抗羊肚菌菌丝特异性抗原单克隆抗体的制备与初步鉴定

为利用免疫学技术研究羊肚菌[Morchella esculenta(L.)Pers.]菌丝生长发育规律,以菌株代号为51589的羊肚菌菌丝破碎液为抗原免疫Balb/c鼠,将致敏Balb/c鼠的B细胞与骨髓瘤细胞SP2/0进行细胞融合,经镜检和ELISA间接法检测,筛选出一分泌抗羊肚菌菌丝抗原单克隆抗体的杂交瘤细胞株,命名为W2F3。经传代培养和亚克隆纯化,得到稳定分泌抗羊肚菌菌丝抗原单克隆抗体的细胞株,经鉴定W2F3单克隆抗体抗体亚类为Ig G1,轻链亚型为Kappa,初步确定该单抗对应的抗原具有羊肚菌属特异性。     

不同氮源对羊肚菌菌丝生长的影响

为了获得野生羊肚菌(Morchella spp.)资源,并探讨其菌丝生长的最适氮源,本试验以羊肚菌属QYY、XF、 FS菌株为供试菌株,以菌丝长势、密度、均匀度、菌丝生长速率和菌丝生长指数为指标,考察6种氮源对3个羊肚菌菌株菌丝生长的影响。结果表明,除尿素外,羊肚菌3个菌株均能够很好地利用其他5种氮源,不同产地的羊肚菌菌株对氮源的利用各不相同,羊肚菌菌株QYY、 XF、 FS的最适氮源分别为硝酸钾、硝酸铵、硝酸钾,菌丝生长速率分别为4.56 mm·d~(-1)、 9.92 mm·d~(-1)、 6.63 mm·d~(-1),菌丝生长指数分别为22.83、 49.58、 26.50。     

五株羊肚菌碳、氮源及适合发酵菌株的筛选

以菌丝体生物量和胞外多糖量为指标,分别采用不同碳源和氮源对5株羊肚菌[Morchella angusticeps、M.conica、Morchella sp.(50647)、Morchella sp.(50648)和M.esculenta]进行液体发酵,筛选出这5株羊肚菌的最佳碳源、氮源以及最适合液体发酵的菌株.实验结果表明:5株羊肚菌最佳碳源均为可溶性淀粉,最佳氮源分别为:硫酸铵、蛋白胨、酵母膏、硝酸钠、硝酸钠.最适合液体发酵的菌株为Morchella angusticeps,其生长到第7天时菌丝体生物量最高,可达5.76 g/L;第12天左右时胞外多糖产量最高,可达2.17 g/L.     

羊肚菌的培养研究

自古以来,羊肚菌(Morchella spp.)在许多国家中都是人们所喜爱的食用菌。人们很早就想栽培羊肚菌供食用,但是至今还没有研究出商业栽培的方法。我在查谟和克什米尔调查期间,找到了五种羊肚菌。然而,有一种倾向,认为本属没有那么多种。现已知道,羊肚菌对营养条件的反应、培养特征和组织结构,对于羊肚菌属中种的划分是很有参考价值的。现将八株羊肚菌的培养特征报导如下: 材料和方法 (一)研究用的菌株:尖顶羊肚菌(Morchellaconica),小羊肚菌(M.deliciosa),杂种羊肚菌(M.hybrida),羊肚菌(M.esculenta),小顶羊肚菌(黑     

羊肚菌衰老特性的测定分析

应用野生高羊肚菌(Morchella elata)及3种可驯化栽培的梯棱羊肚菌(M. importuna)、六妹羊肚菌(M.sextelata)和七妹羊肚菌(M. eximia)的15个样品共计52个分离株,包括相同子囊果不同部位和相同部位的多个组织分离株,以及单孢分离株,采用继代培养的方法测定这些菌株的生长曲线和形态特征,分析这些菌株的菌丝生长速度、菌核和色素产生及菌落形态等培养性状。结果为:除了两个组织分离菌株在继代培养5 320 h仍然存活以外,其余供试菌株均在不同时间段内老化死亡,其中寿命最长者为连续继代培养3 814 h,继代培养总生长距离213.7 cm;寿命最短者仅继代培养360 h,总生长距离5.9 cm。羊肚菌的老化表型为菌核产生数量下降、产生色素增加、菌丝生长速度下降和菌落边缘不整齐。表明羊肚菌菌丝的快速老化是一个普遍现象,可能与低等生物遗传物质的快速变异有关。鉴于菌株老化严重影响栽培产量,建议将菌株寿命测定作为羊肚菌优质菌种评判的一个指标。并提出在生产中减缓羊肚菌老化的一些措施。     

