两种大型真菌菌丝体对重金属的耐受和富集特性

用平板培养法检测大型真菌秀珍菇和猪肚菇菌丝体对重金属铬、铅和锰的耐受及富集特性。分别测定了3种重金属不同浓度处理下秀珍菇和猪肚菇的菌落直径、菌丝体干重和菌丝体中的重金属含量。结果表明:秀珍菇和猪肚菇菌丝体对Cr的耐受特性和耐受能力相当,二者的菌落直径和菌丝体干重均随Cr处理浓度的增加先升高后降低,生长抑制率为50%的Cr浓度都约为200 mg/L,对铬的最大耐受浓度都为500 mg/L。秀珍菇菌丝体对Pb敏感,100 mg/L的Pb即可极显著的抑制秀珍菇菌丝的生长,而猪肚菇则直到500 mg/L的Pb,菌丝体的生长都未受显著影响;二者生长抑制率为50%的Pb浓度分别为100 mg/L和700 mg/L,最大耐受浓度分别为1000 mg/L和2000 mg/L;因此,猪肚菇对Pb的耐受能力比秀珍菇强。秀珍菇不耐锰,而猪肚菇对锰表现出相对较高的耐受能力,生长抑制率为50%的Mn浓度约为1000 mg/L,最大耐受能力为6000 mg/L。秀珍菇菌丝体对Cr和Pb、猪肚菇菌丝体对Cr和Mn均没有达到超富集。但猪肚菇菌丝体中Pb的含量可达1125.56 mg/kg(干重),达到超富集水平,暗示猪肚菇可能是铅超富集大型真菌。     

秀珍菇粉对H22荷瘤小鼠体内抗肿瘤作用研究

目的本试验旨在研究秀珍菇抗肿瘤作用及其抑瘤机制。方法选用ICR雄性小鼠60只,取10只作为空白组,剩余50只小鼠通过腋部皮下接种H22小鼠腹水构建荷瘤小鼠模型,造模后随机分为模型组、阳性药(CTX)组、秀珍菇低、中、高剂量组,每组10只。模型组小鼠灌服生理盐水,阳性药组小鼠按20 mg/kg体重隔天腹腔注射注射环磷酰胺(CTX)生理盐水溶液,秀珍菇低、中、高剂量组小鼠分别按750、1500、3000 mg/kg体重每天剂量灌服秀珍菇生理盐水混悬液。给药10 d后,测定各组小鼠平均瘤重,肿瘤抑制率;测定免疫器官脏器指数、血清免疫球蛋白和细胞因子含量;测定肝、肾抗氧化指标;观察肿瘤和脾组织HE染色病理切片。结果 (1) CTX及秀珍菇低、中、高剂量组小鼠的平均瘤重均极显著低于模型组(P0. 01),四组小鼠肿瘤抑制率分别为55. 18%,29. 06%、47. 47%和48. 80%。(2)与空白组比较,模型组小鼠脾指数、血清Ig A和TNF-α含量及肝MDA含量显著升高(P0. 05,P0. 01),血清IL-6含量有上升趋势(P 0. 05),而血清IL-2含量、肝CAT和GSH-Px活性及肾SOD、GSH-Px和CAT活性显著降低(P0. 05,P0. 01);同时,CTX组小鼠胸腺指数显著低于空白组(P0. 05)。(3)秀珍菇对荷瘤小鼠免疫和抗氧化功能异常改变具有逆转作用:与模型组比较,秀珍菇各剂量组小鼠血清Ig A和TNF-α含量及高剂量组小鼠IL-6水平显著降低(P 0. 05,P 0. 01),低剂量组小鼠血清IL-2水平显著升高(P 0. 01);秀珍菇处理还显著提高了各剂量组荷瘤小鼠肝CAT活性及肾SOD和CAT活性(P 0. 05,P 0. 01),显著升高中、高剂量组肝GSH-Px活性和低、中剂量组肾GSH-Px活性(P 0. 05,P 0. 01),并显著降低中剂量组的肝MDA含量(P 0. 05)。(4)与CTX组比较,秀珍菇高剂量组小鼠脾指数显著提高(P 0. 05),而血清IL-6水平显著降低(P 0. 05)。(5)肿瘤组织病理切片显示CTX组和秀珍菇各剂量组的肿瘤坏死面积明显增加。结论秀珍菇可抑制H22实体移植瘤生长,其机制与其具有较强的免疫调节和抗氧化作用有关。     

