基因芯片在细菌学研究中的应用

目前基因芯片技术广泛应用于生物学研究的各个领域,在细菌学研究中基因芯片技术突破了以往单个或数个基因研究的局限,在细菌功能基因组学、药物作用靶点、细菌-宿主细胞间相互作用以及细菌进化等方向发挥着重要作用,初步揭示了细菌生物功能多样性受到多个基因的调控,但相关的研究尚有待深入.     

脯氨酸代谢与植物抗渗透胁迫的研究进展

脯氨酸被认为是植物和细菌内的一种相容渗透剂,有助于植物和细菌抵御渗透胁迫。本文就近年来有关植物体内脯氨酸合成和代谢、脯氨酸含量受渗透胁迫的影响情况、脯氨酸合成降解有关的酶及其基因、脯氨酸在细胞中的运输和定位、ABA与脯氨酸的诱导合成以及脯氨酸和植物抗渗透胁迫关系的研究进展作了简要综述。     

植物内生细菌的研究

植物内生细菌是能够定殖在植物细胞间隙或细胞内,并与寄主植物建立和谐联合关系的一类微生物I’］。自本世纪初开始,人们不断地从植物的根、叶、甚至茎和种子上分离并鉴定出多种微生物,但其在寄主植物中的生物学作用却未能引起研究者的重视。近年来,由于一些研究结果表明内生细菌能够作为外源基因的载体,又具有植物保护剂的功能,可以产生植物促生物质或可作为联合固氮菌剂,由此有关内生细菌的研究才引起植物学家和微生物学家的兴趣。l植物内生细菌的研究现状有关植物内生细菌的研究主要涉及两方面的内容：（l）生态学研究：内生细菌…     

脯氨酸代谢与植物抗渗透胁迫的研究进展

脯氨酸被认为是植物和细菌内的一种相容渗透剂,有助于植物和细菌抵御渗透胁迫。本文就近年来有关植物体内脯氨酸合成和代谢、脯氨酸含量受渗透胁迫的影响情况、脯氨酸合成降解有关的酶及其基因、脯氨酸在细胞中的运输和定位、ＡＢＡ与脯氨酸的诱导合成以及脯氨酸和植物抗渗透胁迫关系的研究进展作了简要综述。     

PLD基因的基本功能及在植物中的利用研究现状

磷脂酶D(PLD)是一种重要的磷脂水解酶,它广泛分布于各种植物、动物和微生物(细菌、真菌)中。近几年的研究发现它在植物细胞信号转导、膜运输及降解、脂降解、细胞分化和生殖、细胞防御反应中均发挥重要作用。本文从PLD的基本特性、功能、应用等方面综述了其最新研究进展并讨论了其在植物中的应用前景。     

甾醇在调节植物生长发育中的研究进展

甾醇是一种异戊二烯类化合物,在生物的生长发育中起着重要作用。甾醇不仅是真核细胞膜的结构成分,而且也是甾醇激素生物合成的前体,在植物细胞分裂、胚胎发生和发育、参与逆境胁迫中起着关键作用。在植物中,甾醇衍生的油菜素内酯(brassinosteroids,BRs)在生长发育中的多种功能已被广泛研究,BRs作为一类植物激素,协同其他激素在植物生长发育中发挥多种功能,从细胞分裂、细胞扩张、气孔导度和根系发育,BRs在植物生命周期的各个方面都发挥着重要的作用。除了这些功能外,BRs作为植物甾醇合成途径的重要产物,作为一种重要的信号分子响应逆境胁迫及调控植物的形态建成。本文对BRs合成途径中的相关基因的研究进展进行了概述,并且综述了油菜素内酯的生物合成及其在调节植物生长发育中的研究进展,最后对油菜素内酯的研究前景进行了讨论和展望。     

吲哚乙酸跨界信号调节植物与细菌互作

吲哚-3-乙酸(indole-3-acetic acid,IAA)作为植物体内普遍存在的内源生长素参与调节植物生命活动的诸多方面。研究发现,自然界中不仅植物可以合成IAA,许多微生物(包括植物病原菌或益生菌)同样具有分泌IAA的能力,可以诱发植物病害,或促进植物生长。有趣的是IAA不仅作为细菌的次生代谢物干扰寄主植物的激素稳态,也作为信号分子影响细菌基因表达和生理活动,通过整合进入细菌复杂代谢网络,调节植物与细菌的相互作用。通过讨论植物相关细菌IAA的生物合成途径及其调控,以及参与调节细菌基因表达、影响细菌生理和行为及其与寄主植物的互作等,概述该领域的研究动态与进展,揭示IAA不仅调节植物生长发育和防御,也作为跨界信号在调控植物与微生物互作中发挥重要作用,旨在为深入研究和更好地了解IAA跨界信号机制,通过遗传操纵细菌IAA信号通路以改善植物生长发育及其胁迫耐力提供新思路。     

