Batch solvent extraction of flavanolignans from milk thistle (Silybum marianum L. Gaertner)

Seeds of milk thistle (Silybum marianum L. Gaertner) contain silymarins and ca. 25% (w/w) of oil. A pre-treatment step involving refluxing with petroleum ether is usually performed before extraction of the silymarins using organic solvents. This paper compares the extraction of whole and defatted milk thistle seeds in various solvents as a function of temperature. The extraction of whole seeds of milk thistle with water at 50, 70 and 85 degrees C was also examined: the yield of silymarin increased with increasing water temperature. In most cases, ethanol at 60 degrees C recovered the largest quantities of silymarins. However, boiling water proved to be an efficient extraction solvent for the more polar silymarins such as taxifolin and silychristin, even when using whole seeds. Extractions of defatted seed meal with boiling ethanol returned maximum yields of 0.62, 3.89, 4.04, and 6.86 mg/g defatted seed of taxifolin, silychristin, silybinin A and silybinin B, respectively. When extracting defatted seed meal with ethanol, yields of taxifolin, silybinin A and silybinin B were, respectively, 6.8-, 0.95-, 1.7- and 1.6-fold higher than when extracting whole seeds. When extracting with boiling water, the yields of silychristin, silybinin A, and silybinin B were 380, 47 and 50% higher for whole seeds compared with defatted seeds.     

Effect of different carbon sources on cellulase production by Hypocrea jecorina (Trichoderma reesei) strains

The ascomycete Hypocrea jecorina, an industrial (hemi)cellulase producer, can efficiently degrade plant polysaccharides. At present, the biology underlying cellulase hyperproduction of T. reesei, and the conditions for the enzyme induction, are not completely understood. In the current study, three different strains of T. reesei, including QM6a (wild-type), and mutants QM9414 and RUT-C30, were grown on 7 soluble and 7 insoluble carbon sources, with the later group including 4 pure polysaccharides and 3 lignocelluloses. Time course experiments showed that maximum cellulase activity of QM6a and QM9414 strains, for the majority of tested carbon sources, occurred at 120 hrs, while RUT-C30 had the greatest cellulase activity around 72 hrs. Maximum cellulase production was observed to be 0.035, 0.42 and 0.33 µmol glucose equivalents using microcrystalline celluloses for QM6a, QM9414, and RUTC-30, respectively. Increased cellulase production was positively correlated in QM9414 and negatively correlated in RUT-C30 with ability to grow on microcrystalline cellulose.     

Comparative sequences of two type 1 dengue virus strains possessing different growth characteristics in vitro

The complete genome sequences of two dengue-1 virus strains having different growth characteristics (Mochizuki and A88) were compared with other published strains. The sequence analysis indicated several unique amino acid changes throughout the coding region of Mochizuki strain, mostly in envelope (E) protein. A unique amino acid, Ile-69 for Mochizuki strain at E protein resulted in the loss of an Asn-67-linked glycosylation site. A Thr substitution for Ala-114 at C protein and amino acid changes found in E, non-structural NS3, NS4a, and NS5 proteins were unique for A88 strain. These substitutions might be correlated to their different growth characteristics in vitro.     

接种木霉菌对黄瓜幼苗生长和根际土壤AM真菌侵染的影响

在盆栽试验中分别接种3株长枝木霉菌株Trichoderma longibrachiatum MF-1、MF-2和MF-3,以不接种木霉菌处理作为对照,研究木霉菌接种对土著AM真菌和黄瓜幼苗生长的影响。结果表明,菌株MF-1和MF-2显著提高了AM真菌侵染率和根外菌丝密度,与对照相比,AM真菌侵染率分别提高了26.85%和54.66%,根外菌丝密度分别是对照的3.94和3.76倍。接种菌株MF-2使植株地上部生物量显著提高了39.07%。菌株MF-3显著提高土壤pH和土壤有效磷含量。Pearson相关分析发现,添加木霉菌后,AM真菌侵染率与根外菌丝密度和孢子密度均呈显著正相关关系,土壤pH与植株地上部生物量和土壤有效磷含量均呈显著正相关关系。研究表明,3株长枝木霉与土著AM真菌的联合作用效果有明显差异,菌株MF-1和MF-2显著促进AM真菌生长,菌株MF-2更有利于黄瓜幼苗生长,而菌株MF-3主要改善土壤pH和有效磷含量。将几种木霉菌复合应用,有助于达到联合促生和改善土壤环境的综合效果。     

