Study On The Technique Of Tissue Culture Of New Guinea Impatiens

To explore the efficient way in vitro culture ,the study aims at establishing a rapid and effective technique system of tissue culture of New Guinea Impatiens.Through direct inducting of adventitious buds,the stems with axillary buds were used as explants.In addition,the leaves and the stems were utilized to induce callus which can induce cluster buds to multiply. Experiment was designed primarily by single factor and orthogonal design. Through variance analysis and multiple comparisons,we foud out a series of technical parameter combination for the tissue culture of New Guinea Impatiens which provided the scientifical and technical support for its factorization production.The main results were as follows:(1)The suitable explant of tissue culture of New Guinea Impatiens was the top of the new stem with axillary buds which showed lower pollution rate and higher differentitation rate.(2)The pretreated explants immersed in 2.5% NaCIO for 10-15 minutes or in 0.1% HgCl₂ for 5-7 minutes were sterilized well. (3)NAA and 6-BA could promote the differentiation of adventitious buds and showed the highest differentiation rate in the treatment of 6-BA 2.0 mg-L^-1+ NAA 0.1 mg-L^-1. (4)pH 5.8-6.4 was suitable for the differentiation of adventitious buds.The differentitation rate was low in the neutral condition. (5)MS culture medium was the optimum medium for multiple culture. (6)NAA and 6-BA were the suitable auxins and cytokines respectively in vitro culture of New Guinea Impatiens. In addition, the organic accretion had certain effects on multiple culture of the adventitious buds, and the best multiplication efficiency was obtained in the treatment of 6-BA 2.0 mg-L^-1+NAA 0.2 mg-L^-1+LH 200 mg-L^-1. (7)The multiplication cycle for about 45 days, can both guarantee the quantity and the quality of the adventitious buds. (8)The cane sugar could improve the multiplication coefficient significantly, and the suitable concentration range were 30-40 g-L^-1 .(9)The cooperation of NAA and cane sugar had prominent effects on the adventitious bud height. NAA 0.1 mg-L^-1 cane sugar 5 % was the best treatment for making the weak buds stronger. (10)Even in the MS medium without any hormones, test tube shoots of New Guinea Impatiens could rooted easily, but the roots showed thin and weak.Adding the auxin in the MS medium could improve the quality of the plant roots. MS+0.1 mg-L^-1BA and 1/4MS+0.5 mg-L^-1IBA were suitable for the culture of rooting.The root length and thick degree were moderate and the amount ofthe roots were large in the two treatments. ODThe suitable acclimatization time were 5-7 days.The well-penetrated substance adapted to the plant transplantation in the initial stage and pure vermiculite was the most suitable transplantation substance. (12)The treatment of 2,4-D0.2mg-L^-1+6-BA0.1 mg-L^-1 which could guarantee the quality and induction rate of the callus, was the most suitable hormone matching of callus inducement. (13)Dark culture was superior to light or week light culture for the inducing of blade callus. (14)To overcome the brown phenomenon,the method of scratching explants could promote the callus inducement and improve the effects of callus inducement.Scratching explant is favorable to the callus inducement. (15) In practical tissue culture production of New Guinea Impatiens,the sand sugar and soft sugar could replace the cane sugar as the source on the multiplication growth stage and tap water could replace distilled water to reduce the culture cost.