卫星灵芝液体发酵富集硒锌的研究

研究不同浓度的硒、锌对卫星灵芝菌丝体生长的影响,初步探讨卫星灵芝菌丝体生物富集硒、锌的效应。采用平板培养法及液体发酵法研究锌、硒对卫星灵芝菌丝体生长的影响及富集效应。培养基中不同浓度的亚硒酸钠对菌丝体生长均具有不同程度的抑制作用,但灵芝菌丝体的富硒量随着硒浓度的增加而提高,当亚硒酸钠浓度为40 mg/L时,菌丝体中的生物量、富硒量及富硒转化率最高,分别为1.54%、2 131.55 mg/kg、32.91%;培养基中硫酸锌浓度低于150 mg/L的范围内对卫星灵芝菌丝体生长有明显的促进作用,硫酸锌浓度为60 mg/L时菌丝体中的锌含量和富锌转化率最高,分别为1 142.91 mg/kg、1.76%。培养基中同时添加40 mg/L的亚硒酸钠和60 mg/L硫酸锌,菌丝体生长量1.60%,富硒量301.85 mg/kg,富硒率4.84%;富锌量为540.41 mg/kg,富锌率为5.72%。     

不同培养条件下杏鲍菇对硒的耐受性与硒形态分析

为了明确不同培养条件下杏鲍菇对外源硒的耐受性,分别在固体培养基、液体发酵培养基和栽培基质中添加不同量的亚硒酸钠,并检测杏鲍菇子实体中富集硒的存在形态。结果表明：固体培养条件下,硒浓度低于160 mg/L时,对菌丝的生长没有显著影响;液体培养条件下,4 mg/L的硒元素即可对杏鲍菇菌丝体的生长产生显著抑制;栽培模式下,基质中补充10~600 mg/kg的硒元素,不会影响杏鲍菇菌丝体的生长,且子实体中的硒含量会随基质中硒浓度的增加而增加。子实体硒形态分析表明,富集硒以硒代蛋氨酸、甲基硒代半胱氨酸、硒代胱氨酸和亚硒酸盐［Se（IV）］四种形式存在,其中硒代蛋氨酸是硒与氨基酸的主要结合形式。     

蛹虫草对锌的耐性与富集特征

在培养基内添加不同量的锌,研究其对蛹虫草子实体的形成、子实体和菌丝体生物量、子实体多糖含量和葡萄糖含量的影响,以及蛹虫草子实体和菌丝体对锌的富集能力。结果表明锌对上述各项都有影响。液体培养条件下,锌浓度在453906mg/L范围内可以促进菌丝体生长,锌浓度超过4077mg/L时,菌丝生长受到抑制。培养基锌的浓度在4077mg/L以下时,蛹虫草菌丝体锌的富集量随着液体培养基锌浓度的提高而提高。固体培养条件下,锌含量在226453mg/kg范围内可以促进蛹虫草子实体生长,并且在此含量范围内,蛹虫草子实体中葡萄糖含量较高。培养基锌含量在680906mg/kg时,子实体多糖含量较高。培养基锌含量在2038mg/kg以下时,蛹虫草子实体中锌的富集量随着培养基锌含量的提高而提高,在培养基锌含量为2038mg/kg时,子实体中锌的含量达到28570mg/kg(干重)。     

张家界市部分农作物可食部位硒含量研究

采用国家标准( GB/T5009.93-2003),用荧光分光光度计对张家界市22种农作物40个样本可食部位中的硒含量进行了分析.结果表明,测定的22个种类40个样本均属于富硒及高硒作物,以茶叶的硒平均含量最高为2.0102 mg/kg,其次是豆类平均含量为1.8518 mg/kg,薯类含量最低为1.0240 mg/kg.所测定的各种作物可食部位富硒能力依次为:茶叶2.0102 mg/kg＞豆类(干)1.8518 mg/kg＞水稻、玉米1.5326 mg/kg＞蔬菜类1.3438 mg/kg＞水果类1.0783 mg/kg＞薯类1.0240 mg/kg.该研究确定了部分农作物硒含量的水平及富硒作物品种.     

