高压静电场对贯叶连翘种子休眠破除及药用成分的影响

将高压静电场(HVEF)技术应用于贯叶连翘种子休眠破除及组培苗培养,结果显示,贯叶连翘种子在静电场(100 kv/m)中处理1 h,其发芽势(SPO)和发芽率(SPT)是对照组(无静电处理)的5.69倍和2.45倍;贯叶连翘组培苗在静电场(135 kv/m～150 kv/m)中处理1 h,其药用成分总金丝桃素含量和贯叶金丝桃素含量是对照组的1.35～1.53倍;静电场(150 kv/m)处理后,POD活性是其对照的1.30倍.研究表明,一定强度的静电场作用,能有效提高贯叶连翘种子发芽率,促进总金丝桃素和贯叶金丝桃素代谢含量.     

贯叶连翘醇提条件的多指标优化

本研究采用超声波提取法,对贯叶连翘中总黄酮、金丝桃素类和贯叶金丝桃素等主要有效成分在醇提过程中的提取条件(溶媒种类、浓度、提取时间等)进行了考察。结果表明,以总黄酮和金丝桃素类化合物为指标,用65％～80％的乙醇水溶液超声提取30min,提取效率较高;以贯叶金丝桃素为指标,则以用75％～90％的甲醇提取30min效果较好。     

茉莉酸甲酯(MeJA)对贯叶连翘悬浮细胞生长和贯叶金丝桃素产量的影响

培养基中添加100 μm01·L-1MeJA后,贯叶连翘悬浮细胞生物量增加量和贯叶金丝桃素(HF)的产量分别是未经MeJA处理的1.3倍和1.73倍;培养3 d时,培养基中添加100 μmol·L-1 MeJA能提高HF产量,是未经MeJA处理的2.4倍.     

贯叶连翘愈伤组织中分泌细胞群的发生与金丝桃素类物质的积累

贯叶连翘(Hypericum perforatum L．)是一种传统草药,在欧洲被广泛用于治疗抑郁症。其重要的活性成分,金丝桃素类物质储存在茎、叶和花瓣的分泌细胞团中。本文应用组织化学及电子显微镜技术,研究体外培养的贯叶连翘叶肉细胞脱分化产生愈伤组织以及细胞发育过程中金丝桃素类物质的积累、运输的情况,进一步探讨细胞的生长发育与次生代谢产物的关系。发现金丝桃素类物质产生于愈伤组织培养后期,在愈伤组织表面所形成的分泌细胞群中,最初在细胞质中形成,之后运输至液泡中积累,内质网参与了金丝桃素类物质的合成过程。这些结果为利用组织培养技术提高金丝桃素类物质含量提供了理论基础和依据。     

贯叶连翘野生转家化前后生物学性状特征观察

野生贯叶连翘 (HypericumperforatumL .)在江苏经过了 3a的引种栽培 ,其生物学特性检测结果表明 ,栽培贯叶连翘的生物量、开花的一致性和生长的适应性等明显优于野生贯叶连翘 ;良好的繁殖方式使贯叶连翘的收获期提前了 1a ;成熟后 3个月采收的种子萌发率较高 ,不同浓度赤霉素处理能明显提高其种子的萌发率 ;贯叶连翘的黑色腺体即分泌细胞球数量与金丝桃素间也基本存在着正相关关系 ,贵州贵阳居群和陕西丹凤居群黑色腺体明显多于其他居群 ,其金丝桃素含量在所有居群中为最高 ,而甘肃武都居群黑色腺体最少 ,其金丝桃素含量在 6个居群中最低。 6个野生居群栽培后生物量相近 ,在 4个不同土壤pH的基地大田中均能良好生长 ,说明江苏地区环境条件完全适合贯叶连翘引种栽培。     

贯叶连翘抗抑郁研究新进展

贯叶连翘,又名圣约翰草,是传统中药材之一,圣约翰草在德国用于抗抑郁症已有几百年的历史。贯叶连翘提取物对轻度和中度抑郁症患者和动物模型都有很好疗效,最新药理学研究表明贯叶连翘提取物中抗抑郁的主要成分是贯叶金丝桃素。贯叶金丝桃素是神经递质5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NE)的非竞争性重吸收抑制剂,贯叶金丝桃素还可以抑制突触体对γ氨基丁酸(GABA)和L-谷氨酸(L-glu)的重吸收,其作用机理至今还不甚明了,研究表明它的作用很可能是通过提高突触体细胞内钠离子浓度或通过降低突触体内突触小泡的跨膜pH梯度实现的。     

