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Title: Agrobacterium rhizogenes mediated-transformation of Asimina triloba L. seedling cuttings

Author
item Ayala-Silva, Tomas
item BEYL, CAULA - AL AGRIC & MECH UNIV
item DORTCH, GREGG - AL AGRIC & MECH UNIV

Submitted to: International Journal of Biological Sciences
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/25/2006
Publication Date: 10/1/2006
Citation: Ayala Silva, T., Beyl, C., Dortch, G. 2006. Agrobacterium rhizogenes mediated-transformation of Asimina triloba L. seedling cuttings. International Journal of Biological Sciences.

Interpretive Summary: Cuttings from pawpaw trees can be difficult to root. Innoculation with Agrobacterium rhizogenes at the base of cuttings could improve rooting of some fruit trees and ornamental species. This research compared rooting of pawpaw, with softwood cuttings from mature trees, cuttings from seedlings, and from shoots of rooted cuttings from seedlings. Four treatments were used: Agrobacterium rhizogenes strain MSU-1 (A4 wild type), A.rhizogenes strain MT232 (TR105 mutant), indole-3-butiric acid at 20,000 mg 1 -1 and control. Only the A.rhizogenes treatments induced rooting and only of the seedling cuttings. Among the bacterial strains, no statistical differences in rooting percentage were noted. The MSU-1 strain produced more roots, but these were shorter and thinner than those produced by MT232. No roots resulted from the other treatments. Roots obtained from infected seedlings were evaluated for transformation using polymerase chain reaction. Roots confirmed as transgenic were 100% for both rolB, and rolC. Index words: bacteria, annonacea, hairy root, fruit, DNA.

Technical Abstract: Cuttings from full-grown pawpaw (Asimina triloba) trees can be difficult to root. Innoculation with Agrobacterium rhizogenes at the base of cuttings in vivo/vitro can improve rooting efficiency of some tree fruit and ornamental species. The current research compared rooting of pawpaw with softwood cuttings from mature stands, cuttings from the terminal 3-4 inches of seedlings, and cuttings taken from shoots of rooted cuttings from seedlings. Four treatments were used: Agrobacterium rhizogenes strain MSU-1 (A4 wild type), A. rhizogenes strain MT232 (TR105 mutant), indole-3-butiric acid at 20,000 mg 1 -1 and control. Only the A. rhizogenes treatments induced rooting and only of the seedling cuttings. No statistical differences in rooting percentage were found among the bacterial strains. While the MSU-1 strain produced more roots, these were shorter and thinner than those produced by MT232 strain. No roots resulted from any other treatment. roots obtained from infected seedlings were analyzed for transformation using polymerase chain reaction targeting the folB and rolC vir genes. Roots confirmed as transgenic were 100% for both rolB and rolC. Index words: Pawpaw, Asimina triloba, transformation, IBA, DNA, rolB, rolC