转基因改良植物的胁迫耐性

干旱、盐碱和低温等逆境是严重影响栽培植物生产的非生物胁迫因素。导入外源目的的基因已发展成为改良作物对逆境胁迫耐性的新途径。现今已应用于植物胁迫改良的基因包括编码活性氧清除酶类、膜修饰酶类、胁迫诱导蛋白和渗调物质合成酶等基因。     

Development of selection marker-free transgenic potato plants with enhanced tolerance to oxidative stress

A binary vector devoid of a plant selection-marker gene (designated as pSSA-F) was constructed to overcome bio-safety concerns about genetically modified plants. This vector carried chloroplast-targeted superoxide dismutase (SOD) and ascorbate peroxidase (APX) genes under the control of an oxidative stress-inducible(SWPA2) promoter, and was utilized to transform potato (Solanum tuberosum L.). Integration of these foreign genes into transgenic plants was primarily performed via PCR with genomic DNA. Twelve marker-free transgenic lines were obtained by inoculating stem explants. The maximum transformation efficiency was 6.25% and averaged 2.2%. Successful integration of the SOD and APX genes rendered transgenic plants tolerant to methyl viologen-mediated oxidative stress at the leaf-disc and whole-plant levels. Our findings suggest that this technique for developing selection marker-free transgenic plants is feasible and can be employed with other crop species.     

Genetic engineering of polyamine and carbohydrate metabolism for osmotic stress tolerance in higher plants

Plant growth and productivity are greatly affected by various stress factors. The molecular mechanisms of stress tolerance in plant species have been well established. Metabolic pathways involving the synthesis of metabolites such as polyamines, carbohydrates, proline and glycine betaine have been shown to be associated with stress tolerance. Introduction of the stress-induced genes involved in these pathways from tolerant species to sensitive plants seems to be a promising approach to confer stress tolerance in plants. In cases where single gene is not enough to confer tolerance, metabolic engineering necessitates the introduction of multiple transgenes in plants.     

植物非生物胁迫相关转录因子研究方法

转录因子是一类能够与启动子区域顺式作用元件特异性结合的蛋白质,是一大类转录调控因子,也是植物中最大的基因家族之一。转录因子可以调节众多下游基因的表达,对植物的生长发育、形态建成、激素调节,以及抵抗多种生物和非生物胁迫具有重要作用。结合近年来转录因子的研究进展,归纳总结了植物非生物胁迫相关转录因子研究的主要策略和方法,包括转录因子结构域、亚细胞定位、转录激活作用、转录因子复合体以及转录因子功能的研究,为植物转录因子的相关研究提供理论和方法的参考。     

Heterologous expression of the AtDREB1A gene in chrysanthemum increases drought and salt stress tolerance

DNA cassette containing an AtDREB1A cDNA and a nos terminator,driven by a cauli- flower mosaic 35S promoter,or a stress-inducible rd29A promoter,was transformed into the ground cover chrysanthemum(Dendranthema grandiflorum)'Fall Color'genome.Compared with wild type plants,severe growth retardation was observed in 35S:DREB1A plants,but not in rd29A:DREB1A plants.RT-PCR analysis revealed that,under stress conditions,the DREB1A gene was over-expressed constitutively in 35S:DREB1A plants,but was over-expressed inductively in rd29A:DREB1A plants.The transgenic plants exhibited tolerance to drought and salt stress,and the tolerance was significantly stronger in rd29A:DREB1A plants than in 35S:DREB1A plants.Proline content and SOD activity were increased inductively in rd29A:DREB1A plants than in 35S:DREB1A plants under stress conditions.These results indicate that heterologous AtDREB1A can confer drought and salt tolerance in transgenic chrysanthemum,and improvement of the stress tolerance may be related to enhancement of proline content and SOD activity.     