猴头菌菌丝富硒特性及富硒深层发酵的研究

为获得高产的富硒猴头菌菌丝体,采用平板法,通过Na_2SeO_3浓度梯度实验,从H2、H99、NK 3种猴头菌中筛选出耐硒品种;以猴头菌菌丝干重为指标,采用单因素试验结合正交试验优化培养基的碳源、氮源、无机盐和维生素,并进一步通过均匀设计试验确定各营养元素的配比;以菌丝干重和有机硒含量为指标,确定深层发酵培养基中Na_2SeO_3的浓度。结果表明:3种猴头菌中H2最耐硒,在含6μg/mL的Na_2SeO_3平板培养基中培养的菌丝适合作为后续研究的菌种;优化的液体培养基成分为5.8%可溶性淀粉、5.8%黄豆粉、0.04%CaCl_2、0.024%VB_1+VB_2+VB_6,此时菌丝干重理论值可达2.37 g/100 mL;当液体培养基含12μg/mL的Na_2SeO_3时,菌丝生长不受抑制,有机硒含量最高,达136.8 mg/kg菌丝。该研究为富硒猴头菌产品的开发和利用奠定了基础。     

黄粉虫幼虫对硒的生物积累

在饲料中添加含硒化合物喂养黄粉虫Tenebrio molitor L.幼虫,测定幼虫硒含量、粪便硒含量和体重的变化,计算黄粉虫幼虫特定生长率及幼虫对硒生物积累系数,分析黄粉虫有效积累硒的条件。结果表明,饲料硒含量在15~20mg/kg时,幼虫硒含量明显提高,对硒的生物积累系数高于其它试验组水平,饲料硒含量过高,幼虫硒含量降低,正常生长受到抑制。黄粉虫幼虫特定生长率、取食量、排粪量、干物质含量随着饲料硒含量的增加而降低,死亡率、粪便硒含量随着饲料硒含量的增加而增大。饲料硒含量为15～20mg/kg时黄粉虫幼虫对硒的生物积累效果最好。     

灵芝菌丝体富硒条件优化及其硒多糖抗氧化活性研究

利用正交试验对灵芝菌丝体液体发酵富硒条件进行优化,研究培养基中不同碳源、氮源和亚硒酸钠浓度对菌丝体生物量、硒含量、富硒率以及胞外酶活性的影响,此外还对硒多糖的抗氧化活性进行了初步研究。结果表明:当培养基中碳源为20%马铃薯、氮源为2%麸皮、Na_2SeO_3浓度为5 mg/L时,培养基中菌丝生物量最高达到3.86 g/L,硒含量最高为0.576 mg/g,富硒率也达到最高为44.47%。Na_2SeO_3对菌丝生长的影响大于碳源和氮源;同样的Na_2SeO_3对胞外酶活性的影响最大,最优组的POD和PPO酶活性都较其他组高,且随着Na_2SeO_3浓度的升高,其酶活性逐渐降低。富硒灵芝菌丝体粗多糖都具有较强的体外抗氧化活性,通过相关性分析发现,其粗多糖的抗氧化活性与硒含量、富硒率具有显著的正相关。显示硒多糖在食品和药品等领域具有良好的开发利用前景。     

富硒螺旋藻培养技术研究

采用富硒技术对印项螺旋藻培养进行强化,对硒（IV）浓度和亚硫酸盐的影响,以及硒的生物富集及其对藻细胞分子官能团结构的影响等进行了较为详细的研究,并对相关的可能机理进行了讨论。研究发现,硒对印顶螺旋藻生长具有刺激或抑制的双重作用。在0.02mg/L-411.00mg/L浓度范围内,硒不仅可以加快印顶螺旋藻的生长,而且还可以提高其生物量;同时,钝顶螺旋藻对硒的事集随着硒浓度的增加而增加,较为缓慢的生长利于钝顶螺旋藻对硒的富集。研究还证实,NaSO3会减轻高浓度Na2SeO3对印顶螺旋藻生长的毒性,富硒培养不会对藻细胞分子官能团结构产生损害。实验得出钝顶螺旋藻富硒培养较佳的硒处理浓度在10mg／L－40mg／L。     