土壤微生物膜生理生态功能研究进展

土壤微生物膜是由土壤细菌及其分泌物积聚形成的生物群落,是生物土壤结皮的初始形态和重要组成部分。作为土壤细菌生命过程中最典型的生存形式,土壤微生物膜不仅能保护基质内细胞生存,还可黏附于土壤颗粒和植物根系表面,发挥重要的生态功能。本文在解析土壤微生物膜结构与组成的基础上,从土壤质量与植物健康两个方面总结分析了土壤微生物膜生理生态功能:土壤微生物膜代谢活性高于游离细胞,可高效分泌胞外聚合物并且具有更强的有机物质转化速率,在提升土壤肥力,吸附、固持和降解土壤污染物和促进土壤团聚体形成方面具有重要意义;土壤微生物膜可通过多种微生物间协同作用、促进分泌多种促生物质与胞外聚合物以发挥固持作用等改善植物养分利用状况,增强植物抗逆性。揭示土壤微生物膜生态功能的微观机制、筛选和应用功能性土壤微生物膜是未来重要的发展方向。     

细菌中I型毒素蛋白与细胞膜相互作用机制的研究进展

I型毒素-抗毒素（TA）系统在细菌基因组中广泛存在,在细菌的生长、生存中发挥多种生物学功能,包括抗菌、红细胞毒性、促进持留菌形成、抑制细菌生长或导致细菌休眠等。绝大部分I型毒素蛋白以细胞膜作为靶标,目前已知的一种作用机制是在细胞膜上形成孔洞结构,造成膜电位的下降或细胞膜的破坏,从而抑制ATP的合成或导致细菌死亡;另一种可能的作用机制是毒素蛋白作用在细胞膜上,改变细胞的形状,导致细胞进入休眠状态。I型毒素蛋白-细胞膜作用机制的复杂性和生物功能的多样性远超预期。因此,解析I型毒素蛋白在不同细胞膜中的组装机制及其所形成结构特征就变得非常重要,这也是揭示其结构-功能关系的关键。本文通过综述已报道的I型TA系统的结构特征与生物学功能,结合对其跨膜结构域的预测,探讨了其可能在细胞膜中形成的不同结构及其对功能的影响,分析了影响作用机制的关键因素。这些研究既给耐药细菌的治疗带来机遇,又为新型抗菌药物的研发带来思路。     

生长素和细胞分裂素调控植物根和微生物互作的研究进展

植物在与其生长环境中的微生物长期共同进化过程中形成了复杂而微妙的共生体系。生长素和细胞分裂素等植物激素不仅调控植物的自身发育,而且在植物-微生物互作中发挥重要的调节作用。综述了生长素和细胞分裂素调控植物根和土壤微生物包括有益菌或病原细菌和真菌间相互作用的最新研究进展,旨在揭示生长素和细胞分裂素调控功能的共性和特异性,为提高农作物的产量和抵抗微生物病害能力提供理论和实践指导。     

脯氨酸在植物生长和非生物胁迫耐受中的作用

脯氨酸是生物界分布最广的渗透保护物质之一,干旱、高盐、高温及重金属等非生物胁迫条件都会导致植物体内脯氨酸含量的增加,其作用是防止渗透胁迫对植物造成的伤害、清除自由基,还可以作为氮、碳以及NADPH的重要来源。近年来,在转化脯氨酸代谢相关基因提高植物胁迫抗性方面也取得了很大进展。本文概要介绍了脯氨酸在植物生长和耐受非生物胁迫中的作用、与植物脯氨酸累积有关的信号转导、胁迫条件下脯氨酸的吸收和器官间的运输途径,以及通过转基因技术过量表达脯氨酸提高植物胁迫耐性的代谢工程的进展。     