哈茨木霉与绿色木霉对杉木种子萌发和幼苗生长的影响

以杉木种子为材料,研究不同浓度(0.003、0.03、0.3、3、30、300 mg·L^-1)哈茨木霉和绿色木霉溶液对杉木种子萌发和幼苗生长的影响.结果表明: 各浓度木霉溶液处理对杉木种子萌发和幼苗生长均有一定的促进作用,其促进效果随着处理浓度的增加均呈先升后降的趋势.与对照相比,0.03 mg·L^-1哈茨木霉和绿色木霉处理对提高杉木种子的发芽率、发芽势、发芽指数、活力指数、根长、苗高和鲜质量效果最佳,分别提高了57.6%、125.0%、51.0%、209.2%、114.3%、16.1%、24.6%和42.7%、76.7%、43.9%、185.4%、113.8%、8.6%、22.6%;0.3 mg·L^-1哈茨木霉和绿色木霉显著提高杉木幼苗超氧化物歧化酶和过氧化物酶活性,分别增加了157.6%、179.9%和127.5%、116.2%,而丙二醛含量降低了86.1%和72.4%. 0.03～0.3 mg·L^-1浓度的哈茨木霉和绿色木霉不仅能显著促进杉木种子的萌发和幼苗生长,而且能够提高其抗氧化酶活性,增强杉木幼苗的抗逆性.     

黄绿木霉菌代谢产物对大豆菌核病核盘菌的抑菌能力研究

利用室内实验的方法,研究了黄绿木霉对大豆菌核病菌的抑菌能力。黄绿木霉在与核盘菌（Sclerotinia sderotiorum）对峙培养过程中,5-7d即可将菌核病菌完全覆盖,在形成的菌核上会有黄绿木霉孢子出现,扫描电镜中观察到核盘菌的菌丝体发生变形;其代谢产物在经121℃处理25min后,抑菌作用仍可高达100％;在对黄绿木霉发酵过程中,利用PD培养基即可达到有效的控制大豆菌核病菌菌丝生长的目的,而且利用PD培养基发酵3-4d的黄绿木霉代谢产物抑菌效果最好,抑菌中浓度最低,发酵时间过长与过短,均不利于对病原菌的抑制。经黄绿木霉代谢产物处理的菌核菌电导率与呼吸强度均发生变化,在光学显微镜下观察到菌核菌菌体发生变化,且处理后的菌核菌致病力明显降低。     

Providing the plant extract silymarin to lactating sows: effects on litter performance and oxidative stress in sows

Silymarin is an extract from the plant milk thistle that was shown to have antioxidant and hyperprolactinemic properties. Taking into account the essential role of prolactin for lactating sows and the systemic oxidative stress occurring during lactation, it is of interest to investigate the potential beneficial effects of silymarin on lactating sows. A study was therefore carried out to determine the effects of providing either 1 or 8 g/day of the plant extract silymarin to lactating sows. Sows in first, second or third parity were fed conventional diets during gestation and, at farrowing, were assigned as controls (CTL, n=33), or were fed 1 g/day (SYL1, n=33) or 8 g/day (SYL8, n=33) of silymarin. The silymarin was provided in two equal amounts per day, and was fed throughout a 20-day lactation. The performance of sows and their litters was assessed and circulating concentrations of prolactin (days 7 and 18), urea (days 7 and 18) and oxidative status, via protein carbonyls and superoxide dismutase activity (day 18), were measured in sows. Milk samples were obtained on day 18 to measure standard composition. There was no effect of silymarin (P>0.10) on circulating prolactin or urea, or on oxidative damage to proteins or antioxidant potential in sows. Lactation feed intake, backfat and BW of sows were unaffected by treatment (P>0.10) as was the case for milk composition and piglet growth (P>0.10). Results demonstrate that providing up to 8 g/day of the plant extract silymarin to lactating sows had no beneficial effects in terms of circulating prolactin concentrations or oxidative status of sows, or in terms of performances of sows and their litters.     

