| Lilium martagon var.pilosiusculum Freyn is a perennial bulbous flower belonging to the genus Lilium,the family Liliaceae.L.martagon var.Pilosiusculum is the whorled leaf of lily,and which also includs L.paradoxum,L.tsingtauense and L.distichum in China.The flowers of L.martagon var.pilosiusculum are purple and drooping with a high ornamental value.Its bulbs are edible with a high medicinal value.And the L.martagon var.pilosiusculum has a broad market prospect.However,the L.martagon var.pilosiusculum distributed in a narrow range.The environments of its originated are getting worse and worse.Because of the man-made over-destruction and collection,the number of the precious of the wild resources has been reduced year by year.At present,the studies of L.martagon var.pilosiusculum are focused on karyotype analysis and medicinal value,there is no systematic research and analysis on in vitro regeneration technology.In order to establish the in vitro regeneration system of L.martagon var.pilosiusculum,the scales of L.martagon var.pilosiusculum and the leaves of aseptic seedling were selected as explants,the key technologies such as of induction,proliferation,rooting and transplanting were discussed in vitro in this paper.The establishment of the in vitro regeneration system of L.martagon var.pilosiusculum could effectively protect the precious wild lily resources of L.martagon var.pilosiusculum,meanwhile,it could also establish the basis for tissue culture breeding,hybrid breeding and molecular breeding.The main test results are as follows:For scales of L.martagon var.pilosiusculum as the explants.The results showed that the best disinfection method to the bulb scale explants was 70%ethanol for 30 s,followed by 0.1%HgCl2 for 15 min;The scale induction rate ranked with the orders of inner layer>middle layer>outer layer and middle part>basal part>upper part;The suitable medium for induction was MS + 1 mg·L-1 6-BA+ 1 mg·L-1 NAA,with 78.66%induction rate.For tube plantlet leaf of L.martagon var.pilosiusculum as the explants.The result showed that the leaf induction rate ranked with the orders of basal part>middle part>upper part.The placement of sterile leaves directly affects the late growth of tube plantlet,The induction rate of the orientation of the asepic leaves was higher than of distal axis.The suitable medium for induction was MS + 1 mg·L-1 6-BA +0.5 mg·L-1 NAA,with 75.89%induction rate.Through proliferation culture for adventitious buds of L.martagon var.pilosiusculum,We found that MS + 0.5 mg·L-1 6-BA + 0.05 mg·L-1 NAA was the optimal subculture medium with 3.46 multiplication coefficient;Every bud could multiply about 4 and the rate of the differentiation was higher as the subcuture times was three;When sucroseon concentration was 60 g·L-1,fresh weight increased 10.49 times,bulb diameter increased to 1.15 cm;When NAA concentration was 2 mg·L-1,fresh weight increased 3.82 times,bulb diameter increased to 1.00 cm;When IBA concentration was 2 mg·L-1,fresh weight increased 3.81 times,bulb diameter increased to 0.90 cm;Compared with light treatment,dark treatment had a significant effect on the formation of small bulbs in vitro.For the tube plantlet of L.martagon var.pilosiusculum,we conducted root culture.The result showed that the suitable rooting medium was 1/2 MS + 0.2 mg.L-1 NAA + 0.2 mg·L-1 IB A with a rooting rate of 100.0%;0.5 g·L-1 activated carbon can promote rooting of tube plantlet,and the root quality is good;The component of peat and perlite?vermiculite(1:2:1)is good for L.martagon var,pilosiusculum growth after transplant,promote seeding growth and their survive ratio was 71.12%. |
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