液体发酵蜜环菌培养条件的优化及菌体多糖的分离

以四因素三水平的正交实验设计培养基中碳源、氮源浓度，对蜜环菌进行液体发酵，通过对生长的菌丝体干重的测定得到该菌株生长最适培养基配方，并证明氮源为其生长的最重要因素，同时对该菌株的培养优化条件进行了研究，并提取蜜环菌菌体多糖，发现液体发酵菌体的多糖含量远高于人工栽培的蜜环菌菌素。     

蜜环菌的菌索

人工栽培天麻的成败,产量的高低与蜜环菌菌索生长情况密切相关,掌握蜜环菌菌索的生物学特性和侵染原理,以使在生产中造成利于菌索生长的环境条件。蜜环菌的菌丝形成很粗的索状物,菌丝之间分泌有由α1—4葡聚糖,β1—4葡聚糖组成的粘液,这些物质在菌索表面受氧化颜色变深,形成坚固外层一“皮层”。菌丝的“分生组织”代替菌丝正常地顶端生长。蜜环菌菌索的生长点在呈根冠状结构尖端后25     

蜜环菌对硒的耐受能力及硒对其多糖产率的影响

采用不同的硒浓度对蜜环菌进行了补硒培养,研究了硒对蜜环菌生长、蜜环菌胞外多糖和胞内多糖产率的影响。结果表明:在硒处理浓度为20 mg／L以下时,能够促进蜜环菌菌丝体的生长,超过该浓度之后,硒对蜜环菌菌丝体的生长产生阻碍;对于蜜环菌多糖来说,在试验所涉及的浓度范围内,硒能够促进胞外多糖的分泌,但对胞内多糖合成的影响不如胞外多糖明显。     

蜜环菌YN3862菌株的高密度发酵工艺

以蜜环菌YN3862菌株为试材,采用固体培养和液体深层发酵相结合的方法,研究蜜环菌的最优母种培养基和最佳发酵条件,以期达到高密度培养蜜环菌目的。结果表明:该菌株最适母种培养基成分为木屑100 g·L^-1,麦麸100 g·L^-1,葡萄糖20 g·L^-1,琼脂15 g·L^-1,牛肉膏5 g·L^-1;最佳液体培养基成分为葡萄糖1%,蔗糖2%,蚕蛹粉3%,MgSO₄·7H₂O 0.075%,KH₂PO₄ 0.15%,乙醇1%;液体深层发酵采用200 L发酵罐,装入100 L液体培养基,以5%接种量接入匀质后的液体菌种,发酵温度25℃,溶解氧含量(DO)为55%左右,在发酵的0～24、24～36、36～48、48～144 h,转速分别设置为60、80、120、150 r·min^-1,在发酵时间为120 h时,生物量(干质量)最高,达到2.5 g·(100mL)^-1     

伴栽天麻蜜环菌的适宜培养基筛选

<正>天麻生长离不开蜜环菌(Armillaria mellea),蜜环菌是天麻无性繁殖生长阶段的营养物质基础。蜜环菌属担子菌亚门、伞菌目、白蘑科、蜜环菌属。据现有报道,蜜环菌纯培养的母种培养基为PDA或改良PDA。实践表明,蜜环菌虽能在PDA或改良PDA基质上生长,但菌丝灰白、细弱,易于核化,菌索生长势差,一般需要20~     

蜜环菌的培养I.菌种培养

蜜环菌 (Armillaria mellea)是一种兼性寄生菌 ,以腐生为主 ,兼营寄生生活。其子实体是一种美味可口的食用菌。蜜环菌可在60 0多种树上生存 ,在自然界多生长在树根和枯木桩上。当它侵染在活树上时常引起根腐病。与此相反 ,有些植物被蜜环菌侵染后生长旺盛 ,例如天麻 (Gastrodia elata )及猪苓 (Grifolaumbellata)必须靠蜜环菌侵染提供营养才能生长繁殖。因此为了推广、发展天麻和猪苓生产提供优良的菌种及菌材。本文就蜜环菌的人工培养技术简要介绍如下 :1　菌种的分离蜜环菌菌种是在自然界中从长有蜜环菌索的树根中将其分离下来或用野生…     

