Protoplast isolation of callus in Echinacea augustifolia.

Preparations from Echinacea are among the most widely used herbal medicines. Most uses of Echinacea are based on the reported immunological properties. In this paper, we used callus of Echinacea augustifolia for isolation and researched the factors influencing the process of protoplasts preparation, the result indicated it was easy to isolate protoplast from buff-green Callus in E. augustifolia which was looked like granule with symmetrical character. The result showed that the best enzyme solution concentration is composed of cellulase 2.0% (w/v), pectinase1.0% (w/v), hemicellulase 0.5% (w/v), 0.7 mol/L mannitol and 50.0 x 10(4) gFW(-1) protoplasts were obtained after 8 h later. The yield of protoplasts was significantly influenced by cellulase concentration and incubation time. Finally, we summarized an integrated approach including callus induction, callus culture, isolation, purification, identification and calculation of protoplasts.

A method for isolating functional RNA from callus of Dendrobium candidum contented rich polysaccharides.

Isolation of high-quality RNA from Dendrobium candidum is particularly difficult. D. candidum contains considerable amounts of polysaccharides that coprecipitate with RNA, which render RNA unsuitable for either cDNA synthesis and/or PCR amplification. In this paper, a rapid and efficient method was described for functional RNA isolation from the callus of D. candidum. The procedure included: (i) an extraction with phenol and isopropyl alcohol, to remove proteins and polyphenols; (ii) purifications by lithium chloride, pre-cooled (-20 degrees C) ethanol successively to remove polysaccharides. The method resulted in high-quality RNA suitable for DDRT-PCR and cDNA library analysis finally.

Linear alkyl benzene sulphonate (LAS) degradation by immobilized Pseudomonas aeruginosa under low intensity ultrasound.

We studied the LAS degradation of immobilized Pseudomonas aeruginosa with low-intensity ultrasonic and the influence of original LAS concentration, pH, rotary velocity and different conditions of low-intensity ultrasonic irradiation on the degradation of LAS. In our experiment, the degradation rate of LAS was the main index. We found that low-intensity ultrasonic irradiation could improve the metabolism of microorganism cells and promote the LAS biodegradation of immobilized cells. In the experiment, 50 mg/l LAS were used to simulate wastewater, and low-intensity ultrasonic was considered. We found the influence was obvious, and the optimal degradation rate was acquired when the conditions of ultrasonic were frequency 24 kHz, power 8 W, stimulation time 5 s, intermissive time 30 s, and total time 10 min. The LAS degradation rate of immobilized cells with ultrasonic were respectively 40% and 9.5% higher than that of the suspending cells and immobilized cells without irradiation.

Study of mRNA expression of Arabidopsis thaliana under stimulation using modified differential display RT-PCR with silver staining.

We studied differential expressions of Arabidopsis thaliana seedlings under sound stimulation using modified differential display RT-PCR with silver staining in this paper. Six differentially expressed cDNA fragments were obtained. Molecular weight of fragments was between 200 and 600 bp, respectively. On the base of the experimentation, we considered sound stress would have been a positive or negative influence on plant growth. The experimental results indicated some genes was activated, meanwhile some were restrained under after stimulation under sound stress.

Effect of chemical and physical factors to improve the germination rate of Echinacea angustifolia seeds.

Seeds of Echinacea angustifolia are known for their deep dormancy. In this paper, we studied the responses of E. angustifolia seeds to some chemical and physical factors, such as scarification, chilling (5 degrees C) period, light and applied BA (6-benzylaminopurine), GA3 (gibberellic acid) and sound stimulation. When the seed coat layers were removed, the germination rate grew up from 6 to 20% (incubated in light) and the mean time germination (MTG) was reduced from 18 to 6.6 days. On the basis of layers-removed, chilling and continuous light gave significantly higher germination rate (up to 70%). Compare the data of seeds chilled by 0, 6, 12, 18 and 24 days, the maximum germination rate (up to 70%) achieved at 18-days chilling treatment. Further increases in the chilling period could slightly improve germination. Exogenous application of 0.1, 0.2, 0.3 mg/L GA3 or BA in the previous pretreatment increased germination to 78, 90 and 84% or 76, 86 and 84%, respectively. Obviously, the best concentration of GA or BA is 0.3 mg/L. And the GA3 or BA treatment shortened the MTG to about 4 days. The influence of sound stimulation was also tested in the experiment. The result showed that one 100 dB and 1000 Hz sound wave (sine-wave) was beneficial to the germination of E. angustifolia seeds.

