|关键词||新疆雪莲 高频再生 条件优化 瓶外生根|
This study focused on different relevant factors on adventitious shoots formation, multiplication, rejuvenation and rooting of Saussurea involucrata Kar. et Kir. In the experiments we established a stable plantlet regeneration system of this endangered plant. Furthermore, we succeeded in out test-tube rooting in this plant for the first time. Our established regeneration system simplified the process of operation, shortened the period of rapid propagation and increased the survival rate of transplant seedlings. This research laid the technical foundation for large scale artificial propagation of S. involucrata Kar. et Kir. The establishment of high frequency adventitious bud regeneration system of S. involucrata Kar. et Kir was stable and repetitive. The effects of ratio of differnent hormones, explant types, culture conditions on regeneration time and regeneration frequency were analyzed. The results were as follows: different parts of 25-day old seedlings served as explants and were cultured on MS medium with 1.0mg/L 6-benzyladenine (6-BA), 0.1mg/L α-naphthalene acetic acid (NAA) and 0.1mg/L 2,4-Dichlorophenoxyacetic acid (2,4-D) for 30 days; then the explants were transferred to MS medium with 1.0mg/L 6-BA and 0.1mg/L 2,4-D for another 15~20 days. Large quantities of adventitious buds were induced in the 45-day terms. The budding ratio was at the highest frequency when using leaf as explant (up to 66%) and each explant could get 6.1 adventitious buds. Through the research on effects of different environmental factors, we determined the best conditions for proliferation and rejuvenation of S. involucrata Kar. et Kir. The buds were proliferated on MS medium supplemented with 0.5mg/L 6-BA and 0.05mg/L NAA. The best inoculation density was 12~16 buds each bottle. The materials should be transferred to MS medium supplemented with 0.05mg/L paclobutrazol (PP333) to release hormone and rejuvenation every 2~3 generations. The proliferation multiple of adventitious buds reached 5.2 at 20±1℃ and the buds grew well. Out Test-tube rooting of Saussurea involucrata Kar. et Kir was realized for the first time. Compared with inner Test-tube rooting rate, out Test-tube rooting rate was a bit lower but the livability after transferring-planting was obviously higher. The quality of Test-tube seedlings was a crucial factor for out Test-tube rooting. Our results indicated that the perfected culture process for out Test-tube rooting was as follows: Firstly, the strong buds was brought out from bottle after trained for 3 days and washed thoroughly clean, then immersed for 30 min in the solution supplemented with 1mg/LNAA and 1g/L chlorothalonil. Secondly, the treated strong buds were cuttaged in the sterilized vermiculite. The volume ratio of vermiculite and solution supplemented with half-strength MS macroelementsand was between 2:1 to 3:1. Out Test-tube rooting ratio reached 80% after 30d at 20±1℃ and 3500lx. The regenerated plants was transplanted into artificial soils in greenhouse, the transplanting survival rate was 100% with shading and moisture retention.
|傅晓春. 新疆雪莲组培快繁体系优化的研究[D]. 北京. 中国科学院研究生院,2007.|
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