液体培养蜜环菌对碳氮源的利用试验

天麻为兰科天麻属植物天麻的根茎.《本草纲目》谓天麻“治语多恍惚,善惊失忘”.我国古代医学家均认为天麻有广泛的治疗功效,是镇痉熄风,治疗痫搦惊悸的重要药物.天麻需与蜜环菌共生,没有蜜环菌共生,天麻早期就不能生长发育.近年来研究证明,蜜环菌的固体培养物和发酵液都具有与天麻相似的药理作用和临床功效.由于野生天麻较难得,人工栽培天麻周期较长,因此,寻求天麻的替代物——蜜环菌发酵物很有意义.本文就蜜环菌液体发酵对碳氮源的利用特性进行了报道,为蜜环菌深层发酵的进一步研究提供依据.     

蜜环菌的培养--Ⅱ菌枝、菌材、子实体的培养

5　菌枝的培养蜜环菌枝是人工栽培猪苓 (直接伴栽 )和天麻培养菌材最好的菌种。用树枝杆中较幼嫩的枝段培养而成。5 .1　菌种准备　培养菌枝所采用的菌种主要是人工培养的蜜环菌纯菌种。也可选用野生的蜜环菌菌种。若选用野生蜜环菌时 ,应挖取无杂菌污染 ,带有生长旺盛粗壮的蜜     

2株优良天麻共生蜜环菌生长条件筛选

筛选出2株昭通本地天麻共生蜜环菌(SNA03、SNA04)菌索生长的最佳条件。采用单因素试验和正交试验法进行筛选,以生物量为考察指标。2株菌菌索生长的最适温度为25℃,暗培养下菌索生长速度最快;单糖(葡萄糖)为2菌株生长的最佳碳源,有机氮中的酵母膏和蛋白胨为最佳氮源,最适无机元素为K2SO4(SNA03)和KH2PO4(SNA04),最适维生素为VB2;最佳营养条件组合为葡萄糖∶酵母膏∶K2SO4(SNA03)或KH2PO4(SNA04)∶VB2=15g·L-1∶3g·L-1∶2g·L-1∶0.005g·L-)1,氮源(酵母膏)是影响菌索生长的最主要因素。来自昭通的2株天麻共生蜜环菌对营养和环境条件的要求基本一致。最佳生长条件的确定,对昭通蜜环菌快速工业化制种及天麻的大规模生产将起到一定的推动作用。     

天麻袋栽技术

用塑料袋栽天麻能较好利用空间 ,节约费用和提高单位产量 ,同时蜜环菌生长处于密封环境 ,栽培前又经有效杀菌能减少病原菌和害虫的发生。现将该技术介绍如下 :1　栽培季节　常法栽种天麻一般宜选择当年 11～ 12月和来年 3～ 4月 ,在年底栽种可让天麻在沙土中度过 2个月的休眠期 ,以减少保藏的麻烦。袋栽天麻宜在 3～ 4月栽种 ,开春后蜜环菌在适温下快速生长 ,天麻可直接放菌索上 ,蜜环菌能很快在种麻上着生 ,提高种麻的接种率 ,促进天麻早期生长 ,此时栽种产量高。2　生产种的制备　主料碎木屑 84 % ,米糠 10 % ,石膏 1% ,土豆 3% ,蔗糖 1% …     

Effects of radiation from 188Re on smooth muscle cell proliferation

AIM: Although endovascular radiotherapy inhibits neointimal hyperplasia, the exact alterations induced by β-particles irradiation remain to be elucidated. The objective of this study was to investigate the ability and the cellular mechanism of local β^-particles emission from ¹⁸⁸Re to inhibit vascular smooth muscle cells (SMCs). METHODS: The SMCs in vitro were irradiated by ¹⁸⁸Re with single doses of 2.6 Gy-25.8 Gy. The effects of β-particles on SMCs, such as effective irradiate doses, the period of inhibition for SMCs proliferation, the changes of cell proliferation rate and DNA synthesis rate, cell cycle progression and related gene expression, were investigated by cell count, [³H]-TdR incorporation, cell cycle progression analysis, cell viability and immunocytochemistry, respectivecy. RESULTS: β-particles irradiation with dose of 5.2 Gy could inhibit significantly SMCs proliferation. At dose of 20.6 Gy DNA synthesis inhibitory rate was 92%, SMCs proliferation rate was only 3%. Renoval of ¹⁸⁸Re did not abolish the inhibitory effects of β-particles on SMCs proliferation. The expression of P53 was up regulation and PCNA was down regulation after irradiation. CONCLUSION: β-particles from ¹⁸⁸ Re was significantly effective and permanent in inhibiting SMCs proliferation, and inhibitory effect was in dose-dependet manner ED50was 5 Gy, the best dose to inhibit SMCs proliferation was 20 Gy. β-particles irradiation induced SMCs to occur G₀/G₁ arrest, damaged the ability of SMCs reproliferation and led to cell clonogenic death. P53 and PCNA had regulatiory effects on SMCs proliferation after β-particles irradiation.     