Soundwave stimulation triggers the content change of the endogenous hormone of the Chrysanthemum mature callus.

Hormones have been suggested to play a prominent role in the control of callus growth. In this paper, with the method of indirect enzyme-linked immunosorbent assays (ELISA), we investigated the induction effect of soundwave on the dynamic change of endogenous indole-3-acetic acid (IAA) and abscisic acid (ABA) in vitro during the differentiation process of Chrysanthemum synchronized mature Callus. These experiments showed that groups treated by optimal soundwave (1.4 kHz, 0.095 kdb) had significantly higher IAA levels and lower ABA than that of the control, which had been implicated activation of endogenous IAA and inhibition of ABA. Through the biochemical analysis, it revealed that the increased level of IAA as well as decreased levels of ABA correlated with soundwave stimulus. High rate of IAA/ABA was favorable to development of the callus and differentiation of mature callus. We conclude that soundwave contributes to endogenous hormone as well as the control of callus growth.

Extraction of total RNA from Chrysanthemum containing high levels of phenolic and carbohydrates.

Some standard methods are available in total RNA extraction of plant tissue, which are effective for the ordinary plants. But it is difficult to extract the total RNA from the plants with high levels of phenolic compounds, carbohydrates, or other compounds that bind and/or coprecipitate with RNA. In this paper, a method was described that used the soluble polyvinylpyrrolidone (PVP), ethanol precipitation, phenol extraction and LiCl precipitation. By this method, RNA capable of reverse-transcription and polymerase chain reaction (PCR) amplification was isolated from Chrysanthemum that contains high levels of phenolic compounds and carbohydrates. The OD260/OD280 absorbance ratio is 2, and the RNA is intact. Other specialized RNA extraction methods failed to deliver suitable product. Bands of PCR products are clearly appearing on the polypropylene acyl gel electrophoresis (PAGE) gel, which indicates that the purity, concentration and integrity of total RNA were suitable for the sequent researches.

Relationship between the minute structure and the lodging resistance of rice stems.

The aim of this experiment is to measure the morphological and mechanical properties, and to analyze the relationship between the morphology and lodging resistance of several different kinds of rice stem. Choose the typical species of tall, mid and short stem and hybrid rice, ten individual plants each species (all should be caulis), and when paddies are full heading, intercept the third internode above the ground. The minute structure characteristics include the size of outer diameter, Culm wall thickness, and the number of big and small vascular bundles. Mechanical properties include the tensile breaking point and the tensile elastic modulus. Based on the above characteristics it is concluded that the middle stem has the highest lodging resistance. Thus when we select the high-yield and lodging resistance species, the middle and rigid stem should be considered.

Effect of low frequency ultrasonic stimulation on the secretion of siboflavin produced by Ecemothecium Ashbyii.

The structure of the cell membrane of Ecemothecium ashbyii is considered to be one of the main obstacles in the improvement of riboflavin productivity, which impedes the release of riboflavin from the cell into the fermentation broth. The results of the present study show that stimulation on growth and riboflavin biosynthesis phase, the content of riboflavin was most obviously enhanced, as compared with the control and the total riboflavin quantity released by ultrasonic treatment. The optimal stimulation time was from 104 to 112 h and ultrasonic should be loaded every 1.5 h.

Research on Solid-state Fermentation on Rice Chaff with a Microbial Consortium / 微生物学通报

A microbial consortium of Trichoderma reesei AS3 3711, Aspergillus niger AS3 316 and Saccharomyces cerevisiae AS2 399 was constructed to decomposed rice chaff on the basis of the characters of each microorganism and the mechanism of cellulases In this experiment, rice chaff was pretreated with NaOH before fermentation so that the lignin structure of rice chaff was degraded and hemi cellulose was dissolved partly, which remove the protection of lignin and hemi cellulose on cellulose and demolish its special crystal structure After pretreatment, rice chaff can be degraded more easily with the microbial consortium The optimal technical paths and technological methods were achieved for intenerating rice chaff with the microbial consortium perfectly through orthogonal experiment According to the technological methods, some experiments were done at 30℃ with pH 4 5 It was found that the highest filter paper enzyme activity (FPA) was 5 64U/g and the ratio of cellulose degradation (RCD) was 28 05%