[背景]由病原菌 Sclerotium denigrans 侵染引起的平贝母菌核病是其主要的鳞茎病害之一,给平贝母种植产业带来了巨大的损失.[目的]筛选出对平贝母菌核病具有拮抗效果的木霉菌株.[方法]以平贝母菌核病作为靶标菌,采用平板对峙试验、平板对扣法、圆盘滤膜法与发酵液抑菌试验筛选对平贝母菌核病具有拮抗效果的木霉菌株.采用顶空固相微萃取的方法检测拮抗效果较好的木霉菌挥发性成分;二硝基水杨酸(dinitrosalicylic acid,DNS)比色法测定木霉菌的 β-1,3葡聚糖酶的活性;室内防效试验验证其对平贝母菌核病的防治效果.[结果]平板对峙试验发现木霉菌F1、F2 和D6 对平贝母菌核病菌的生长具有较强的抑制作用,其抑菌率分别为 91.06%、87.00%和 86.12%;平板对扣法发现木霉菌 E17 和 A26 对菌核病菌的抑制效果最为明显,抑菌率分别为74.96%和 75.86%;圆盘滤膜法发现菌核病菌在F2、C6、D3、F4、A26、B30、D4 和D6 的琼脂培养基上均不生长,抑菌率达 100%;发酵液抑菌试验表明木霉菌D3 抑制效果最强,可完全抑制菌核病菌的生长,抑菌率为 100%;对A26、D4、E8、E17 和D3 这 5 株木霉进行GC/MS挥发性产物分析,在E17 发现了具有抗真菌活性的 6-戊基-2H-吡喃-2-酮等活性物质;DNS比色法发现β-1,3葡聚糖酶活性最高的木霉菌为F1;室内防效试验测定发现D3能明显抑制平贝母鳞茎菌核病的病变,对平贝母菌核病具有潜在的生防活性.[结论]木霉菌D3 在防治平贝母菌核病中是极具开发价值的菌种.

为优化大叶冬青瞬时高温灭菌工艺,分析其对大叶冬青灭菌效果及质量的影响,采用正交试验法,研究灭菌前后大叶冬青1,1-二苯基-2-苦基肼(1,1-Diphenyl-2-picrylhydrazyl,DPPH)自由基清除率及多酚类化合物和皂苷类化合物含量的变化,以灭菌温度、灭菌时间、药材粒度为考察因素,以灭菌率、DPPH自由基清除率、多酚类、皂苷类化合物含量为评价指标,通过权重分析法(criteria importance though intercrieria correlation,CRITIC)确定指标相关性权重并计算综合评分.结果表明,按照综合加权法确定的权重系数是多酚类化合物含量25.78%、皂苷类类化合物含量47.22%、DPPH·清除率26.99%,最佳灭菌工艺条件为:灭菌温度160或170℃,灭菌时间10 s,粉碎粒度不大于24目.按优化工艺灭菌后的样品微生物限度检查均符合药典规定;灭菌前后样品的多酚、皂苷类化合物含量及DPPH自由基清除率均无明显变化,且样品多酚、皂苷类化合物的指纹图谱相似度≥0.9.综上所述,瞬时高温灭菌能有效杀灭大叶冬青中的微生物,且对质量无影响.