不同植物在水分胁迫条件下脯氨酸的累积与抗旱性的关系

Ca植物春小麦幼苗叶片累积脯氨酸的数量随水分胁迫时间和强度递增,累积的数量与品种的抗旱性没有相关性。C_4植物玉米和高粱累积脯氨酸的数量低于小麦,但高粱多于玉米。CAM植物落地生根脯氨酸含量略有变化。旱生植物细枝岩黄芪的同化枝累积脯氨酸的数量高于中生植物钻天杨和槐的叶片,而梭梭的同化校则低于槐而高于钻天杨。表明这些植物累积脯氨酸的数量与它们的抗旱性无夫。因此似不宜用脯氨酸数量的多少作为植物抗旱性的生理指标。     

Proline as a stress protectant in yeast: physiological functions, metabolic regulations, and biotechnological applications

Proline is an important amino acid in terms of its biological functions and biotechnological applications. In response to osmotic stress, proline is accumulated in many bacterial and plant cells as an osmoprotectant. However, it has been shown that proline levels are not increased under various stress conditions in the yeast Saccharomyces cerevisiae cells. Proline is believed to serve multiple functions in vitro such as protein and membrane stabilization, lowering the T _m of DNA, and scavenging of reactive oxygen species, but the mechanisms of these functions in vivo are poorly understood. Yeast cells biosynthesize proline from glutamate in the cytoplasm via the same pathway found in bacteria and plants and also convert excess proline to glutamate in the mitochondria. Based on the fact that proline has stress-protective activity, S. cerevisiae cells that accumulate proline were constructed by disrupting the PUT1 gene involved in the degradation pathway and by expressing the mutant PRO1 gene encoding the feedback inhibition-less sensitive γ-glutamate kinase to enhance the biosynthetic activity. The engineered yeast strains successfully showed enhanced tolerance to many stresses, including freezing, desiccation, oxidation, and ethanol. However, the appropriate cellular level and localization of proline play pivotal roles in the stress-protective effect. These results indicate that the increased stress protection is observed in yeast cells under the artificial condition of proline accumulation. Proline is expected to contribute to yeast-based industries by improving the production of frozen dough and alcoholic beverages or breakthroughs in bioethanol production.     

植物脯氨酸合成酶基因工程研究进展

逆境条件下植物细胞积累脯氨酸是植物适应逆境的一种反应,有利于减轻逆境对植物的伤害。简要回顾了生物体脯氨酸代谢过程和逆境下植物积累脯氨酸的作用,重点总结了植物体内脯氨酸合成酶基因吡咯啉-5羧-酸合成酶(P5CS)的克隆、表达和转基因研究进展。研究认为,P5CS是一个逆境胁迫应答基因,通过基因工程方法调节P5CS基因的表达有利于提高植物体脯氨酸的积累量,改善植物的抗逆性。因此,目前可以在大田植物中开展利用提高脯氨酸来改善植物抗逆性的研究。     

Proline metabolism and transport in plant development

Proline fulfils diverse functions in plants. As amino acid it is a structural component of proteins, but it also plays a role as compatible solute under environmental stress conditions. Proline metabolism involves several subcellular compartments and contributes to the redox balance of the cell. Proline synthesis has been associated with tissues undergoing rapid cell divisions, such as shoot apical meristems, and appears to be involved in floral transition and embryo development. High levels of proline can be found in pollen and seeds, where it serves as compatible solute, protecting cellular structures during dehydration. The proline concentrations of cells, tissues and plant organs are regulated by the interplay of biosynthesis, degradation and intra- as well as intercellular transport processes. Among the proline transport proteins characterized so far, both general amino acid permeases and selective compatible solute transporters were identified, reflecting the versatile role of proline under stress and non-stress situations. The review summarizes our current knowledge on proline metabolism and transport in view of plant development, discussing regulatory aspects such as the influence of metabolites and hormones. Additional information from animals, fungi and bacteria is included, showing similarities and differences to proline metabolism and transport in plants.     

Proline as a biochemical marker in relation to the ecology of two halophytic Juncus species