秀珍菇乙烯受体基因克隆及其在1-MCP保鲜处理下的表达

本研究通过生物信息与分子生物学手段,对秀珍菇乙烯受体基因进行克隆,并用荧光定量PCR检测1-MCP处理前后秀珍菇乙烯受体基因的表达情况。本研究克隆到1个秀珍菇可靠的乙烯受体基因PpuETR1,该基因编码的氨基酸序列含有保守的组氨酸激酶结构域,与双孢蘑菇的组氨酸蛋白激酶感应蛋白和植物的同源乙烯受体同源。该基因在1-MCP处理后表达量受到了明显的抑制,在贮藏期间均呈下调表达,这从分子水平上说明1-MCP抑制了乙烯受体基因的表达,从而减缓了乙烯信号通路对秀珍菇成熟衰老作用。实验结果为1-MCP对秀珍菇的保鲜作用机理研究提供了一定的数据支持。     

60Coγ辐射对秀珍菇子实体蛋白质营养水平的影响

采用国际上通用的非生物评价方法,研究了不同剂量60Coγ辐射对秀珍菇子实体蛋白质营养价值的影响,以期为秀珍菇辐射育种研究提供科学依据.结果表明：在相同培养料以辐射剂量为1.00处理的栽培的秀珍菇子实体蛋白质营养价值评价的六项指标中氨基酸评分、化学评分、必需氨基酸指数、氨基酸比值系数分和生物价分别为74.3、58.5、78.3、66.9和73.6,均居5种剂量处理之首,营养指数为25.8,均居第2位.综合评价的结果显示,以辐射剂鼍为1.00 kGy处理的相同培养料栽培秀珍菇子实体其蛋白质营养价值最高.     

60Co辐射对秀珍菇子实体蛋白质营养水平的影响

采用国际上通用的非生物评价方法,研究了不同剂量^60Co辐射对秀珍菇子实体蛋白质背养价值的影响,以期为秀珍菇辐射育种研究提供科学依据。结果表明：存相同培养料以辐射剂量为1．00kGy处理的栽培的秀珍菇子实体蛋白质营养价值评价的六项指标中氨琏酸评分、化学评分、必需氦基酸指数、氨基酸比值系数分种生物价分别为74．3、58．5、78．3、66．9和73．6,均居5种剂量处理之首,营养指数为25．8,均居第2位综合评价的结果显示,以辐射利量为1．00kGy处理的相同培养料栽培秀珍菇子实体其蛋白质营养价值最高.     

一株野生秀珍菇的鉴定及其生物学特性的分析

本研究以野外采集获得的1株野生侧耳属真菌为对象,对其进行rDNA-ITS序列扩增测序并分析后初步确定其为肺形侧耳(Pleurotus pulmonarius),并与本实验室中现有的秀珍菇菌株序列进行随机扩增多态性DNA(RAPD)、ISSR、相关序列扩增多态性(SRAP)的综合分析后进一步确定该野生菌株与其他菌株间具有一定的遗传距离,是一株新的秀珍菇菌株。以福建省认定的秀珍菇品种秀迪1号为对照菌株,对该菌株进行生物学特性进行研究,结果表明,其菌丝可在温度为5~45℃的范围内生长,最适生长温度为30℃;菌丝可生长的pH范围为pH 3~10,最适pH值为7。在含可溶性淀粉、酵母粉为碳氮源的培养基中,菌丝生长速度快,长势好。该野生菌株生长所需的最适碳、氮源及生长条件与对照菌株相似,这为其进一步的驯化栽培提供一定的理论基础。     

60Co-γ辐照在秀珍菇中的应用研究

采用不同剂量的^60 Co-γ射线辐照秀珍菇进行贮藏保鲜试验,结果表明：常温下210辐照,并采用聚乙烯塑料袋分装,可以延长鲜菇的保鲜期约10天。通过对维生素C和氨基酸含量的测定试验表明：辐处理前后无明显变化。辐照灭菌与常规高温灭菌相比,秀珍菇鲜菇产量高于对照。菌丝在经辐照灭菌的培养料上生长快,污染率下降,而且出菇快。     

自交选育秀珍菇新菌株“申秀1号”

