Preliminary Study on in Vitro Germination of Yangmei Pollen

[Objectives]To explore the conditions for in vitro culture of Yangmei pollen.[Methods]Experiments were conducted on the germination characteristics of three types of Yangmei pollen using in vitro culture and germination method.[Results]The suitable medium ratio for the germination of Yangmei pollen was 10%sucrose+0.01%borax+1%agar;the cultivation temperature of 30℃was more suitable for the germination of Yangmei pollen than 25℃;through analysis of variance,among the three types of Yangmei pollen,pollen of male 1 had the strongest viability,and it was the better pollination type.[Conclusions]The research could provide certain basis for introduction and cultivation of Yangmei and improving hybrid breeding effect.

Comparison of electrochemical dissolution of chalcopyrite and bornite in acid culture medium

The electrochemical dissolution process of chalcopyrite and bornite in acid bacteria culture medium was investigated by electrochemical measurements and X-ray photoelectron spectroscopy（XPS） analysis. Bornite was much easier to be oxidized rather than to be reduced, and chalcopyrite was difficult to be both oxidized and reduced. The relatively higher copper extraction of bornite dissolution can be attributed to its higher oxidation rate. Covellite（CuS） was detected as the intermediate species during the dissolution processes of both bornite and chalcopyrite. Bornite dissolution was preferred to be a direct oxidation pathway, in which bornite was directly oxidized to covellite（CuS） and cupric ions, and the formed covellite（CuS） may inhibit the further dissolution. Chalcopyrite dissolution was preferred to be a continuous reduction-oxidation pathway, in which chalcopyrite was initially reduced to bornite, then oxidized to covellite（CuS）, and the initial reduction reaction was the rate-limiting step.

Effects of Genotypes and Basic Medium on Culture of Maize Mature Embryos

[Objective]This study was to screen out suitable genotypes and basic medium for the culture of maize mature embryos.[Method]Using mature embryos of nine maize genotypes as explants,the effects of genotypes and basic medium on callus induction and subculture were investigated.[Result]The genotypes performed better in callus induction and subculture were found in turn 853-35,853-209,Dan 34 and 81162.MS medium is better than N6 medium in the callus induction from maize embryos,while N6 medium is more suitable for callus subculture.[Conclusion]Our study further improved the tissue culture system in maize with mature embryos as explants.

Effects of Four Culture Media on Callus Induction Rate of Wheat Anther

Effects of four culture media including MS, N6, C17 and K on wheat anther callus induction in vitro culture were studied. The results showed that the callus in- duction rate of four kinds of culture medium was in the order of K〉C17〉N6〉MS.

Propagation of Abies beshanzuensis by Water Cultured Medium

[Objective] The experiment aimed to explore the influences of phytohormones (ABT and IAA) and nutrient solution on rooting of Abies beshanzuensis M.H.Wu by water cultured medium. [Method] The Abies beshanzuensis M.H.Wu were treated by water (CK), 10 mg/L ABT+ water, 10 mg/L IAA+ water, 10 mg/L ABT+ hoagland solution, 10 mg/L IAA+ hoagland solution, then the rooting process was observed and the formation rate of callus, rooting rate, number of rooting, and root length were investigated and analyzed. [Result] ABT and IAA had obvious influences on callus induction, rooting rate and the number of root of Abies beshanzuensis M.H.Wu by water culture, so they were suitable to be used in water propagation of Abies beshanzuensis M.H.Wu. The treatments of phytohormones had no regular influences on the longest root length and average root length. The nutrient solutions would not generate obvious influence on propagation of Abies beshanzuensis M.H.Wu at firstly stage, but they generated influence on root growth after rooting. [Conclusion] The research provided new ideas for propagation of Abies beshanzuensis M.H.Wu, which could make it out of endangerment situation quickly.

Screening of High-quality Substrate for Soilless Culture of Begonia cucullata Willd.

