

年卷期: 2019 年 41 卷 4 期

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收录情况: SCI

摘要: To isolate protoplast, a pre-treatment was completed with the order to reduce and identify the phenolic contents round the year to encourage the isolation of protoplasts. Protoplasts from in vivo mesophyll leaves of apple cultivar "Anna" was isolated from 15 days old leaves by plasmolying in medium containing 90 g L-1 mannitol for half hour, then 130 g L-1 mannitol for half hour. Then using enzymatic mixture involving (1.5% cellulose + 0.5% pectianase + 1.5% Macrozyme) Prior to isolation. Anyhow, diverse factors, for example, Osmotic pressure, incubation period, sieve pore size, centrifugation period and hormonal balance was estimated using the techniques for isolation. The quantity of cells was computed as the quantity of cells per square on haemocytometer. A considerable higher yield of protoplast formation was noted in the CPW medium using a pore size of 25 mu m with using incubation period for 20 hours. Moreover, the best protoplast regeneration with using of protoplast density of 2.0 x 10(5) in MS medium supplemented by 1.0 mg L(-1)NAA and 0.3 mg L(-1)BAP. We believed that our protocol might encourage the plant recovery using in apple somatic hybridization programs.

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