秦巴山区富硒蛹虫草有效成分及硒存在形态分析

为探究秦巴山区富硒蛹虫草有效成分及硒存在形态,以秦巴山区蛹虫草CM-1518为研究对象,研究不同质量分数亚硒酸钠(0～500 mg/kg)对蛹虫草生长发育的影响,并对其有效成分及硒存在形态进行分析。试验结果表明当亚硒酸钠质量分数为100 mg/kg时,蛹虫草鲜质量最高,为293.41 g/盒。当亚硒酸钠质量分数为200 mg/kg时,虫草素、虫草酸含量最高,分别为1.06 mg/g和2.10 mg/g,表明硒与虫草素和虫草酸可协同增效,但虫草多糖含量变化规律不明显,亚硒酸钠的添加不利于腺苷的合成积累。经计算,富硒蛹虫草中有机硒所占百分比均高达99.9%,低浓度的亚硒酸钠可促进可溶性蛋白和可溶性多糖中硒的合成,但高浓度的硒却降低其合成,且富硒蛹虫草中可溶性多糖中硒含量高于可溶性蛋白硒含量。试验表明适宜浓度的亚硒酸钠可促进蛹虫草生长发育及有效成分合成积累。     

生物富硒对灵芝营养成分的影响

在人工栽培灵芝中,通过外施硒盐以生物转化法获得富硒灵芝,以不同含硒量的富硒灵芝子实体为试样,系统研究了灵芝生物富硒后对其主要营养成分多糖、蛋白质、氨基酸和矿物质等的影响。结果表明：硒可以明显提高灵芝中多糖、蛋白质以及氨基酸的含量,但并不改变灵芝蛋白质的分布和蛋白质中氨基酸的组成配比,而对矿质元素则有不同的影响,如富硒灵芝子实体中人体必需微量营养元素硒有极明显地增加,而铜和钼等人体必需微量元素的含量有一定量的增加,但铁、钙、锶等矿质元素的含量则有所下降。     

Bioeffects of selenite on the growth of Spirulina platensis and its biotransformation

The bioeffects of selenium on the growth of Spirulina platensis and the selenium distribution were investigated. S. platensis was batch cultured in Zarrouk medium containing increasing concentrations of sodium selenite. The biotransformation characteristic of selenium was analysed by the determination of the detailed selenium distribution forms. At 35 degrees C, 315.2 microEm(-2) x s(-1), sodium selenite concentrations below 400 mg x l(-1) were found to stimulate algal growth, especially in the range of 0.5-40 mg x l(-1). However, above 500 mg x l(-1) sodium selenite was toxic to this alga with the toxicity being related to the sulfite level in the medium. S. platensis was found to resist higher selenite by reducing toxic Se(IV) to nonsoluble Se(0). Selenium was accumulated efficiently in S. platensis during cultivation with accumulated selenium increasing with selenite concentration in the medium. It was demonstrated that inorganic selenite could be transformed into organic forms through binding with protein, lipids and polysaccharides and other cell components. The organic selenium accounted for 85.1% of the total accumulated selenium and was comprised of 25.2% water-soluble protein-bound, 10.6% lipids-bound and 2.1% polysaccharides-bound selenium. Among the organic fractions lipid possessed the strongest ability to accumulate Se (6.47 mg x kg(-1)). The 14.9% inorganic selenium in S. platensis was composed of Se(IV) (13.7%) and Se(VI) (1.2%).     