【目的】对自交子代群体主要栽培和商品性状进行综合分析,并选育综合性状优良的秀珍菇新菌株。【方法】以秀珍菇商业菌株“3108”为亲本进行自交,综合分析其自交子代群体的菌丝生长速度、产量、菇型、抗杂、抗逆性和栽培周期等性状以选育新菌株,利用ISSR (Inter-sample sequence repeat)鉴定新菌株和亲本菌株之间遗传差异性。【结果】自交子代中,菌丝生长速度和产量分别有19.5%和8.9%的菌株优于亲本;子代群体中菌丝生长速度和子实体产量之间无相关性(R=0.102,P>0.05);37%的菌株栽培周期比亲本短;菇盖平展、厚且不易碎分别占53%、11%,菌柄直径和长度变化幅度分别为5?13 mm和21?75 mm;发生黄枯病,出现萎缩、死菇现象以及畸形分别占23%、19%、5%。【结论】通过对子代群体各性状综合分析,从中选育出生长快、产量高、菇盖厚且不易碎、柄粗长、抗杂和抗逆性强、栽培周期短等优良性状的秀珍菇新菌株“申秀1号”,前三潮菇平均单袋产量334 g,生物学效率高达74%,比亲本菌株增产7.1%,且种性稳定。经ISSR鉴定表明,该菌株具有自身特异性。     

秀珍菇辐射新株系蛋白质营养价值的主成分和聚类分析

对18个秀珍菇辐射新株系子实体中的必需氨基酸指数、生物价、营养指数、氨基酸比值系数分、化学评分和氨基酸评分的6项蛋白质营养价值指标进行主成分分析.结果表明,必需氨基酸指数和生物价因子、氨基酸比值系数分和化学评分因子、营养指数因子,这3个主要因子对变异的累积贡献率达95.88%.依据主成分向量将18个秀珍菇辐射新株系聚为4类.     

我国肺形侧耳栽培菌株交配型分析

为分析我国栽培肺形侧耳的遗传多样性,对肺形侧耳菌株的交配型进行测定,测试的肺形侧耳菌株包括俗称为秀珍菇和凤尾菇的菌株。结果显示,我国栽培的秀珍菇与凤尾菇菌株间可以发生性亲和,属于同一生物学物种,但它们之间的交配型特异性低,36个菌株中只含4种A交配型和3种B交配型,暗示我国栽培的肺形侧耳种质资源基因匮乏。     

A Review of: “Progress in Separation and Purification Vol. 3, E. S. Perry and C. J,van Oss,eds., Willy/Intersciencc,New York, 1970; xi + 316 pares; hard cover; 515.95”

The optimal culture conditions of exopolysaccharides (EPS) production in submerged culture medium by Pleurotus geesteranus 5 ^# were determined using an orthogonal matrix method. The optimal defined medium (per liter) was 60.0 g maltose, 5.0 g tryptone, 1 mM NaCl, 5 mM KH₂PO₄, and initial pH 6.0 at 28 °C. In the optimal culture medium, the maximum EPS production was 16.97 g/L in a shake flask. Two groups of EPSs (designated as Fr-I and Fr-II) were obtained from the culture filtrates by size exclusion chromatography (SEC), and their molecular characteristics were examined by a multiangle laser-light scattering (MALLS) and refractive index (RI) detector system. The approximate weight-average molar masses of the Fr-I and Fr-II of EPS were determined to be 3.263 × 10⁴ and 5.738 × 10³ g/mol, respectively. The low values of polydispersity ratio (1.176 and 1.124 for Fr-I and Fr-II, respectively) of EPSs mean that these EPS molecules exist much less dispersed in aqueous solution without forming large aggregates. Furthermore, the experiments in vitro indicated that P. geesteranus 5^# EPS exhibit high antitumor and antioxidative effects.     

A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa based on ligninolytic enzymes