贯叶连翘中金丝桃素的合成与积累研究进展

金丝桃素是贯叶连翘的主要药理活性成分。本文概述了金丝桃素的化学与生物合成途径,介绍了金丝桃素在贯叶连翘个体发育过程中的积累以及利用贯叶连翘的细胞和组织培养技术生产与积累金丝桃素的研究进展。最后,指出分子生物学和电子显微镜技术的发展为深入研究金丝桃素的产生和积累提供了有利工具。     

从贯叶连翘中提取金丝桃素的工艺探讨

对从贯叶连翘中提取金丝桃素的工艺条件进行了优化研究 ,结果表明 ,金丝桃素提取较佳工艺为 :45℃的水浸泡 2 h后 ,加入 0 .5 % Na OH,用 75 %甲醇在 0 .0 5 Mpa下 78℃提取5次 ,每次 1 .5 h,溶媒量为 5倍 ,浓缩温度与干燥温度为 78℃     

用贯叶金丝桃残渣栽培香菇的研究

用提取金丝桃素后的贯叶金丝桃残渣和锯木屑作培养基分别栽培香菇 ,比较了由两种培养基栽培香菇的生物学效率、香菇的可溶性蛋白含量和超氧化物歧化酶 (SOD)活力。结果表明 :用贯叶金丝桃残渣栽培的香菇有较高的生物学效率、其香菇可溶性蛋白含量和SOD活力亦高 ,说明贯叶金丝桃残渣适合栽培香菇 ,为栽培香菇开发了一种新的生物资源。     

NO对金丝桃悬浮细胞生长及金丝桃素生物合成的促进作用研究

一氧化氮 (NO)是近年来发现的一种新型植物信号分子。以硝普钠 (Sodiumnitroprusside ,SNP)为一氧化氮 (NO)的供体 ,研究外源NO对金丝桃悬浮细胞生长及金丝桃素生物合成的影响。试验结果表明 ,金丝桃悬浮细胞在含 0 5和 15 0mmol LSNP的培养基中培养 2 0d后 ,细胞的干重分别为对照组的 140%和50% ;细胞中金丝桃素的含量分别为对照组的 98%和210%。试验结果表明 ,低浓度SNP处理有利于金丝桃悬浮细胞生长 ,而高浓度SNP可以促进金丝桃素的合成。在细胞培养初期 (0d)加入 0.5mmol LSNP并在指数生长后期 (14d)加入15.0mmol LSNP的金丝桃悬浮细胞在培养 2.5d后 ,细胞的干重和金丝桃素的含量分别为对照组的1.4和1.8倍 ,金丝桃素的产量达15.2mg/L ,比对照高3.2倍。SNP对金丝桃悬浮细胞生长及金丝桃素含量的影响可以被NO专一性淬灭剂CPITO(2-4-carboxyphenyl-4 ,4 ,5 ,5-tetramethylimidazoline-1-oxyl-3-oxide)所抑制,说明SNP是通过其分解产物NO影响细胞生长和金丝桃素的合成。试验结果同时表明,在15.0mmol/L的SNP处理下,金丝桃悬浮细胞中的苯丙氨酸解氨酶(PAL)的活性显著升高,推测NO可能通过触发金丝桃悬浮细胞的防卫反应,激活了细胞中金丝桃素的生物合成途径。     

<Emphasis Type="Italic">In vitro</Emphasis> regeneration of <Emphasis Type="Italic">Melothria maderaspatana</Emphasis> via indirect organogenesis