To screen antagonistic endophytic bacteria isolated from the plant of Anemarrhena asphodeloides, of which could inhibit six common diseases of Panax ginseng and three ones of Asarum sclerotium, to provide a base for biological control of some Chinese medicinal materials. The antagonistic endophytic bacteria were isolated and screened by tissue slicing, tissue grinding and hyphal growth rate. The strains with significant bacteriostatic activity（P＜0.05） were identified according to 16S rDNA sequence and phylogenetic tree analysis, and the bacteriostatic rate of fermentation broth of active strains was detected by hyphal growth rate method. The fermentation conditions of the strain were optimized by orthogonal test. From the 359 endophytic bacteria isolated from the plant of A. asphodeloides three endophytic bacteria were screened, they are strains of BR10, BR11 and BR26, and all of them have had antagonistic effect and strong bacteriostatic activities to the nine pathogens, there was a highest bacteriostatic rate 92.41% of them. The results of phylogenetic analysis of 16S rDNA sequence showed that the BR11 had the closest relationship with many strains of Rahnella aquatilis, and was in the same branch of the phylogenetic tree, so the BR11 strain was identified as R. aquatilis. The optimum medium formula and culture conditions of BR11 strains were determined: beef paste 1.5%, glucose 1.5%, yeast extract 0.5%, MgSO 4 0.15%, initial pH value 7.5, temperature 29 ℃, fermentation time 72 h. The endophytic bacteria BR11 from A. asphodeloides is an excellent candidate for the control of ginseng and Asarum diseases.

In the present study, the inhibitory potential of 14 Trichoderma strains (isolated from Asarum rhizosphere) was investigated against Sclerotinia asari using the plate dilution method. The activity of antioxidant enzymes viz; catalase (CAT), peroxidase (POD), superoxide dismutase (SOD), and malondialdehyde (MDA) in S. asari treated with the two Trichoderma strains was also evaluated. Untargeted metabolomic analysis by using LC/MS analysis was carried out to determine differential metabolites in T. hamatum (A26) and T. koningiopsis (B30) groups. Moreover, transcriptome analysis of S. asari during the inhibition of S. asari by B30, and A26 compared with the control (CK) was performed. Results indicated that inhibition rates of T. koningiopsis B30, and T. hamatum A26 were highest compared to other strains. Similarly, non-volatile metabolites extracted from the B30 strain showed a 100% inhibition of S. asari. The activity of CAT, SOD, and POD decreased after treatment with A26 and B30 strains while increasing MDA content of S. asari. Antifungal activity of differential metabolites like abamectin, eplerenone, behenic acid, lauric acid, josamycin, erythromycin, and minocycline exhibited the highest inhibition of S. asari. Transcriptome analysis showed that differentially expressed genes were involved in many metabolic pathways which subsequently contributed toward antifungal activity of Trichoderma. These findings suggested that both Trichoderma strains (B30 and A26) could be effectively used as biocontrol agents against Sclerotinia disease of Asarum.

收集吉林省人参主产区锈腐病病样,对其进行病原菌的分离鉴定,确定优势病原菌.采用菌丝生长速率法、孢子萌发法、菌丝干质量测量法3种方法相结合,测定Rg1不同质量浓度条件下对人参锈腐病菌(Ilyonectria ro-busta)的致病力影响.通过测定施加不同质量浓度Rg1后人参粗壮柱孢菌(I.robusta)侵染相关酶活力测定试验确定最佳Rg1添加量.开展了Rg1最适质量浓度条件下,pH值以及碳氮源对人参锈腐病菌菌丝生长的影响.研究结果表明,当Rg1质量浓度为2 mg/L时对粗壮柱孢菌的生长具有较强的促进作用.在此条件下,病原菌最适生长pH值为5.0,最适碳源为葡萄糖,最适氮源为硝酸钾.明确了Rg1对人参锈腐病菌(I.robusta)具有明显的趋化性.

新农科建设背景下,专业课程的改革与建设应该以高校培养应用型人才为导向.药用植物栽培是培养现代化中药材生产方向人才的专业核心课程,是研究植物类中药材生产的理论与关键技术体系的科学.为培养学生较强的实践能力,我们从社会需求、理论教学、实践教学和传统文化教育等4个方面着手,认真研究了课程设计理念与思路,通过多维教学方法体系的实施,满足了人才培养的目标.

