Purification of an Aminopeptidase Preferentially Releasing N-terminal Alanine from Cucumber Leaves and Its Identification as a Plant Aminopeptidase N

In this study, a highly active foliar aminopeptidase preferentially releasing N-terminal alanine from artificial substrates was purified and characterized from cucumber (Cucumis sativus L. suyo). The enzyme had a molecular mass of 200 kDa consisting of two subunits of 95 kDa. It was a metalloprotease the pH optimum of which was 8 to 9. It cleaved Ala-, Gly-, Met-, Ser-, Leu-, Lys-, and Arg artificial substrates. An internal amino acid sequence was similar to those of aminopeptidase N (clan MA, family M1) of microorganisms, and was very similar to that of a putative aminopeptidase N of Arabidopsis thaliana. From these results, the highly active aminopeptidase in cucumber leaves was identified to be a plant aminopepitdase N.     

Thermal Dissection of Lentil Seedling Amine Oxidase Domains by Differential Scanning Calorimetry

The relationships between the structural and energetic domains of lentil seedling amine oxidase (LSAO) were investigated using modifiers that target the active site and the carbohydrate moiety of the enzyme. An irreversible inhibitor, aminoguanidine, specifically modified the active site of the lentil enzyme, whereas sodium metaperiodate cleaves carbohydrate moieties covalently bound to the native enzyme. Differential scanning calorimetry (DSC) measurements were made on the modified LSAOs. Deconvolution of the reversible thermal DSC profiles of the modified enzyme gave three subpeaks (energetic domains), each of which was assigned to one of the three structural domains of the native protein. Our results led us to conclude that deglycosylation of LSAO has no effect on thermal stability, whereas binding of the inhibitor imparts more stability to the enzyme.     

Photolysis of 3-Hydroxyisoxazoles

Photolysis of 3-hydroxy-5-phenylisoxazole in methanol with a low-pressure mercury lamp afforded 5-phenyl-4-oxazolin-2-one, together with small amounts of benzoic acid and benzoylacetamide. Similarly, 3-hydroxy-5-methylisoxazole in distilled water afforded 5-methyl-4-oxazolin-2-one as the major product. Both isoxazoles were stable in sunlight for up to 20 days.     

Purification and Properties of a Pectin trans-Eliminase in Erwin aroideae Formed in the Presence of Nalidixic Acid

A strain of Erwinia aroideae produces a remarkable amount of pectolytic enzyme when the organism was induced by nalidixic acid for the bacteriocin production. This pectolytic enzyme was purified approximately 60-fold from the induced medium by carboxymethyl-cellulose and Sephadex G–75 gel column chromatographies after batchwise treatment with carboxymethyl- and diethylaminoethyl-celluloses. The purified enzyme was almost homogeneous on sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis, a molecular weight of about 28,000 to 32,000 was determined for this enzyme. The optimum pH of the enzyme activity was about 8.0 to 8.2. The purified enzyme produced reaction products from pectin and methoxylated pectic acid which had a strong absorption at 235 nm indicating a trans-eliminase reaction. Pectin or pectic acid with higher methoxyl content was a good substrate for this enzyme, while no significant activity was observed when pectic acid was a substrate. The limit of degradation of pectin and pectic acid with higher methoxyl content (90% esterified) by the enzyme were 6.5% and 43%, respectively. It was concluded that the enzyme is a new endo-pectin trans-eliminase from bacterial origin.     

Carpology,germination and seedling performance in Digitalis thapsi L. (Scrophulariaceae)

Summary

We analyse the fruiting phase of the life cycleof Digitalis thapsi L. We studied the production and size of seeds and capsules, and the interpopulational variability and inter-relationships of these parameters. The capsule volume, number of seeds per capsule, of seeds per individual, and of capsules per individual were inter-related and fairly affected by the medium, whereas seed weight was found to bea more independent character. A germination study showed that the species has agreat germinative capacity and has positive photoblasty. It is non-dormant especially if incubated under experimental conditions close to the climatic conditions of the site of provenance of the samples. An increase in temperature speeds up the process without increasing germinability. The morphological and biometric characteristics of the first stages of development of the species following germination were analysed. It was found that seed weight is not positively correlated with cotyledon length nor with any of the other parameters con-sidered, and that the length of the fifth leaf is a useful parameter to express inter-populational genetic variability. Seedling production in spring seems to be influenced more by physical than by chemical factors.     

Functional compatibility in cucumber mycorrhizas in terms of plant growth performance and foliar nutrient composition

Functional compatibility in cucumber mycorrhizas in terms of plant and fungal growth, and foliar nutrient composition from all possible combinations of six cucumber varieties and three species of arbuscular mycorrhizal (AM) fungi was evaluated. Measurements of foliar nutrient composition included N, P, K, Mg, Ca, Na, Fe, Zn, Mn and Cu. Growth of AM fungi was measured in terms of root colonisation, as examined with microscopy and the AM fungus biomarker fatty acid 16:1ω5 from both phospholipids and neutral lipids. Different responses of plant growth and foliar nutrient profiles were observed for the different AM symbioses examined. The AM fungus Claroideoglomus claroideum caused growth depression in association with four out of six cucumber varieties; Rhizophagus irregularis caused growth promotion in one of six cucumber varieties; whereas Funneliformis mosseae had no effect on the growth performance of any of the cucumber varieties examined. All three AM fungi markedly altered host plant shoot nutrient composition, with the strongest contrast observed between cucumber–R. irregularis symbioses and non‐mycorrhizal cucumber plants, independent of cucumber variety. On the other hand, AM fungal growth in roots differed between the three AM fungi, but was unaffected by host genotype. Strong build‐up of storage lipids was observed for R. irregularis, which was more moderate in the two other AM fungi. In conclusion, strong differential responses of cucumber varieties to inoculation with different AM fungi in terms of growth and shoot nutrient composition revealed high functional diversity in AM symbioses in cucumber plants.     