Transgenic plants: An insight into oxidative stress tolerance mechanisms

Environmental stresses considerably limit plant productivity. At the molecular level the negative effect of stress is often mediated by reactive oxygen species-initiated oxidative damage. Hence, it was hypothesised that increased tolerance to several environmental constraints could be achieved through enhanced tolerance to oxidative stress. In recent years much effort has been undertaken to improve oxidative stress tolerance by transforming plants with native or bacterial genes coding either for reactive oxygen species-scavenging enzymes or for enzymes modulating the cellular antioxidant capacity. This review deals with data on transgenic plants with altered antioxidant capacity and focuses on the new insight into the antioxidant defence mechanism given by this type of experimental model.     

DREB转录因子在植物非生物胁迫中的作用及应用研究

转录因子也称反式作用因子,是能够与真核生物基因启动子区域中顺式作用元件发生特异性相互作用的DNA结合蛋白。DREB转录因子作为植物特有的转录因子,通过与DRE调控元件特异结合,能促进许多与低温、高盐和干旱相关基因的表达。本文综述了近年DREB转录因子的研究进展,并对其结构和生物学功能、表达调控和信号传递途径以及DREB基因在改良植物抗逆胁迫中的应用进行了讨论,同时对该领域的发展前景进行了展望。     

Heterologous expression of the AtDREB1A gene in chrysanthemum increases drought and salt stress tolerance

DNA cassette containing an AtDREB1A cDNA and a nos terminator, driven by a cauliflower mosaic 35S promoter, or a stress-inducible rd29A promoter, was transformed into the ground cover chrysanthemum (Dendranthema grandiflorum) ‘Fall Color’ genome. Compared with wild type plants, severe growth retardation was observed in 35S:DREB1A plants, but not in rd29A:DREB1A plants. RT-PCR analysis revealed that, under stress conditions, the DREB1A gene was over-expressed constitutively in 35S:DREB1A plants, but was over-expressed inductively in rd29A:DREB1A plants. The transgenic plants exhibited tolerance to drought and salt stress, and the tolerance was significantly stronger in rd29A:DREB1A plants than in 35S:DREB1A plants. Proline content and SOD activity were increased inductively in rd29A:DREB1A plants than in 35S:DREB1A plants under stress conditions. These results indicate that heterologous AtDREB1A can confer drought and salt tolerance in transgenic chrysanthemum, and improvement of the stress tolerance may be related to enhancement of proline content and SOD activity.     

Improvement of plant abiotic stress tolerance through modulation of the polyamine pathway

Polyamines(mainly putrescine(Put),spermidine(Spd),and spermine(Spm))have been widely found in a range of physiological processes and in almost all diverse environmental stresses.In various plant species,abiotic stresses modulated the accumulation of polyamines and related gene expression.Studies using loss-of-function mutants and transgenic overexpression plants modulating polyamine metabolic pathways confirmed protective roles of polyamines during plant abiotic stress responses,and indicated the possibility to improve plant tolerance through genetic manipulation of the polyamine pathway.Additionally,putative mechanisms of polyamines involved in plant abiotic stress tolerance were thoroughly discussed and crosstalks among polyamine,abscisic acid,and nitric oxide in plant responses to abiotic stress were emphasized.Special attention was paid to the interaction between polyamine and reactive oxygen species,ion channels,amino acid and carbon metabolism,and other adaptive responses.Further studies are needed to elucidate the polyamine signaling pathway,especially polyamine-regulated downstream targets and the connections between polyamines and other stress responsive molecules.     

