Inorganic nitrogen assimilation in the non-N2-fixing cyanobacterium Phormidium laminosum. I. Cellular levels of glutamine synthetase and NADPH-dependent glutamate dehydrogenase

Cell-free extracts of nitrate-grown as well as of ammonium-grown cells of the filamentous non-nitrogen-fixing cyanobacterium Phormidium laminosum (strain OH-1-p.Cl₁) showed detectable levels of both glutamine synthetase (GS, EC 6.3.1.2) and NADPH-dependent glutamate dehydrogenase (GDH, EC 1.4.1.4) activities. The GS level of nitrate-grown cells was higher than that of ammonium-grown cells, whereas the GDH level was higher in ammonium-grown cells and depended on the external ammonium concentration. When nitrate-grown cells were transferred to an ammonium-containing medium, a decrease of GS and an increase of GDH specific activities occurred, even in the presence of nitrate. Conversely, when ammonia-grown cells were transferred to a nitrate-containing medium, an increase of GS and a decrease of GDH-specific activities took place. Both these effects were inhibited by chloramphenicol and were probably mediated by de novo protein synthesis. When either cell type was transferred to a medium without nitrogen source, the specific activities of both enzymes increased. When nitrate-grown cells were transferred to nitrate medium with L-methionine-DL-sulphoximine (MSX) added, the specific activity of GDH also increased. Here we present some evidence that, under certain conditions of nitrogen availability, GDH would play a minor role in ammonium assimilation.     

Escherichia coli relA strains as hosts for amplification of pBR322 plasmid DNA

Abstract We have proposed that guanosine tetraphosphate produced in Escherichia coli cells subjected to an isoleucine limitation inhibits pBR322 DNA replication [1]. In E. coli relA which cannot synthesize guanosine tetraphosphate (ppGpp) upon amino acid limitation pBR322 DNA is amplified after arginine starvation. The yield of plasmid DNA amplified either by chloramphenicol (Cm) or by arginine limitation is compared. The plasmid yield per cell is equal in amino acid-starved cells and in cells treated with Cm. To increase the plasmid content per ml of cell suspension the growth medium was supplemented with increasing amounts of nutrients. Plasmid DNA can be isolated in large quantities by this procedure. This simple method can be used for the enrichment of pBR325 DNA which cannot be amplified by Cm treatment. Our results indicate that E. coli relA strains might be suitable hosts for the amplification of pBR322 and related plasmids in E. coli .     

Effects of a stay-green mutation on plant nitrogen relations in Lolium perenne during N starvation and after defoliation

The stay-green mutation of the nuclear gene sid results in inhibition of chlorophyll degradation during leaf senescence in grasses, reducing N remobilization from senescing leaves. Effects on growth of Lolium perenne L. were investigated during N starvation (over 18 d) and after severe defoliation, when leaf growth depends on the remobilization of internal N. Rates of dry mater production, partitioning between shoots and roots, and re-partitioning of N from shoots to roots were very similar in stay-green and normal plants under N starvation. Km and Vmax for net uptake of NH4+ were also similar for both genotypes, and Vmax increased with the duration of N deprivation. The mutation had little effect on recovery of leaf growth following severe defoliation, but stay-green plants recommenced NO3- and K+ uptake 1 d later than normal plants. Import of remobilized N into new leaves was generally similar in both lines. However, stay-green plants remobilized less N from stubble compared with normal plants. It was concluded that the sid locus stay-green mutation has no significant adverse effect on the growth of L perenne during N starvation, or recovery from severe defoliation when plants are grown under an optimal regime of NO3- supply both before and after defoliation. The absence of any effect on leaf dry matter production implies that the difference in foliar N availability attributable to this mutation has little bearing on productivity, at least in the short to medium term.     

Carbohydrate metabolism in the stick insect, Carausius morosus

Age-related changes in some parameters related to carbohydrate metabolism in the stick insect, Carausius morosus, were investigated during the 6th instar and up to day 37 of adult life. Total haemolymph carbohydrate concentration and the fat body glycogen content are low and may be related to the low activity of this insect. Trehalose constitutes about 75–80% of the total blood carbohydrate pool. During the moult, total blood carbohydrate, fat body glycogen and haemolymph volume, decrease while glycogen phosphorylase activity of the fat body is slightly activated. The effects are brought about mainly by reduced feeding activity, but may also be influenced by the shedding and replacement of the cuticle. During starvation, blood homeostasis is maintained at the expense of fat body glycogen via an activation of phosphorylase. During reproduction, although no dramatic changes in fat body glycogen levels occur, blood carbohydrates are maintained and fat body phosphorylase is slightly activated. The possibility is discussed that during moulting and reproduction, blood sugar homeostasis is maintained by a hormonal mechanism controlling glycogen phosphorylase. No circadian rhythm in any parameter investigated is observed.     