从健康防风中分离筛选对人参病害有拮抗作用的内生细菌,为人参病害的生物防治提供理论依据.采用生长速率法筛选对人参病害具有拮抗作用的防风内生细菌菌株,利用16S rDNA序列分析鉴定获得的具有拮抗作用的菌株.在防风中共分离得到83株内生细菌,通过初筛和复筛,从中筛选到5株对人参灰霉病菌等6种人参的病原菌拮抗作用较好的菌株,分别为SSBR8、SSBR11、SSBR42、SSBR59和SSBR64,其中SSBR8和SSBR59 为嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia);SSBR11(Agrobacterium tumefaciens)和 SSBR64(Agrobacterium arsenijevicii)为 土壤杆菌属;SSBR42(Achromobacter spanius)为无色杆菌属.SSBR8 对人参立枯病菌、灰霉病菌、疫病菌的抑菌效果较好,抑菌率均＞76％.说明防风内生菌资源丰富,并且存在高活性拮抗菌株,值得深入开发.

Background Asarum heterotropides Fr. Schmidt var. mandshuricum (Maxim.) Kitag. is an important medicinal and industrial plant, which is used in the treatment of various diseases. The main bioactive ingredient is the volatile oil having more than 82 identified components of which methyleugenol, safrole, myristicin, and toluene account for about 70% of the total volume. As a sciophyte plant, the amount of light it absorbs through leaves is an important factor for growth and metabolism. Results We grew Asarum plants under full, 50, 28, and 12% sunlight conditions to investigate the effect of different light irradiances on the four major volatile oil components. We employed de novo transcriptome sequencing to understand the transcriptional behavior of Asarum leaves regarding the biosynthetic pathways of the four volatile oil components, photosynthesis and biomass accumulation, and hormone signaling. Our results demonstrated that the increasing light conditions promoted higher percent of the four components. Under full sunlight conditions, cinnamyl alcohol dehydrogenase and cytochrome p450719As were upregulated and led the increased methyleugenol, safrole, and myristicin. The transcriptomic data also showed that Asarum leaves, under full sunlight conditions, adjust their photosynthesis-antenna proteins as a photoprotective response with the help of carotenoids. Plant hormone-signaling related genes were also differentially expressed between full sunlight and low light conditions. Conclusions High light induces accumulation of major bioactive ingredients A. heterotropides volatile oil and this is ascribed to upregulation of key genes such as cinnamyl alcohol dehydrogenase and cytochrome p450719As. The transcriptome data presented here lays the foundation of further understanding of light responses in sciophytes and provides guidance for increasing bioactive molecules in Asarum .

[背景]菌核病是北细辛根部主要病害之一,木霉菌作为目前应用最广泛的生物防治真菌,利用木霉菌防治北细辛菌核病是目前研究的热点.[目的]通过稀释分离法对健康北细辛植株根际土壤进行菌株分离,以期筛选出有效拮抗北细辛菌核病的生防木霉菌.[方法]以北细辛菌核病菌为靶标菌,采用平板对峙培养、挥发性与非挥发性物质抑菌的方法对分离得到的木霉菌进行筛选,采用生长速率法对筛选出的木霉菌的发酵液进行抑菌效果测定,并采用硫代巴比妥酸法测定筛选出的木霉对北细辛菌核病菌的丙二醛(Malondialdehyde,MDA)含量、紫外吸收法测定过氧化氢酶(Catalase,CAT)活性、氮蓝四唑法测定超氧化物歧化酶(Superoxide Dismutase,SOD)活性、愈创木酚法测定过氧化物酶(Peroxidase,POD)活性的影响.[结果]从土壤中分离出木霉菌共14株,通过形态学和ITS-RPB2双基因联合构建系统发育树,鉴定其为哈茨木霉(Trichoderma harzianum)、钩状木霉(Trichoderma hamatum)、拟康氏木霉(Trichoderma koningiopsis)、深绿木霉(Trichoderma atroviride)、短密木霉(Trichoderma brevicompactum)和装絮木霉(Trichoderma tomentosum).对峙培养试验表明,钩状木霉A26、拟康氏木霉B30、钩状木霉C6、哈茨木霉A17对北细辛菌核病菌抑制率均在90％以上,挥发性物质抑制测定结果显示钩状木霉C6抑制率最高,为53.73％±0.07％,木霉菌的非挥发性物质抑菌作用较强,哈茨木霉A17、钩状木霉A26、钩状木霉C6的非挥发性物质对细辛菌核病菌的抑制率均在75％以上,而拟康氏木霉B30抑制率可达100％.因此,筛选出的哈茨木霉A17、钩状木霉A26、拟康氏木霉B30、钩状木霉C6为拮抗效果较强的生防木霉菌,这4株木霉菌的发酵液对北细辛菌核病菌的抑制率分别为56.33 ％±0.12％、77.22％±0.06％、82.28％±0.03％、46.20％±0.04％.经这4株木霉菌的非挥发性物质处理7d后,菌核病菌MDA含量显著增加,钩状木霉A26是对照组的7.7倍,最为显著;菌核病菌抗氧化酶活性均降低,与对照组相比,CAT、SOD、POD活性分别下降了19.67％-75.84％、4.71％-68.71％和3.57％-67.86％.[结论]从北细辛健康植株根际土壤中分离的木霉菌株哈茨木霉A17、钩状木霉A26、拟康氏木霉B30、钩状木霉C6对北细辛菌核病菌均有较好的抑制效果,可用于北细辛菌核病的生物防治.