An effective protocol was developed for in vitro regeneration of the Melothria maderaspatana via indirect organogenesis in liquid and solid culture systems. Organogenesis was achieved from liquid culture calluses derived from leaf and petiole explants of mature plants. Organogenic calluses (98.2?±?0.36 and 94.8?±?0.71%) were induced from both leaf and petiole explants on Murashige and Skoog (MS) liquid medium containing 6.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 µM thidiazuron (TDZ); and 6.0 µM 2,4-D and 1.0 µM benzyladenine (BA) combinations, respectively. Adventitious shoot regeneration (68.2?±?0.06 shoots per explant) was achieved on MS medium supplemented with 2.0 µM BA, 4.0 µM TDZ, 10% v/v coconut water and 0.06 mM glutamine from leaf-derived calluses. Petiole-derived calluses produced adventitious shoots (45.4?±?0.09 shoots per explant) on MS medium fortified with 2.0 µM BA, 4.0 µM TDZ, 10% v/v coconut water, and 0.08 mM glutamine. Elongation of shoots occurred in MS medium with 2.0 µM gibberellic acid (GA₃). Regenerated shoots (2–3 cm in length) rooted (74.2?±?0.38%) and hardened (85?±?1.24%) when they were transferred to 1/2-MS medium supplemented with 3.0 µM indole-3-butyric acid (IBA) followed by garden soil, vermiculate, and sand (2:1:1 ratio) mixture. The elongated shoots (4–5 cm in length) were exposed simultaneously for rooting as well as hardening (100%) in moistened [(1/8-MS basal salt solution with 5 µM IBA and 100 mg l^?1 Bavistin® (BVN)] garden soil, vermiculate, and sand (2:1:1 ratio) mixture. Subsequently, the plants were successfully established in the field. The survival percentage differed with seasonal variations.     

生姜茎尖组织培养和快速繁殖研究

以生姜茎尖为外植体进行培养,筛选诱导愈伤组织和生根的最佳培养基。结果表明,在茎尖培养中,合理的消毒处理对茎尖成活率影响很大;以MS+6-BA 1.5mg/L+NAA 0.1mg/L为最适茎尖诱导培养基,可一次诱导形成愈伤组织,并直接形成带根幼苗,月增殖倍数为6.9倍,成活率76.9%。生姜腋芽继代培养基MS+6-BA 2.0mg/L+NAA 0.1mg/L+KT 1.0mg/L,以腐殖质土:菜园土=1:1基质可促进小苗后期生长,加速成苗。     

Molecular mass sorting of proteome using hollow fiber flow field-flow fractionation for proteomics

Hollow fiber flow field-flow fractionation (HF FlFFF) has been demonstrated as a tool for pre-fractionating proteomes by differences in molecular mass (Mr), where the resulting protein fractions are subsequently digested and analyzed by shotgun proteomics using two-dimensional liquid chromatography-electrospray ionization-tandem mass spectrometry (2D-LC-ESI-MS/MS). HF FlFFF is a separation device capable of fractionating proteins or cells by hydrodynamic radius, and protein fraction can be readily collected as intact conditions in aqueous buffer solutions. In this study, HF FlFFF was applied to fractionate the proteome of Corynebacterium glutamicum, a well known soil bacterium that has been widely used in bioindustry due to its remarkable ability to secrete high amounts of glutamic acid. The collected HF FlFFF fractions of different MW intervals were enzymatically digested for protein identification by 2D-LC-ESI-MS/MS. Experiments showed improvements in protein identification when HF FlFFF pre-fractionation was applied, due to decreases in the ionization suppression effect and the MS exclusion effect by spectral congestion. Pre-fractionation of C. glutamicum proteome allowed us to find 90 additional proteins by 2D-LC-ESI-MS/MS that were not found by a direct shotgun analysis without pre-fractionation. A total of 415 proteins were found overall with 203 proteins commonly found from experiments with and without pre-fractionation.     

Root and needle litter decomposition responses to enhanced supplies of N and S in a Norway spruce forest in southwest Sweden

The effects of additions of ammonium sulfate (NS) on the decomposition of litter derived from Norway spruce roots (< 2 and 2 - 5 mm in diameter) in the humus and mineral soil layers (0 - 15 cm) of a Norway spruce stand in southern Sweden were investigated over a 6-year period. To this purpose, litterbags were incubated in the humus layer and in the mineral soil in June 1996, with roots collected from NS and control (C) plots incubated in the NS and C plots, respectively. The N concentrations in fine roots (< 2 mm) in the NS- plots were higher than those in 2 - 5 mm roots in both humus and mineral soil layers. In the humus layer, N concentrations in the fine roots in the C- and NS- plots were 12.8 and 15.7 mg g ^{? 1}, respectively. By the end of the fifth year the < 2 mm roots in humus layer had lost 48.5 and 50% of their mass in the C and NS plots, respectively, while the corresponding values for the 2 - 5 mm diameter class were 44 and 54%. The fresh root litter may be a sensitive indicator to responses to enhanced N and S deposition, although decomposition rates of both litter types are affected.     

