濒危植物翅果油树愈伤组织诱导中褐变问题的研究

针对翅果油树愈伤组织诱导的褐变问题,从外植体、碳源、抗氧剂的类型及浓度等的筛选进行了研究。结果表明;幼龄的茎段是诱导愈伤组织的最佳材料。葡萄糖为碳源的培养基能有效地防止褐变。抗氧化剂Vc和Na2S2O3能减轻褐变程度,其中0．15g／L的Vc效果最佳,其次为0．1g／L和0．2g／L。Na2S2O3对褐变的抑制作用不及Vc,3种浓度无明显区别。吸附剂AC不能防止外植体褐变,且抑制愈伤组织的诱导。     

马蹄莲的组织培养

TissueCultureofZantedescltiaaethiopicaQILIWang,HANSu－Ying,YANGYun－Long,ZANGMeng－Yi,ZHANGJing－Tao（fort7wizlofmp,sleZlirzA叶如仪份功U——ndg,了～卯03080至）1植物名称马蹄莲de川b如旧办班规彻仲山）。2材料类别球茎。3培养条件基本培养基为改良MS（其中：NH4NO。,KNO3减半,加KH。PO;25mg／L,生根的MS全量）。附加物（mg／L）：（）6－BA2＋NAA005＋IBA0．05＋LH50＋Vc5;（2）6－BA0．75＋NAA0．2＋IBA02＋LH50＋Vc5;（3）6BA3＋NAA0．2＋LH80＋Vc40;（4）6．BAI．5＋NAA0…     

水蕨的组织培养和快速繁殖

1植物名称水蕨[Ceratopteris thalictroides（L．）Brongn]。 2材料类别营养叶、孢子叶。 3培养条件基本培养基为MS培养基。（1）诱导愈伤组织培养基：MS＋6．BA2mg．L^-1（单位下同）＋NAA0.5;（2）诱导芽培养基：MS＋6．BA0．25＋IBA0．25＋活性炭（AC）0．5g．L^-1;（3）生根培养基：MS＋6．BA0．5＋IBA0．25＋AC0．5g．L^-1。     

手性拆分环氧氯丙烷菌株的筛选、鉴定及产酶条件研究

从土壤中筛选到5株环氧化物水解酶生产菌,并通过ITS序列鉴定了其中的C375菌,结果为黑曲霉（Aspergillus nigerZJB-09103）。考察了培养基不同碳源、氮源、金属离子和pH等对产酶的影响,得到了较佳的培养基条件：淀粉16g／L,豆饼粉3g／L,蛋白胨3g／L,KH2PO4 0．4g／L,K2HPO4 0．8g／L,MgSO4 0．2g／L,ZnSO4 0．03g／L,pH6．5。采用优化后的培养基条件,酶活力达到156．1U／L,比优化前初始发酵培养条件下的酶活提高了252％,当环氧化物水解酶催化时间为10h时,（s）-环氧氯丙烷的对映体过量值（e．e．）可达99．0％。产率为18．6％。     

响应面法优化酶法转化合成维生素C糖苷（AA-2G）条件的研究

采用Design—Expert软件的Central Composite Design（CCD）响应面设计对环糊精葡萄糖苷转移酶转化合成糖基抗坏血酸（AA-2G）的五个主要因素（转化时间、转化温度、pH、Vc浓度、β-环糊精浓度）进行了研究。采用降维分析方法对pH与转化时间、转化温度、Vc浓度、β-环糊精浓度以及反应温度与反应时间的交互作用对酶法转化合成AA-2G的影响进行了分析。建立了影响因素与响应值之间的回归方程,根据回归方程优化得到最佳转化条件为：转化时间25h,温度36．5℃,pH5．4,Vc72dL,β-环糊精55g／L。在此条件下,AA-2G的理论产量为10．06g／L,在验证实验中AA-2G的产量为9．76g／L,与预测的理论产量接近,比优化前提高了33％。     

山葵的离体培养和植株再生

CInvitroCultureandPlangtletRegenerationofEutremawasabiWANGGuang-Dong,LIShi-Jun(DepartmentofHorticulture,NanjingAgriculturalUniversity,Nanjing210095)1植物名称山葵（Eutremawasabi）,又名山嵛菜。2材料类别带侧芽的根状茎。3培养条件（1）诱导培养基：1／2MS＋6－BA0．2mp·L－1（单位下同）＋NAA0．05;（2）增殖培养基：MS＋6-BA0．2＋KT0．2＋NAA0．05;（3）生根培养基：1／2MS＋NAA0．05。培养基（1）附加琼脂粉5．0g·L－1,其余附加6.0g·L－1。（1）和（2）培养基加蔗糖30g·L－1,（3）加20g·L－…     