A novel breeding strategy for new strains of Hypsizygus marmoreus and Grifola frondosa using ligninolytic enzymes as markers was evaluated with the detection and analysis of activities and composition of 15 edible fungi. The results showed that the activity and composition of ligninolytic enzyme system varied in response to changes of fungal strains. By analyzing the growth rate of mycelia and their ability to produce ligninolytic enzymes, H. marmoreus and P. geesteranus, G. frondosa and P. sajor-caju were screened for further study. Three colonies of 26 regenerated colonies of H. marmoreus and P. geesteranus protoplast fusion and one colony of 48 regenerated colonies of G. frondosa and P. sajor-caju were selected respectively. At the same time, these four strains were identified using RAPD and ISSR molecular markers. The results showed that the strains HM5G1and PS7F1are new strains and have low similarity to parental strains H. marmoreus and G. frondosa. These results are supported by the results of antagonism tests. These two fusants were significantly higher in their ligninolytic enzyme activity than H. marmoreus and G. frondosa. The growth rates of strains HM5G1and PS7F1 were also noticeably higher than those of H. marmoreus and G. frondosa, by 1.36 and 1.5 times respectively. The biological efficiency of the strain HM5G1 was 11.5 % higher than that of the parental strain H. marmoreus. This work suggests that it is an efficient way of breeding new strains to use the decolorization of ligninolytic enzymes as a preliminary screening marker.     

Requirement for P granules and meiosis for accumulation of the germline RNA helicase CGH-1

In Caenorhabditis elegans, lack of the conserved germline RNA helicase CGH-1 causes infertility and excessive levels of physiological germline apoptosis, a process that normally claims about half of all developing oocytes. In yeast the CGH-1 ortholog is a key component of degradative "processing (P) bodies," which may share some properties with germline protein-RNA complexes such as P granules. During oogenesis CGH-1 associates with P granules, but also accumulates to high levels in additional cytoplasmic particles. Here we show that appropriate levels and localization of CGH-1 depends on some P granule components and on mechanisms that establish meiotic development. At the same time, germ cell death is not increased by various abnormalities in P granules or meiosis. We conclude that in developing oocytes CGH-1 particles accumulate specifically in response to meiotic development and have distinct functions from P granules, and may be dynamic protein-mRNA structures.     

Developmental regulation of the Inositol 1,4,5-trisphosphate phosphatases in Dictyostelium discoideum

The cellular slime mold Dictyostelium discoideum is a microorganism in which growth and development are strictly separated. Starvation initiates a developmental program in which extracellular cAMP plays a major role as a signal molecule. In response to cAMP several second messengers are produced, including cAMP, cGMP and inositol 1,4,5-trisphosphate, (Ins(1,4,5)P3). Ins(1,4,5)P3 levels are controlled by the activation of phosphoinositidase C and the activity of the Ins(1,4,5)P3-degrading phosphatases. In Dictyostelium discoideum two major routes for the dephosphorylation of Ins(1,4,5)P3 are present: a 5-phosphatase, which hydrolyses Ins(1,4,5)P3 at the 5-position producing Ins(1,4)P2 as in vertebrate cells, and a 1-phosphatase which removes the 1-phosphate, giving Ins(4,5)P2, as in plants. In this paper we show that at the onset of development both the 1-phosphatase and the 5-phosphatase are present in equal amounts. During development the 5-phosphatase disappears leaving the 1-phosphatase as the single enzyme to remove Ins(1,4,5)P3. We conclude that during development Dictyostelium discoideum switches from a mixed type of Ins(1,4,5)P3 degradation to a more plant-like degradation pathway.     

Four decades of post-agricultural forest development have caused major redistributions of soil phosphorus fractions

Fertilisation of agricultural land causes an accumulation of nutrients in the top soil layer, among which phosphorus (P) is particularly persistent. Changing land use from farmland to forest affects soil properties, but changes in P pools have rarely been studied despite their importance to forest ecosystem development. Here, we describe the redistributions of the P pools in a four-decadal chronosequence of post-agricultural common oak (Quercus robur L.) forests in Belgium and Denmark. The aim was to assess whether forest age causes a repartitioning of P throughout the various soil P pools (labile P, slowly cycling P and occluded P); in particular, we addressed the time-related alterations in the inorganic versus organic P fractions. In less than 40 years of oak forest development, significant redistributions have occurred between different P fractions. While both the labile and the slowly cycling inorganic P fractions significantly decreased with forest age, the organic fractions significantly increased. The labile P pool (inorganic + organic), which is considered to be the pool of P most likely to contribute to plant-available P, significantly decreased with forest age (from >20 to <10% of total P), except in the 0-5 cm of topsoil, where labile P remained persistently high. The shift from inorganic to organic P and the shifts between the different inorganic P fractions are driven by biological processes and also by physicochemical changes related to forest development. It is concluded that the organic labile P fraction, which is readily mineralisable, should be taken into account when studying the bioavailable P pool in forest ecosystems.     