In order to investigate a better soilless culture medium and the possibility of grass clippings as alternative substrate for peat moss to grow Begonia cucullata Willd. in pots, the seedlings of widely cultivated B. cucullata were grown in pots with eight types of soilless substrates： T1（peat： perlite=1：1）, T2（peat： perlite： pine bark=1：1：1）, T3（corn stover： perlite： pine bark=1：1：1）, T4（corn stover： perlite： pine bark=2：1：1）, T5（grass clipping： perlite： pine bark=1：1：1）, T6（grass clipping： perlite：pine bark=2：1：1）, T7（vermiculite： perlite： pine bark=1：1：1） and T8（vermiculite： perlite： pine bark=2：1：1）, respectively, in a shade house of Shanghai Chenshan Botanical Garden to evaluate plant growth performance. The results showed that the stem diameter of B. cucullata increased significantly from day 30 to day 60 after potting,and the plant height, leaf number and leaf area increased largest from day 60 to day 90 after potting in all treatments. The growth of plants performed best in T6,with the highest branch number, leaf thickness, flower number, aboveground fresh and dry weights and relative chlorophyll content. Therefore, the substrate with grass clipping： perlite： pine bark=2：1：1 was the best culture medium for growing B. cucullata in this study, and the grass clippings could replace peat moss as alternative substrate for growing B. cucullata in containers.

COMPARISON OF POLYSACCHARIDES ISOLATED FROM THE MYCELIA OF A CULTIVATED STRAIN OF PORIA COCOS GROWN IN DIFFERENT LIQUID CULTURE MEDIA

Mycelium of a cultivated strain of Poria cocos was grown by submerged fermentation in a liquid mediumcontaining corn steep liquor with orbital shaking. Six polysaccharides coded as ac-PCM1, ac-PCM2, ac-PCM3-Ⅰ andⅡ, ac-PCM4-Ⅰand Ⅱ were isolated from the myelium by extracting with 0.9% NaCl aqueous solution, hot water, 0.5 mol/L NaOHaqueous solution and 88% formic acid. Exo-polysaccharide was obtained from the culture medium and coded as ac-PCM0.The monosaccharide composition and molecular weights of these polysaccharides were characterized by using infraredspectroscopy, gas chromaography, elemental analysis, ^(13)C-NMR, viscometry and light scattering. The results indicated thatac-PCM0, ac-PCM1 and ac-PCM2 are heteropolysaccharides containing glucose, galactose, mannose and fucose, and ac-PCM3-Ⅰ and ac-PCM3-Ⅱ mainly consist of D-glucose. The content of the glucose in the polysaccharides increased with theisolation progress. Remarkably, α-glucan and β-glucan coexisted in the extract by NaOH aqueous solution (ac-PCM3), andcould be separated by chemical methods. The protein in the ac-PCM polysaccharides cultured from the medium containingcorn steep liquor was higher than that in the ab-PCM from the medium with bran extract. Therefore, the polysaccharidesfrom Poria cocos mycelia cultured in different media have different chemical composition, molecular weights and conformations.

Improved elastase production by Bacillus sp. EL31410—further optimization and kinetics studies of culture medium for batch fermentation

An efficient culture medium producing a bacterial elastase with high yields was developed further following preliminary studies by means of response surface method. Central composite design (CCD) and response surface method-ology were applied to optimize the medium constituents. A central composite design was used to explain the combined effect of three medium constituents, viz, glucose, K2HPO4, MgSO47H2O. The strain produced more elastase in the completely optimized medium, as compared with the partially optimized medium. The fitted model of the second model, as per RSM, showed that glucose was 7.4 g/100 ml, casein 1.13 g/100 ml, corn steep flour 0.616 g/100 ml, K2HPO4 0.206 g/100 ml and MgSO47H2O 0.034 g/100 ml. The fermentation kinetics of these two culture media in the flask experiments were analyzed. It was found that the highest elastase productivity occurred at 54 hours. Higher glucose concentration had inhibitory effect on elastase production. At the same time, we observed that the glucose consumption rate was slow in the completely optimized medium, which can explain the lag period of the highest elastase production. Some metal ions and surfactant additives also affected elastase production and cell growth.