硒对水稻的生态毒理效应及临界指标研究

通过土壤添加硒盆栽试验,研究了黄棕壤不同浓度Se对水稻生物量、叶绿素含量、抗氧化酶系统的影响.结果表明,低浓度Se(<8mg·kg^-1)对水稻根和地上生物量、叶绿素a含量、叶绿素a/b有促进作用,并整体提高了作物抗氧化酶系统,使MDA下降,SOD、CAT、POD、GSHPx活性相应提高.而高浓度Se(>16mg·kg^-1)对水稻根和地上生物量、叶绿素a含量、叶绿素a/b有明显的抑制作用,对抗氧化酶系统产生胁迫效应,使GSHPx明显上升,MDA上升,SOD、CAT、POD酶活性明显下降.各项生理指标与土壤Se处理浓度之间相关分析表明,土壤Se浓度与地上部干重相对百分率、根干重百分率、叶绿素a/b、MDA、SOD、CAT、POD之间有显著相关性.比较胁迫效应10%值的临界指标EC10值表明,作物地上部干重抑制率作为土壤临界值制定的依据更为可靠,EC10为21mg·kg^-1.     

农杆菌介导的碱性成纤维细胞生长因子（bFGF）转化金针菇的研究

构建了金针菇表达载体p139035S-bFGF,并将重组质粒转入到根癌农杆菌EHA105中。以白金针菇Flammulina velutipes菌丝球为受体材料,用根癌农杆菌介导转入碱性成纤维细胞生长因子（bFGF）。金针菇菌株对潮霉素（Hyg）抗性测验结果表明,在PDA固体培养基上的潮霉素筛选浓度为9μg/mL,在液体培养基中为6μg/mL。探索头孢霉素（Cefotaxime）对菌丝的毒性实验、农杆菌的菌液浓度、侵染时间、乙酰丁香酮（AS）的添加及其浓度的控制、共培养的时间等因素对转化效率的影响。结果表明,Cef对金针菇菌丝几乎无抑制作用,最佳抑菌浓度为400μg/mL;农杆菌的菌液浓度OD600为0.5,侵染时间在30min左右,在AS为200μmol/L的共培养基上共培养72h,转化率最高。PCR与Southern杂交结果证明bFGF基因已整合到金针菇的基因组中。在无选择压力的PDA固体培养基上继代培养5次后仍检测到bFGF基因的存在,表明bFGF基因在转基因金针菇中稳定遗传。     

施硒对两种类型玉米硒元素分配及产量、品质的影响

通过盆栽试验,以普通玉米品种郑单958(ZD958)和糯玉米品种京紫糯218(JN218)为试验材料,研究了不同硒水平(0、10、25、50 mg·kg-1)下,玉米植株各器官对硒的分配和转运差异以及硒对玉米产量和籽粒品质的影响.结果表明： 低含量(≤10 mg·kg-1)硒促进了玉米生长,植株生物量和籽粒产量均显著增加;高含量(≥25 mg·kg-1)硒抑制了玉米生长,植株干物质积累量减少,籽粒产量和品质下降.施硒显著提高了玉米植株各器官的硒含量,硒在各器官的分配为根系>叶片>茎秆>叶鞘,两种类型玉米各器官硒含量均与土壤硒含量呈显著正相关.JN218在自然低硒土壤环境中具有较强的硒富集能力,而ZD958在10 mg·kg-1 硒水平下硒积累量高于JN218.如果以籽粒和地上部营养器官的硒积累量为评价标准,自然低硒(025 mg·kg-1)或高硒(25 mg·kg-1)土壤适宜种植JN218,而富硒(10 mg·kg-1)或硒污染(50 mg·kg-1)土壤适宜种植ZD958.     

Selenium Pretreatment Enhances the Radioprotective Effect and Reduces the Lethal Toxicity of Wr-2721