Accumulation of glycinebetaine in rice plants that overexpress choline monooxygenase from spinach and evaluation of their tolerance to abiotic stress

BACKGROUND AND AIMS: Glycinebetaine (GB), a quaternary ammonium compound, is a very effective compatible solute. In higher plants, GB is synthesized from choline (Cho) via betaine aldehyde (BA). The first and second steps in the biosynthesis of GB are catalysed by choline monooxygenase (CMO) and by betaine aldehyde dehydrogenase (BADH), respectively. Rice (Oryza sativa), which has two genes for BADH, does not accumulate GB because it lacks a functional gene for CMO. Rice plants accumulate GB in the presence of exogenously applied BA, which leads to the development of a significant tolerance to salt, cold and heat stress. The goal in this study was to evaluate and to discuss the effects of endogenously accumulated GB in rice. METHODS: Transgenic rice plants that overexpressed a gene for CMO from spinach (Spinacia oleracea) were produced by Agrobacterium-mediated transformation. After Southern and western blotting analysis, GB in rice leaves was quantified by (1)H-NMR spectroscopy and the tolerance of GB-accumulating plants to abiotic stress was investigated. KEY RESULTS: Transgenic plants that had a single copy of the transgene and expressed spinach CMO accumulated GB at the level of 0.29-0.43 micromol g(-1) d. wt and had enhanced tolerance to salt stress and temperature stress in the seedling stage. CONCLUSIONS: In the CMO-expressing rice plants, the localization of spinach CMO and of endogenous BADHs might be different and/or the catalytic activity of spinach CMO in rice plants might be lower than it is in spinach. These possibilities might explain the low levels of GB in the transgenic rice plants. It was concluded that CMO-expressing rice plants were not effective for accumulation of GB and improvement of productivity.     

Altered physiology in trehalose-producing transgenic tobacco plants: Enhanced tolerance to drought and salinity stresses

Transgenic tobaccoNicotiana tabacum L. var. SR1) plants that over-express theEscherichia coli trehalose-6-phosphate synthase (TPS) gene(otsA) synthesized small amounts of trehalose (<400 μg g^-1 leaf) while non-transformants produced no detectable trehalose. Some transgenic plants expressing a high level ofotsA exhibited stunted growth and morphologically altered leaves. We tested F₂2 homozygous plants devoid of phenotypic changes to determine their physiological responses to dehydration and salinity stresses. All transgenic plants maintained better leaf turgidity under a limited water supply or after treatment with polyethylene glycol (PEG). Furthermore, fresh weight was maintained at higher levels after either treatment. The initial leaf water potential was higher in transgenic plants than non-transformants, but, in both plant types, was decreased to a comparable degree following dehydration. When grown with 250 mM NaCl, transgenic plants exhibited a significant delay in leaf withering and chlorosis, as well as more efficient seed germination. Our results suggest that either trehalose or trehalose-6-phosphate can act as an osmoprotective molecule without maintaining water potential, in contrast to other osmolytes. Furthermore, both appear to protect young embryos under unfavorable water status to ensure subsequent germination.     

非生物胁迫相关NAC转录因子的结构及功能

NAC是植物特有的一类转录因子,参与植物多个生长发育过程,还参与植物对逆境胁迫的响应。本文对非生物胁迫相关NAC转录因子的结构特征、功能预测、表达特性、在转基因植物中的作用及调控路径进行综述。非生物胁迫相关NAC转录因子具有典型的NAc胁迫亚家族结构特征,根据这些结构特征可以预测其功能;非生物胁迫相关NAc转录因子能响应多种非生物胁迫,其转基因过表达大多能使转基因植物提高一种或几种胁迫耐受性;非生物胁迫相关NAc转录因子有着复杂的调控路径。这些NAc转录因子可用于提高转基因植物的逆境耐受性。     

NAC转录因子在植物抗病和抗非生物胁迫反应中的作用

NAC转录因子是植物特有的一类转录因子,其共同特点是在N端含有一段高度保守、由约150个氨基酸组成的NAC结构域,而C端为高度变异的转录调控区。研究表明,NAC转录因子不仅参与植物生长发育的调控,而且在植物抗逆反应中具有重要的调控作用。文章着重介绍NAC转录因子在植物抗逆反应中的作用及其调控机制,并简要讨论NAC转录因子生物学功能的研究方向。