Development And Anthocyanidin Accumulation Of Bulbil On Lilium Lancifolium Tunb.

Lilium lancifolium have edible and medicinal value.Bulbils are tiny bulbs which grow on leaf axils.Bulbils are important reproductive organs of Lilium lancifolium.It is important to study the growth and development of bulbils and anthocyanin accumulation in bulbils.Bulblets and bulbils of Lilium lancifolium were used as materials in this experiment.We studied the morphology changes during the development process of bulbils,the development of buds and rootbefore and after dormancy release,and the effect of different stratification treatments on the germination of bulbils and growth of bulblet of Lilium lancifolium.The conditions of high efficiency liquid chromatography(HPLC)for Lilium lancifolium bulbil anthocyanins were screened and optimized.To know the basic rules of anthocyanin formation and accumulation parttern,the content of anthocyanin during the formation of bulbils was determined and analyzed.This study aims at providing a theoretical suppport for evaluation of bulbils of Lilium lancifolium as prodution ‘seed' as well as development and utilization of anthocyanins in bulbils.The conclusions of this study are shown as follows:1.The annual average temperature of qinghai is 5.5 ? to 8.0 ?,open-field-cultivation of Lilium lancifolium can grow normally.The growth period of aboveground part is about 180 to 190 days,and the dormant bulb can live through the winter safely at-15.0?.45 to 60 days after germination,bulbils began formation when average plant height reached 44.9 cm and plant presented 12 ~ 14 leaf nodes.The development of bulbils lasted about 13 weeks.bulbils mainly distributed in 35 leave nodes between 45 cm to 100 cm of the plant.The formation process of bulbil can be divided into three stages: ‘torpedo',‘fetal' and ‘spherical',in which bulbils can be harvested as seed after entering the sphere stage.Buibils growed rapidly before 7 Weeks(After abbreviated as W),the growth rate was highest at 7W and weight was largest at 9W.The results showed that the number of scales and the number of pigmentation scales reached the maximum at 13 W.For morphology matured buds,92.5% scale was purple obviously.The ratio of bud length to bulbil height decreased during the process of bulbil development,morphology variation of bulbil presented negative correlation to bud volume.2.In the process of stratification treatment under 5??10??15? and the room temperature(17?~20?),central bud and root growed continuously,and elongation rate increased as the growth of treatment temperature however,lower temperature treatment helps breaking of bud dormancy and meanwhile improving germination rate and speed.The lower of the treatment temperature,the effect of dormancybreaking was more significant and the germination rate and seedling emergence potential was higher.It is more conducive to use bulbils as "seed" forLilium lancifolium reproduction.There was no significant effect on the transformation of bulbils to bulbs under different temperature treatment.The results also showed that 5? treatment could not only break the dormancy of the buds,but also improved the early emergence and increased the seedling emergence rate and the yield of small bulbs per unit area.3.Themethods of HPLC was screened and optimized for determinationof anthocyanins in bulbils ofLilium lancifolium,Agilent ZORBAX SB-C18;mobile phase A(0.04% formic acid): B(methanol)=95:5;velocity:0.5mL/min;detection wavelength 520nm;column temperature of 35 ?;automatic sampling,sample volume 10?L;gradient elution program: 0 ~ 40 min,A is 95% ~ 0,B is 5% ~ 100%;40 ~ 60 min,A is 0,B is 100%.4.The anthocyanin content in bulbils of each growth stage increased as the development of bud.The content of anthocyanin showed a double peak pattern.During the period from 8W to 14 W,the anthocyanin content increased sharply,and the accumulation of anthocyanin in the shoots accelerated with the decrease of temperature.The content of anthocyanin was no longer increasing at 14 W,which indicated that this is the best time to collect bulbils for anthocyanin extraction.At the same time,the content of cyanidin-3-O-glucoside in each grain of bulbil was 344.76 mg/kg,and each grain of bulbil could contain 16.5760mg/kgcyanidin-3-O-glucoside.The content of cyanidin-3-O-glucosidein each plant was up to 485.1813mg/kg.