采用组织块法,从健康辽藁本中共分离出64株内生细菌,经初筛和复筛,菌株ZHAB63对11种常见中药材病原菌拮抗率均在50.00％以上(P<0.01),根据菌株形态特性、16SrDNA基因序列测定以及系统发育树初步鉴定发现,菌株ZHAB63为解淀粉芽孢杆菌(Bacillus amyloliquefaciens);采用L9(34)正交设计试验优化该菌株发酵培养基及发酵条件,确定液体发酵最适培养基为:蔗糖15.0 g·L-1,酵母粉20.0 g·L-1,蛋白胨20.0 g·L-1,硫酸镁5.0 g·L-1;最佳发酵条件:温度30℃,pH 6.5,转速180 r·min-1,培养时间48 h.研究表明,辽藁本中内生细菌资源丰富,以其研制新配方生物农药有待深入开发.

目的 研究47份不同来源、不同表型正品细辛样品的有效成分含量,评价正品细辛样品的质量,揭示有效成分形成与遗传和地域的关系,为细辛优良品种选育与开发利用提供理论依据.方法 冷凝回流法提取挥发油、乙醇热浸法提取醇溶性浸出物,HPLC法测定细辛脂素含量、气质联用(GC-MS)分析挥发油组分,组间联接法进行聚类分析.结果 47份细辛样品挥发油量0.81％～3.32％.有5份样品挥发油量超过3.0％.醇溶性浸出物量为9.87％～29.40％,含量超过20％的样品占总数的63.8％.细辛脂素质量分数为0.118％～0.424％,有26份样品细辛脂素质量分数超过0.25％;47份细辛挥发油可分离出的化合物种类48～77种,化合物种类和含量有较大差异.基于有效成分的聚类分析将不同来源地、不同表型的细辛分为10类,聚类方式基本是按照来源地为基础进行聚类,同一来源地的汉城细辛和北细辛聚在一类中.结论 在同一栽培环境和条件下,不同来源地、不同植物学表型的样品个体间有效成分含量间存在差异,新品种选育时可依照植物表型选择优良育种材料.大部分样品聚类与来源地域有关而与植物表型以及植物学分类无关.