矮牵牛的离体培养和快速繁殖

InvitroCultureandRapidPro阴切nonofCommonpetunianHua,WENChun－Xu（PbeedeyIfes～lrchIndddeofHdri,SAsfor050061）1植物名称矮牵牛（Petun。htwde）,又名碧冬茄。2材料类别茎尖、带芽的茎段。3培养条件（回）诱导培养基：l／3MS＋6－BAImg／L（单位下同）＋IBA0．02＋GellanGum（进口琼脂）2g／L;（2）增殖培养基：MS＋6．BA0．5～1＋IBA0．2;（3）生根培养基：l／2MS＋IBA0．5。培养基含蔗糖25g／L,（2）和（3）附加普通琼脂55g／L。PH58,高压灭菌。培养温度22～25”C,光照度1500IX,照光10～12h／d。毛生…     

高浓度糖发酵制备乙醇

以树干毕赤酵母为发酵菌株,混合糖（木糖、葡萄糖）为发酵底物,通过培养基和培养条件的改变来确定树干毕赤酵母高糖浓度发酵时所需的条件。研究结果表明：在24h发酵周期内初始木糖质量浓度为63．0g／L较适宜;在36h发酵周期内初始木糖质量浓度为72．0g/L较适宜。24h发酵周期内,在36．0g／L木糖中添加的葡萄糖质量浓度以54．0g/L为最佳,发酵结束乙醇质量浓度达32．9g／L;36h发酵周期内,添加的葡萄糖质量浓度以72．0g／L为最佳,发酵结束乙醇质量浓度为36．9g／L。以（NH4）2SO4为N源时较适合戊糖发酵制备乙醇,（NH2）2SO4的最佳质量浓度为1．1g／L。发酵前8h摇床转速为90r／min,后16h为150r／min,乙醇质量浓度较高,可达17．5g/L。     

TNF—α对哮喘模型大鼠气道平滑肌细胞增殖及ERK1／2活性的影响

目的探讨TNF—α对哮喘大鼠气道平滑肌细胞（ASMCs）增殖及对ASMCs上ERK1／2mRNA、p-ERK1／2表达水平的影响。方法通过对哮喘模型大鼠ASMCs培养,分别以0．2μg／L、1．0μg/L、20μg/L TNF-α干预ASMCs生长。采用流式细胞仪、MTT法检测ASMCs增殖情况,观察不同浓度TNF—α对ASMCs增殖的影响。RT-PCR检测ASMCs上ERK1／2mRNA表达,免疫细胞化学染色法检测磷酸化ERK1／2蛋白的表达及定位。结果哮喘组ASMCsS期比例、A值、ERK1／2mRNA、p-ERK1／2蛋白的表达量分别为（34．45±2．08）％、（0．550±0．010）、（0．995±0．118）、（130．77±4．16）,与对照组（11．17±0．96）％、（0．292±0．008）、（0．576±0．098）、（163．82±1．38）比较均显著增高（均P〈0．01）。各TNF—α干预组ASMCs的S期比例、A值、ERK1／2mRNA和p-ERK1／2蛋白表达量与哮喘组比较均显著降低（均P〈0．01）,0．2μg/L和1．0μg／LTN-α组p-ERK1／2蛋白表达量高于对照组（P〈0．01）,20μg／L TNF-α组p-ERK1／2蛋白表达量与对照组比较无差异（P〉0．05）。结论与正常鼠相比,慢性哮喘大鼠气道平滑肌细胞增殖明显,处于S期的细胞比例明显增高。经TNF—α干预后,慢性哮喘大鼠气道平滑肌细胞处于S期的细胞比例减少,增殖减弱,TNF-α可能抑制慢性哮喘大鼠气道平滑肌细胞增殖。TNF—α可下调慢性哮喘大鼠气道平滑肌细胞上ERK1／2mRNA及p-ERK1／2表达,TNF-α可能通过抑制ERK信号转导通道的活性对气道平滑肌细胞的增殖进行调控。     

