Plant stem cell line derived from cambium of herbaceous plant with storage root and method for isolating the same
The present invention relates to a cell line derived from the cambium of an herbaceous plant having a storage root and a method for isolating the same. More specifically, relates to a cambium-derived homogeneous cell line having the ability to divide, which is obtained from the cambium-containing storage tissue of an herbaceous plant having a storage root without a separate dedifferentiation process, and to a method for isolating the same. The cell line derived from the cambium of an herbaceous plant having a storage root has active division ability and is homogeneous. Also, it is stable during culture, because it has not undergone a dedifferentiation process. Thus, through the optimization of proliferation thereof, the cell line can be allowed to proliferate in a large amount within a short time. Accordingly, the cell line derived from the cambium of an herbaceous plant having a storage root makes it possible to produce large amounts of useful plants which are difficult to cultivate outdoor due to various problems associated with the period of cultivation, the selection of cultivation land, cultivation cost and the like.
1. A method for isolating a cell line derived from the cambium of an herbaceous plant having a storage root, the method comprising:
(a) obtaining storage root tissue of an herbaceous plant having a storage root, wherein the tissue comprises cambium;
(b) treating the tissue such that differentiated, non-cambium cells lose the ability to divide, by subjecting the tissue to an agent which produces an osmotic stress;
(c) inducing cambium cell proliferation by culturing the obtained cambium-containing storage root tissue in an induction medium comprising a cambium-division-specific hormone, wherein the cambium-division-specific hormone is indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), or a combination thereof and;
(d) collecting the induced cambium-derived cell line.
2. The method of claim 1 , wherein the herbaceous plant is selected from the group consisting of Codonopsis lanceolata, Ostericum koreanum, Platycodon grandiflorum, Pueraria thunbergiana, Aralia contonentalis, Ledebouriella seseloides, Angelica gigas , carrot, sweet potato, Maca, and cassava.
3. A method for isolating a cell line derived from the cambium of a ginseng root, the method comprising:
(a) obtaining root tissue of a ginseng plant, wherein the tissue comprises cambium;
(b) treating the tissue such that differentiated, non-cambium cells lose the ability to divide, by subjecting the tissue to an agent which produces an osmotic stress;
(c) inducing cambium cell proliferation by culturing the obtained cambium-containing root tissue in an induction medium comprising a cambium-division-specific hormone, wherein the cambium-division-specific hormone is indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), or a combination thereof; and
(d) collecting the induced cambium-derived cell line.
4. The method of claim 3 , wherein the ginseng plant is selected from the group consisting of cultivated ginseng and true wild ginseng.
5. The method of claim 1 , wherein said induction medium comprises IAA or IBA at a concentration of about 0.1 to about 5 mg/l.
6. The method of claim 1 , wherein the treatment of step (b) comprises subjecting the tissue to an agent which produces osmotic stress wherein the agent is sucrose.
7. The method of claim 1 , further comprising transferring the cambium-derived cell line to a maintenance medium which supports continued proliferation of said cambium-derived cell line.
8. The method of claim 7 , wherein said maintenance medium comprises auxin.
9. The method of claim 8 , wherein said maintenance medium comprises one or more of 2,4-dichlorophenoxyacetic acid (2,4-D), picloram, or IBA.
10. The method of claim 9 , wherein said maintenance medium comprises 2,4-D, picloram or IBA at a concentration of about 1 to about 5 mg/L.
11. The method of claim 3 , further comprising preserving said cambium-derived cell line by freezing, wherein said cambium-derived cell line can be thawed and recultured.