Expression of biotransformation genes in woodrat (Neotoma) herbivores on novel and ancestral diets: identification of candidate genes responsible for dietary shifts

The ability of herbivores to switch diets is thought to be governed by biotransformation enzymes. To identify potential biotransformation enzymes, we conducted a large-scale study on the expression of biotransformation enzymes in herbivorous woodrats ( Neotoma lepida ). We compared gene expression in a woodrat population from the Great Basin that feeds on the ancestral diet of juniper to one from the Mojave Desert that putatively switched from feeding on juniper to feeding on creosote. Juniper and creosote have notable differences in secondary chemistry, and thus, should require different biotransformation enzymes for detoxification. Individuals from each population were fed juniper and creosote diets separately. After the feeding trials, hepatic mRNA was extracted and hybridized to laboratory rat microarrays. Hybridization of woodrat samples to biotransformation probes on the array was 87%, resulting in a total of 224 biotransformation genes that met quality control standards. Overall, we found large differences in expression of biotransformation genes when woodrats were fed juniper vs. creosote. Mojave woodrats had greater expression of 10× as many biotransformation genes as did Great Basin woodrats on a creosote diet. We identified 24 candidate genes that may be critical in the biotransformation of creosote toxins. Superoxide dismutase, a free radical scavenger, was also expressed to a greater extent by the Mojave woodrats and may be important in controlling oxidative damage during biotransformation. The results are consistent with the hypothesis that biotransformation enzymes limit diet switching and that woodrats in the Mojave have evolved a unique strategy for the biotransformation of creosote toxins.     

Mixed function oxidases and herbivore polyphagy: the devil's advocate position

Abstract. 1. Evidence used to support the theory that mixed function oxidases (MFOs) are important in the survival of polyphagous herbivores is briefly reviewed. This evidence includes data on patterns of variation in MFO activity among species and among developmental stages within species. It also includes data on induction of MFO activity by plant compounds and metabolism of plant cornpounds by MFOs.
2. It is argued that the evidence presently available does not offer strong support of the theory because key pieces of information are lacking. Evidence which tends to refute the theory is reviewed and discussed.
3. Experiments are proposed which could more rigorously test the theory.     

Chemical Properties of the Diets of Two Lemur Species in Southwestern Madagascar

Seasonal dietary variations demonstrate the importance of certain plant parts during the year. A parallel analysis of their nutritional constituents provides further information on underlying patterns of consumption of the plant parts and the relative importance of key nutrients. I studied the diets of Lemur catta (ring-tailed lemurs) and Propithecus verreauxi verreauxi (sifakas), for 9 mo over a 13-mo period in the highly seasonal tropical dry forest site of Beza Mahafaly in southwestern Madagascar. I tested dietary plant parts for nutrients—protein, free amino acids, and sugars—and for 2 potential deterrents, phenolics and tannins, using plant extracts prepared in the field. I compared consumption of nutrients and secondary compounds throughout the year and between seasons. Nutrients are balanced throughout the year. The 2 lemur species do not appear nutrient-starved in either season, though actual quantities of nutrients and contributing food parts differ for each species. Lemur catta consumes high levels of sugar throughout the year, whereas Propithecus takes in higher levels of protein. The effects of phenolics and tannins are quantitative, and they appear to deter consumption of plant parts only past a certain threshold. Sifakas consume them in greater quantities than those of ring-tailed lemurs, which appear more sensitive to their effects. Sifakas may have a higher tolerance for secondary plant metabolites, which is consistent with reports for other folivores. The overall stability of nutrients throughout the year indicates no lean period that coincides with the decline in food abundance during the dry season, though actual caloric intake probably decreases.     

植食性哺乳动物与植物协同进化研究进展

从动物-植物协同进化模式,植物对动物采食反应及动物对植物防卫的适应对策等方面综述了以植物次生化合物为媒介的植食性哺乳动物-植物协同进化的研究进展,动物与植物的协同进化模式包括成对协同进化,扩散协同进化,躲避-辐射协同进化,多样性的协同进化,平行分枝进化,互惠进化等模式,植物不仅以超补偿反应,物理防卫作为对植食性动物采食的应答,延长植食性动物的觅食时间,降低植食性动物的觅食效率,更能以其派生的次生化合物抑制动物的摄食,进而影响其消化,代谢及生长等生理生态特征,动物通过改变觅食行为,调整对各食物项目的相对摄入量,减少次生化合物的摄入量,动物还通过氧化,还原,络合,改变消化道内环境,形成相应的降解酶,改变代谢率等途径降低次生化合物对其的负作用。     

Glycosyltransferases:key players involved in the modification of plant secondary metabolites

