Regulation of the amylase secretion by the yeast Filobasidium capsuligenum and a 2-deoxy-d-glucose resistant mutant

Summary The regulation of extracellular amylase production by the basidiomycetous yeast Filobasidium capsuligenum CCY 64-5-1 was characterized using growing and resting cells. A basal level of amylolytic activity was produced with various carbon sources including glucose. Amylase secretion was repressed by glucose and, more severely, by 2-deoxy-d-glucose, whereas compounds with -1,4-linked glucose, such as methyl glucoside, maltose, -cyclodextrin and soluble starch, served as inducers. Repression was not relieved by exogenously added cAMP. The effects of several metabolic inhibitors on amylase secretion were studied. Following UV-mutagenesis a mutant strain (FC-5) capable of growing in a 2-deoxy-d-glucose supplemented corn starch medium was selected for further characterization. This strain produced more amylase, had acquired an increased resistance against repression by glucose, and retained a growth rate comparable to the wild type. FC-5 was also characterized by a reduced glucokinase activity and an increased hexokinase activity.     

Secretion of alpha-amylase and multiple forms of glucoamylase by the yeast Trichosporon pullulans

Trichosporon pullulans IGC 3488 produced extracellular alpha-amylase and glucoamylase activities when grown in batches in a medium containing corn steep liquor and soluble starch or corn starch. alpha-Amylase, unlike glucoamylase activity, was secreted biphasically. For both amylases the maximum concentration was found in stationary phase cultures. The amylolytic enzymes, previously concentrated by ammonium sulfate precipitation, were separated into a glucoamylase fraction and an alpha-amylase fraction by Ultrogel AcA 54 gel filtration. Pullulanase activity was located in the glucoamylase fraction, whereas cyclodextrinase activity was restricted to the alpha-amylase fraction. Isoamylase and alpha-glucosidase were not detected. Electrophoretic analysis showed that alpha-amylase activity was due to a single protein. Glucoamylase, however, occurred in multiple forms. The four glucoamylases and the alpha-amylase were glycoproteins.     

Isolation and characterization of fluorescent Pseudomonas associated with the roots of rice and banana grown in Sri Lanka

Bacterial populations in different parts of the rhizosphere of rice and banana in Sri lanka were examined. On rice, the number of aerobic bacteria and the population of fluorescent bacteria were higher in the rhizoplane as compared to the exorhizosphere. However, the opposite was observed with banana. Percentage of fluorescent bacteria was significantly higher on banana (10.8%) than on rice from the wet and dry zones of Sri Lanka (4.3% and 2.7%, respectively). In the endorhizosphere fraction of rice, bacterial populations were very low. Fluorescent bacteria were absent.Based on 33 phenotypical tests, 89 fluorescent isolates were grouped into 5 clusters. The three major clusters covered the isolates belonging to the Pseudomonas fluorescens-putida group, whereas the remaining small clusters contained other UV-fluorescent bacteria. SDS-PAGE of total cell proteins enabled classification of the isolates into one of 12 different protein-polymorphic types. Only a partial correlation was found between the latter classification and the phenotypical one. Cyanogenesis was observed with strains of P. fluorescens only. Isolates P. fluorescens RW9S1 and P. cepacia RW5P1 displayed a potent antagonism against several fungi.     

17个建兰品种光合特性分析

以17个建兰(Cymbidium ensifolium)品种为材料,采用改良的丙酮法提取叶绿素,再通过Arnon丙酮法公式计算光合色素含量,利用捷克FluorCam开放式叶绿素荧光仪测定不同品种的叶绿素荧光参数。结果表明,17个建兰品种的光合色素和叶绿素荧光参数具有不同程度的差异,其中‘铁骨素’(C. ensifolium ‘Tiegusu’)、‘逸红双娇’(C. ensifolium ‘Yihongshuangjiao’)和‘闽南黄蝶’(C. ensifolium ‘Minnanhuangdie’)的光合色素含量高于其他品种,表明这3个品种具有良好的光合效率,吸收光能的能力较强;‘铁骨素’最大荧光产量(Fm)、Kautsky诱导效应最大荧光(Fp)、PS Ⅱ原初光能转化效率(Fv/Fm)和非光化荧光淬灭系数(NPQ)均为最高。综上可知,‘铁骨素’的光合生理特性优于其他品种,可作为优良建兰品种进行种植推广。     