木霉对植物的促生作用及其机制

简要回顾了木霉促进植物生长作用及其机制的研究进展,探讨了木霉促生作用的存在问题及对策,并对木霉促生作用的应用前景进行了展望。     

绿色木霉原生质体诱变筛选纤维素酶高产菌株

以绿色木霉F264为出发菌株制备其原生质体,对其进行紫外线诱变处理,筛选出一株纤维素酶高产突变株绿色木霉F-UV264,其滤纸酶活由出发菌株的5.2IU/g干料提高到15.78IU/g干料,产酶能力提高了3倍。经过10代PDA斜面继代培养及发酵试验,表明该菌株是比原菌株更优秀的稳定高产株。     

不同生长调节物质对水稻生长及镉积累的影响

比较脱落酸(ABA)、乙烯利(ETH)、水杨酸(SA)和茉莉酸甲酯(MeJ A)4种植物生长调节物质(PGR)对水稻生长及籽粒镉(Cd)积累的影响差异。试验采用重金属污染土种植水稻,于分蘖期、灌浆期各进行1次PGR叶面喷施处理,分析灌浆期叶片光合指标,丙二醛(MDA)含量以及收获期各部位生物量和Cd含量。结果表明:(1)低浓度ABA(5mg/L)可维持水稻正常产量;高浓度ABA(15mg/L)则导致产量下降。ETH对水稻地上部生长和单株产量有显著抑制作用,SA和MeJ A(0.56mg/L)均可保证地上部正常生长,维持正常产量。(2)外施4种PGR均抑制灌浆期叶片气孔开放,降低蒸腾速率和光合速率,抑制效果最明显的是高浓度MeJ A(0.56mg/L)。(3)在供试浓度范围内SA、低浓度ABA(5mg/L)以及高浓度MeJ A均可降低灌浆期叶片MDA含量,减少质膜过氧化水平。(4)4种PGR均可降低水稻籽粒Cd含量,其中低浓度ABA(5mg/L)抑制籽粒Cd积累的效应最佳。相关性分析结果表明,PGR抑制籽粒积累Cd的效应与地上部向籽粒转运Cd的调控机制有关,与蒸腾速率无显著相关关系。(5)综上所述,低浓度ABA(5mg/L)处理对水稻产量无影响,且籽粒Cd含量降低程度最大。适当浓度的PGR可降低水稻籽粒Cd含量,在中低度重金属污染农田生态修复实践中具有一定的应用前景,但必须精确控制PGR的处理时间和处理浓度,避免出现抑制生长和降低产量的负效应。     

毒死蜱降解木霉菌对几种重要植物病原真菌的生防活性

木霉菌既是广泛应用的防治植物病害的生防菌,又是一类很有应用潜力的环境污染修复菌。针对分离筛选出的6株高效降解毒死蜱的木霉菌株,进行了土传植物真菌病害的生防活性试验。结果表明,在对峙培养条件下,供试木霉菌株对几种病原真菌均具有较为显著的抑制率,发酵滤液对多数病原真菌具有明显的抑菌作用。所有供试木霉菌株能在立枯丝核菌、灰霉、终极腐霉菌落上着生,并逐渐覆盖全部菌落;但不能在茄腐镰孢菌、尖孢镰孢菌、大丽轮枝菌上生长。真菌重寄生现象观察结果表明,供试木霉菌仅对立枯丝核菌具有明显的重寄生现象。研究结果表明,筛选出的高效降解毒死蜱的木霉菌菌株可对多种土传植物病原真菌具有良好的生防潜力。     

不同施氮量对乌龙茶生长和生理的影响

以武夷肉桂为研究对象,研究不同施氮量对乌龙茶幼龄茶树生长和生理的影响。结果表明,幼龄茶树对氮肥的需求不强烈,其新梢生物量、根生物量和总生物量以及茶叶产量随施氮量的增加而下降。茶树新梢全氮、叶绿素、游离氨基酸、茶多酚和咖啡碱的含量随施氮量的增加而增加,而茶树碳氮比随着施氮量增加而下降;但施氮并没有影响茶树总碳含量。老叶叶绿素含量、根全氮和硝态氮含量、新梢总糖含量与施氮量呈二次曲线回归关系,适度施氮促进根对氮的吸收、老叶叶绿素合成和新梢总糖代谢,过度施氮则相反。新梢生物量与其硝态氮含量和游离氨基酸总量显著负相关;根生物量与根碳氮比和新梢咖啡碱含量显著负相关;茎叶生物量和总生物量与根含氮量显著正相关,但与新梢硝态氮和氨基酸含量显著负相关。过度施氮造成茶树生产力下降的主要原因是因为过度施氮极显著提高了茶树氨基酸代谢水平,使用于茶树生长的碳代谢产物（如总糖）减少,进而影响茶树的生长。     