There's no place like home: seedling mortality contributes to the habitat specialisation of tree species across Amazonia

Understanding the mechanisms generating species distributions remains a challenge, especially in hyperdiverse tropical forests. We evaluated the role of rainfall variation, soil gradients and herbivory on seedling mortality, and how variation in seedling performance along these gradients contributes to habitat specialisation. In a 4‐year experiment, replicated at the two extremes of the Amazon basin, we reciprocally transplanted 4638 tree seedlings of 41 habitat‐specialist species from seven phylogenetic lineages among the three most important forest habitats of lowland Amazonia. Rainfall variation, flooding and soil gradients strongly influenced seedling mortality, whereas herbivory had negligible impact. Seedling mortality varied strongly among habitats, consistent with predictions for habitat specialists in most lineages. This suggests that seedling performance is a primary determinant of the habitat associations of adult trees across Amazonia. It further suggests that tree diversity, currently mostly harboured in terra firme forests, may be strongly impacted by the predicted climate changes in Amazonia.     

Frequency‐dependent selection for rare genotypes promotes genetic diversity of a tropical palm

Negative frequency‐dependent selection among species is a key driver of community diversity in natural systems, but the degree to which negative frequency‐dependent selection shapes patterns of survival and genetic diversity within species is poorly understood. In a 5‐year field experiment, we show that seedlings of a tropical palm with rare genotypes had a pronounced survival advantage over seedlings with common genotypes, with effect sizes comparable to that of light availability. This ‘rare genotype advantage’ led to an increase in population‐wide genetic diversity among seedlings compared to null expectations, as predicted by negative frequency‐dependent selection, and increased reproductive success in adult trees with rare genotypes. These results suggest that within‐species negative frequency‐dependent selection of genotypes can shape genetic variation on ecologically relevant timescales in natural systems and may be a key, overlooked source of non‐random mortality for tropical plants.     

农林废弃物混配基质的理化性质及其对油茶幼苗生长效应的综合评价

以腐熟的甘蔗渣( SB)、木薯皮( CP)、花生壳( PS)和火炭灰( BA)及园土( GS)为原料,按照不同体积比配制成9种混配基质,并以等体积比泥炭和园土混配基质为对照,对各基质的理化性质及基质中油茶( Camellia oleifera Abel)幼苗的生长状况进行比较;在对基质理化指标和油茶幼苗的壮苗指数进行线性回归分析和通径分析的基础上,采用主成分分析和综合评价法对各基质的育苗效果进行综合评价。结果表明：9种农林废弃物混配基质的容重、总孔隙度和毛管孔隙度分别为0.23~0.47 g·cm-3、60.90%~67.23%和46.53%~58.27%, pH 6.72~pH 7.14,各基质的速效氮含量、速效磷含量、速效钾含量、pH值、电导率和通气孔隙度均高于或显著高于对照,容重则显著低于对照。在不同基质中油茶幼苗的茎粗、单株叶片数、叶长、叶宽、叶绿素相对含量( SPAD)、单株茎叶干质量、单株根干质量和壮苗指数均存在一定差异,而株高和根冠比却无显著差异;其中S9混配基质也V( SB)：V( CP)：V( BA)=2：1：1页中幼苗的大部分生长指标较高,表现出明显的生长优势。线性回归和通径分析结果显示：除通气孔隙度外,基质的其他8个理化指标基本涵盖了影响幼苗壮苗指数的关键因素;其中,速效钾含量是对壮苗指数直接影响最大的负相关因子,而速效磷含量、电导率和总孔隙度对壮苗指数则有较大的正向直接作用,并且三者通过速效钾含量对壮苗指数产生较大的负向间接作用;此外,基质容重对壮苗指数也有一定的负向直接作用。综合评价结果显示：S9和S7混配基质也V( SB)：V( BA)：V( GS)=4：3：3页对油茶育苗效果的综合得分较高,分别为14.363和14.337,建议将S9混配基质作为油茶育苗的首选替代基质,S7混配基质作为备选基质。     

不同BR施用方式诱导黄瓜幼苗对Ca(NO_3)_2胁迫抗性的研究

为探讨外源油菜素内酯(brassinosteroid,BR)诱导黄瓜幼苗对Ca(NO3)2胁迫抗性的效果,研究了3种外源BR施用方法(0.01mg·L-1 BR浸种、0.1mg·L-1 BR喷叶及其二者结合施用)对Ca(NO3)2胁迫(60mmol·L-1)下黄瓜幼苗生长、生理活动以及光合作用的影响。结果表明:(1)3种外源BR方法处理后,Ca(NO3)2胁迫下的黄瓜幼苗株高、茎粗、展开叶片数、叶面积、干重含水量均显著提高,同时其叶片游离脯氨酸和可溶性糖含量上升,过氧化物酶活性提高,而其丙二醛(MDA)含量趋于无Ca(NO3)2胁迫对照的水平;(2)外源BR处理还提高了Ca(NO3)2胁迫下黄瓜幼苗的净光合速率、蒸腾速率和气孔导度,却抑制了Ca(NO3)2胁迫下胞间CO2浓度的升高。研究认为,适宜浓度的外源BR浸种和喷叶处理均可有效增强黄瓜幼苗渗透调节能力,降低细胞膜质过氧化伤害程度,提高抗氧化酶活性和光合效率,从而表现出对Ca(NO3)2胁迫的抗性,并以操作简便、用量极低的0.01mg·L-1 BR浸种方法效果最佳。