Glycosyltransferases are members of the multigene superfamily in plants that can transfer single or multiple activated sugars to a range of plant molecules,resulting in the glycosylation of plant compounds.Although the activities of many glycosyltransferases and their products have been recognized for a long time,only in recent years were some glycosyltransferase genes identified and a few functionally characterized in detail.Glycosylation is thought to be one of the most important modification reactions towards plant secondary metabolites,and plays a key role in maintaining cell homeostasis,thus likely participating in the regulation of plant growth,development and in defense responses to stress environments.With advances in plant genome projects and the development of novel technologies in analyzing gene function,significant progress could be made in gaining new insights into the properties and precise biological roles of plant secondary product glycosyltransferases,and the new knowledge will have extensive application prospects in the catalytic synthesis of glycoconjugates and metabolic engineering of crops.In this review,we summarize the current research,highlighting the possible biological roles,of plant secondary metabolite glycosyltransferases and discuss their potential applications as well as aspects to be further studied in the near future.     

Glycosyltransferases: key players involved in the modification of plant secondary metabolites

Glycosyltransferases are members of the multigene superfamily in plants that can transfer single or multiple activated sugars to a range of plant molecules, resulting in the glycosylation of plant compounds. Although the activities of many glycosyltransferases and their products have been recognized for a long time, only in recent years were some glycosyltransferase genes identified and a few functionally characterized in detail. Glycosylation is thought to be one of the most important modification reactions towards plant secondary metabolites, and plays a key role in maintaining cell homeostasis, thus likely participating in the regulation of plant growth, development and in defense responses to stress environments. With advances in plant genome projects and the development of novel technologies in analyzing gene function, significant progress could be made in gaining new insights into the properties and precise biological roles of plant secondary product glycosyltransferases, and the new knowledge will have extensive application prospects in the catalytic synthesis of glycoconjugates and metabolic engineering of crops. In this review, we summarize the current research, highlighting the possible biological roles, of plant secondary metabolite glycosyltransferases and discuss their potential applications as well as aspects to be further studied in the near future.     

Engineering the plant cell factory for secondary metabolite production

Plant secondary metabolism is very important for traits such as flower color, flavor of food, and resistance against pests and diseases. Moreover, it is the source of many fine chemicals such as drugs, dyes, flavors, and fragrances. It is thus of interest to be able to engineer the secondary metabolite production of the plant cell factory, e.g. to produce more of a fine chemical, to produce less of a toxic compound, or even to make new compounds, Engineering of plant secondary metabolism is feasible nowadays, but it requires knowledge of the biosynthetic pathways involved. To increase secondary metabolite production different strategies can be followed, such as overcoming rate limiting steps, reducing flux through competitive pathways, reducing catabolism and overexpression of regulatory genes. For this purpose genes of plant origin can be overexpressed, but also microbial genes have been used successfully. Overexpression of plant genes in microorganisms is another approach, which might be of interest for bioconversion of readily available precursors into valuable fine chemicals. Several examples will be given to illustrate these various approaches. The constraints of metabolic engineering of the plant cell factory will also be discussed. Our limited knowledge of secondary metabolite pathways and the genes involved is one of the main bottlenecks. This revised version was published online in August 2006 with corrections to the Cover Date.     

The relationship between diet and the size of the midgut caeca in grasshoppers (Insecta: Orthoptera: Acridoidea)

A survey of the size and form of the midgut caeca in relation to diet has been carried out on 173 species from 21 families and subfamilies of Acridoidea (grasshoppers). Although differences exist in the size of the anterior caecal arms relative to body length, these differences are not related to the type of food eaten. Assuming that the anterior arms have a key role in digestive and absorptive processes, this suggests that different foods make similar demands on these processes. The posterior caecal arms are smaller in graminivorous species than in species eating other types of plants as a whole or part of their diet. This is true across all the taxa, including those families and subfamilies that are predominantly forb-feeding. It is suggested that the posterior caecal arms have a special role in the detoxification of plant secondary compounds and that the requirement for this is reduced in graminivorous species because of the lower levels of toxic secondary compounds in grasses. A specialized pocket region is present in the posterior caecal arms of some forb-feeding species. Its occurrence across the taxa is spasmodic. It may be concerned with the removal of phenolic compounds.     

Methyl jasmonate-induced nicotine production in Nicotiana attenuata: inducing defenses in the field without wounding

The functional significance of herbivore-induced plant traits known to directly or indirectly influence herbivore performance remains largely untested under field conditions due to the difficulty of uncoupling the response to herbivory from the act of herbivory. The signals that activate many of the induced responses in plants are endogenously produced in response to wounding, unlike many of the predator-induced responses found in aquatic invertebrates (which are activated by exogenous cues derived from predators). Jasmonates, endogenously-produced damage signals, activate diverse wound-induced responses in plants including induced nicotine production in Nicotiana sylvestris. The results presented here are from two experiments which illustrate the use of jasmonates to uncouple induced nicotine production in Nicotiana attenuata (Torrey ex. Watson) from wounding. The exogenous addition of methyl jasmonate (MJ) in small quantities (11 g for a 1.4 g dry mass plant) to roots of hydroponically-grown plants induces de novo nicotine synthesis and increases whole-plant nicotine concentrations just as wounding does. The MJ-induced changes were proportional to the quantity of MJ given. Moreover, the effects of MJ were additive to the effects of damage. Applications of MJ to shoots were less effective. Root treatments also worked with plants growing in a field plot. The application of MJ represents a promising tool for examining the functional significance of induced nicotine responses in plants growing in their native environments.     

