Biosynthesis and Catabolism of L-Ascorbic Acid in Plants

L-Ascorbic acid (AsA) is a vital antioxidant compound that plays a critical role in the cellular metabolism of plants and animals. Research on plant AsA metabolism experienced a significant resurgence after 1998 following the identification of AsA-deficient Arabidopsis mutants and the elucidation of a biosynthetic pathway accepted by the overwhelming majority of the plant science community. The identification and cloning of novel biosynthetic genes and the ensuing metabolic engineering of plant AsA content has however revealed a more complex picture. Additional biosynthetic routes have been identified and unexpected biochemical phenotypes were observed upon expression of animal AsA biosynthetic genes. The isolation of novel AsA conjugates from plant tissues and the evidence for long distance transport of AsA in plants have provided additional facets to its functionality. Although some progress has been made regarding the impact of AsA recycling on pool size, we still do not have a clear picture of the biochemistry of AsA degradation. This communication comprehensively reviews new developments in the AsA metabolic system and prompts directions for future research.     

Natural products – modifying metabolite pathways in plants

The diversity of plant natural product (PNP) molecular structures is reflected in the variety of biochemical and genetic pathways that lead to their formation and accumulation. Plant secondary metabolites are important commodities, and include fragrances, colorants, and medicines. Increasing the extractable amount of PNP through plant breeding, or more recently by means of metabolic engineering, is a priority. The prerequisite for any attempt at metabolic engineering is a detailed knowledge of the underlying biosynthetic and regulatory pathways in plants. Over the past few decades, an enormous body of information about the biochemistry and genetics of biosynthetic pathways involved in PNPs production has been generated. In this review, we focus on the three large classes of plant secondary metabolites: terpenoids (or isoprenoids), phenylpropanoids, and alkaloids. All three provide excellent examples of the tremendous efforts undertaken to boost our understanding of biosynthetic pathways, resulting in the first successes in plant metabolic engineering. We further consider what essential information is still missing, and how future research directions could help achieve the rational design of plants as chemical factories for high-value products.     

6. Chasing the enzymes of secondary metabolism: Plant cell cultures as a pot of gold

The development of plant cell cultures for the study of the biosynthesis of secondary metabolises in the 1970s revolutionized the field. It became possible to identify, characterize and ultimately, in specific cases, to purify the biocatalysts involved in the individual transformations. The precise knowledge of the biosynthetic pathways provided by the identification of these enzymes, of the stereo- and substrate- specificity of lhe reactions they catalyse and of the sequence of these reactions in situ opened new fields for study in plant sciences. We now have detailed knowledge of light and elicitor regulation of flavonoid genes, of the molecular mode of action of certain herbicides, and are beginning to form an understanding of the regulation of alkaloid biosynthesis. It is without question that plant cell cultures have become a central, indispensable vehicle in secondary metabolic research.     

植物次生代谢物途径及其研究进展

植物次生代谢是植物在长期进化过程中与环境相互作用的结果,由初生代谢派生。萜类、生物碱类、苯丙烷类为植物次生代谢物的主要类型,其代谢途径多以代谢频道形式存在,具有种属、生长发育期等特异性。从植物次生代谢物的分类、代谢途径及代谢调控基因工程等方面展开论述,重点介绍了次生代谢物的生物合成途径,以及利用基因工程等技术对植物次生代谢途径进行遗传改良等方面的研究进展,为全面认识植物代谢网络、合理定位次生代谢及其关键酶、促进野生植物资源可持续利用等提供理论依据。     

Gibberellin biosynthesis: its regulation by endogenous and environmental signals

The hormone gibberellin (GA) plays an essential role in many aspects of plant growth and development, such as seed germination, stem elongation and flower development. In recent years, exciting progress has been made in understanding how the biosynthesis of this hormone is regulated by endogenous and environmental factors. This has resulted from isolation of genes encoding enzymes involved in GA biosynthesis and metabolism, which also enabled us to manipulate the pathway by modifying the expression of these genes in transgenic plants. In addition, new GA response mutants provided information about how signaling components are involved in feedback regulation of the GA biosynthetic pathway.     

The genetic manipulation of medicinal and aromatic plants

Medicinal and aromatic plants have always been intimately linked with human health and culture. Plant-derived medicines constitute a substantial component of present day human healthcare systems in industrialized as well as developing countries. They are products of plant secondary metabolism and are involved in many other aspects of a plant's interaction with its immediate environment. The genetic manipulation of plants together with the establishment of in vitro plant regeneration systems facilitates efforts to engineer secondary product metabolic pathways. Advances in the cloning of genes involved in relevant pathways, the development of high throughput screening systems for chemical and biological activity, genomics tools and resources, and the recognition of a higher order of regulation of secondary plant metabolism operating at the whole plant level facilitate strategies for the effective manipulation of secondary products in plants. Here, we discuss advances in engineering metabolic pathways for specific classes of compounds in medicinal and aromatic plants and we identify remaining constraints and future prospects in the field. In particular we focus on indole, tropane, nicotine, isoquinoline alcaloids, monoterpenoids such as menthol and related compounds, diterpenoids such as taxol, sequiterpenoids such as artemisinin and aromatic amino acids.     

