| Lily is an important economic crop with ornamental,edible and medicinal value.Lilium davidii var.unicolor Salisb is a kind of lily with important edible value.Lilium regale Wilson is a good material as parents for resistance breeding activities.Lilium lancifolium Thunb.is the only natural triploid lily with edible and medicinal value.Oriental hybrid which is of high value and easy-buying is an excellent material suitable for cuttings of scales.Polyploid breeding is the main breeding method to restore plant fertility,improve resistance and increase plant value of edible and medicinal value.The current research about lily polyploid breeding is mainly focused on the improvement of ornamental characteristics,however the studies about the lily varieties with edible and resistant value are rarely few.In order to obtain the tetraploid germplasm with restored fertility,improved resistance,increased edible and medicinal value,we used the soaking method to induce the tissue culture of the lily outer scales to conduct the chromosome doubling research,which taking the Lilium davidii var.unicolor Salisb,Lilium regale Wilson,Lilium lancifolium Thunb.,‘Sorbonne' and ‘Siberia' as the materials.The main results are as follows:1.Using the colchicines and Oryzalin as the inducers soaked the tissue culture of Lilium davidii var.unicolor Salisb,Lilium regale Wilson and Lilium lancifolium Thunb.and successfully identified and obtained tetraploid Lilium davidii var.unicolor Salisb,but did not obtain the Lilium lancifolium Thunb.and Lilium regale Wilson.The suitable inducer treatment is 0.05 % colchicine treatment 48 h,which the maximum induction rate is 33.3%.The suitable lysis solution for lily flow cytometry detection is Galbraith's buffer.The genotype size of the diploid lily was 35.3 ± 0.9 G and the genotype size of the tetraploid genome was 70.2 ± 0.6 G.The karyotype formula of the diploid is 2n = 2x = 24 = 2m + 2sm + 6st + 14 t,the tetraploid karyotype formula is 2n = 4x = 48 = 4m + 4sm + 12 st + 28 t.The identification of cytology and nutrient components showed the stomatal density was significantly lower than that of diploid,but the guard cell length was longer than diploid.The content of polysaccharide in tetraploid scale was 50% higher than that of diploid.The tetraploid plants of Lilium regale Wilson and Lilium lancifolium Thunb.were not obtained.2.In the chromosome doubling by cutting method,using colchicine and Oryzalin soaked ‘Sorbonne' and ‘Siberia' bulb outer scales,although the doubled plants were not obtained,it was found that reagent A was able to induce the bulb scales regeneration in a short period of time and significantly increased the reproductive coefficient of the cuttings.3.Reagent A makes the optimal combination of rapid induction of small bulb scales by cuttings: 4% of the reagent A soaked the outer bulb scales for 12 h.The propagation coefficient of the ‘Sorbonne'cuttings treated with reagent A increased to 2.25,‘Siberia'cuttings treated with reagent A increased to 2.60,Lilium tigrinum cuttings treated with reagent A increased to 2.50.4.Establishment of the propagation system for tetraploid L.davidii var.unicolor Salisb.Tissue culture techniques and bulb scale cutting techniques were used to propagate the obtained tetraploid lily of L.davidii var.unicolor Salisb.Tissue culture method: the suitable differentiating medium for tetraploid L.davidii var.unicolor Salisb is 6-BA(1 mg/L)?NAA(0.1 mg/L),the pH is 6.0,the concentration of sucrose is 30 g/L.The appropriate culture medium for bulb scales is the MS basic medium,with pH 6.0 and the concentration of sucrose is 90 g/L.Scale cutting propagation method: The cutting propagation coefficient of the diploid L.davidii var.unicolor Salisb is increased to 2.12 by 4% of the reagent A soaked the outer bulb scales for 12 h.The treatment was applied to the outer scale of the tetraploid lily until the tetraploid lily was expanded to a certain scale to establish the propagation system of the tetraploid germplasm. |
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