神经元蛋白3.1结合蛋白的筛选与鉴定

哺乳动物肺组织发育的基本模式总体分为上皮分支的形态发生和分隔膜的形成两个部分．基因p311是克隆到的一个在分隔膜形成阶段特异表达的基因,可能在肺泡发育中起重要作用．为进一步探讨神经元蛋白3.1(P311)对肺发育过程的影响,构建了小鼠肺组织 cDNA文库,以融合Gal4 DNA结合结构域的重组P311蛋白为诱饵,利用酵母双杂交技术从文库中筛选P311结合蛋白．通过免疫共沉淀和双分子荧光互补等技术进一步验证, SPARC(secreted protein, acidic and rich in cysteine)被确定为P311 相互作用蛋白．进一步的研究发现,SPARC在肺组织中具有与P311相似的表达时序特征,双重免疫组织化学染色显示SPARC和P311在小鼠肺组织中共定位于肺泡上皮细胞和肌成纤维细胞中．提示P311可能通过与SPARC的相互作用影响肺泡发育．     

东方蜜蜂背单眼的结构和胚后发育

本研究通过形态解剖和BrdU免疫组织化学方法对东方蜜蜂Apis cerana Fabricius背单眼的胚后发育过程进行了比较研究,结果表明:东方蜜蜂的每一个背单眼都包括角膜晶体、角膜生成细胞、小网膜细胞以及后部的单眼神经。蜜蜂的背单眼起源自头壳上皮;其胚后发育的高峰期集中在蛹发育的前3d;其新细胞主要来源于上皮细胞和圆锥形单眼囊周围细胞的有丝分裂;单眼同脑的联系在P1期前后就已经建立;角膜晶体的形成在P5以后。说明单眼的结构和发育同其功能密切相关。     

Developmental competence of oocytes from prepubertal Bos indicus crossbred cattle

The aim of the present study was to evaluate the effect of age on embryogenic competence of oocytes recovered from Bos indicus crossbred calves and heifers. Cumulus-oocyte complexes (COCs) were collected from 4- to 7-month-old calves (experiment 1) and from 9- to 14-month-old heifers (experiment 2) during processing at an abattoir. In both experiments cow COCs were used as control. COCs were in vitro matured and fertilized, and the presumptive zygotes co-cultured with cumulus cells until 224 h post insemination (hpi). In experiment 1, the development rate during the first 68-72 hpi was similar (P > 0.05) between embryos derived from calves and cows. Fewer embryos from calves developed to the blastocyst stage, resulting in a lesser blastocyst production as well as lesser hatching rate (P < 0.05). The embryo development after blastocyst stage was, nevertheless, similar (P > 0.05) between blastocysts derived from calves and cows, suggesting that the development after blastocoele formation is not compromised in embryos derived from calves. In experiment 2, there were no differences (P > 0.05) on cleavage, blastocyst and hatching rates between embryos derived from prepubertal heifers and cows. The rate of blastocyst development until hatching was also similar (P > 0.05). These results indicate that oocytes from 9- to 14-month-old B. indicus crossbred heifers have the same developmental competence as oocytes derived from cows, while ocytes derived from 4- to 7-month-old B. indicus crossbred calves are less competent in developing to the blastocyst stage in vitro. It suggests that oocyte competence in B. indicus crossbred cattle is achieved around 9-14 months of age.     

“双季稻-鸭”共生生态系统稻季磷循环

稻鸭共生是对中国传统农业稻田养鸭的继承与发展。在长江流域双季稻主产区湖南布置了稻田养鸭田间对比试验,以常规稻作为对照,采用投入产出法,分析"稻鸭共生"生态系统P的输入输出及循环情况。结果表明:"早稻-鸭"共生生态系统P输出为52.28kg·hm-2,其中鸭产品P为1.71kg·hm-2;"晚稻-鸭"共生生态系统P输出为83.24kg·hm-2,其中鸭产品P为6.17kg·hm-2;在早、晚稻两季,"稻鸭共生"系统在目前的P养分投入水平下,降低了土壤P的亏缺量;鸭子系统P输入主要来自系统外投入的饲料P;鸭粪P在系统内循环利用,两季循环率分别为12.5%和13.5%;两季稻作后,"稻鸭共生"土壤截存的P量为195.41kg·hm-2,比常规稻作显著提高了36.9%;"稻鸭共生"可对稻田土壤增加鸭粪P为17.75kg·hm-2。