Modulating the root elongation by phosphate/nitrogen starvation in an OsGLU3 dependant way in rice

Previously, we showed that OsGLU3, a β-1,4-endoglucanase, can affect the cellulose synthesis for root elongation in rice. And the phosphate starvation induced root elongation in rice depends on the function of OsGLU3. Here, we further showed that OsGLU3 is also dispensable for nitrogen starvation induced root elongation in rice.     

Detection of mRNAs correlated with proliferation of cells in suspension cultures of Catharanthus roseus

Gene expression correlated with proliferation was investigated in Catharanthus roseus (L.) G. Don. cells. Polyadenylated RNAs were isolated from cells in proliferative states or in nonproliferative states and the variations in the population and levels of mRNA were analyzed by in vitro translation followed by separation of the corresponding polypeptides by two-dimensional gel electrophoresis. Levels of two mRNAs increased in the actively cycling cells, while they were hardly detected in cells in stationary phase or in cells arrested by starvation of phosphate, sucrose or nitrogen. The molecular masses of the translated products of these two mRNAs were 53 and 60 kDa. No mRNAs were specifically synthesized in common among the various cultures of cells whose growth was arrested by nutrient starvation.     

植物氮素营养与病害发生关系研究进展

氮素营养不仅影响植物的正常生长发育,还会影响植物的感病性或抗病性。该文主要综述了氮素营养及其代谢对植物病害发展的影响、诱导病原菌侵染的寄主氮营养信号和氮素营养对植物与病原菌互作相关基因表达的影响,尤其是氮素受限(饥饿)对病原菌基因的诱导表达、植物衰老基因和抗病基因的关系、植物衰老过程中防御基因的表达等国际研究的热点领域所取得的成果和进展,并讨论了有待深入研究的问题。     

TOTAL LIPID PRODUCTION OF THE GREEN ALGA NANNOCHLOROPSIS SP. QII UNDER DIFFERENT NITROGEN REGIMES1

The green alga Nannochloropsis sp. QII was cultivated in media with sufficient and growth-limiting levels of nitrogen (nitrate). Nitrogen deficiency promoted lipid synthesis yielding cells with lipids comprising 55% of the biomass. The major lipids were triacylglycerols (79%), polar lipids (9%) and hydrocarbons (2.5%). The polar lipids consisted of a broad range of phospholipids, glycolipids and sulfolipids. Other lipids identified were pigments, free fatty acids, saponifiable and unsaponifiable sterol derivatives, various glycerides, a family of alkyl-1, 4-dioxane derivatives and a series of alkyl- and hydroxyalkyl-dimethyl-acetals. Experiments in which ¹⁴CO₂ was provided at different times in the growth cycle demonstrated that enhanced lipid biosynthesis at low nitrogen levels resulted principally from de novo CO₂ fixation.     

沉水植物无机氮利用策略研究进展

沉水植物在水体生态系统中发挥着关键作用,能通过直接和间接途径缓解水体中的氮负荷,促进湖泊生态系统的良性运转。沉水植物在无机氮的利用策略上与陆生植物差异显著,为了适应水环境,沉水植物地上和地下部分能够同时获取环境中的氮,并将其进行向上或向下的运输。为了减少高浓度氮的毒害,沉水植物地上和地下部分在氮利用中存在一定的权衡关系。沉水植物氮同化的主要场所是叶片,主要通过谷氨酰胺合成酶/谷氨酸合成酶（GS /GOGAT）循环和谷氨酸脱氢酶（GDH）途径进行氮的同化。目前针对沉水植物的相关研究还远不及陆生植物,仅涉及到生理响应层面,仍需深入探索沉水植物氮素利用的机理,开发合适的遗传转化体系,并利用基因编辑等技术对基因功能进行验证,对关键蛋白质的结构与功能展开深入研究。     

不同形态N素对水曲柳幼苗生长的影响

在温室内用砂培的方法研究了ＮＯ＾－３－Ｎ、ＮＨ＾＋４－Ｎ及其不同配比对水曲柳（Ｆｒａｘｉｎｕｓ ｍａｎｄｓｈｕｒｉｃａ）幼苗生长的影响。结果表明，水曲柳幼苗在营养液ＮＯ＾－３－Ｎ：ＮＨ＾＋４－Ｎ为７５：２５时生长最好，营养液中ＮＨ＾＋４－Ｎ比例继续增加则生长下降。过量的ＮＨ＾＋４－Ｎ可抑制水曲柳幼苗根系生长，降低幼苗的地下／地上比。营养液中ＮＨ＾＋４－Ｎ比例增加，水曲柳幼苗的净光合速率下降，体内Ｐ     

