| Castanea henryi , both of its seed and wood being useful , has very high economic worth , for example, its fruit tasted well and enjoyed for a long time, its timber for making furniture, its flower as nectar source, etc.. Its fruit is a kind of rare woody grain, contain a large number of starch , protein and various kinds of vitamin in the pulp. But the cloning is uneasy to make Castanea henryi rooting. At present, there are many problems in the research of Castanea henryi, such as the confusing cultivar series , the serious disease, etc., and some studys on Castanea henryi are nearly in the macroscopic field , little in the microcosmics field.This research, for setting up the regeneration system of Castanea henryi by means of tissue culture offer the basic means and theoretical foundation for stock breeding, orientation fosters and heredity transform and gene cloning, etc..The material adopted in this research came from Shuiyuang village of Jianou town in Fujian Province. There are 13 cultivars of Castanea henryi, such as Youtongzi, Youzhen, HuangZhen, DaJianZhui, YouLiZi, BaiLuZi, ZhongJianZhui ,WuZhen, WuKeChangMang, OuNingZi ,ChuSuHong, ShuiZhen, from which the embryo, axillary bud, bast etc. were taken as explants for being cultured and regenerated into plantlets, by means of callus culturing, enrich callus culturing , differentiation callus culturing, induced adventitious bud culturing, enrich adventitious bud culturing and rooting culturing . Various elements and hormones, hormones concentrations, varietys of explants, the inoculation way the browning sucrose kinds and concentrations the vitamin B1. the gibberellin and other organic nutrition in culture medium, such as the agar, the pH and the external environmental condition etc., were tested in tissue culture of Castanea henryi. The following results had been received:.1. The different concentrations of mercuric chloride and sodium hypochlorite disinfected to the explants of embryo , axillary bud, basts under different time, worked not well. But the embryos which directly draw from the kernels disinfected by 75% of the alcohol are better to controlling pollution rate.2. Taking care of 13 cultivars of Castanea henryi as material , YouZhen is regarded as first-class ones, HuangZhen and DaJianZhui etc. as second, ChuSuHong etc. as third, YouTongZi, ShuiZheng and BaiLuZi etc. as the fourth. When different organs of Castanea henryi were taken as explants, the embryo was regarded as the best. In inoculation way, the radicle insertingculturing was regarded as the best.3. The basic culture medium of callus culture is MS. When callus was induced , 2,4-D was better than NAA and IBA. while enriching callus, the growth hormone is better than the mitosin, and 2,4-D is the best kind of the growth hormone. Differentiating callus showed that BA is superior to KT, IBA is superior to IAA. The differentiating effect changed from good state to bad one as the decrease of the density of B A. The callus had hardly vitality and was likely browning to death when the concentration of BA reach to 5 mg/1. Considering the function of mitosin and the growth hormone , when the 3 mg/1 of BA matched with the 0.5 mg/1 of NAA and the 1.5 mg/1 of BA matched with the 0.2 mg/1 of NAA0.1 or the 0.5 mg/1 of NAA ,the result of differentiation was good. It is obvious that different kinds of density mitosin matched with the different kinds of density growth hormone. AgNO3 1mg/1 had not function on the callus defferentiation.4. The basic culture medium of Castanea henryfs adventitious bud is M medium. In preliminary test of induced adventitious bud, BA with low density as 11.5mg/l was better than other higher density treatment. IBA is the best kind of growth hormone, the second is NAA, and the third is IAA. In the experiment of hormone matching, when the proportion of the mitosin and the growth hormone reached 8:1, the normal adventitious bud could be brought out. But there were certain accommodations, and the experiment tested out three kinds of good treatments: BA+NAA=2m... |
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