生长调节剂对黄绿蜜环菌菌丝生长的影响

选用三十烷醇(triacontanol)和肌醇(inositol)两种生长调节剂对黄绿蜜环菌(Armillaria luteo-virens)菌丝生长的影响进行研究。结果表明:三十烷醇和肌醇对菌丝生长都有较明显的影响,但肌醇效果不如三十烷醇。在培养基中添加1.5ⅹ10-7mg.mL-1三十烷醇,可明显提高菌丝生长速度,缩短培养周期。     

Changes in fetal mannose and other carbohydrates induced by a maternal insulin infusion in pregnant sheep

Background

The importance of non-glucose carbohydrates, especially mannose and inositol, for normal development is increasingly recognized. Whether pregnancies complicated by abnormal glucose transfer to the fetus also affect the regulation of non-glucose carbohydrates is unknown. In pregnant sheep, maternal insulin infusions were used to reduce glucose supply to the fetus for both short (2-wk) and long (8-wk) durations to test the hypothesis that a maternal insulin infusion would suppress fetal mannose and inositol concentrations. We also used direct fetal insulin infusions (1-wk hyperinsulinemic-isoglycemic clamp) to determine the relative importance of fetal glucose and insulin for regulating non-glucose carbohydrates.

Results

A maternal insulin infusion resulted in lower maternal (50%, P < 0.01) and fetal (35-45%, P < 0.01) mannose concentrations, which were highly correlated (r² = 0.69, P < 0.01). A fetal insulin infusion resulted in a 50% reduction of fetal mannose (P < 0.05). Neither maternal nor fetal plasma inositol changed with exogenous insulin infusions. Additionally, maternal insulin infusion resulted in lower fetal sorbitol and fructose (P < 0.01).

Conclusions

Chronically decreased glucose supply to the fetus as well as fetal hyperinsulinemia both reduce fetal non-glucose carbohydrates. Given the role of these carbohydrates in protein glycosylation and lipid production, more research on their metabolism in pregnancies complicated by abnormal glucose metabolism is clearly warranted.     

Role of IP3R in LH-EGFR-induced mouse oocyte meiotic resumption

AIM: To explore the effect of inositol 1, 4, 5-trisposphate receptor (IP₃R) in luteinizing hormone-epidermal growth factor receptor (LH-EGFR)-induced oocyte meiotic resumption. METHODS: Models of mouse cumulus-oocyte complexs (COCs) culture and follicle culture in vitro were generated to study the effects of 2-aminoethyl diphenyl borate (2-APB) and heparin (IP₃R specific inhibitors) on LH/EGF-induced oocyte meiotic resumption and EGF-induced cumulus cell expansion. Real-time PCR was used to detect the mRNA expression of cumulus expansion-related factors. The changes of the intracellular calcium level were monitored using Fluo 3-AM, and the cGMP level was measured by ELISA. RESULTS: The inhibitors of IP₃R, 2-APB and heparin, dramatically reversed EGF-induced oocyte maturation (P<0.05) and decreased cGMP levels in COCs (P<0.05). In addition, 2-APB and heparin reversed EGF-induced cumulus expansion, and significantly inhibited EGF-induced cumulus expansion-related factor expression (P<0.05). The activation of IP₃R increased intracellular calcium level, and the study found that 2-APB and heparin dramatically reversed EGF-induced elevation of calcium level in cumulus cells (P<0.05). Follicular culture in vitro showed that 2-APB and heparin significantly reversed the LH-induced oocyte maturation (P<0.05). CONCLUSION: LH-EGFR signaling pathway increases calcium level in cumulus cells through IP₃R, resulting in meiotic resumption.     

金花茶松散型愈伤组织诱导与悬浮细胞系建立

采用组织培养及显微观察技术,以金花茶植株茎段、叶片、花药及离体培养的金花茶体细胞胚为试验材料,对松散型愈伤组织的获得及悬浮细胞系的建立进行了研究。结果表明:在黑暗条件下,以金花茶体细胞胚切块为外植体,在MS+KT 0.5 mg/L+NAA 8.0 mg/L+硝酸银5mg/L+蔗糖30 g/L培养基上诱导出愈伤组织后,将其转接到该诱导培养基上连续培养3个月,最后从培养物中挑选状态良好的松散型愈伤组织在分别含有肌醇与硝酸银的2种培养基上交替培养,可以获得金花茶松散型愈伤组织;方差分析表明:蔗糖添加量、肌醇添加量及硝酸银添加量均对金花茶悬浮培养液细胞密度有显著影响且后两者分别与光照条件存在相互作用;将松散型愈伤组织在分别添加100 mg/L肌醇与2.5 mg/L硝酸银的液体增殖培养基上交替培养,可以建立并保持金花茶悬浮细胞系。该研究结果可为金花茶大规模细胞培养提供科学依据。     

Analysis of total inositol phosphates in municipal solid waste compost-treated soils by two extraction methods

