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Intl. Res. J. Appl. Basic. Sci. Vol., 3 (3), 642-649, 2012ableaves with leaves stem rootpetiols without petiolsFig 2: a) Effect of different explants on production of transformed hairy roots, b) effect of age ofexplants on production of transformed hairy roots. Comparison of mean were done by LSD (differentletter on bars refers to significant differences ( p< /5).High percentage of transgenic leaves can be related to the high sensitivity of leaf explants to bacterialthan others considered ,which this sensitivity depends on the physiological status of tissues (Pavar &Maheshvari 2003). Hamill 1993 reported the key effect of rol genes on the inactive auxin and cytokninpathway become active due to the difference between the amount of a transgenic explants of a plant.Alsomany studies showed that the explants age can have many effects on the abundance production of hairyroots because of explants age affect on physiological properties of cells and thus affects its ability to betransgenic (Mehrotra et al. 2008). Meristem tissue and younger parts are usually the high proliferation andtherefore are suitable for being transgenic, the older plants are usually lower amplification . Increasingplant age and thus plant organ has led to further differentiation of tissues and cells. And consequently onthe ability of the cell proliferation and its ability to deal with the factors causing the disease is effective(Bageri & Safari 1387).Determine the best strain for infection and the root production in addition to using AR15834, the strainsA4, 9435 and C318 were also used. Results based on the fig 3- a showed that the highest transgenic ratewith 66% frequency obtained from use of strain AR15834, the statistics from the rest of the strainsshowed significant differences at 5% level.In different concentrations of bacteria the results showed that bacterial suspension with a concentration of0/6 was the best concentration of bacterial suspensions for infection of explants which showed thehighest percentage of transformation. Bacterial concentration of 0/6, with other concentration showedsignificant difference at 5% level (fig 3-b). aaabbbbbcbc
Intl. Res. J. Appl. Basic. Sci. Vol., 3 (3), 642-649, 2012Fig 3: a) effect of different strains of A. rhizogenes on production of transformed hairy roots, b) effect ofdifferent concenteration of A. rhizogenes on production of transformed hairy roots. (different letter onbars refers to significant differences ( p< 0/05).The results obtained indicate the same results of Kamada et al (Kamada et al.1986) and Manu et al (Manoet al.1986) studies,who for the first time to produce tropane alkaloids using Agrobacterium rhizogenesstrain ATCC15834 in Atropa belladonna and Scopolia japonica plants ,the hairy roots were successfullyestablished. According to these studies, the success of the transgenic plants formation by strain AR15834is likely because of the high sensitivity of this plant to the bacterial strain. The results of transgenic hairyroots induction showed that a wide variety of root growth were observed after co-cultivation withA.rhizogenes. Batra and colleagues reported (Batra et al.2004) that entering both the TL and TR or any ofthese sections alone can cause changes in growth and morphology of produced roots because they canchange the balance of hormones in these roots. Tiwari and colleagues (Tiwari et al.2007) reported in theirstudy of hairy root that, there is a direct relationship between rolB expression, root growth and lateralbranches of hairy roots. Advantages, including rapid growth, low doubling time, ease of maintenance andability to synthesize a range of chemical compounds of hairy roots has become this technique as animportant and permanent source of valuable secondary metabolite. Hairy roots can grow indefinitely onhormone-free medium and also due to the distinctive nature have genetic stability and produce highlevels of secondary metabolites (Pavar & Maheshvari 2003).Overall results showed that Agrobacterium rhizogenes strain AR15834 with the concentration of 0/6 wassuccessful for contamination and induction of hairy roots in 14-day old leaf explants of P.oleracea (fig 4).
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