Modelling the growth of Trichoderma virens with limited sampling of digital images

AIMS: Using limited digital image sampling, a model of fungal growth in soil that considers both hyphal production and lysis was constructed for two strains of Trichoderma virens over a range of four temperatures. MATERIALS AND METHODS: A growth model was developed by fitting the radial cross sectional data with a modified form of the Ratkowsky equation to determine maximum growth rate and a modified Arrhenius equation to determine maximal rate of decrease in area covered by mycelia. The parameters obtained from a combined equation were then verified by using the data obtained from the whole colony to determine the appropriateness of the model. CONCLUSIONS: Using a limited data set and a combination of the Ratkowsky and Arrhenius equations, the mycelial coverage of the T. virens colony was determined, relating microscopic hyphal growth to macroscopic colony growth. This model was sufficiently robust to predict growth across four temperatures for a genetically modified and wild-type strain of T. virens. SIGNIFICANCE AND IMPACT OF STUDY: By using simple assumptions for the increase and eventual decline in fungal growth on a resource-limited medium, this model constructs an initial framework onto which additional parameters such as nutrient consumption could be incorporated for prediction of fungal growth.     

光照强度对广西地不容光合特性和生长的影响

以盆栽当年生广西地不容为材料,研究不同光照强度(100%、50%、30%和15%自然光强)对其光合特性和生长的影响。结果表明:随着光照强度的降低,广西地不容最大净光合速率(P_(max))、光饱和点(LSP)、光补偿点(LCP)先减小,而后稍有增大;表观量子效率(AQY)在30%和50%光强下显著高于100%和15%光强处理;叶片叶绿素总量(Chl)、叶绿素a(Chla)、叶绿素b(Chlb)、类胡萝卜素(Car)含量随光强的减弱而增大,Car/Chl随光强的减弱而减小,Chla/Chlb比值在各处理间无显著差异;单叶面积随生长光强的减弱而增大,比叶重(LMA)则随着生长光强的减弱而减小;30%光强下广西地不容块根生物量最高,光照过强和过弱都不利于其生物量的积累。广西地不容对光强的适应范围较广,但光照过强或过荫均对生长造成不良影响,光合速率降低,生长减缓,块根生物量积累下降,30%光强是其当年生苗生长的最佳光强。     

不同生长状态珊瑚光谱特征

珊瑚礁生态系统迅速退化是目前重要的生态环境问题之一,应用遥感技术监测大范围珊瑚礁的结构组成和变迁有很大的潜力。珊瑚光谱响应特征受珊瑚生态习性影响,在光学上相似而容易造成混淆误判。采集了西沙群岛大量石珊瑚样品的光谱,对其光谱特征进行分析及成因探讨。通过导数光谱、主成分分析研究了不同生长状态珊瑚的光谱差异,并建立珊瑚生长状态高光谱遥感判别准则。结果表明,珊瑚的光谱特性及其变化均较为复杂,受珊瑚种类和生长环境影响,光谱形状主要由共生藻色素吸收决定的。结合520—530 nm、564—574 nm和600—605 nm的导数光谱可以区分健康珊瑚、白化珊瑚和藻类覆盖的死珊瑚。总体判定准确度优于80%,误判的主要来源是种内珊瑚反射率差异。研究表明珊瑚礁环境高光谱遥感可以定量评估珊瑚状态的变化。     

Antagonistic assessment of Trichoderma spp. by producing volatile and non-volatile compounds against different fungal pathogens