应用改良滴冻法超低温保存两种柿属植物的研究

以君迁子（Diospyros lotus L．）和柿（D．kaki Thunb．）组培苗茎尖为试材,对影响超低温保存效果的主要因素,如低温锻炼方式、预培养条件、PVS：（30％甘油＋15％乙二醇＋15％二甲基亚砜＋0．4mol／L蔗糖）处理时间等进行了研究。建立了2种柿属植物的超低温保存程序：（1）切取1cm左右试管苗梢段继代到1／2MS（KNO3和NH4NO3减半）培养基中,交替低温[昼（25±1）℃、夜（4±1）℃]锻炼6周;在含0．5mol／L蔗糖的1／2MS培养基上预培养5d,20℃下装载液（2．0mol／L甘油＋0．4mol／L蔗糖）过渡10min,0℃下PVS2处理1．5h;（2）投入液氮保存;（3）40℃水浴化冻,洗涤5～6次后接种于含1．0mg／LTDZ、0．6g／L可溶性PVP、30g／L蔗糖和7．0g／L琼脂的培养基（作者在优化柿属植物离体培养体系试验中获得）上暗培养1周,转入25℃,1500lx培养室。按照上述程序培养,‘鄂柿1号’、‘湘西甜柿’和君迁子的成活率分别为79．6％、67．4％和60．9％。     

Charcoal and activated carbon as adsorbate of phytotoxic compounds – a comparative study

This study compares the potential of natural charcoal from Scots pine ( Pinus sylvestris L.) and activated carbon to improve germination under the hypothesis that natural charcoal adsorbs phytotoxins produced by dwarf-shrubs, but due to it's chemical properties to a lesser extent than activated carbon. Activated carbon has been used in many bioassays as an adsorbate to clean aqueous solutions.
We used aqueous extracts from young leaves of Calluna vulgaris (L.) Hull and Vaccinium myrtillus (L.) as phytotoxin sources in two different concentrations (10 and 14 gr. of dried leaves in 100 ml distilled water). Germination of pine seeds was prevented by the higher concentration of both species, while the lower ones did not show significantly reduced germination. Both ericaceous species showed a very similar potential to prevent germination of Scots pine seeds.
Supplemented carbon (activated carbon, powdered or granulated pine charcoal) restored germination in strong extracts. Adding activated carbon resulted in germination of almost 100%. With pine charcoals added, lower germination percentages were observed. The charcoal powder was more effective (60% for C. vulgaris ; 28% for V. myrtillus ) than the charcoal granulate (30% and 16%, respectively) in restoring germination.
Chemical and surface analysis of the three carbon supplements revealed that activated carbon had by far the biggest active surface area (641 m² g⁻¹), and thus many more cavities to bind phytotoxins than natural charcoal (total surface area of 142 m² g⁻¹).
We conclude, that charcoal produced by forest fires can have a positive effect on seed germination, but to a much lesser extent than activated carbon. Previous studies, which used activated carbon as an equivalent for charcoal, overestimated the effect of charcoal on germination.     

Detoxification of sugarcane bagasse hydrolysate improves ethanol production by Candida shehatae NCIM 3501

Sugarcane bagasse hydrolysis with 2.5% (v/v) HCl yielded 30.29g/L total reducing sugars along with various fermentation inhibitors such as furans, phenolics and acetic acid. The acid hydrolysate when treated with anion exchange resin brought about maximum reduction in furans (63.4%) and total phenolics (75.8%). Treatment of hydrolysate with activated charcoal caused 38.7% and 57.5% reduction in furans and total phenolics, respectively. Laccase reduced total phenolics (77.5%) without affecting furans and acetic acid content in the hydrolysate. Fermentation of these hydrolysates with Candida shehatae NCIM 3501 showed maximum ethanol yield (0.48g/g) from ion exchange treated hydrolysate, followed by activated charcoal (0.42g/g), laccase (0.37g/g), overliming (0.30g/g) and neutralized hydrolysate (0.22g/g).     