小叶锦鸡儿天然居群叶形态性状变异研究

为揭示小叶锦鸡儿(Caragana microphylla)天然居群叶形态性状的变异规律及其生态适应性特征,该研究以10个小叶锦鸡儿天然居群为对象,通过多重比较、巢式方差分析、相关性分析、聚类分析和主成分分析等方法,对7个叶形态性状进行分析。结果表明：(1)小叶锦鸡儿叶形态性状在居群内和居群间均存在极显著差异(P < 0.01),平均变异系数为10.13%,不同性状的变异幅度为6.23%~12.78%;平均叶形态性状的表型分化系数为43.62%,居群内变异(30.09%)大于居群间变异(24.91%),说明居群内是其叶形态性状变异的主要来源。(2)相关性分析表明,环境因子对小叶锦鸡儿的叶形态性状变异有很大的影响,在地理空间上主要呈现出沿海拔梯度的变异模式;主成分分析的结果显示,小叶宽、叶柄宽和叶柄长对小叶锦鸡儿叶形态变异起主导作用;利用欧式距离对小叶锦鸡儿居群进行UPGMA聚类分析结果显示,基于叶形态性状和环境因子可分别将小叶锦鸡儿10个居群分为3类和2类,Mantel检验结果表明,小叶锦鸡儿的叶形态性状变异不存在地理连续性。研究结果为小叶锦鸡儿的适应性进化和开发利用提供了理论依据。     

H2S作为下游信号参与SA对低温弱光下黄瓜幼苗光合作用的调控

水杨酸(SA)和硫化氢(H₂S)在调控非生物胁迫下植物生长发育和生理代谢中均起着非常重要的作用,但二者作为信号分子在调控低温弱光下黄瓜光合作用中的互作关系还不清楚。本试验以黄瓜幼苗为试材,分别用SA和硫氢化钠(NaHS,H₂S供体)及其清除剂或抑制剂喷撒叶面,以适宜温光下去离子水处理为对照(CK),研究低温(8 ℃/5 ℃,昼/夜)弱光(100 μmol·m^-2·s^-1)下SA和H₂S对黄瓜幼苗光合作用的调控及互作关系。结果表明: SA可明显增强L-/D-半胱氨酸脱巯基酶(LCD、DCD)活性及其mRNA表达,促进内源H₂S产生;NaHS对苯丙氨酸解氨酶和异分支酸合成酶活性、mRNA表达量及内源SA含量影响不大。SA和NaHS可使低温弱光下黄瓜幼苗的光合速率、气孔导度和蒸腾速率明显提高,胞间CO₂浓度显著降低;同时增强核酮糖-1,5-二磷酸羧化酶、Rubisco活化酶、景天庚酮糖-1,7-二磷酸酯酶和果糖-1,6-二磷酸醛缩酶活性及其mRNA表达,促进光合碳同化;提高光下PSⅡ实际光化学效率和暗下PSⅡ最大光化学效率,从而减轻低温弱光胁迫对黄瓜幼苗的光合机构的损伤和生长量的影响。H₂S清除剂次牛磺酸(HT)可使SA对低温弱光下黄瓜幼苗的光合作用和生长促进效应明显减弱,而SA抑制剂多效唑和氨基茚磷酸对H₂S诱导的黄瓜幼苗光合机构对低温弱光的耐受性无显著影响,说明H₂S作为SA的下游信号,参与调控低温弱光下黄瓜幼苗的光合作用。     

HylA,an Alternative Hydrolase for Initiation of Catabolism of the Phenylurea Herbicide Linuron in Variovorax sp. Strains

Variovorax sp. strain WDL1, which mineralizes the phenylurea herbicide linuron, expresses a novel linuron-hydrolyzing enzyme, HylA, that converts linuron to 3,4-dichloroaniline (DCA). The enzyme is distinct from the linuron hydrolase LibA enzyme recently identified in other linuron-mineralizing Variovorax strains and from phenylurea-hydrolyzing enzymes (PuhA, PuhB) found in Gram-positive bacteria. The dimeric enzyme belongs to a separate family of hydrolases and differs in K_m, temperature optimum, and phenylurea herbicide substrate range. Within the metal-dependent amidohydrolase superfamily, HylA and PuhA/PuhB belong to two distinct protein families, while LibA is a member of the unrelated amidase signature family. The hylA gene was identified in a draft genome sequence of strain WDL1. The involvement of hylA in linuron degradation by strain WDL1 is inferred from its absence in spontaneous WDL1 mutants defective in linuron hydrolysis and its presence in linuron-degrading Variovorax strains that lack libA. In strain WDL1, the hylA gene is combined with catabolic gene modules encoding the downstream pathways for DCA degradation, which are very similar to those present in Variovorax sp. SRS16, which contains libA. Our results show that the expansion of a DCA catabolic pathway toward linuron degradation in Variovorax can involve different but isofunctional linuron hydrolysis genes encoding proteins that belong to evolutionary unrelated hydrolase families. This may be explained by divergent evolution and the independent acquisition of the corresponding genetic modules.