الفهرس 
Material and methodsOnly 14 pages are availabe for public view
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Abstract
This investigation was carried out at the Tissue Culture Unit.
Horticulture Department, Faculty of Agriculture, Moshtohor, Benha
University during the period from 2003 to 2006.
New leaves from in vitro and sterilized in vivo pineapple
(Ananas comnosus cv. Smoth Cauun) and banana spp. (Musa cv.
Grand Naine) were taken and divided into small sections 1-2 mm.
These sections of both in vitro and in vivo explant source were
immersed in different enzyme mixtures in small Petri-dish (6 em in
diameter). Also, different shaking periods and speeds were used
during protoplast isolation stage. Meanwhile, different pore size of
mesh sieves and centrifugation speeds were concerned in filtration
stage. Moreover, different medium types, protoplast densities and
different concentrations of auxin and cytokinin as well as different
antibiotis were added to culture media, during protoplast culture
stage.
The obtained results can be summarized as follow:
5.1. Pineapple:
S.l.a. Protoplast isolation:
1- In vitro explant source surpassed in VlVO explant ill
increasing protoplast isolation.
2- Enzyme mixture consists of 1% cellulase + 0.5%
Macerozyme was more superior in protoplast isolation as
compared with the other enzyme mixtures used.
3- The highest protoplasts were obtained when in vitro explant
source was immersed in solution supplemented with sucrose
as osmotic pressure factor.
4- Incubating in vitro explant source for 20 hours in enzyme
mixture solution succeeded in maximizing protoplast yield.
5- Shaking the incubated mixtures of enzyme solution and in
vitro explant for 15 min. with speed rate 75 rpm encouraged
the highest protoplast yield.
S.l-b. Filtration:
1- 25 /lID pore size mesh sieve was superior in increasing the
protoplast yield of in vitro explant.
2- By increasing pore size mesh sieve had reduced protoplast
yield for both explants source used hence they increased cell
residues.
3- Centrifugation at the rate of 1000 rpm maximized the
protoplast yield of in vitro explant source.
S.l.c. Protoplast culture:.
1- Kao and Michayluk medium was more preferable in
increasing protoplast development. While, Gambourge (B5)
medium failed to be effective in protoplast development.
2- Culturing protoplast at density rate 2.5 x 104 was preferable
in maximizing protoplast development of in vitro explant
source.
3- The highest protoplast was obtained when in vitro explant
source was immersed in solution supplemented with
mannitol as osmotic pressure factor.
3- Addition of both 3.0 mg/L auxm ( AA) and’ 0.2 mgIL
cytokinin (BAP) to culture medium succeeded in inducing
the highest protoplast of in vitro explant source.
4- The best development of protoplsat as a result of reducing
contamination was noticed when adding the combination of
0.4 giL Ampicilin + 0.1 giL Gentamycin + 0.1 gIL
tetracycline was supplemented to the culture medium.
5.2. Banana:
S.l.a. Protoplast Isolation:
1- In vitro explant source was more superior than tn vivo
explant in increasing protoplast isolation.
2- Enzyme mixture consist of 1% cellulase + 1% Macerozyme
+ I% pectinase was more preferable an increasing protoplast
isolation.
4- Incubating in vitro explant source for 24 hours in enzyme
mixture solution induced an increase in protoplast yield.
5- Shaking the incubated mixtures of enzyme solution and in
vitro explant source for 15 min. with speed rate 75 rpm
caused the highest protoplast yield.
S.l.b. Filtration:
1- 25 /lID pore SIze mesh SIeve gave a good response
concerning protoplast yield of in vitro explant source.
2- Increasing pore size mesh sieve gave negative result III
reducing protoplast yield for both explant sources used since
they increased cell residues.
3- Centrifugation at the rate of 1000 rpm maximized protoplast
yield of in vitro explant source.
S.2.c. Protoplast culture::
1- Murashige and Skooge medium encouraged best protoplast
development. While KM medium was inferior in this
respect.
2- Culturing protoplast at density rate 2.5 x 104 increased
protoplast development of in vitro explant source.
3- Adding the combination of 3.0 mg/L Auxin (NAA) and OJ
mg/L cytokinin (BAP) to the culture medium induced the
highest protoplast development of in vitro explant source.
4- Best development of protoplast as a result of reducing
contamination was obtained when adding the 0.4 giL
Ampicilin + 0.1 g/L Gentamycin + 0.1 gIL tetracycline to
the culture medium.