植物多胺代谢途径研究进展

多胺是一类小分子生物活性物质,广泛存在于生物体内,与植物的生长发育、衰老及抗逆性都有着密切的联系。目前,在植物中的多胺合成途径已经基本揭示,其生理作用在分子水平上逐步得到阐明。对多胺合成突变体和各种转基因植物的研究也使得人们更深入地了解了多胺以及其合成代谢相关酶在植物生长发育等生理过程中的重要作用。以下概述了植物多胺代谢途径,重点综述了代谢途径中各基因的功能及遗传操作的最新进展,并对将来的研究方向尤其是相关基因在植物抗逆境 (包括生物和非生物逆境) 基因工程方面的应用作了讨论。     

植物苯丙烷代谢及其对重金属胁迫的响应研究进展

苯丙烷代谢途径是植物中最重要的次生代谢途径之一,在植物抵抗重金属胁迫中直接或间接发挥了抗氧化作用,并能够提高植物对重金属离子的吸收与胁迫耐性。本文就苯丙烷代谢途径核心反应与关键酶系进行了总结,同时分析了木质素、类黄酮及原花青素等关键代谢产物的生物合成过程及相关机制,并以此为基础探讨了苯丙烷代谢途径关键产物响应重金属胁迫的相关机制。此外,结合当前研究现状,就苯丙烷代谢参与植物防御重金属胁迫的相关研究提出展望,以期为重金属污染环境的植物修复提供理论依据。     

Identification of Cytokinin Biosynthesis Genes in Arabidopsis: A Breakthrough for Understanding the Metabolic Pathway and the Regulation in Higher Plants

The primary biosynthetic reaction of cytokinin is thought to be the isopentenylation of an adenine nucleotide such as AMP with dimethylallylpyrophosphate. For many years, the nature of the enzyme catalyzing this reaction in higher plants had not been identified despite the physiological importance of these compounds. However, the completion of the genomic sequence of Arabidopsis thaliana, a model plant for genetic research, has provided us with new opportunities to solve these problems. Recent studies have revealed the cytokinin biosynthesis enzyme is encoded by a small multigene family that is structurally related to both bacterial adenylate isopentenyltransferase and tRNA isopentenyltransferase. Interestingly, biochemical studies of some of the gene products indicate that ADP and ATP, rather than AMP, are preferentially used as substrates for this biosynthetic reaction. These findings require reconsideration of the currently accepted cytokinin biosynthetic pathway. In addition, there is an increasing body of evidence suggesting that the expression of these cytokinin synthesis genes is affected by the availability of nutrients.     

Regulation of the biosynthesis of plant hormones by cytochrome P450s

 Cytochrome P450 monooxygenases are a large group of heme-containing enzymes, most of which catalyze hydroxylation reactions. Since the discovery of cytochrome P450 in plants, more than 500 forms have been found, and they appear to be involved in the biosynthetic pathways of a large variety of primary and secondary metabolites. In particular, cytochrome P450s are involved in the biosynthesis of plant hormones, and play important roles in the regulation of plant growth and development. Recent genetic and functional analyses of cytochrome P450s in plants have significantly improved our understanding of not only the biosynthetic pathways themselves, but also of plant development from the perspective of hormonal control of morphogenesis. This review summarizes the present status of research on cytochrome P450s' roles in regulating the biosynthesis of plant hormones. Received: January 30, 2002 / Accepted: March 4, 2002     

The effects of season and water availability on chemical composition,secondary metabolites and biological activity in plants

Plants react towards changes in their environment, which can be a result of biotic or abiotic activities. Numerous studies have investigated the effects of abiotic stress on plants, and how it affects the primary as well as secondary metabolism. Generally it is accepted that plants react to environmental stress by increasing secondary metabolites. This is however a very broad and simplified explanation and often inaccurate. Various examples are provided where plants react positively, and often negatively towards seasonal variation and water availability, resulting in a lowering of certain secondary metabolites concentration, while others are increased. Furthermore species differences, cultivars and interaction of other environmental factors such as temperature complicates a simple conclusion from the effect of stress on plants. The differential expression of genes in different species and in different metabolic pathways ensures a complex and very specific reaction of a plant to environmental stress. Overall the paper provides support for a complex and intricate response system which differs for each plant species, and could be explained by understanding and studying the different metabolic pathways responsible for secondary metabolite production.     

