Method for rapidly propagating Phellodendron amurense by utilizing callus regeneration pathway

Abstract

The invention provides a method for rapidly propagating Phellodendron amurense by utilizing a callus regeneration pathway. The method disclosed by the invention comprises the following steps of: adopting an aseptic seedling stem of Phellodendron amurense as an explant, and culturing the explant on a culture medium containing 1.5 mg / l of MS+BA, 0.5 mg / l of NAA, 20 g / l of glucose and 6.0 g / l of agar, wherein the inductivity of callus is 100%, the adventitious bud incidence rate of the callus can be up to 75.4%, and the number of average regeneration adventitious buds of each explant is 14.7; continuously culturing regeneration adventitious buds on a successive multiplication culture medium containing 0.5 mg / l of MS+BA, 20 g / l of cane sugar and 6.0 g / l of agar, wherein the average multiplication multiple after one month is 3.4; carrying out rooting culture of regeneration seedlings on a culture medium containing 0.8 mg / l of 1 / 4MS+IBA, 20 g / l of cane sugar and 6.0 g / l of agar, wherein the incidence of adventitious roots is 90.6%; and the number of average regeneration adventitious roots is 4.3; and transplanting strong regeneration seedlings, wherein the transplanting survival rate is 100%. According to the invention, a regeneration system for regenerating the adventitious buds of explanted stems of the Phellodendron amurense has high-frequency property and repeatability; the culture period is short; a very good foundation is laid for rapid propagation of Phellodendron amurense tissue culture and improvement of Phellodendron amurense by using genetic engineering.

Description

Technical field: [0001] The invention is a method for rapidly propagating Phellodendron amurense through a callus regeneration approach, which is simple in operation, short in cycle, high in adventitious bud regeneration frequency, and fast in propagation speed. The invention belongs to the field of agricultural biotechnology. Background technique: [0002] Amur cork (Phellodendron amurense Rupr.) belongs to Rutaceae (Rutaceae) Phellodendron genus, is one of the three precious broad-leaved tree species in Northeast my country, is an important medicinal plant and timber tree species, is a relic plant of the ancient tropical flora in the third century, and is the second largest plant in the country. Level protected endangered species (Fan Jihong, 2006). Berberine, an important secondary metabolite in Phellodendron amurense, is the main active ingredient of Coptis chinensis, a traditional Chinese medicine, which has anti-inflammatory, anti-tumor, and immune-enhancing effects. ...

AI Technical Summary

Problems solved by technology

[0003] Research on the tissue culture of Cortex Phellodendron is relatively few at present, adopting the seed of Cortex Phellodendron as explant (Zhao Bo, master's degree thesis of Northeast Forestry University, 2007) has seen research report, but adopting seed to be that explant handles more trouble, Because each explant needs to be sterilized, if the sterilization is not complete, it is easy to cause a large number of explants to be polluted