Culture medium and culture method for inducing callus differentiation through peony leaves

Abstract

The invention relates to a culture medium and culture method for inducing callus differentiation through peony leaves. The method comprises the following steps: in late March, selecting tender peony leaves; after sterilizing the leaves, inoculating the sterilized leaves to an induction medium, wherein the temperature is 25+ / -2 DEG C; culturing in dark for 2 months; inducing to form callus; transferring the callus to a differential medium, wherein the temperature is 25+ / -2 DEG C; the illumination time is 12 h.d<-1>; the illumination intensity is 1200+ / -1001x; culturing for 2 months to obtain adventitious buds. According to the method, the callus initiation rate of peony leaves can reach up to 98.60%, the vitrification ratio is 0%, and the differentiation ratio of the adventitious buds can reach 58.99%; each callus block can form 4-10 adventitious buds, so that the technical problem that the ratios of adventitious bud differentiation is low when peony leaves are adopted as explants to induce callus is solved; the method provides technical support for carrying out peony gene engineering, and is wide in market prospect.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and in particular relates to a culture medium and a culture method for inducing callus differentiation through paeoniae officinalis leaves. Background technique [0002] Peony is a traditional famous flower in my country. Except for some areas in South China where the weather is hot and not suitable for growth, it is almost all over the gardens of the country, but most of them are conventional cultivated varieties, and there are few new varieties with novel characters. Therefore, Chinese peony growers And scientific research institutions are actively carrying out the work of peony variety improvement and new variety cultivation. The improvement of peony varieties and the cultivation of new varieties are mainly carried out through cross breeding. This traditional method has the disadvantages of long cycle, low efficiency, and inability to carry out directional improvement, which delays the pace ...

AI Technical Summary

Problems solved by technology

Wang Jifeng et al. (Wang Jifeng, Li Qing, Meng Hui. Research on callus induction and differentiation of five peony varieties. Journal of Beijing Forestry University, 2011,32(3):213-216.) published a paper 'The callus of five peony varieties Callus was successfully induced and adventitious buds were differentiated in "Research on Induction and Differentiation of Wound Tissue", but the callus was severely browned, vitrified, and brittle, and the bud differentiation rate was low, only 7.95%.

CN1675992A discloses a method for asexual reproduction of shoot tips, but the shoot tips do not go through the dedifferentiation process and cannot form root systems, so they have no application value in production and scientific research

CN104304029A also discloses a kind of asexual reproduction approach of rhizomes, which is still unsuitable for transgenic research and application

Sun Xiaomei et al. (Sun Xiaomei, Cheng Wan, Liu Ping, Zhou Wenqiang, Wang Dan, Yang Hongguang. Research on callus induction and adventitious bud differentiation of peony. Journal of Shenyang Agricultural University, 2014-02,45(1):24-27 .) In the published paper "Study on Callus Induction and Adventitious Bud Differentiation of Paeoniae Alba" it was stated that the callus was induced from the leaves of aseptic peony seedlings as explants, but the callus induction rate was low, only 32.2 %, which is far from the requirements of the peony genetic transformation regeneration system

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