植物萜类次生代谢及其调控

植物次生代谢在植物生长发育、环境适应、抵御病虫害等方面发挥着重要作用,这些天然产物组成地球上最丰富的有机化合物的宝库．萜类是植物代谢产物中种类最多的一类,具有重要的生理和生态功能,一些成分还有应用价值．近十几年来,人们在萜类化合物的分离、鉴定、应用、生物合成、相关基因与基因族、酶蛋白结构和功能、代谢调控以及代谢工程等各方面取得了重大进展．本文概述了植物萜类化合物代谢及其调控领域的研究进展与发展趋势．     

Metabolic Engineering of Tropane Alkaloid Biosynthesis in Plants

Over the past decade, the evolving commercial importance of so-called plant secondary metabolites has resulted in a great interest in secondary metabolism and, particularly, in the possibilities to enhance the yield of fine metabolites by means of genetic engineering. Plant alkaloids, which constitute one of the largest groups of natural products, provide many pharmacologically active compounds. Several genes in the tropane alkaloids biosynthesis pathways have been cloned, making the metabolic engineering of these alkaloids possible. The content of the target chemical scopolamine could be significantly increased by various approaches, such as introducing genes encoding the key biosynthetic enzymes or genes encoding regulatory proteins to overcome the specific rate-limiting steps. In addition, antisense genes have been used to block competitive pathways. These investigations have opened up new, promising perspectives for increased production in plants or plant cell culture. Recent achievements have been made in the metabolic engineering of plant tropane alkaloids and some new powerful strategies are reviewed in the present paper.     

Lessons learned from metabolic engineering of cyanogenic glucosides

Plants produce a plethora of secondary metabolites which constitute a wealth of potential pharmaceuticals, pro-vitamins, flavours, fragrances, colorants and toxins as well as a source of natural pesticides. Many of these valuable compounds are only synthesized in exotic plant species or in concentrations too low to facilitate commercialization. In some cases their presence constitutes a health hazard and renders the crops unsuitable for consumption. Metabolic engineering is a powerful tool to alter and ameliorate the secondary metabolite composition of crop plants and gain new desired traits. The interplay of a multitude of biosynthetic pathways and the possibility of metabolic cross-talk combined with an incomplete understanding of the regulation of these pathways, explain why metabolic engineering of plant secondary metabolism is still in its infancy and subject to much trial and error. Cyanogenic glucosides are ancient defense compounds that release toxic HCN upon tissue disruption caused e.g. by chewing insects. The committed steps of the cyanogenic glucoside biosynthetic pathway are encoded by three genes. This unique genetic simplicity and the availability of the corresponding cDNAs have given cyanogenic glucosides pioneering status in metabolic engineering of plant secondary metabolism. In this review, lessons learned from metabolic engineering of cyanogenic glucosides in Arabidopsis thaliana (thale cress), Nicotiana tabacum cv Xanthi (tobacco), Manihot esculenta Crantz (cassava) and Lotus japonicus (bird’s foot trefoil) are presented. The importance of metabolic channelling of toxic intermediates as mediated by metabolon formation in avoiding unintended metabolic cross-talk and unwanted pleiotropic effects is emphasized. Likewise, the potential of metabolic engineering of plant secondary metabolism as a tool to elucidate, for example, the impact of secondary metabolites on plant–insect interactions is demonstrated.     

异戊二烯类植物激素赤霉素和脱落酸的代谢调控

赤霉素和脱落酸在植物生理过程中具有重要的调控作用,其生物合成途径迄今已基本阐明。赤霉素与类胡萝卜素的生物合成途径具有共同前体牻牛儿基牻牛儿基二磷酸,而脱落酸则直接来自于类胡萝卜素。参与这两种植物激素和类胡萝卜素代谢过程的大多数酶基因已经从不同植物中获得克隆;各种调控方式也随着分子生物学的研究工作而得到鉴定。本文就近年来对赤霉素和脱落酸等代谢调控机制及其与植物类胡萝卜素代谢之间关系的研究工作做简要回顾。     

植物天然产物微生物重组合成研究进展

植物天然产物是小分子药物、营养品、化妆品、香精香料等的主要来源之一,在国民经济中发挥重要的作用。目前植物天然产物主要依赖于植物提取,这种生产方式占用耕地、生长周期长,而且植物活性成分往往含量低、生产成本高。通过解析植物天然产物生物合成途径,在微生物细胞中重构,创建细胞工厂,实现利用可再生原料发酵合成,为植物天然产物的供给提供了新的路线。本文重点介绍了中国科学院天津工业生物技术研究所在萜类、黄酮类、苯丙素类等重要类型植物天然产物微生物重组合成方面的研究进展,简要探讨了当前研究面临的挑战及未来前景。