Although WR-2721, S-2-(3-aminopropylamino)ethylphosphorothioic acid, is an effective radioprotector, its use is limited by its toxicity. Combining WR-2721 with other agents might decrease its toxicity and/or increase its effectiveness. The effect of selenium (Se) pretreatment on the acute toxicity and radioprotective effect of WR-2721 was studied in male CD2F1 mice. Injection of 1.6mg/kg Se 24 hr before WR-2721 (800-1200 mg/kg, IP) decreased the lethality of WR-2721 significantly. Lower doses of Se were also effective, but simultaneous administration was not effective. Se injection alone (1.6 mg/kg) 24 hr before cobalt-60 irradiation increased the survival (dose reduction factor, DRF = 1.1) significantly. A synergistic effect on post-irradiation survival was observed when Se was injected 24 hr before WR-2721 (200-600 mg/ kg IP before irradiation). For example, after exposure to 22 Gy (1 Gy/min), 30-day survival was 100% when mice were treated with both Se and 600mg/kg WR-2721, and was 13% with WR-2721 alone. The DRF after 400 mg/kg WR-2721 was 2.6 with Se compared to 2.2 without Se pretreatment. Alkaline phosphatase activity in bone marrow cells and serum was significantly depressed after treatment with 1.6 mg/kg Se, suggesting that a retardation of conversion of WR-2721 to its active free sulfhydryl form through the action of alkaline phosphatase might be partly responsible for the effects of Se. Other possible mechanisms related to the antioxidant properties of Se are under investigation.     

Selenium Pretreatment Enhances the Radioprotective Effect and Reduces the Lethal Toxicity of Wr-2721

Although WR-2721, S-2-(3-aminopropylamino)ethylphosphorothioic acid, is an effective radioprotector, its use is limited by its toxicity. Combining WR-2721 with other agents might decrease its toxicity and/or increase its effectiveness. The effect of selenium (Se) pretreatment on the acute toxicity and radioprotective effect of WR-2721 was studied in male CD2F1 mice. Injection of 1.6mg/kg Se 24 hr before WR-2721 (800-1200 mg/kg, IP) decreased the lethality of WR-2721 significantly. Lower doses of Se were also effective, but simultaneous administration was not effective. Se injection alone (1.6 mg/kg) 24 hr before cobalt-60 irradiation increased the survival (dose reduction factor, DRF = 1.1) significantly. A synergistic effect on post-irradiation survival was observed when Se was injected 24 hr before WR-2721 (200-600 mg/ kg IP before irradiation). For example, after exposure to 22 Gy (1 Gy/min), 30-day survival was 100% when mice were treated with both Se and 600mg/kg WR-2721, and was 13% with WR-2721 alone. The DRF after 400 mg/kg WR-2721 was 2.6 with Se compared to 2.2 without Se pretreatment. Alkaline phosphatase activity in bone marrow cells and serum was significantly depressed after treatment with 1.6 mg/kg Se, suggesting that a retardation of conversion of WR-2721 to its active free sulfhydryl form through the action of alkaline phosphatase might be partly responsible for the effects of Se. Other possible mechanisms related to the antioxidant properties of Se are under investigation.     

Tolerance of wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) to high soil and solution selenium levels

The fertilisation of wheat crops with Se is a cost-effective method of enhancing the concentration of organic Se in grain, in order to increase the Se intake of animals and humans. It is important to avoid phytotoxicity due to over-application of Se. Studies of phytotoxicity of Se in wheat grown in Australia, where rainfall and grain yield are usually relatively low, have not been reported previously, and overseas studies have had varied results. This study used trials conducted in the field, glasshouse and laboratory to assess Se phytotoxicity in wheat. In field trials that used rates of up to 120 g ha^–1Se as selenate, and in pilot trials that used up to 500 g ha^–1 Se soil-applied or up to 330 g ha^–1 Se foliar-applied, with soils of low S concentrations (2–5 mg kg^–1), no Se toxicity symptoms were observed. In pot trials of four weeks duration, the critical tissue level for Se toxicity was around 325 mg kg^–1 DW, a level attained by addition to the growth medium of 2.6 mg kg^–1 Se as selenate. Solution concentrations above 10 mg L^–1 Se inhibited early root growth of wheat in laboratory studies, with greater inhibition by selenite than selenate. For selenite, Se concentrations around 70 mg L^–1 were required to inhibit germination, while for selenate germination % was unaffected by a solution concentration of 150 mg L^–1 Se. Leaf S concentration and content of wheat increased three-fold with the addition of 1 mg kg^–1 Se as selenate to the growth medium. This effect is probably due to the induction of the S deficiency response of the main sulphate transporter. This study found wheat to be more Se-tolerant than did earlier studies of tobacco, soybeans and rice. We conclude that Se phytotoxicity in wheat will not be observed at the range of Se application rates that would be used to increase grain Se for human consumption (4–200 g ha^–1 Se as selenate, which would result in soil and tissue levels well below those seen in the above studies), even when – as is common in Australia – soil S concentration and grain yield are low.     

