Assessment Research For Seed Quality Control, Priming Mechanism And Medicine Collecting, Processing Of Gentiana Macrophylla Pall

This study has preliminarily proposed a method to test and classify the quality of Gentiana macrophylla L. seeds by researching the seed biology; We also proposed a sound system to improve the ability of seed germination by selecting the best initiator and the optimal condition for priming. At the same time, we tried to uncover the physiological and molecular mechanism of seed priming by study the changes of material metabolism, energy metabolism, and signal transduction in priming seed. In addition, we took Gentiana macrophylla L. as medical herb in this study, analyzing the differences of the effective ingredients’concentration between different years, harvest time and different sections of the plant; we also tested the effects of5different dry methods on several physiological indexes. Here are the main results as below:1. Setting up the preliminary standards applied to the G. macrophylla seed quality test by analysing the germination, viability and other indexex. Using these methods, we analyzed the seeds from multiple locations in the past2years, and set up the priciples for classifying the seed quality.2. Establishing a priming method for G. macrophylla seed. The germination rate of the treated seeds was one or two days ahead the untreated seeds. The proportion of the treated seed germination is up to94.8%, increased15.6%, and compared with the untreated seed. On the6th day, the germination potential has been increased49.5fold, reaching up to75%; the germination index also has been increased90.6%.3. The priming has changed the metabolisms of seed internal storage, inducing the seed germination. Compared with untreated seeds, the electric conductivity of the FeSO4treated seeds has been decreased; the content of glucose and sucrose have been reduced byl2.8%and9.6%respectively, and the content of soluble sugar, soluble proteins and raw proteins have been increased1.9%,49.7%and2.6%.The myristic acid and methyl lignocerate contents increased by4.9%and9.0%, the oleic acid, linoleic acid, docosanoic acid and linolenic acid content decreased by8.1%,8.4%,6.7%and6.7%, respectively;4. The priming treatment also changed the energy materials, enzymes and hormones in the seeds. Compared with the untreated seeds, the activity of acid phosphatase and isocitrate lyase have been increased by18.6%and7.3%respectively; the internal ABA content has been reduced by64.9%; there were21.6folds increase in GA content; the ATP content reached up to2.87E-05μmol/g, increasing2.17folds; the activity of COX and MDH in mitochondrion have been increased by68.1%and72.3%respectively. Meanwhile, the activity of CAT, SOD, APX, GSH, GR, GSH/GSSG and ASC in the peroxisome has also been increased.5. The priming caused the protein changes in the seeds. We found13differentially expressed proteins which relative intensity reached more than1.5times,6of which have been confirmed the names and functions. They were respectively Thioredoxin h, Translation initiation factor5A, Glutathione peroxidase, Pyruvate dehydrogenase E1component, UDP-glucose pyrophosphorylase, Heat shock protein70isoform3, which related to the regulation of reserve substance, growth and development of organisms, environmental resistance, and so on.6. Unigene bank of G. macrophylla seed was established. Average length of81492unigenes was721kb in this bank. Gene functions of35722unigenes were annoted in data banks of NR, Swissportm, COG, CO and KEEG, offering better understandings of molecular biological study for G. macrophylla. H2O and FeSO4priming caused914and2050differential expression genes compared to control, respectively. These differential expression genes designed20and23COG functional classifications,40and49GO functional classifications,66and90metabolic pathways, respectively. According to Gene function annotation,36enzymes of substences metabolism genes regulation,3enzymes of energy metabolism genes regulation,5enzymes of resistant gene regulation, and7enzymes of hormone metabolism genes regulation were screened due to FeSO4priming.7. The dry matter quality and the effective ingredients of plant roots, stems, leaves and flowers always change with the growth cycles and years. The triennial plants have much more dry matters and effective ingredients than the biennial ones. In biennial and triennial flowers, the contents of gentiopicroside were7.47%,7.54%, respectively; the content of swertiamarin were1.98,2.25times compared to withering period root at the same year; sweroside were7.15,4.28times respectively; longanic acid took up87.0%,42.4%. Further researches would be launched to explore the medicinal values of the flowers.8. The dry methods would affect the medicine quality. There were more soluble sugars and proteins left in the plants under vacuum frozen condition or40℃heat treatment; the starch degradability was slower, gelatinization degree increased, the bulk density decreased and porosity increased; color were more close to fresh samples; the loss of gentiopicroside content reduced to the minimum size, the content of swertiamarin, sweroside, longanic acid were increased compared to fresh samples; at the same times, the process decreased the content of element.