Effect of L-Arg on plasma content of endothelin and expression of c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy

AIM:To study the effect of L-Arg on plasma content of endothelin (ET) and the expression of proto-oncogene c-fos mRNA in the left ventricle of rats with renovascular hypertensive hypertrophy. METHODS: The level of c-fos mRNA were measured by in situ hybridization. The ET in plasma were measured by radioimmunoassay. RESULTS:After eight weeks of treatment with L-Arg, the expression of c-fos decreased markedly (P＜0.01). The ET content in plasma also decreased significantly by L-Arg(P＜0.01).CONCLUSION: Plasma ET content and the expression of c-fos in the left ventricle of rats with renovascular hypertensive hypertrophy could be decreased by L-Arg administration.     

Ergebnisse der Leistungsprüfungen der Süßkirschensorte ‚Stella‘ auf Gisela- und Weiroot-Unterlagen in Bulgarien

Zusammenfassung Die Leistungsprüfungen wurden im Zeitraum 1997 bis 2003 mit den Unterlagen Gisela 4 und 5, den Klonnummern 195/20 und 497/8 aus der Gisela-Serie sowie Weiroot 10, 13, 53, 72 und 158 durchgeführt. Dabei dienten Sämlinge von P1 (bulgarische Selektion aus Prunus mahaleb) als Kontrolle. Alle Unterlagen waren mit der Sorte Stella veredelt und im Dezember 1996 in der Versuchsanlage der Agraruniversität in Plovdiv, Bulgarien, im Abstand von 6 m×4,5 m gepflanzt worden. Dabei erfolgte ein Pflanzschnitt. Nach Abschluss der natürlichen Kronenentwicklung wurde jedes Jahr ein Winterschnitt vorgenommen. Der Boden wurde durch mechanische Bearbeitung offen gehalten und nach dem 4. Standjahr wurden die Baumstreifen mit Herbiziden behandelt. Die Wasserversorgung erfolgte durch eine dem natürlichen Gefälle folgende Überflutung, allerdings nicht immer zum optimalen Zeitpunkt, da keine eigene Wasserquelle zur Verfügung stand.Basierend auf den Ergebnissen bis zum Anfang des 7. Standjahres können die untersuchten Unterlagen in zwei Gruppen differenziert werden: starkwüchsig—Weiroot 10, P1 und Weiroot 13; mittelstarkwachsend bis schwachwüchsig—Gi 497/8, Gisela 4, Weiroot 53, Weiroot 158, Gi 195/20, Weiroot 72 und Gisela 5. Letztere zeichnete sich durch besondere Schwachwüchsigkeit aus. Die meisten Wurzelschosser bildeten Gisela 4, Weiroot 10 und Weiroot 13. Weiroot 53, Weiroot 72 und Weiroot 158 entwickelten deutlich weniger und P1, Gisela 5, Gi 195/20 sowie Gi 497/8 keine Wurzelschosser. Den frühesten Blühbeginn induzierte Gisela 4. Die anderen Unterlagen führten, in Abhängigkeit von den Temperaturbedingungen des jeweiligen Jahres, zu einer Verspätung der Blüte: P1 und Weiroot 10 um 1–2 Tage; Gi 497/8, Weiroot 13 und Weiroot 158 um 2–4 Tage; Weiroot 72 um 2–7 Tage; Gi 195/20 um 3–6 Tage; Weiroot 53 um 3–8 Tage und Gisela 5 um 3–10 Tage. Die Reifezeit der Früchte war bei den Bäumen auf Gisela 5 im Vergleich zu den anderen Varianten um 2–3 Tage verspätet. Gisela 5, Weiroot 72 und Gisela 4 induzierten bei der aufveredelten Sorte die höchsten Ertragsleistungen, P1 die geringsten. Bei den Bäumen auf Gisela 5 war die Fruchtgröße geringer als bei den anderen Unterlagen. Bäume auf Gisela 5 brauchen intensive Pflege. Nur wenn alle Produktionsfaktoren und kulturtechnischen Maßnahmen optimiert werden, kann das hohe Ertragspotenzial dieser Unterlage ausgeschöpft werden.     

