| As one kind of protected and rare and endangered plants in second class, the natural regeneration of population on Chosenia arbutifolia (Pall.) A. Skv has been disturbed seriously by human beings. It is necessary to strengthen the research in the characteristic of seed germination since its breeding strategy is mainly by seedlings, providing the basis for the whole species’expanded reproduction. This article studies the effect of seed germination by the storage date and three environmental factors (light, temperature and water), then gets the conclusion of various conditions, storage 3 weeks in 4℃; 70% light intensity; 25℃; 2mm in moisture.In order to set up the high efficient tissue culture and rapid propagation system then provide theoretical guidance for the conservation, axillary buds from stem segments which derived from the healthy cuttings seedlings in the greenhouse as the experimental materials are used to screen the best disinfection method, medium of induction and differentiation. Axillary buds from stem segments which collected in the greenhouse were used as explant, the impact of disinfection methods on its germination of axillary buds were studied in this paper by ultrasound-assist method L9(32):2 factors(70% alcohol and 0.1% HgCl2) and 3 levels (different times of two disinfectants). The best disinfection combination is the time of 70% alcohol disinfection is 10 seconds and the time of 0.1% HgCl2 disinfection was 8 minutes by comparing the death rate, having the lowest polluting rate, the highest germination rate and having the latest initial germination time. Range analysis results showed that the time of alcohol treatment is the main factor of inducing germination of axillary buds. But the HgCl2 treatment time is the main factor of disinfection effect.In order to determine the best induction medium through comparing the germination rate and growth condition of axillary bud by using 5 kinds of basic culture medium (WPM,DCR, MS,N-68 and ACM).The results show that the growth condition of axillary buds is the best under WPM culture medium condition, in which the leaves were deep green and expansion, and stem were stronger and the germination rate is also the highest. After the induction, The 4 kinds of basic mediums(MS,WPM, ACM, DCR),3 kinds of hormone(IBA,6-BA and GA3) and 4 levels are applied for the four factors-four levels (L16 (44)) orthogonal experiment to study the differentiation of axillary buds. The result show that The DCR medium containing 0.40mg/L 6-BA,0.30mg/L IBA and 0.40mg/L GA3(91.67%)and DCR medium containing 0.30mg/L 6-BA,0.30mg/L IBA and 0.50mg/L GA3(91.84%), the effect of multiplication is the best.)are screened as the optimum differentiation medium, and the basic medium and GA3 are the main factors affecting the differentiation rate, whereas IBA and 6- BA are the secondary factors. Continuing the multiplicating proceed to screen the best multiplicating medium then the result is DCR medium containing 0.30mg/L 6-BA,0.30mg/L IBA and 0.50mg/L GA3,and the effect of multiplication is the best, when the subculture cycle is 35d, the multiplication can ever reach the highest and even the seeding growth is the best.This research studies the characteristic of seed germination and rapid propagation technology, laying a foundation for preserving germplasm resource efficiently and propagating well about Chosenia arbutifolia, also providing information for the protec-biology research on other rare, endangered plants. |
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