Influences of Lead (II) Chloride on the Nitrogen Metabolism of Spinach

Lead (Pb²⁺) is a well-known highly toxic element. The mechanisms of the Pb²⁺ toxicity are not well understood for nitrogen metabolism of higher plants. In this paper, we studied the effects of various concentrations of PbCl₂ on the nitrogen metabolism of growing spinach. The experimental results showed that Pb²⁺ treatments significantly decreased the nitrate nitrogen absorption and inhibited the activities of nitrate reductase, glutamate dehydrogenase, glutamine synthase, and glutamic–pyruvic transaminase of spinach, and inhibited the synthesis of organic nitrogen compounds such as protein and chlorophyll. However, Pb²⁺ treatments increased the accumulation of ammonium nitrogen in spinach cell. It implied that Pb²⁺ could inhibit inorganic nitrogen to be translated into organic nitrogen in spinach, thus led to the reduction in spinach growth.     

The effect of nitrogen starvation on the ultrastructure and pigment composition of chloroplasts in the acidothermophilic microalga Galdieria sulphuraria

We studied the effect of nitrogen starvation on growth indices, vitality, ultrastructure, and the photosynthetic apparatus of unique acidothermophilic microalga Galdieria sulphuraria (Galdieri) Merola. Long-term nitrogen starvation ceased G. sulphuraria growth and cell division. During the first days of starvation, phycobiliproteins degraded first, then the content of chlorophyll and carotenoids decreased to trace amounts, chloroplast reduced, cell wall became thinner, and storage compounds accumulated. However, the cells were alive. A comparison with the effects of nitrogen starvation on other photosynthesizing organisms showed that suppression of cell division, reduction of the photosynthetic apparatus to some minimum, and accumulation of storage compounds are a universal response to this stress.     

氮源对C.vulgaris油脂积累的影响

实验室条件下,考察了在发酵过程中不同氮源对小球藻的生物量和油脂积累的影响,确定了小球藻的最佳氮源;并对比分析了含氮培养与缺氮培养的生物量、油脂含量、氮消耗量、生物量氮消耗比率和油脂氮消耗比率的不同。结果表明:小球藻在1.6 g/L Na NO3时获得最大生物量,为562.2 mg/L,在0.8 g/L Na NO3时获得最大相对油脂含量为12.01%;以油脂含量为考察指标时,培养小球藻的最佳氮源为0.8 g/L Na NO3;缺氮培养时,最大油脂含量为13.49%,比含氮培养高约15%;含氮培养时,最高生物量为626.3 mg/L,比缺氮培养高约1.9倍。氮源对生物量,相对油脂含量,生物量氮消耗比率和油脂氮消耗比率具有明显的影响。藉此,提出了通过改变培养方式,达到调控小球藻细胞内生理代谢组分的可行性。     

Possible role of adenylates in the engagement of the cyanideresistant pathway in nutrient-starved Catharanthus roseus Cells

In Cathuranthus roseus (L.) G. Don cells the cyanide-resistant pathway is engaged after phosphate or nitrogen starvation. Re-addition of these nutrients disengaged it again. Re-addition of phosphate leads to a transient disengagement which becomes only permanent after a second addition of phosphate. Disengagement after re-addition of nitrogen is slow: it takes 9 days before the activity has disappeared. In this system the mechanism of engagement of the cyanide-resistant pathway was studied. Addition of phosphate to phosphate-starved cells induced cell division within 24 h. The disengagement of the cyanide-resistant pathway was probably only an indirect effect of phosphate because the cellular P, content, which increased rapidly after addition, was low again before the cyanide-resistant pathway was disengaged. A better correlation was observed between high ADP and adenylate content of the cells and disengagement of the cyanide-resistant pathway. In addition it appeared that the engagement of the cyanide-resistant pathway was not the result of a limited carrier capacity of the cytochrome pathway. It is tentatively concluded that the engagement of the cyanide-resistant pathway in phosphate-starved cells was the result of a limited adenylate content. After nitrogen addition to N-starved cells, it took 5 days until the first growth occurred. Before the cyanide-resistant pathway was disengaged, its activity increased with the increased respiration rate which preceded growth. Within 72 h a higher ADP content was observed, which was still high after 10 days. The stimulation of the cytochrome pathway by uncoupler was small and more or less the same with and without added nitrogen, as long as the cyanide-resistant pathway was engaged. After disengagement the stimulation by uncoupler was significantly larger. It is suggested that the engagement during N-starvation was the result of a limited carrier capacity of the cytochrome pathway. Stimulation of the metabolism by re-addition of phosphate, nitrogen or sucrose resulted in a rapid increase in the levels of uracil nucleotides and uridine diphosphoglucose (UDPG) which are involved in sucrose metabolism.