多效唑对猕猴桃离体试管苗生长及内源激素的影响

多效唑（ＰＰ３３３）处理猕猴桃试管苗，降低了其生长强度；植株体内的ＧＡ３、ＩＡＡ和ＺＴ含量下降，ＡＢＡ的含量上升，乙烯释放率增加；并且能降低外源的ＧＡ３和ＩＡＡ促进生长的作用，而外源的ＧＡ３和ＩＡＡ又能不同程度地逆转多效唑的抑制作用，使植株恢复生长。     

Proteomic analysis between pathological scars and normal skin

AIM: To investigate and screen the sensitive proteins in the formation mechanism of pathological scars by comparing the results of differential proteomic analysis between pathological scars and normal skin.METHODS: Two-dimensional gel electrophoresis was used to detect the protein expression profiles in 8 keloid patients, 8 hypertrophic scar patients and 3 matched normal skin patients.The proteins that showed differential expression of over 4-fold change were cut and analyzed by MALDI-TOF/TOF mass spectrometry.RESULTS: A two-dimensional protein profiling comparison between pathological scars and normal skin was successfully established.On average, 2 978 spots in keloid, 2 975 spots in hypertrophic scar and 3 053 spots in normal skin were identified using gel analysis software.Compared with normal skin, there were totally 36 differentially-expressed proteins in keloid and hypertrophic scar identified from the spots of over 4-fold change, including 16 proteins in both keloid and hypertrophic scar (8 up-regulated and 8 down-regulated), 11 only in keloid (9 up-regulated and 2 down-regulated) and 9 only in hypertrophic scar (4 up-regulated and 5 down-regulated).CONCLUSION: Proteomic analysis can identify the proteins with variance of pathological scars versus normal skin, thus providing probable new clues to reveal the formation mechanism of pathological scars.     

Compositional and Functional Properties of Saskatoon Berry and Blueberry

Abstract

Saskatoon berry (Amelanchier alnifolia Nutt., Rosaceae) and blueberry (Vaccinium corymbosum L., Ericaceae) are substantially equivalent in all characteristics that are important to the consumer, including fruit color, shape, size, nutrition, texture, and uses. In addition, both fruits are native to North America and they have practically identical historical uses and known health benefits. Their composition, processing, nutritional value and metabolism, intended uses, and levels of undesirable substances are compared.     

Cryopreservation of shoot apices of hawthorn in vitro cultures originating from East Asia

The objective of this study was to establish a cryopreservation protocol for hawthorn shoot apices (Crataegus pinnatifida Bge.). Cryopreservation was carried out via encapsulation–dehydration, vitrification, and encapsulation–vitrification on shoot apices excised from in vitro cultures. We began by showing that cold-acclimation enhanced the regrowth of cryopreserved apices from 10.0 to 65.5% in encapsulation–dehydration. We then decided that the encapsulation–dehydration method was an optimal cryopreservation method for hawthorn shoot apices in terms of its high recovery after cryopreservation as well as its ease of use compared with vitrification and encapsulation–vitrification. In encapsulation–dehydration, the protocol leading to optimal regrowth was as follows: after cold-acclimation at 5 °C in the dark for 2 weeks, excised shoot tips were pretreated for 24 h at 25 °C on hormone-free Murashige and Skoog [Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15, 473–497] (MS) basal medium with 0.4 mol/L sucrose, then encapsulated and precultured in liquid MS medium with 0.8 mol/L sucrose for 16 h at 25 °C. Precultured beads were dehydrated for 6 h at 25 °C in the dessicator containing 50 g silica gel to a moisture content of 15.3% (fresh-weight basis) before cryostorage for 1 h. In addition, we examined the effect of adding glycerol to both the alginate beads and loading solution to enhance regrowth after cryopreservation in encapsulation–dehydration. In the present study, it was shown that adding 0.5 mol/L glycerol resulted in high regrowth percentages (82.5–90.0%) in four Crataegus species.     

