Clones, hairy root

The "hairy root" clones that are produced can be cultivated to produce metabolites (Hjortso and Flores, 1987). 

After 3 weeks from the co-culture with bacteria, survival rate of the leaf explants, frequency of hairy root formation and number of hairy roots per explant were determined. Figs. (39A and 40). The bacteria concentration and duration of co-culture greatly affected the induction rate the highest rate of 50% was obtained when the leaf explants were co-cultured for 2 days with 10 bacteria, Fig. (40B). In this condition, two hairy root clones per explant were obtained, Fig. (40C). 

A. Induction of hairy roots by co-culture method, Infected segments were cultured on 1/2 MS solid medium containing 500 mg/1 Claforan at 25 °C in the dark. B The selected hairy root clone that demonstrated typical hairy root morphology. The hairy roots were cultured in MS liquid medium at 25 C in the dark for 4 weeks. C The regenerated shoots from the hairy (left) and adventitious (right) roots. The shoots were cultured on MS solid medium at 25 C under 16 hr/day light. 

Tropane alkaloid production of hairy root clones... 

Fig. (49). Hyoscyamine concentration and yield found in the different hairy root clones of A. belladonna (n=2-5)...

Organized growth cultures maintain their original organ structure such as root cultures and embryo cultures. Root cultures can be established from root tips taken from many plants. The hairy root clones that are produced can be cultivated to produce metabolites. Embryo cultures may be established from embryos removed from sterilized seeds, ovules, or fruits. Embryo cultures can be employed for the rapid production of seedlings from seeds that have a protracted dormancy period. 

Five hairy root clones of D. lanata were established by infection of leaf... 

Using this protocol about 20 "hairy root" clones of... 

In our laboratory, in order to study the influence of plant ecotypes in tropane alkaloids profile, we analyzed and compared the alkaloid production in different organs of Argentinean and Colombian B. Candida in vitro -germinated seedlings as well as in hairy root clones obtained fi-om both ecotypes [27]. In addition, we scale-up one of the Argentinean hairy root clones obtained in a 1.5L bioreactor [28]. 

The results reported by Zarate et al. [62] about the clone 19 of baetica hairy roots were coincident to the results that we found for Colombian hairy root clones. In this case, the scopolamine content predominated over the 6P-hydroxyhyoscyamine one [27]. 

Figure 1. Hyoscyamine, 6p-hydroxyhyoscyamme and scopolamine content in (A) hairy root clones (HR). Argl-Arg4 HR obtained from Argentinean 5. cawJzt/a plants. C0II-C0I6 HR obtained from Colombian B. Candida plants. (B) B. Candida stem, leaf and root samples from Argentinean and Colombian plants. L leaves R roots S steams A Argentinean ecotype C Colombian ecotype [27].

Several reports have determined that the overexpression of /j(5/jgene resulted in the accumulation of higher amounts of tropane alkaloids. Among them, Hashimoto et al. [85] introduced this gene in. belladona hairy roots. The resulting roots contained almost 5-fold scopolamine concentration than wild type hairy roots [85], In addition, Jouhikainen et al. [14] overexpressed the H6H in H. muticus. The results showed that the best hairy root clone produced a 100-fold higher alkaloids concentration than the others [14]. 

Sevon N, Hiltunen R, Oksman-Caldentey KM (1998) Somaclonal variation in transformed roots and protoplast-derived hairy root clones of Hyoscyamus muticus. Planta Med 64(1) 37 1... 

Isolation of individual polyploid hairy roots clones... 

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