Aims Osmolytes, used for maintaining osmotic balance and as 'osmoprotectants', are synthesized in plants as a general, conserved response to abiotic stress, although their contribution to stress-tolerance mechanisms remains unclear. Proline, the most common osmolyte, accumulates in many plant species in parallel with increased external salinity and is considered a reliable biochemical marker of salt stress. We have measured proline levels in two halophytic, closely related Juncus species under laboratory and field conditions to assess the possible relevance of proline biosynthesis for salt tolerance and therefore for the ecology of these two taxa.Methods Proline was quantified in plants treated with increasing NaCl concentrations and in plants sampled in two salt marshes located in the provinces of Valencia and Alicante, respectively, in southeast Spain. Electrical conductivity, pH, Na + and Cl ? concentrations were measured in soil samples collected in parallel with the plant material.Important findings Treatment with NaCl inhibited growth of J. acutus plants in a concentration-dependent manner, but only under high salt conditions for J. maritimus. Salt treatments led to proline accumulation in both species, especially in the more salt-tolerant J. maritimus. The results, obtained under laboratory conditions, were confirmed in plants sampled in the field. In all the samplings, proline contents were significantly lower in J. acutus than in the more tolerant J. maritimus growing in the same area. No direct correlation between soil salinity and proline levels could be established, but seasonal variations were detected, with increased proline contents under accentuated water deficit conditions. Our results suggest that proline biosynthesis is not only an induced, general response to salt stress but also an important contributing factor in the physiological mechanisms of salt tolerance in Juncus, and that it therefore correlates with the ecology of both species.     

Proline and hydroxyproline metabolism: implications for animal and human nutrition

Proline plays important roles in protein synthesis and structure, metabolism (particularly the synthesis of arginine, polyamines, and glutamate via pyrroline-5-carboxylate), and nutrition, as well as wound healing, antioxidative reactions, and immune responses. On a per-gram basis, proline plus hydroxyproline are most abundant in collagen and milk proteins, and requirements of proline for whole-body protein synthesis are the greatest among all amino acids. Therefore, physiological needs for proline are particularly high during the life cycle. While most mammals (including humans and pigs) can synthesize proline from arginine and glutamine/glutamate, rates of endogenous synthesis are inadequate for neonates, birds, and fish. Thus, work with young pigs (a widely used animal model for studying infant nutrition) has shown that supplementing 0.0, 0.35, 0.7, 1.05, 1.4, and 2.1% proline to a proline-free chemically defined diet containing 0.48% arginine and 2% glutamate dose dependently improved daily growth rate and feed efficiency while reducing concentrations of urea in plasma. Additionally, maximal growth performance of chickens depended on at least 0.8% proline in the diet. Likewise, dietary supplementation with 0.07, 0.14, and 0.28% hydroxyproline (a metabolite of proline) to a plant protein-based diet enhanced weight gains of salmon. Based on its regulatory roles in cellular biochemistry, proline can be considered as a functional amino acid for mammalian, avian, and aquatic species. Further research is warranted to develop effective strategies of dietary supplementation with proline or hydroxyproline to benefit health, growth, and development of animals and humans.     

Activation of a new proline transport system in Salmonella typhimurium.

Proline uptake can be mediated by three different transport systems in wild-type Salmonella typhimurium: a high-affinity proline transport system encoded by the putP gene and two glycine-betaine transport systems with a low affinity for proline encoded by the proP and proU genes. However, only the PutP permease transports proline well enough t allow growth on proline as a sole carbon or nitrogen source. By selecting for mutations that allow a putP mutant to grow on proline as a sole nitrogen source, we isolated mutants (designated proZ) that appeared to activate a cryptic proline transport system. These mutants enhanced the transport of proline and proline analogs but did not require the function of any of the known proline transport genes. The mutations mapped between 75 and 77.5 min on the S. typhimurium linkage map. Proline transport by the proZ mutants was competitively inhibited by isoleucine and leucine, which suggests that the ProZ phenotype may be due to unusual mutations that alter the substrate specificity of the branched-chain amino acid transport system encoded by the liv genes.     

Proline oxidase in cultured mammalian cells

We sought a cultured cell line with Proline Oxidase activity to study the regulation and physiologic role of the enzyme in mammalian tissues. Among the cell lines tested, only LLC-RK1 cells, derived from rabbit kidney, had significant Proline Oxidase activity; the Km for proline of the enzyme from these cells was similar to that for the liver enzyme. LLC cells, Proline Oxidase positive, were able to convert proline to CO2. In contrast, CHL cells, Proline Oxidase negative, did not have this capability. The presence of Proline Oxidase in LLC cells and the absence of the enzyme in fibroblasts suggest that Proline Oxidase may serve as a marker enzyme for distinguishing parenchymal kidney cells from fibroblasts in culture. Cells transformed by SV40 virus and cells transformed by methylcholanthrene had activities higher that the parent cell line, but this effect of transformation could not be generalized to all transformed cells. Finally, L-hydroxy proline at 100-fold greater concentration than substrate L-proline failed to decrease proline oxidation. This finding suggests distinct degradative enzymes for these two amino acids.