梯棱羊肚菌M83和M101菌株培养特性差异

以可正常出菇的梯棱羊肚菌M83和不能正常出菇的梯棱羊肚菌M101作为供试菌株,对2种菌株在不同的温度、p H、光照、碳源、氮源上的菌丝生长速度、长势及菌核生长情况等进行分析和比较。结果表明,2种菌株菌丝的最适生长温度为15℃~25℃,但M83菌丝在5℃~30℃能正常生长,而M101菌丝在30℃不能正常生长;2种供试菌株菌丝生长最适p H为7~8,M83的生长速度明显快于M101;2种菌株菌丝在黑暗条件下的生长速度、长势均优于光暗交替条件;2种菌株可利用多种碳源和氮源,但M83对麦芽糖和可溶性淀粉的利用能力显著强于M101;2种菌株菌丝生长最适氮源为蛋白胨,对硝态氮的利用能力均强于对铵态氮的利用。     

不同培养基上羊肚菌菌株产无性孢子观察

利用光学显微镜观察并研究5株羊肚菌菌株在3种培养基上产生无性孢子的情况.结果 ,粗腿羊肚菌(Morchella crassipes)在3种不同培养基上均未产生无性孢子,尖顶羊肚菌(M.esculenta)和梯棱羊肚菌(M.importuna)在腐殖土培养基上产生无性孢子;表明无性孢子产生与否并非是羊肚菌属真菌的共有特征...     

不同碳源、氮源对羊肚菌菌丝生长及菌核形成的影响

以羊肚菌六妹菌株6611和梯棱菌株M2018为供试菌株,采用菌丝培养的方法,研究了11种碳源、7种氮源对2种菌株菌丝生长、菌核形成的影响,以期为羊肚菌的菌种制备、液体培养条件优化以及相关功能性保健产品的开发利用提供参考依据.结果 表明:2种菌株的菌丝在供试的碳源、氮源中均能生长.其中菌株6611,在麦芽糖、葡萄糖、甘露醇、山梨醇、D-半乳糖、硝酸钠、酒石酸铵培养基中菌丝长势较好;在山梨醇、乳糖、D-半乳糖、淀粉、硝酸钠、酒石酸铵培养基中菌核形成较多且以近边缘分布的黄色、褐色小颗粒的形态居多.菌株M2018,在麦芽糖、淀粉、葡萄糖、乳糖、硝酸钠、磷酸氢二铵、酒石酸铵培养基中菌丝长势较好;在山梨醇、D-核糖、D-半乳糖、硝酸钠、磷酸氢二铵、酒石酸铵培养基中菌核形成较多且以分散分布、近边缘分布的黄色、褐色球状或块状的形态居多,在尿素培养基中无菌核形成.     

羊肚菌液体菌种培养配方的研究

近年来,我国已开展了羊肚菌培养的研究,但通常是采用固体培养法,对其菌丝球液体培养的研究尚不多见。为此,我们进行了羊肚菌液体菌种培养配方的研究,以期为羊肚菌菌种的生产提供依据。 一、材料和方法 (一)供试菌株 羊肚菌5.69(Morchellaesculenta),和羊肚菌5.70(Morchella conica),引自广东省微生物研究所。     

羊肚菌不同菌株生物学特性及优良品种选育

通过对8个羊肚菌不同菌株间的拮抗性、菌丝体生长特性及菌核形成试验的分析比较,结果表明羊肚菌不同菌株间的差异属于种类或地理分布差异,其导致了生物学特性和外形上的差别。美味羊肚菌菌株M918与尖顶羊肚菌菌株M928亲缘关系及生物学特性相近,M316、M318、M320、M326各有差异。菌株M918、M928的菌丝生长速度及浓密程度明显优于其它菌株,生长旺盛,抗杂能力强,日平均生长速度可达1cm·d-1,是一种菌丝生长较快、菌核形成较多的羊肚菌优良品种,适宜羊肚菌人工栽培和营养菌丝发酵使用的菌种。     

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.     

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.     

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.