Relationship Between Differential Expression of Bax and Bcl-2 Genes and Developmental Differences of Porcine Parthenotes Cultured in PZM-3 and NCSU-23

The developmental competence of porcine parthenotes cultured in porcine zygote medium-3 （PZM-3） and North Carolina State University-23 （NCSU-23） media was investigated. After in vitro maturation oocytes were electro-activated, and the subsequent developmental competence, rates of apoptotic, fragmented and arrested embryos from the two media were evaluated. Also, the ratio of mRNA expression of Bcl-2 and Bax gene was determined. Results demonstrated that cleavage, blastocyst, hatched blastocyst rates, and blastocyst cell numbers were significantly higher in PZM-3 than in NCSU-23. The rate of fragmented embryos in PZM-3 was lower than in NCSU-23 on d 1 and 3 （P〈0.05） while the percentage of arrested embryo was lower in PZM-3 than in NCSU-23 on d 4 and 5 （P〈0.05）. TUNEL positive signals were higher in NCSU-23 than in PZM-3 from d 3 to 7 （P〈0.05）. The ratios of Bcl-2 and Bax mRNA expression in embryos were higher on d 5 than on d 3 and 1 in PZM-3 （P〈0.05）. In contrast, the ratios of Bcl-2 and Bax mRNA expression in embryos on d 1 were higher than on d 3 and 5 in NCSU-23 （P〈0.05）. Additionally, the ratios of Bcl-2 and Bax mRNA expression in embryos in PZM-3 were higher than in NCSU-23 on d 3 and 5 （P〈0.05）. In conclusion, lower apoptotic embryo rates and down-regulating Bax together with up-regulating expression of Bcl-2 transcripts may be responsible for the better developmental competence of porcine parthenotes in PZM-3.

Screening and Characterization of Nitrite-Degrading Bacterial Isolates Using a Novel Culture Medium

In this study,a novel culture medium that simulates shrimp pond conditions was established to screen nitrite-degrading isolates.The medium was supplemented with nitrite as a nitrogen source and shrimp feed as the major carbon source,to achieve the high nitrogen and low carbon nutritional status found in shrimp farming ponds.Screening using this medium identified potent denitrifying Bacillus isolates,among which Bacillus subtilis M7-1 was considered best.M7-1 was able to completely degrade nitrite-N in 24 h without much consumption of dissolved oxygen.Efficient denitrification activity took place in liquid cultures within a set of non-stringent ranges of pH(5.0–9.0),salinity(0–30)and temperature(25–35℃).The denitrifying enzyme gene was amplified,sequenced and further identified as nirS type.In biosecurity assessments,M7-1 had no negative effects on shrimps at a dose of 106 cfu mL−1.M7-1 could therefore be used in aquaculture to reduce and control the nitrogen concentration,and to promote the development of sustainable and healthy culture systems.

Effects of different culture media on the growth of Indian sandalwood(Santalum album L.) seedlings in Zhanjiang,Guangdong,southern China

We studied the effects of different culture media on the growth of India sandalwood （Santalum album L.） seedlings in Zhanjiang, Guangdong Province in southern China. Five different growth substrates, lateritic subsoil, burnt soil, agricultural soil, peaty soil and coconut dust, were used as the basic culture materials and seven different treatments of composition were used as potting media. Kuhnia rosmarinifolia Vent. was used as a primary host plant for all treatments. Statistically significant differences were found between treatments in respects of survival rate （p 〈 0.001）, height （p 〈 0.001）, ground diameter （p 〈 0.001） and biomass （p = 0.002）, as well as for quality index （p 〈 0.001） of S. album seedlings after 6-month growth in containers with different culture media. Among all treatments, the treatment combining burnt soil, peat and coconut dust in a weight ratio of 1：1：1 plus 2% calcium super-phosphate as basal manure achieved the best performance for most of the seedling growth parameters, including survival rate （98%）, height （35.81 cm）, ground diameter （0.56 cm）, biomass （4.46 g） and quality index （0.65）, followed by the treatment using only burnt soil plus 2% calcium super-phosphate as the culture medium （survival rate 86%, height 29.23 cm, ground diameter 0.48 cm, biomass 3.36 g and quality index 0.52）, while the treatment using only lateritic subsoil plus the basal manure as the medium obtained the poorest results in survival rate （38%）, height （12.04 cm）, ground diameter （0.19 cm）, biomass （0.26 g） and quality index （0.043）. Increasing the proportion of lateritic subsoil in the medium when mixed with peat and coconut dust did not show statistically significant differences in survival, height, ground diameter, biomass nor in quality index. In consideration of cost, using burnt soil plus 2% calcium super-phosphate as basal manure may be the optimum culture medium for large-scale production of Indian sandal- wood seedlings in Guangdong, southern China.