Coupling past management practice and historic landscape change on John F. Kennedy Space Center,Florida

Historic landcover dynamics in a scrubby flatwoods (Tel-4) and scrub landscape (Happy Creek) on John F. Kennedy Space Center were measured using aerial images from 1943, 1951, 1958, 1969, 1979, and 1989. Landcover categories were mapped, digitized, geometrically registered, and overlaid in ARC/INFO. Both study sites have been influenced by various land use histories, including periods of range management, fire suppression, and fire management. Several analyses were performed to help understand the effects of past land management on the amount and spatial distribution of landcover within the study sites. A chi-squared analysis showed a significant difference between the frequency of landcover occurrence and management period. Markov chain models were used to project observed changes over a 100-year period; these showed current management practices being effective at Tel-4 (restoring historic landscape structure) and much less effective at Happy Creek. Documenting impacts of past management regimes on landcover has provided important insight into current landscape composition and will provide the basis for improving land management on Kennedy Space Center and elsewhere.     

Metallothionein inhibits homocysteine-induced proliferation of rat vascular smooth muscle cells

AIM:To investigate the effect of metallothionein(MT) on proliferation of rat vascular smooth muscle cells (VSMCs) stimulated by homocysteine and its mechanism. METHODS:VSMCs proliferation was measured by [³-H]-TdR incorporation, mitogen-activated protein kinase(MAPK)activity were determined by immunoprecipitation method, the intracellular contents of MT and malondialdehyde (MDA)were assayed by -hemoglobin saturation method and TBA reaction, respectively, and lactate dehydrogenase (LDH) leakage was measured by NADH oxidation. RESULTS:Hcy(10^-6-10^-4 mmol/L) stimulated [³-H]-TdR incorporation by the VSMCs in a concentration-dependent manner. Compared with control, [³-H]-TdR incorporation in VSMCs treated with 0.1 mmol/L Hcy was increased by 4.2 fold (P＜0.01). Meanwhile, Hcy enhanced MAPK activity, MDA formation and LDH release (P＜0.01)in a concentration-dependent manner. Treatment of VSMCs with MT alone did not change above parameters, compared with control. However, MT (10^-6-10^-4 mol/L)attenuated significantly Hcy-stimulated proliferation of VSMCs (P＜0.01)in a concentration-dependent manner. And MT inhibited obviously Hcy-induced activation of MAPK activity, MDA formation and LDH release. Preincubation of VSMCs with 0.5 mmol/L ZnCl₂ for 6 h induced an increase cellular MT content by 5.7-fold (P＜0.01). The MT-overexpressed VSMCs resisted Hcy-stimulating action on MAPK activity, MDA formation and LDH leakage (P＜0.01). CONCLUSION:These results show that MT has an inhibitory effect on Hcy-induced VSMCs proliferation, and that MT could inhibit Hcy-stimulated MAPK activity and lipid peroxidation.     

XBP-01 accelerated apoptosis induced by oxidized low density lipoprotein in vascular smooth muscle cells

AIM: Previous studies performed with XBP-01 in vitro indicated that XBP-01 could inhibit vascular smooth muscle cells from being transformed into foam cell and could eliminate the atherosclerotic plaque in C57BL/6J mouse. This experiment is to investigate its mechanism of eliminating plaques in vitro. METHODS: The cultured porcine artery smooth muscle cells incubated with XBP-01 of 0.1 mg/L for 24 h after preincubated with oxidized low density lipoprotein of 15 mg/L for 72 h in vitro. The samples were analyzed by fluorescence microscope, confocal microscope system and flow cytometry. RESULTS: Apoptosis was triggered by being incubated with oxidized low density lipoprotein and this process was accelerated additionally by being incubated with XBP-01. CONCLUSION: XBP-01 can be effective in eliminating atherosclerotic plaque by accelerating the process in which oxidized low density lipoprotein induced smooth muscle cell apoptosis.