Trichoderma spp. are well-known biological agents that have significant antagonistic activity against several plant pathogenic fungi. In the present study, Trichoderma spp. were tested in vitro for their antagonistic activity against different spp. of Fusarium and Alternaria viz. Alternaria alternata, A. brassicae, A. solani, Fusarium oxysporum and F. solani using dual plate assay and by the production of volatile and non-volatile compounds. The results obtained revealed that Trichoderma harzianum and T. viride effectively inhibited the growth and spore production of different spp. of Fusarium and Alternaria. The highest growth inhibition was found in A. alternata 62.50% and 60.00% by non-volatile compounds of T. harzianum and T. viride, respectively. Similarly, the volatile compounds inhibit the maximum growth of A. alternata 40.00% and 35.00% by T. harzianum and T. viride, respectively. Volatile and non-volatile compounds of Trichoderma spp. were analysed by GC-MS technique and the properties of distinguished compounds showed antifungal, antimicrobial and antibiotic activities. Volatile compounds of T. harzianum and T. viride showed highest percent abundance for glacial acetic acid (45.32%) and propyl-benzene (41.75%), respectively. In case of non-volatile compounds, T. harzianum and T. viride showed D-Glucose, 6-O-α-D-galactopyranosyl- (38.45%) and 17-Octadecynoic acid (36.23%), respectively. The results of present study confirmed that T. harzianum can be used as a promising biological control agent against Alternaria and Fusarium spp. that cause diseases in various vegetables and crops.     

不同诱导方法对农杆菌介导的木霉菌遗传转化效率的影响

[目的]比较不同诱导方法对根癌农杆菌EHA105介导的哈茨木霉T88遗传转化体系转化效率的影响,以确定最佳的诱导方法.[方法]通过对复铺培养基法、转膜培养法和液体共培养法转化效率的比较来确定3种方法的优劣.[结果]复铺培养基法转化效率约为10个转化子/107个木霉孢子,转膜培养法转化效率约为20个转化子/107个木霉孢子,液体共培养法转化效率达到100个转化子/107个木霉孢子.[结论]在农杆菌介导的木霉菌遗传转化体系中,液体共培养法的转化效率最高,是常用的复铺培养基法及转膜培养法转化效率的5-10倍.     

Factors affecting the production of Trichoderma harzianum secondary metabolites during the interaction with different plant pathogens

Aims: Strains of Trichoderma spp. produce numerous bioactive secondary metabolites. The in vitro production and antibiotic activities of the major compounds synthesized by Trichoderma harzianum strains T22 and T39 against Leptosphaeria maculans, Phytophthora cinnamomi and Botrytis cinerea were evaluated. Moreover, the eliciting effect of viable or nonviable biomasses of Rhizoctonia solani, Pythium ultimum or B. cinerea on the in vitro production of these metabolites was also investigated. Methods and Results: T22azaphilone, 1‐hydroxy‐3‐methyl‐anthraquinone, 1,8‐dihydroxy‐3‐methyl‐anthraquinone, T39butenolide, harzianolide, harzianopyridone were purified, characterized and used as standards. In antifungal assays, T22azaphilone and harzianopyridone inhibited the growth of the pathogens tested even at low doses (1–10 μg per plug), while high concentrations of T39butenolide and harzianolide were needed (>100 μg per plug) for inhibition. The in vitro accumulation of these metabolites was quantified by LC/MS. T22azaphilone production was not enhanced by the presence of the tested pathogens, despite its antibiotic activity. On the other hand, the anthraquinones, which showed no pathogen inhibition, were stimulated by the presence of P. ultimum. The production of T39butenolide was significantly enhanced by co‐cultivation with R. solani or B. cinerea. Similarly, viable and nonviable biomasses of R. solani or B. cinerea increased the accumulation of harzianopyridone. Finally, harzianolide was not detected in any of the interactions examined. Conclusions: The secondary metabolites analysed in this study showed different levels of antibiotic activity. Their production in vitro varied in relation to: (i) the specific compound; (ii) the phytopathogen used for the elicitation; (iii) the viability of the elicitor; and (iv) the balance between elicited biosynthesis and biotransformation rates. Significance and Impact of the Study: The use of cultures of phytopathogens to enhance yields of Trichoderma metabolites could improve the production and application of novel biopesticides and biofertilizers based on the active compounds instead of the living microbe. This could have a significant beneficial impact on the management of diseases in crop plants.