Asarum heterotropoides Fr. var. mandshuricum (Maxim) Kitag (Chinese wild ginger) is an important medicinal herb. Essential oil extracted from its roots is the key ingredient and is mainly composed of phenylpropanoid compounds. As a skiophyte plant, light is a crucial factor for A. heterotropoides var. mandshuricum growth and metabolism. To investigate the effects of light irradiation on the essential oil biosynthesis in A. heterotropoides var. mandshuricum, the plants were cultivated in four light irradiation treatments (100, 50, 24 and 12% full sunlight). The photosynthetic capacity, essential oil content and composition, activities of several enzymes and levels of some secondary metabolites involved in the shikimic acid and cinnamic acid pathways were analyzed. The leaf mass per area, average diurnal net photosynthetic rate, and the essential oil content increased significantly with increasing light intensity. Phenylalanine, cinnamic acid, and p-coumaric acid in the cinnamic acid pathway were at their highest levels in plants cultivated in 100% full sunlight. The highest content of shikimic acid in the shikimic acid pathway was obtained in plants grown in 50% sunlight transmittance. The activity of the enzymes 3-Deoxy-D-arabino-heptulosonate-7-phosphate synthase, phenylalanine ammonia lyase, cinnamate-4-hydroxylase and 4-coumarate:CoA ligase increased proportionally with light intensity. Overall, we conclude that high light irradiation promotes high net photosynthetic rate, high activity of enzymes and high amounts of phenylpropanoid precursor metabolites leading to significant biosynthesis of essential oil in A. heterotropoides var. mandshuricum.

研究人参内生菌B69菌株在人参根、茎、叶及其根际土壤中的定殖规律,以及B69菌株对人参根腐病的防效,以期为新型人参根腐病生防制剂的开发提供理论依据.采用抗生素标记法,筛选出最佳标记菌株;采用盆栽和田间试验测定自然土与无菌土处理下B69菌株在人参植株内和根际土壤中的定殖能力.结果表明,标记菌株与未标记菌株在形态特征、拮抗能力、防病效果等方面无显著差异;B69菌株能够高效定殖在人参根部;B69菌株对人参根腐病有较好的防治效果,田间试验结果表明该菌处理后的病情指数为25.41,对根腐病的相对防效为58.56％.说明B69菌株是有防治人参根腐病潜力的生防菌.

为了解干旱胁迫对菘蓝光合特性的影响,采用盆栽法,利用大型移动防雨棚开展了菘蓝干旱胁迫试验,研究干旱胁迫对菘蓝净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)、蒸腾速率(Tr)、气孔限制值(Ls)和水分利用率(WUE)日变化的影响.结果 表明,在适宜水分和轻度干旱胁迫下,气孔限制是菘蓝Pn降低的主要因素;中度和重度干旱胁迫下,非气孔限制是菘蓝Pn降低的主要因素.干旱胁迫会导致菘蓝的Tr降低和Tr峰值的前移.随着干旱胁迫程度的加剧,菘蓝为适应干旱缺水的条件会使叶片的Pn、Tr和Ge均显著下降,从而提高干旱情况下的WUE,缩小与充足供水情况下的差距.

研究人参内生细菌B69菌株对人参根腐病病原菌的作用机制,通过硫酸铵沉淀、低温乙醇沉淀等方法获得菌株发酵液中活性成分,对其抑菌物质粗提液进行稳定性测定,并对其抑菌成分进行分离,采用平板扩散法测定提取成分的抑菌活性.结果表明,从B69菌株发酵液得到的提取液具有抑菌活性;对抑菌物质粗提物进行高温、酸碱度、紫外线、反复冻融和蛋白酶K处理,抑菌活性均未见明显变化.从B69发酵液中初步分离出具有抑制根腐菌活性的活性肽和胞外多糖.表明B69菌株能产生多种抑菌活性成分,抑菌成分对胁迫条件不敏感,值得进一步开发利用.

