| Strobilanthes cusia (Nees) Ktze is a perennial herbaceous plant, which belongs to Acanthaceae Strobilanthes. Strobilanthes cusia (Nees) Ktze’root and leaf have very high medicinal value, and in the clinical application it played a very good treatment effect. This paper has a further study on the Strobilanthes cusia (Nees) Ktze’s tissue culture system, in order to be able to find the optimal culture formulation of the Strobilanthes cusia (Nees) Ktze’s vitro rapid propagation. While in the process of Strobilanthes cusia (Nees) Ktze’s regeneration system, the tissue culture seeding appeared vitrification phenomenon. Therefor, the test explored the influencing factor of the cause vitrification as well as to study how to make the vitrification tissue culture seeding recover the normal seeding. The research results are as follows.(1) Selection of the best explants:4-6months is the best time to choose explants. The explants taken in this time period are low pollution rate, high survival rate and bud quickly. And the average contamination rate was41.88%, average survival rate was44.29%. The secondly optimum time of taking explants is1-3months, which average contamination rate and average survival rate were61.93%,26.97%.7-9months’s average contamination rate and survival rate(64.15%,24.40%) were not far from the10-12months(64.04%,24.03%). Terminal and axillary buds are the best explants,which carry less bacterial and sprout quickly.(2) The the best explant disinfection process:disinfected20s with75%alcohol, rinsed twice by sterile water, then with0.1%mercuric chloride+3-4drops of Tween disinfecting6min,at last with sterile water rinsing5times. No matter in what time to take the explants, this disinfection process showed lower pollution and higher survival rate(4-6months, the contamination rate and survival rate were10.42%,87.50%;7-9months were34.78%,56.52%;10-12months were39.13%,54.35%;1-3months were37.78%,57.78%).(3) The optimum medium formula of Strobilanthes cusia (Nees) Ktze’s buds start training. Combination of6-BA and NAA would shoot start induction experiments:The mediun MS+6-BA0.5mg/L+NAA0.05mg/L was the best; the average starting speed was6.55d; the germination rate reached96.67%. The combination of KT and IBA had effected on the induction time:The medium MS+KT1.0mg/L of+IBA0.05mg/L was the best, and the average start-up speed was7.42d, bud was95.00%.(4)The optimum medium formula of Strobilanthes cusia (Nees) Ktze proliferation culture was MS+6-BA0.3mg/L+KT1.0mg/L+IBA0.5mg/L, the multiplication coefficient of4.83. In this proliferation culture,6-BA showed the most significant, followed by KT, IBA impacted littlely.(5) In the strong seedling culture, MS+IBA0.5mg/L+GA30.1mg/L was the best. And in this process the numerical value of the auxin/gibberellin is higher, it’s the better for the plant woodythe formation. Thus which also reach the purpose of strong seeding cultivation and which is more conducive to the cultivation of rooting and acclimatization transplanting.(6) The best medium formula of rooting cultivation was1/2MS+IBA0.5mg/L+IAA0.05mg/L, in which the rooting rate reached96.67%. The research of ABT effect on the rooting, ABT1compared with ABT3, ABT1had a better impact effect. The medium of1/2MS+ABT11.5mg/L, the rooting rate reached75.00%. But on the whole, the combination of IBA and IAA had the better effect than the IBA on the rooting culture.(7) The optimum treatment of acclimatization transplanting:Before the tissue seedings were moved into the natural environment, first should be acclimated closely7d and then openly6d, at last transplaned. Just as do that the survival rate was65.88%. The transplanting matrix was sand:sandy soil:peat=1:1:1for the best, after transplanting the survival rate of87.47%.(8) The factors of impacting the Strobilanthes cusia (Nees) Ktze’s vitrification:The different concentration levels of6-BA, sucrose, agar and different pH values, different transit time, the different humidity, which all had the significant impact; While the different concentration levels of NAA and NH4+were not as significant as the above factors, which was not the main influencing factors of causing the vitrification. The range of concentrations of6-BA in this study will be controlled within the range of0-0.5mg/L didn’t cause more serious vitrification, but also conducive to the proliferation in Strobilanthes cusia (Nees) Ktze’s tissue cultivation. Sucrose’s optimum concentration is30g/L and the optimum concentration for the agar is8g/L; While if the both concentrations were too low which could obviously cause aggravation of the vitrification; Excessive concentrations could suppress the vitrification phenomenon, but the plants showed the negative growth. pH value of5.8can play a very good preventive effect on the Strobilanthes cusia (Nees) Ktze’s vitrification; The PH value was too low, the vitrification would serious increase; The PH value was too high, the vitrification rate would be significantly lower; But too high PH value was not conducive to the normal growth of the tissue culture seedlings. Shortened the cultivition time and sealed cultivation bottles after the medium cooled both could have been preventive effect on the vitrification. In this study, NH4+concentration had not great effect on the Strobilanthes cusia (Nees) Ktze’s vitrification, but the slightly higher NH4+concentration could be conducive to seedling growth.(9)The research on the vitrified plantlet’s reverse recovery:Medium supplemented with a certain concentration of activated carbon, AgNO3, polyving akohol and alternating temperature cultivation could play a very good recovery improvement on the tissue cultivation seedlings’ vitrification. Activated carbon concentration of4g/L was the best, the recovery rate of24.1%for the mild vitrification, complete recovery rate of25.74%for normal seedlings.4mg/L AgNO3concentration was the best for the vitrification recovery, which recovery mild vitrification rate reached23.88%, fully restored normal seedling rate reached28.96%, polyving akohol concentration of3g/L could have the best recovery effect, recovery as mild vitrification rate of8.01%, recovery rate of29.58%for normal seedlings;25℃/20℃variable temperature processing enables the vitrification recovery as mild vitrification rate of21.43%, recovery completely normal seedlings rate reached32.14%. |
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