硒镧复合作用对巴西蘑菇子实体形态和数量特征的影响

采用混合培养料试验和电子显微技术,观察了不同硒镧配施水平对巴西蘑菇子实体盖皮和菌褶表面形态及孢子和囊状体数量特征的影响.结果表明,低浓度硒镧配施(Na2SeO3 21.905mg/kg LaCl317.667mg/kg)处理时,孢子数极显著增加,随着硒镧配施浓度升高,孢子数极显著增加,达到峰值后极显著减少;低浓度(Na2SeO 21.905mg/kg LaCl17.667mg/kg)和高浓度(Na2SeO 109.524mg/kg LaCl88.340mg/kg)处理后囊状体数极显著减少,中浓度(Na2SeO365.714mg/kg LaCl353.002mg/kg)处理囊状体数显著增加.研究还发现硒镧配施对巴西蘑菇孢子长、宽、囊状体最大直径和最小直径以及菌盖皮菌丝最大宽度具有促进或抑制作用.扫描电镜观察结果表明,与对照相比,中浓度硒镧配施(Na2SeO365.714mg/kg LaCl3 53.002mg/kg)时,孢子由正常椭圆形变成全部萎缩,菌丝粗壮;囊状体在较低硒镧配施浓度(Na2SeO343.810mg/kg LaCl3 35.335mg/kg)处理时表面纹饰模糊、消失,个别囊状体破裂,表面带有藤蔓状的组织.     

An agglutinin with mitogenic and antiproliferative activities from the mushroom Flammulina velutipes

A hemagglutinin with a molecular mass of 12 kDa was isolated from the fruiting bodies of the mushroom Flammulina velutipes. Its molecular mass is similar to that of the fungal immunomodulatory protein isolated from F. velutipes (FIP-fve) with ice-cold 5% acetic acid and 50 mM 2-mercaptoethanol as extraction medium and to that of the larger 12 kDa subunit of F. velutipes lectin isolated with phosphate buffer as extraction medium. Its hemagglutinating activity cannot be inhibited by a variety of carbohydrates tested. The activity is stable between pH 4 and pH 11. Loss in activity occurred when the temperature is raised to 60 C and 70 C. Activity is indiscernible at and above 80 C. Its N-terminal sequence shows differences from that of FIP-fve. F. velutipes hemagglutinin stimulates [3H-methyl] thymidine uptake by mouse splenocytes. It inhibits proliferation of leukemia L1210 cells with an IC50 of 13 microM.     

芦竹修复镉汞污染湿地的研究

以湿土盆栽方法研究了芦竹在Cd和Hg污染模拟湿地中的富集能力及其在植株中的分布.结果表明,芦竹在101mg·kg^-1Hg污染环境中生长8个月后,对Hg的富集量是根系>茎>叶片,植物地上部分对Hg富集量为200±20mg·kg^-1DW;而在115mg·kg^-1Cd污染环境中生长8个月后,其对Cd的富集量是叶片>根系>茎,芦竹叶片对Cd的富集量在160±26mg·kg^-1DW.重金属在芦竹各器官内的含量随种植时间的延长而增加,8个月生长期富集量比4个月生长期富集量高30%～50%.芦竹生物富集系数(Bio concentrationfactorBCF)随土壤中重金属含量增加而减小.在污染土壤中,芦竹叶、茎对Hg的BCF为1.9和2.1、对Cd为1.5和0.3;在未受污染的空白对照湿土中(含Hg6.8mg·kg^-1,Cd8.5mg·kg^-1),芦竹叶、茎对Hg的BCF为6.8和12.2,对Cd为7.0和2.7,表明芦竹具有生物量大、根系发达、适应性强等特点,对Cd、Hg有较大富集量和较好的耐受性.