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Factors Affecting High-frequency Induction of Hair-like Roots of Reynoutria Japonica Houtt.

YU Shu-hong,ZHAO Li-li,WANG Wei,ZHANG Di-qun~*(College of Pharmacy,Hebei Medical University,Shijiazhuang 050017,China)  
Using Agrobacterium rhizogene ATCC15834,the study transformed Reynoutria japonica to probe the factors affecting the induction of its hair-like roots.The results revealed that the mature leaf blades were the explants with high transformation frequency,which could forms hair-like roots about ten days after the inoculation with their induction rate averaging 55.66% and their root density standing at(4.30).It was proved by PCR test that the genes rolB and tms in Ri plasmid of Agrobacterium rhizogene were completely transformed into the hair-like roots.The inoculum activation by improved YEB culture solution did not significantly affected hair-like root occurrence;the leaf blades directly inoculated without being pre-cultured performed in rooting significantly better than those pre-cultured with MS_0 or TDZ and showed a hair-like root induction rate as high as 81.84% and an induced-root density of 7.04;soaking with diluted inoculum solution,supersonic oscillation or microwave radiation could make the leaf blades form roots early and that 30-second supersonic oscillation showed the most significant root-inducing effect(P0.01);The culture of the inoculated leaf blades with their epicuticles facing upwards was conducive to hair-like root formation.With the different transformation factors optimized,the induction rate of the hair-like roots of Reynoutria japonica could be raised to 97.1% and its hair-like root density could reach 11.3.
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