Chlorophyll content,leaf mass to per area,net photosynthetic rate and bioactive ingredients of Asarum heterotropoides var. mandshuricum,a skiophyte grown in four levels of solar irradiance were measured and analyzed in order to investigate the response of photosynthetic capability to light irradiance and other environmental factors. It suggested that the leaf mass to per area of plant was greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in the decrease of leaf mass to per area at every phenological stage. At expanding leaf stage,the rate of Chla and Chlb was 3. 11 when A. heterotropoides var. mandshuricum grew in full light irradiance which is similar to the rate of heliophytes,however,the rate of Chla and Chlb was below to 3. 0 when they grew in shading environment. The content of Chla,Chlb and Chl( a+b) was the greatest value of four kinds of light irradiance and decreasing intensity of solar irradiance resulted in its decreasing remarkably( P<0. 05). The rate of Chla and Chlb decreased but the content of Chla,Chlb and Chl( a+b) increased gradually with continued shading. The maximum value of photosynthetically active radiation appeared at 10: 00-12: 00 am in a day. The maximum value of net photosynthetic rate appeared at 8: 30-9: 00 am and the minimum value appeared at 14: 00-14: 30 pm at each phenological stage if plants grew in full sunlight. However,when plants grew in shading,the maximum value of net photosynthetic rate appeared at about 10: 30 am and the minimum value appeared at 12: 20-12: 50 pm at each phenological stage. At expanding leaf stage and flowering stage,the average of net photosynthetic rate of leaves in full sunlight was remarkably higher than those in shading and it decreased greatly with decreasing of irradiance gradually( P < 0. 05). However,at fruiting stage,the average of net photosynthetic rate of leaves in full sunlight was lower than those in 50% and 28% full sunlight but higher than those in 12% full sunlight. All photosynthetic diurnal variation parameters of plants measured in four kinds of different irradiance at three stages were used in correlation analysis. The results suggested that no significant correlation was observed between net photosynthetic rate and photosynthetically active radiation,and significant negative correlation was observed between net photosynthetic rate and environmental temperature as well as vapor pressure deficit expect for 12% full sunlight. Positive correlation was observed between net photosynthestic rate and relative humidity expect for 12% full sunlight. Significant positive correlation was observed between net photosynthetic rate and stomatal conductance in the four light treatments. Only,in 12% full sunlight,the net photosynthetic rate was significantly related to photosynthetically active radiation rather than related to environmental temperature,vapor pressure deficit and relative humidity. In each light treatment,a significant positive correlation was observed between environmental temperature and vapor pressure deficit,relative humidity as well as stomatal conductance. Volatile oil content was 1. 46%,2. 16%,1. 56%,1. 30% respectively. ethanol extracts was 23. 44%,22. 45%,22. 18%,21. 12% respectively. Asarinin content was 0. 281%,0. 291%,0. 279% and 0. 252% respectively. The characteristic components of Asarum volatile oil of plant in different light treatments did not change significantly among different groups.

采用盆栽法在薏苡生长季节设3种水分处理(312、468和624 mm),研究干旱胁迫对薏苡净光合速率(Pn)、气孔导度(Gs)、胞间CO2浓度(Ci)、蒸腾速率(Tr)、气孔限制值(Ls)和水分利用率(WUE)日变化的影响.结果表明,3种处理Pn日均值分别为0.56、0.64和0.82μmol/(m2·s);Tr日平均值分别为0.224、0.247、0.355 mmol/(m2·s);WUE日平均值分别为2.773、2.696、2.323μmol/mmol.Pn、Tr、WUE值经统计分析均显示处理间差异显著(P<0.05),表明干旱显著降低了薏苡的光合能力.但是,Gs、Ci和Ls日平均值均未达到显著差异(P>0.05),表明这3项光合参数与干旱关系不密切.