黄土高原典型区域土壤腐殖酸组分剖面分布特征

黄土高原作为典型的气候敏感带和生态环境脆弱区,诸多因素影响着这个区域的土壤有机碳及其组分的分布特征。本文以黄土高原典型区域土壤剖面 0—200 cm土样为对象,分析了土壤腐殖酸、胡敏酸（HA）和富里酸（FA）含量随地理位置及土层深度的分布特征,并进一步探讨了土壤腐殖酸、HA和FA与全氮含量及土壤颗粒组成的关系。结果表明,黄土高原主要类型土壤腐殖酸、HA和FA含量均较低,且存在明显地理位置和土层分异性：从南到北同层次土壤腐殖酸、HA和FA含量均显著降低,同一区域随土层深度增加各组分含量均表现为在 0—40 cm土层明显下降, 40—120 cm土层稍有下降,120 cm土层以下基本稳定;土壤腐殖酸占有机碳比例变化范围为26.6%—54.7%,相对较小,且在整个剖面变化幅度不大,从南向北土壤腐殖酸占有机碳比例有增加趋势;土垫旱耕人为土在 0—40 cm、 40—120 cm和 120—200 cm土层中HA占腐殖酸比例分别为39.8%、49.0%和53.5%,HA/FA分别为0.66、0.96和1.15,黄土正常新成土在以上土层中HA占腐殖酸比例分别为26.3%、33.9%和42.3%,HA/FA分别为0.36、0.51和0.73,干润砂质新成土在以上土层中HA占腐殖酸比例分别为13.4%、37.1%和45.2%,HA/FA分别为0.16、0.59和0.82,说明黄土高原南北主要类型土壤腐殖酸品质总体较差,均属富里酸型土壤,且从南到北腐殖酸品质逐渐下降;土壤腐殖酸、HA和FA均与全氮含量呈极显著线性相关（P<0.01）,土壤有机碳、腐殖酸及HA含量与粘粒及砂粒百分含量亦呈高度线性相关（P<0.01）。     

澳洲青苹组织培养再生体系的研究

通过对澳洲青苹茎尖培养及植株再生体系中无菌培养物的建立、初代培养、继代培养、生根培养和移栽五个技术环节的研究,建立了澳洲青苹组织培养快速繁殖技术体系。结果表明：(1)BA0．3～0．6mg／L NAA0．1～0．3mg／L的MS培养基能促进澳洲青苹茎尖生长或大量侧芽的分化,不定芽生长健壮;(2)无根幼苗在含有IBA0．6～1．0mg／L NAA0．1mg／L的1／2MS培养基上暗培养一周后,再进行自然光培养,30d生根率可达到78%以上;(3)生根苗经50mg／L NAA浸渍1～2h后,移入蛭石：腐殖土：田园土=2：1：1的移栽基质中,保温保湿,成活率达80％以上;(4)进一步建立了自然光下生根、练苗一次完成的同步化体系,缩短了繁育时间,简化了步骤,降低了成本。此技术体系为澳洲青苹优良种苗快速繁殖提供了一条新途径。     

半干旱黄土丘陵区纯林土壤腐殖质异化特征及与其他性质的关系

纯林长期生长或多代连栽必然会导致土壤腐殖质含量和构成发生异化,探究这种异化特征及其与土壤其他性质的关系可以为纯林管理或混交改造提供科学依据。通过对半干旱黄土丘陵区南泥湾林场8种典型纯林土壤腐殖质及其他性质进行系统检测,结果表明:(1)侧柏林土壤腐殖质含量最高(34.61 g/kg),腐殖化程度和稳定性一般;白榆和白桦林土壤的腐殖质含量中等(19.69—23.58 g/kg)、腐殖化程度和稳定性最佳;茶条槭和小叶杨林土壤的腐殖质含量(20.59—22.53 g/kg)和构成均为中等水平;油松、沙棘和刺槐林土壤的腐殖质质量较低(11.77—13.81 g/kg),且腐殖化程度较低,稳定性相对最差;(2)与胡敏酸含量存在显著相互促进作用(P0.05)的土壤性质为CEC、N、微生物量和蛋白酶活性(相关系数0.769—0.926,下同),存在显著相互抑制作用的为有效Cu(-0.793);与富啡酸存在显著相互促进作用的为N、CEC、微生物量、蔗糖酶和磷酸酶活性(0.836—0.955),存在显著相互抑制作用的为有效Cu(-0.822);与胡敏素存在显著相互促进作用的为N、CEC、微生物量、磷酸酶活性和有效Zn(0.766—0.951),存在显著相互抑制作用的为脱氢酶活性(-0.784)。(3)腐殖质构成与其他性质的相关性均不显著(P0.05),其中相对有利于提高胡敏酸/腐殖酸含量之比的土壤性质为蛋白酶、蔗糖酶和过氧化氢酶活性,而不利的是脱氢酶活性;相对有利于提高胡敏酸/富啡酸含量之比的为速效K、CEC和脲酶活性,而不利的是脱氢酶活性。(4)总体而言土壤腐殖质含量较之腐殖质构成与其他性质之间具有更大的相关性;向土壤增施N肥可以促进腐殖质的形成,增加K肥则有利于腐殖质构成的改善,而通过混交改造或增加林下植被是促进纯林土壤腐殖质化过程和解决土壤退化的根本措施。     