‘正午’牡丹微繁殖体系的建立

本研究以‘正午’牡丹腋芽为外植体,建立了高效的牡丹微繁殖体系。腋芽的初始培养基为WPM+0.5mg/L BA+0.2mg/L GA3,培养50d后,一个丛生芽平均可切分为13个繁殖体用于增殖培养;增殖培养基为WPM[1668mg/L Ca(NO3)2·4H2O]+0.5mg/L BA+0.2mg/L GA3,以35d为一个继代培养周期,增殖率为3.0,共继代培养7次;生根培养时,先将无根苗在复壮培养基[1/2MS(296mg/L CaCl2)+0.5g/L活性炭]上培养20d,再转入根诱导培养基[1/2 MS(296 mg/L CaCl2)+1.0 mg/L腐胺+1.0 mg/L IBA]培养30d,最后转入根形成培养基[1/2 MS(296mg/L CaCl2)+4.0g/L活性炭]培养20d,其生根率达77.2%;驯化与移栽基质为珍珠岩∶蛭石∶草炭土=1∶1∶1,组培苗移栽成活率高达92.1%。这表明以‘正午’牡丹腋芽建立的微繁殖体系具备规模化商业生产的价值。     

Effect of nutrient supplementation of crude or detoxified concentrated distilled grape marc hemicellulosic hydrolysates on the xylitol production by Debaryomyces hansenii

Biosynthesis of xylitol using the yeast Debaryomyces hansenii NRRL Y-7426 was carried out using distilled grape marc (DGM) hemicellulosic hydrolysates directly concentrated by vacuum evaporation or after detoxification with activated charcoal. The effect of nutrient supplementation with vinasses, corn steep liquor (CSL) or commercial nutrients was explored. Using crude concentrated hemicellulosic hydrolysates, the maximum xylitol concentration, 11.3?g/L, was achieved after 172?hr (Q ( xylitol )?=?0.066?g/L-hr; Y ( xylitol ) (/SC)?=?0.21?g/g); meanwhile, using detoxified concentrated hydrolysates, the concentration increased up to 19.7?g/L after 72?hr (Q ( xylitol )?=?0.274?g/L-hr; Y ( xylitol ) (/SC)?=?0.38?g/g). On the other hand, using crude or detoxified hydrolysates, the xylose-to-xylitol bioconversion was strongly affected by the addition of nutrients, suggesting that these hydrolysates present essential nutrients favouring the growth of D. hansenii.     

文心兰试管苗分化及育苗技术研究

文心兰原球茎分化苗过渡驯化培养试验表明,改良1号+椰子汁100g/L的培养基有利于原球茎分化成苗,能促进分化苗生长健壮、整齐度好。壮苗与生根培养试验表明,以改良1号为基本培养基,加入香蕉泥100g/L、活性炭1g/L,在较强光照(2 000~3 000 lx)下培养,植株各项生长指标表现较好,有利于后期炼苗成活;以三角瓶为定瓶容器培养,植株接种效率最高、污染率最低;选取3cm高的植株定瓶,可显著提高文心兰工厂化组培育苗的工作效率。     

Effects of manufacturing conditions on the adsorption capacity of heavy metal ions by Makino bamboo charcoal

The objective of this study was to investigate the effects of manufacturing conditions on the adsorption capacity of heavy metal ions by Makino bamboo charcoal. Results show that the specific surface area and iodine number of bamboo charcoal activated at 900 degrees C were larger than those of bamboo charcoal activated at 800 degrees C. The specific surface area of bamboo charcoal activated at 800 degrees C by carbon dioxide was larger than that of charcoal activated by steam. However, a contrary result was observed when the activation temperature was 900 degrees C. The total volume and proportion of micropores in bamboo charcoal activated by carbon dioxide were greater than those in the other sample groups. However, the total volume and bulk volume of meso- and macropores, and average pore diameter for bamboo charcoal activated by steam were greater than those in the other sample groups. Using 5g bamboo charcoal (10-30 mesh) with a soaking time of 24h, a better adsorption effect on Pb2+ (100%), Cu2+ (100%), and Cr3+ (88-98%) was found. However, medium frequencies were observed for the adsorption of Cd2+ (40-80%) and Ni2+ (20-60%). Very limited adsorption of As5+ was detected in this study. For the same charcoal grain sizes, the adsorption capacity of 0.5g of charcoal was better than that of 0.1g. The improved adsorption effect of the sample group activated by steam was compared with the sample group activated by carbon dioxide.     