目的:研究双去甲氧基姜黄素对硫代乙酰胺诱导的小鼠肝纤维化的保护作用及其机制.方法:将ICR小鼠随机分为正常对照组、模型组、水飞蓟素阳性对照组及双去甲氧基姜黄素高、中、低剂量组.除正常对照组外,其余各组均隔天1次连续7 w腹腔注射硫代乙酰胺溶液(200 mg/kg)诱导小鼠肝纤维化模型.双去甲氧基姜黄素高(200 mg/kg)、中(100 mg/kg)、低(50 mg/kg)剂量组1次/d,连续7w分别灌胃给予,正常对照组、模型组给予等体积相应溶剂.治疗结束后,采用生化分析法检测血清丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)、总胆红素(IBI)及肝组织中羟脯氨酸(HYP)含量;酶联免疫吸附法(ELISA)检测血清透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PC-Ⅲ)、Ⅳ型胶原(Ⅳ-C)水平;HE、Masson染色方法观察小鼠肝组织病理变化及肝纤维化的形成程度;蛋白免疫印迹法(Western blot)检测磷酸化的磷脂酰肌醇3-激酶(p-PI3K)、磷酸化的丝氨酸/苏氨酸蛋白激酶(p-Akt)、半胱氨酸天冬氨酸蛋白酶3(Caspase-3)、Bad蛋白的表达.结果:与模型组比较,双去甲氧基姜黄素各剂量组血清AST、ALT、IBI、HA、LN、PC-Ⅲ、Ⅳ-C和肝组织中HYP的水平显著降低(P＜0.05或P＜0.01),肝细胞变性坏死和纤维组织增生明显减轻,肝组织中p-PI3K、p-Akt、Caspase-3、Bad蛋白表达显著下调(P＜0.05或P＜0.01).结论:双去甲氧基姜黄素具有抗肝纤维化作用,其作用机制可能与PI3K/Akt信号通路有关.

目的 探索人参内生菌B69菌株对根腐病的抑制作用,优化人参病害拮抗菌株B69的发酵条件,提高发酵液的活菌含量和抗菌活性.方法 以B69发酵液对病原菌根腐病的抑制作用为活性指标,采用正交试验和单因素试验方法对菌株的最适发酵培养基成分及发酵条件进行优化.结果 在最佳发酵培养基和培养条件下,经检测发酵液的活菌数含量为8.25×108 cfu/mL,较优化前的活菌含量1.82×108cfu/mL增幅达到了显著差异(P<0.05),此含量达到了农业部微生物肥料的技术标准(有效活菌数≥2.0亿/mL).发酵液对人参根腐病的抑菌圈也由优化前的18mm增加到35mm.结论 最佳培养基配方为蔗糖2.0％、淀粉1.0％、牛肉浸膏0.5％、酵母粉0.5％、蛋白胨0.2％和NaCl0.5％;初始pH7.0;最适装液量为100 m L/250 m L;培养条件为35℃、170r/min振荡培养;接种量5％,发酵时间为48h.

Objective To establish and optimize preparation technology of ginsenoside Re liposomes,therefore to improve storage stability.Methods Ginsenoside Reliposomes were prepared by the method of film dispersion-mechanical vibration,which were collected by separating liposome from disclosed free drug by dialysis method.Measure entrapment efficiency by HPLC.Prepare freeze-dried lip osome preparations by freezing-drying technology.Taking entrapment efficiency as the main screening index,optimize liposome formulation and freezing-drying technology by orthogonal test design.Results The entrapment efficiency of ginsenoside Re lipidosomes prepared by the method of film dispersion-mechanical vibration is the highest.The best formulation technology is:Mass ratio of drug and phospholipid is 1 ∶ 30,mass ratio of phospholipid and cholesterol is 16 ∶ 1,ice-water bath ultrasound is 30 min,and double distilled water is hydration solution;The best freezing-drying technology is:Taking sucrose as the freeze-drying protective agent,mass ratio of disaccharide-water is 1 ∶ 10,pre-freezing temperature is-20 ℃,and normal saline of 0.9％ is reconstitution solution.Conclusion The preparation technology of liposome is stable and practicable.The ginsenoside Re liposome prepared by taking the sucrose as the freeze-drying protective agent has good indexes,which can extend the storage period.