Culture Media Influenced Laboratory Outcomes But Not Neonatal Birth Weight in Assisted Reproductive Technology

Summary： Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization （IVF）/intracytoplasmic sperm injection （ICSI） was investigated. A total of 673 patients undergoing IVF/ICSI and giving birth to live singletons after fresh embryo transfer on day 3 from Jan. 1, 2010 to Dec. 31, 2012 were included. Three types of culture media were used during this period： Quinn＇s Advantage （QA）, Single Step Medium （SSM）, and Continuous Single Culture medium （CSC）. Fertilization rate （FR）, normal fertilization rate （NFR）, cleavage rate （CR）, normal cleavage rate （NCR）, good-quality embryo rate （GQER） and neonatal birth weight were compared using one-way ANOVA and Z2 tests. Multiple linear regression analysis was performed to determine the impact of culture media on laboratory outcomes and birth weight. In IVF cycles, GQER was significantly decreased in SSM medium group as compared with QA or CSC media groups （63.6% vs. 69.0% in QA; vs. 71.3% in CSC, P=0.011）. In ICSI cycles, FR, NFR and CR were significantly lower in CSC medium group than in other two media groups. No significant difference was observed in neonatal birthweight among the three groups （P=0.759）. Multiple linear regression analyses confirmed that the type of culture medium was correlated with FR, NFR, CR and GQER, but not with neonatal birth weight. The type of culture media had potential influences on laboratory outcomes but did not exhibit an impact on the birth weight of singletons in ART.

Micropropagation of Eucalyptus grandis×E.urophylla AEC 224 clone

Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis x E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 μM BA and 0.5 μM NAA for another 30 days. In these media, the explant oxidation rate was high （95 %）. Thus, in order to reduce oxidation, different ;culture media were compared： WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 μM TDZ and 0.1μM NAA during 30 days for callus induction and then with 5.0 μM BA and 0.5 μM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA （2.46, 4.90 and 7.35μM）. The highest rooting percentage （35 %） was obtained on medium containing 2.46μM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %.

Study on Microwave Sterilization of Solid Culture Medium

In this paper,microwave sterilization craft of solid culture medium in fungus growth is studied.The results show that microwave not only can be used to sterilize the micro-organisms which is useless for fungus growth in the solid culture medium,but also has positive effect on fungus growth;and the sterilization process is featured with shorter time and higher efficiency compared with the traditional method.

Characteristics of a kHz helium atmospheric pressure plasma jet interacting with two kinds of targets

This study investigates the influence of two types of target,skin tissue and cell culture medium,with different permittivities on a k Hz helium atmospheric pressure plasma jet(APPJ)during its application for wound healing.The basic optical-electrical characteristics,the initiation and propagation and the emission spectra of the He APPJ under different working conditions are explored.The experimental results show that,compared with a jet freely expanding in air,the diameter and intensity of the plasma plume outside the nozzle increase when it interacts with the pigskin and cell culture medium targets,and the mean velocity of the plasma bullet from the tube nozzle to a distance of 15 mm is also significantly increased.There are also multiple increases in the relative intensity of OH(A^(2)Σ→X^(2)Π)and O(3p^(5)S-3s^(5)S)at a position 15 mm away from nozzle when the He APPJ interacts with cell culture medium compared with the air and pigskin targets.Taking the surface charging of the low permittivity material capacitance and the strengthened electric field intensity into account,they make the various characteristics of He APPJ interacting with two different targets together.

Evaluation of Endotoxin in Culture Medium for Human in vitro Fertilization

To determine whether the presence of bacterial endotoxin in the commercial culture media utilized for human in vitro fertilization （IVF）, and evaluate the difference in detecting endotoxin in culture medium between the human sperm motility assay and the 2-cell mouse embryo assay. Methods Thirty-six batches of culture media commonly used in IVF laboratories from 3 manufacturers were determined for thepresence ofendotoxin before using the medium for the assisted reproductive programs （group A）. After being used, 25 specimens among above media were also tested （group B）. The chromogenic limulus amoebocyte lysate （LAL） test was used for quantification the content of endotoxin. In addition, the human sperm motility assay was compared with the 2-cell mouse embryo assay to evaluate the difference in detecting endotoxin in culture medium. Results Endotoxin was not detected in group A. However, 2 samples were positive in group B, Sperm did not show significant change in motility in group A during 24 h of incubation when compared with the control （P〉0. 05）. However, in group A the 2-cell embryo development to blastocyst was suppressed in 3 batches of media. Conclusions Regular screening of each batch of culture medium should be performed if possible although there was no evidence of endotoxin contamination in commercially prepared pre-tested media. Culture environment should be stringently controlled in case the medium is polluted. The sensitivity of the sperm motility assay was lower than that of the mouse embryo assay for detecting low levels of endotoxin or toxic compounds in the medium.

