| In order to find the method that can culture and propagate Liparis nervosa rapidly,we searched for the reserving method using it's seed as experiment material and tried to find out the influence of low temperature at 4? on seed activity and snail enzyme,NaOH on the sprout of seed.The protocorms were used as experiment material with 6-BA,NAA,S-3307 as affecting factors,designing three factors and three standard experiment to select the most appropriate medium that induce lateral bud.The root and plantlets wore used as experiment material for classification and purifying the inner fungus.The fungus was co-existing with seed and procotorm in the saw dust to select the most suitable fungus that can promote the seed sprout and the growth of procotorm.The plantlets were used as the experiment material,choosing bark,saw dust,and cow dung as single matrix or blend matrix to search the various influence of different matrix to the growth.Using N,P,K as three factors,and Box-Behnken Design experiment,search for three different kinds of nutriment solution to the growth of plants.Using the L.nervosa which were cultured for 6,12,18,24 months as materials,the search of the dynamic change of component and the growth in different time were cerrned out.The results show:1.Time is an effect fator for activation of seed,the activation of fresh seed is 93.8%.As the reserving time increased,the activation of seed decreased to 75.2% after two weeks,decreased to 64.4%,and decreased to 56.6% after six weeks.Dealing with snail enzyme,the probability of seed propagating increasing obviously.The W-5 reached highest to 71%.The most suitable combination is 2 mg/ml snail enzyme with 40 mins.After deposing with NaOH,2 mol/L NaOH for 5 mins show the most sprout rate reaching 56.5%,but lower than other groups.So deposing with NaOH solution can't increase the surving probability of seeds.The most suitable one for stem segments propagation is MS + 1.0 mg/L S-3307 +1.5 mg/L 6-BA + 1.0 mg/L NAA + 0.5 g/L Carbon activated.2.Compared with the root grinding,the root segments can get more fungus.The number of fungus decrease with time increaseing.0.13% ClO2 sterilization for 3 mins can extracted fungus,reaching 72.86%,only 19.95% fungus can get with 2% NaClO sterilization for 12 mins.In addition,the sepecres of fungus changed with seasons.Winter can get the most number of fungus,reaching 72.69%.This experiment isolated 45 kinds of fungus,of with one can promote the sprout rate of L.nervosa,eight kinds of fungus can help the differentiation of plants.Protocorm growed slowly after it became a plantlet which indicate the the sprout process,protocorm differentiation and the growth of the plants depend on various kinds of fungus.3.The design of monotypic center of gravity was used and found out the most suitable percentaage of saw dust,bark and cow dung as 36.7%,0% and 63.3%,as a result,the plantlet reached 8.46 cm in height;when matrix were mixed in a percentage of 16.7%,36.7% and 46.7%,the stem diameter reached 8.87 mm;To get the most plantlet height and stem diameter,the best matrix is 19.3%saw dust?30.1% bark and 50.6% cow dung.And the plantlet height was about 8.21 cm and stem diameter was 8.75 mm.To verify the most suitable matrix,the values of 8.19 cm and 8.70 mm were got as expected.N,K showed significant effect on plantlet height,but not P.All there elements have significant influence on stem diometer The most suitable medium is 160.00 mg/L N,150.00 mg/L P and 200.00 mg/L K which promote the plant growth to 8.24 cm in height with stem diameter to 8.91 mm.4.The plant height increased first and then decreased with the increase of cultivation time.There was no significant difference in plant height between 12 months and 18 months in the same cultivation mode.There was no significant difference in plant height between the two cultivars under the same cultivation time.The results showed that there was no significant difference in stem diameter between the two cultivars at the 12 months,18 months and 24 months.Under the same cultivation time,the differences of stem and stem were not significant in different cultivation methods.The content of polysaccharide was the highest for the plants growing in the greenhouse for 18 months,reached 122.97 mg/g,followed by bark cultivation for 12 months,which was 103.28 mg/g,and the lowest in the moss for 24 months,only 45.59 mg/g.There were significant differences in the content of polysaccharides in different cultivation time.Under the same cultivation time,there was significant difference between bark cultivation and moss cultivation.The highest content of cyanobacteria was 8.48mg/g,and the lowest in greenhouse for 24 months was 3.75 mg/g.In the same cultivation mode,with the increase of the time,the content increased and thedecrease.At the same cultivation time,there was significant difference between bark cultivation and moss cultivation for 6 months and 18 months.There was significant difference between wild and artificial cultivation.The content of total flavonoids in bionic cultivation reached highest at 12 months,which was 3.54 mg/g,and 0.99 mg/g was the lowest for 24 hours was.Under the same cultivation time,there was no significant difference in bark cultivation and moss cultivation.There was no significant difference in cultivation between wild and moss cultivation for 6 months,which was significantly different from other cultivation time.The total phenol content in the bark cultivation was the highest,reaching 8.35 mg/g,and the moss cultivation was the lowest in the 6 months,which was 3.85 mg/g.Under the same cultivation time,there was significant difference between bark cultivation and moss cultivation.There was no significant difference between wild and moss cultivation for 6 months,which was significantly different from other cultivation time.The content of Fe and Zn in the greenhouse was ranking in 12 months > 18 months > 6 months > 24 months.The content of Fe and Zn in bark was higher than that of moss cultivation.The content of Se in wildlife was significantly higher than that of the two cultivars,and the Pb was much lower than that of the Green Industry Standard For Medicinal Plants And Preparations.Taking into account the various indicators,combined with cost and time investment,we concluded that the best harvest time of was in 12 months of cultivation. |
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