 Humic substances are the most dynamic component of agricultural soils. In this study, humic substances were extracted from soils based on standard acid/base solubility and by gel filtration using Sephadex G25. Organic P is a component of humic substances, and inositol phosphates are considered to have high prominence in organic P. The objectives of the study were to determine the effects of municipal solid waste (MSW) compost on the amount of inositol phosphates (IP) present in soils, and compare methods of extracting humic substances from soils. Total IP (IP_T) in soil extracts was determined following separation using a AG-1× 8 anion exchange resin, acid digestion and ICAP analysis for P. The percentage of IP (%IP) of the total P in the soil was also determined by colorimetric analysis. The soils were part of an experiment to study the effects of three rates of MSW compost and fertilizers on three crops grown in a Pugwash sandy loam (Humo-Ferric Podzol). The data were compared using ANOVA atP≤0.05; extraction methods, fractions [humic/fulvic, high/low molecular (MW)], and five treatments (three different rates of compost, fertilizer, and untreated soil) were compared. The gel filtration method extracted significantly more IP_T and %IP than the solubility method. In the solubility method, humic and fulvic acids contained the same amounts of IP_T and %IP, while the high MW fraction always contained more IP_T and %IP than the low MW fraction in the gel filtration method. Fulvic acids and the low MW fraction contained similar amounts of IP_T and %IP. There were differences in %IP between compost-treated soils and the non-compost-treated soils, although there were no differences in IP_T due to rate of compost addition. Received: 20 October 1999     

长角豆中肌醇的提取工艺研究

[目的]探讨长角豆中肌醇的最佳提取工艺.[方法]研究不同溶剂、不同提取温度和不同提取时间对长角豆中肌醇提取率的影响,确定粗提工艺后对粗提物采用正交试验法确定最佳精制条件,并用GC法对其中的肌醇进行定量分析.[结果]长角豆中肌醇的粗提工艺为：40％乙醇做溶剂、提取温度为80℃、提取3 h;最佳酶解条件为：pH4.5、植酸酶浓度4.0％、酶解温度40℃,精制后肌醇含量达10.76％.[结论]该研究可为长角豆中肌醇的提取提供参考.     

Inhibitory effect of ATP on proliferative signaling in immortalized human fibroblasts

AIM: To investigate the inhibitory effects of ATP on proliferation signaling in immortalized human fibroblasts. METHODS: Immortalized human fibroblasts were treated with ATP, ATP conbined with calcium or potassium channel antagonists, respectively. The intracelluar-free calcium ([Ca²⁺]i), inositol 1,4,5-trisphosphate(IP₃) levels and cell viability were detected at different time points. RESULTS: ATP significantly increased the [Ca²⁺]i and decreased the IP₃ level in immortalized human fibroblasts, especially at initial stage (P＜0.01) . Compared to ATP alone, the proliferation rates remarkably increased when calcium or potassium channel antagonists were used (P＜0.01, respectively) with ATP. CONCLUSION: The calcium and potassium channels and IP₃ involved in the inhibitory effects of ATP on the proliferative signaling in immortalized human fibroblasts.     

微生物法测定维生素功能饮料中的肌醇含量

[目的]基于葡萄汁酵母菌(ATCC 9080)对肌醇的特异性和灵敏性,定量测定出维生素功能饮料中肌醇的含量。[方法]在含有除肌醇以外所有营养成分的合成培养基中,葡萄汁酵母菌的生长量随着肌醇浓度的增大而增大,且在一定浓度范围内呈线性关系,根据培养后菌悬液的吸光度值与肌醇标准工作曲线进行比较,计算出维生素饮料中肌醇的含量。[结果]微生物法测定肌醇的线性范围为1~10μg,测定3种不同品牌维生素功能饮料中肌醇的方法精密度分别为3.85%、2.46%及2.19%,相对标准偏差(RSD)为1.424%,回收率在95%以上。[结论]微生物法具有快速、简便、检出限低、灵敏度高等优点,是测定维生素饮料中肌醇的有效方法。     

水稻1,3,4-三磷酸肌醇5/6-激酶基因的电子克隆及表达

利用电子克隆的基因克隆策略,以拟南芥1,3,4-三磷酸肌醇5/6-激酶AtITL1的cDNA序列为信息探针,在水稻基因组数据库搜索到高度同源的水稻基因组序列,通过人工序列拼接及RT-PCR方法克隆得到了水稻编码1,3,4-三磷酸肌醇5/6-激酶类似物的基因OsITL1的全长cDNA序列。RNA凝胶印迹分析表明,OsITL1基因的表达在转录水平上受NaCl、干旱及ABA的诱导,OsITL1可能参与了一些非生物胁迫的应答过程,在逆境条件下的基因转录和磷酸肌醇信号转导途径的调控中起重要作用。     

The use of gas‐liquid chromatography to determine the proportions of inositol isomers present as pentakis‐ and hexakisphosphates in alkaline extracts of soils

Abstract

A method for determining the relative proportions of inositol isomers present in soil as their pentakis‐ and hexakisphosphate derivatives (IP₅ + IP₆) is presented. The method involves determination of the isomers as their hexakis‐O‐acetyl derivatives by gas‐liquid chromatography. The only isomers detected in extracts of 4 soils were chiro‐inositol, neo‐inositol, myo‐inositol and soyllo‐to‐inositol. Little variation in the relative proportions of the isomers was found between the 4 soils but the relative proportion of the neo‐isomer was considerably greater than previously published estimates. The method also makes possible the estimation of the (IP₅ + IP₆) fraction, using anion‐exchange chromatography on a aliquot of the extract.