Microbial production of xylitol from D-xylose and sugarcane bagasse hemicellulose using newly isolated thermotolerant yeast Debaryomyces hansenii

A thermotolerant yeast capable of fermenting xylose to xylitol at 40°C was isolated and identified as a strain of Debaryomyces hansenii by ITS sequencing. This paper reports the production of xylitol from D-xylose and sugarcane bagasse hemicellulose by free and Ca-alginate immobilized cells of D. hansenii. The efficiency of free and immobilized cells were compared for xylitol production from D-xylose and hemicellulose in batch culture at 40°C. The maximum xylitol produced by free cells was 68.6 g/L from 100 g/L of xylose, with a yield of 0.76 g/g and volumetric productivity 0.44 g/L/h. The yield of xylitol and volumetric productivity were 0.69 g/g and 0.28 g/L/h respectively from hemicellulosic hydrolysate of sugarcane bagasse after detoxification with activated charcoal and ion exchange resins. The Ca-alginate immobilized D. hansenii cells produced 73.8 g of xylitol from 100 g/L of xylose with a yield of 0.82 g/g and volumetric productivity of 0.46 g/L/h and were reused for five batches with steady bioconversion rates and yields.     

甘蓝型油菜小孢子培养技术的几项改进

本研究在NLN-16和NLN-13的培养基中分别加入0.1mg/L 6-BA和0.05%的活性碳,结果表明6-BA对小孢子再生胚有明显地促进作用,再生胚的频率比对照增加26枚/皿,经分析达到显著水平;而0.05%的活性碳对小孢子再生胚促进作用不显著。对甘蓝型油菜小孢子培养再生植株的染色体加倍及移栽的研究结果表明,在小孢子培养初期加50mg/L秋水仙碱加倍效率最佳,加倍率达到67.6%。小孢子培养的再生苗移栽至大田后,采用遮阳网覆盖小苗,移栽成活率达到87.6%。 Abstract:The application of microspore culture technique was restricted because of its low frequency of embryogenesis and chromosome doubling.Two methods of enhancing the frequency of embryogenesis were employed in the study,namely,activated charcoal treatment in NLN-13 media and 6-BA treatment in NLN-16 media.The treatment with 0.05% activated charcoal produced 24 embryos per plate,which increased 1.7 embryos per plate,as compared with the treatment without activated charcoal.However,the analysis of T-test showed that it was not significant.After adding 0.1mg/L 6-BA in NLN-16 media,the frequency of embryogeny was 38.3 embryos per plate,and it was 26 embryos more per plate than that of CK.Analysis of T-test is significant.This indicates that 6-BA promotes embryogeny in microspore culture.Adding 50mg/L colchicines in NLN-16 media,the doubling frequency was 67.6%.The plantlets transplanted into field with two methods of light-covered net and plastic films were investigated.A survival rate of 87.6% was obtained using light-covered method whereas 57.7% survived using plastic film method.     

Effect of particle size of activated charcoal on separation of Triton X-100 from protein, liver cytosol, and lipoxygenase extracts

Triton X-100 was almost completely removed from bovine serum albumin solutions, BALB/c mouse liver extracts, and avocado peel lipoxygenase extracts by stirring the samples for 30 min in the presence of 250-350 mesh activated charcoal. The procedure did not remove protein significantly and did not reduce enzyme activity. At higher charcoal particle sizes, the efficiency of Triton adsorption was decreased and protein adsorption was increased. ElevateD temperatures enhanced Triton and protein adsorption. Adsorption on activated charcoal of 250-350 mesh is a simple and rapid procedure for Triton removal at a ratio of 0.23 g Triton X-100 per gram of activated charcoal.     

同色兜兰的非共生萌发与快速繁殖研究

以授粉后不同发育时期的同色兜兰种子为材料,观察其形态特征和萌发过程,并探讨建立同色兜兰高效快繁体系的最佳条件。结果表明,种子发育中后期即授粉后210~240d为较适宜的采收期,授粉后210d的种子萌发率最高(达77.79%);1/4 MS和1/2MS为同色兜兰适宜的基本培养基,添加100mL/L椰乳或1g/L蛋白胨对种子萌发及原球茎生长和分化有明显的促进作用;添加1g/L活性炭对原球茎褐化有一定的抑制作用,但添加剂量不宜过大;添加香蕉汁和苹果汁对同色兜兰种子萌发和原球茎生长分化有抑制作用;暗处理对同色兜兰种子萌发无影响;分化后的原球茎在壮苗和生根培养基上培养120d即可得到4~5片叶、高3~5cm的同色兜兰健壮试管苗。