Effects of Culture Media and Light Intensity on in vitro Growth of Oncidium under CO_2 Enrichment Condition

The effects of culture media and light intensity on in vitro growth of Oncidium 慉loha Iwanga were investigated under CO2 enrichment condition. Height, fresh and dry weight of the Oncidium seedlings were measured, and the leaf number per plant, shoot number per plant, leaf width and leaf chlorophyll content were also investigated. The results were as follows: 1) The seedling height, fresh and dry weight, leaf number per plant, leaf width and leaf chlorophyll content of the shoots growing on MS complete culture medium were higher than those on 1/2MS, VW and 1/2VW media. The root number per plant and ratio of dry matter of the seedlings cultured on 1/2MS and 1/2VW media were higher than those on MS and VW; 2) The seedling height, fresh weight, dry weight, dry matter ratio and leaf chlorophyll content, leaf length, leaf width, root length, leaf number per plant, root number per plant of seedlings of Oncidium growing under 4 500 lx and 1 700 lx were higher than those under 750 lx. However, there was no significant difference in those growth parameters mentioned above while dealing with 4 500 lx and 1 700 lx except for the seedling height. Nevertheless, the leaf color of plants under 4 500 lx was lighter and the leaves of the lower parts became yellowish in comparison with those growing under 1 700 lx.

Isolation and Screening of Oenococcus oeni and Its Medium Optimization

[Objectives] This study was conducted to further improve the application of Oenococcus oeni in the wine brewing industry. [Methods] O. oeni was isolated, screened, and identified, and medium optimization was performed to determine the best medium, temperature and oxygen conditions. [Results] O. oeni grew better in mFT80 medium, and the culture conditions were 25 ℃ and an anaerobic condition. [Conclusions] This study provides a reference for the development of China’s wine industry.

Screening of Optimal Differentiation Medium to Lonicera edulis

An excellent Lonicera edulis strain, L1-8 that was bred by Northeast Institute of Geography and Agro-ecology, Chinese Academy of Sciences, was used as material in this research. The axillary buds of its dormant branches were used as explants. A fourfactor and four-level orthogonal test was designed in order to choose the best differentiation medium for providing the technical support of Lonicera edulis micropropagation. The results showed that the culture medium and concentration of 6-BA were the main factors, and the best differentiation condition was MS culture medium containing 2.0 mg · L-1 6-BA, 0.3 mg · L-1 IBA and 1.5 mg · L-1 GA3.

A Preliminary Study on Tissue Culture and Rapid Propagation of Virus-free Plantlets of Benihoppe Strawberry

[ Objective] This study aimed to investigate the tissue culture and rapid propagation techniques of Benihoppe strawberry. [ Method] Shoot tips of new stolons of Benihoppe strawberry were used as experimental materials to analyze the effects of media type, cytokinin type and concentration, carbon source type and concentration, and light intensity on tissue culture and propagation of Benihoppe strawberry. [ Result] MS was the optimal media for plantlet propagation. In the media containing 1.2 ing/L BA （with addition of 0.1 mg/L NAA）, fresh weight, dry weight and propagation coefficient of strawberry plantlets reached the maxi- mum, which were 2.259 g, 0. 221 g and 12.4, respectively. The optimal carbon source was 30 g/L sucrose, and the optimal light intensity was 1 600 lx. [ Conclu- sion] The best media for tissue culture and rapid propagation of Benihoppe strawberry was MS ＋ BA 1.2 mg/L ＋ NAA 0. 1 mg/L ＋ sucrose 30 g/L ＋ agar 8 g/L. This study provided scientific basis for large-scale production of Benihoppe strawberry.