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نويسندگان :(Abbas Yadollahi (Corresponding author     تاريخ انتشار : April 1, 2012
Micropropagation of GF 677 Rootstock
Abstract: The aim of this investigation was to study micropropagation of Almond GF rootstock on in vitro. Explants were cultured in MS medium containing 5 concentrations of 6- Benzylaminoporine (BAP) 0.1, 0.6, 1, 1.5 and 2 mg 1-1 along with control treatment. Each treatment includes 5 replications. After 5 weeks of culture, the results showed that the number and length of shoots in treatment containing 1 mg 1-1 BAP was more than the other treatments

  Introduction
 
GF is a hybrid of Prunus amygdalus × P. persica and is the most commonly used rootstock for peach orchards
(Antonopoulou et al., 2007). This rootstock is tolerant to Fe deficiency and specially suited to soils with poor
fertility, low water availability and high CaCo3 content (Monticelli et al., 2000). It is propagated with vegetative
methods (cutting and tissue culture). Due to showing a varying rooting percentage from year to year and low
efficiency of cutting, tissue culture is a good and fast method for propagation of wealthy and disease-free plants
of GF 667 in large quantity (Dimassi et al., 1995). Hence, a lot of researches about micropropagation of GF
rootstock have been conducted (Antonopoulou et al., 2005). The first investigation about GF micropropagation
was carried out by Kester (1970) and Tabachnik and Kester (Tabachnik and Kester, 1977). One of the main
factor on micropropagation is hormone specially BAP. Furthermore, shoot branching depends on the initiation
and activity of axillary meristems, which are hormonally controlled by cytokinin (Dobranszki and silva, 2010).
The cytokinin BAP promotes cell division, shoot multiplication and axillary bud formation while inhibiting root
development (Sutter, 1996). In most work on shoot multiplication of GF almond rootstock, BA was used as the
cytokinin source mainly in a concentration range between 0.1 and 3 mg 1-1 (Kamali et al., 1990; Ahmad et al.,
2003). It has been reported that 0.6 mg 1-1 BAP produced the highest number of shoot (Ahmad et al., 2003). In
the rooting stage, the induction of roots on explants from in vitro culture is crucial part in any micropropagation
process (Molassiotis et al., 2003/4). The ability of plant tissue to form adventitious roots depends on interaction
of many exogenous and endogenous factors, including hormone. Most reports of adventitious root induction of
woody species have involved treatments with exogenous auxins such as IBA, NAA or IAA (Ainsley et al., 2001).
Exogenous auxins are only required at an early stage to stimulate emergence of new formed roots (Dobranszki
and silva, 2010). It has also been reported the best hormone for rooting was IBA (0.5 mg 1-1) (Vaez-Livari et al.,
2005). Except hormone, other factors that can affect adventitious rooting are included photoperiod, light
intensity and light quality (Rugini et al., 1993). Darkness during the last week of the rooting phase has been
shown to be necessary in stimulating rooting in some woody species (Rugini et al., 1993). The positive effects of
Darkness have also been reported on root number, root length (Vaez-Livari et al., 2005)The objectives of this investigation are included study of a) the effect of Benzylaminoporine on
micropropagation of GF rootstock b) the effect of different rooting-hormones on rooting of GF rootstock on in
vitro condition c) the effect of Darkness on rooting of GF rootstock on in vitro condition
Materials and Metods
  Plant Material Sterilization
Plant explants were collected from controlled plants in greenhouse of faculty of agriculture in winter of 2010. At
first, shoots were excised into 3 cm-long sections, and then for surface disinfection, they were agitated for 5 min
in a solution containing 5 drops of Tween-20 in 100 ml of water, finally they were washed under running water
for 1 hr. For sterilization, firstly explants were agitated in alcohol 70% for 30 sec, then in %0.01 solution of
mercuric chloride for 7 min and finally they were rinsed three times with distilled water
  Basal Medium and Culture Conditions
MS (murashige and SKoog, 1962) medium containing 5 concentrations of BAP along control treatment were
tested (Table 1). The media were supplemented with 30 g l-1 sucrose, 8 g l-1 agar and 50 mg 1-1 citric acid. The
pH of media was adjusted to 5/8 before autoclaving. The explants were maintained at 25± 1 ºc and 16/8 hr
photoperiod for 4 weeks. Data were recorded after 5 weeks.
For rooting, 5 cm-long shoots from previous culture were transferred to 1/2 MS medium containing 200 mg 1-1
Fe-EDDHA, 6 g l-1 agar. Treatments are mentioned in Table 2 and 3. After culturing shoots in different
treatments, jars containing shoots were divided into two groups. Half of each treatment was maintained in the
light and the other half was maintained in the dark for 1 week. The pH of media was adjusted to 5/8 before
autoclaving. The explants were maintained at 25± 1 ºc. Data include percentage of rooting, length and number of
roots were recorded after 1 month
  Data Analysis
The experiment was carried out based on completely randomized design (CRD) with 5 replications per treatment
Statistical analysis of the data was carried out by using SPSS 16 software and difference among treatment means
were compared by using Least significance difference Test (LSD) (Steel et al., 1997)
For rooting, the experiment was carried out based on factorial. Statistical analysis of the data was carried out by
using SPSS 16 software and difference among treatment means were compared by using Least significance
difference Test (LSD) (Steel et al., 1997)



 

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دفتر تهران : خیابان کارگر شمالی_ خیابان شهریور_جنب اورژانس بیمارستان قلب پلاک 25 واحد4
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