Satiety hormone and metabolomic response to an intermittent high energy diet differs in rats consuming long-term diets high in protein or prebiotic fiber

Large differences in the composition of diet between early development and adulthood can have detrimental effects on obesity risk. We examined the effects of an intermittent high fat/sucrose diet (HFS) on satiety hormone and serum metabolite response in disparate diets. Wistar rat pups were fed control (C), high prebiotic fiber (HF) or high protein (HP) diets (weaning to 16 weeks), HFS diet challenged (6 weeks), and finally reverted to their respective C, HF, or HP diet (4 weeks). At conclusion, measurement of body composition and satiety hormones was accompanied by (1)H NMR metabolic profiles in fasted and postprandial states. Metabolomic profiling predicted dietary source with >90% accuracy. The HF group was characterized by lowest body weight and body fat (P < 0.05) and increased satiety hormone levels (glucagon-like peptide 1 and peptide-YY). Regularized modeling confirmed that the HF diet is associated with higher gut hormone secretion that could reflect the known effects of prebiotics on gut microbiota and their fementative end products, the short chain fatty acids. Rats reared on a HF diet appear to experience fewer adverse effects from an intermittent high fat diet in adulthood when rematched to their postnatal diet. Metabolite profiles associated with the diets provide a distinct biochemical signature of their effects.     

花楸腋芽增殖途径快繁技术研究

选用花楸茎节为外植体进行组培腋芽增殖途径研究,结果表明：利于花楸生长的最适宜基本培养基为MS培养基;芽诱导培养基为1/2MS+6-BA 2.5 mg·L^-1+2.4-D 0.5 mg·L^-1;增殖培养基为MS+6-BA（1.600~2.100）mg·L^-1+NAA（0.140~0.230） mg·L^-1,平均繁殖系数达到13倍以上;继代壮苗培养的最佳培养基为MS+6-BA 0.2 mg·L^-1+IBA 0.2 mg·L^-1;在1/2MS+IBA 0.3 mg·L^-1培养基上,平均生根数为5.96,生根率达90.20%。将生根后的组培苗移栽至腐殖土：泥炭土：河沙（比例为3：2：1）的基质中,成活率达95.55%。     

Polyphenolic content and physiological activities of Chinese hawthorn extracts

Hawthorn polyphenol (HP) was prepared by ethyl acetate treatment of the ethanol extract (HE) of Chinese hawthorn fruit. The concentrations of 15 polyphenols in the HP, HE, extraction residue (HJ), and a hawthorn leaf extract (HF) were determined by HPLC. For HP, the total content of the 15 polyphenols was 21.4%, comprised of 19.7% of procyanidins, 1.21% of chlorogenic acid, and 0.48% of flavonoids, compared to 2.55% for the HE. The yields of procyanidin monomer, dimer, trimer, tetramer, and pentamer were 50.5%, 30.3%, 23.0%, 14.6%, and 12.5% respectively, and the mean degree of polymerization was reduced to 1.39 (HP) from 1.65 (HE). Seven different physiological actions of the four extracts were investigated. The HP showed strong O(2)(-) and (.-)OH scavenging capacities (IC(50) values of 6.3 microg/ml and 1.1 microg/ml respectively), as well as selective prolyl endopeptidase inhibition (IC(50) of 60 microg/ml). The active constituents appeared to be procyanidins.