Effect of plant secondary compounds on in vitro methane,ammonia production and ruminal protozoa population

Aims

The objective of this study was to evaluate the potential of secondary plant metabolites from 38 sources to serve as antimethanogenic additives in ruminant diets. The effect of leaf tannins from these different plant sources on rumen fermentation, protozoal populations and methanogenesis was also studied.

Methods and Results

Samples (200 mg dry matter, DM) were incubated without and with polyethylene glycol (PEG)‐6000 (400 mg DM) as a tannin binder during 24‐h incubation in the in vitro Hohenheim gas system. In the leaf samples, total phenol (g kg^?1 DM) was maximum in Pimenta officinalis (312) followed by Oenothera lamarckiana (185) and Lawsonia inermis (105). Of the 38 samples, condensed tannins exceeded 4·0 g kg^?1 in only Alpinia galanga (7·50), Cinnamomum verum (4·58), Pelargonium graveolens (18·7) and Pimenta officinalis (23·2) and were not detected in seven samples. When the bioactivity of the leaf samples was assessed using the tannin bioassay, the percentage increase in the amount of gas produced during incubation of samples with the tannin‐binding agent PEG‐6000 over the amount produced during incubation without the tannin binder ranged from nil (zero) to 367%, with the highest being recorded with A. galanga leaves. The ratio of methane reduction per ml of total gas reduction was maximum with Rauvolfia serpentina (131·8) leaves, followed by Indigofera tinctoria (16·8) and Withania somnifera (10·2) leaves. Total and differential protozoal counts increased with added PEG in twenty‐two samples, maximum being in Pimenta officinalis. Increased accumulation of total volatile fatty acids during incubation with added PEG‐6000 was recorded, and the values ranged from zero to 61%. However, the increase was significant in only 11 of the 38 tannin sources tested indicating noninterference of tannin on in vitro fermentation of carbohydrates by the majority of samples tested. Conversely, in 26 of 38 plant sources, the leaf tannins reduced N‐digestibility as evidenced by increased accumulation of NH₃‐N with added PEG.

Conclusions

Our study unequivocally demonstrated that plants containing secondary metabolites such as Rauvolfia serpentine, Indigofera tinctoria and Withania somnifera have great potential to suppress methanogenesis with minimal adverse effect of feedstuff fermentation.

Significance and Impact of the Study

It was established that methanogenesis was not essentially related to the density of protozoa population in vitro. The tannins contained in these plants could be of interest in the development of new additives in ruminant nutrition.     

植物挥发性物质及其代谢工程

植物挥发性物质在植物之间和植物与昆虫间的化学通讯中起着重要作用.有关这些次生物质的生物合成、代谢调控、生理功能以及与环境相互作用的研究近十多年来取得了重要进展.迄今为止,已经有3 0多种植物挥发性物质的合成酶基因被克隆.这些基因调控着植物萜类、芳香化合物、脂肪酸衍生物这三大类主要挥发性物质的生物合成.由于潜在的应用价值,近几年该领域颇受注目,特别是应用基因工程技术设计植物释放特殊气味物质,诸如特定的驱避剂或者其它控制植物或昆虫行为的特殊气味乃至与人类健康相关的药用气味物质.该文就植物挥发性物质的生物合成、生理和生态功能以及基因工程方面的研究进展作一概述.     

Mountain gorilla geophagy: A possible seasonal behavior for dealing with the effects of dietary changes

Geophageous Rwandan mountain gorillas excavate and eat weathered leucite-rich regolith (subsoil C horizons) from the slopes of Mount Visoke in the Virunga Mountains. In the months of the dry season, the gorillas reportedly ingest a halloysitic natural earth having a chemical composition similar to that of Kaopectate, a pharmaceutical used by humans to treat gastrointestinal upsets. Several plants known to contain potential toxins are consumed more heavily by gorillas in these months. New information from geochemical and mineral analyses suggests that geophagy may alleviate intestinal problems associated with changes in their diet because the ingested weathered regolith,containing halloysitic clay minerals, may act as a pharmaceutical agent that helps to adsorb toxins and to control dehydration in the dry season.     

Metabolic engineering of plant secondary metabolite pathways for the production of fine chemicals

The technology of large-scale plant cell culture is feasible for the industrial production of plant-derived fine chemicals. Due to low or no productivity of the desired compounds the economy is only in a few cases favorable. Various approaches are studied to increase yields, these encompass screening and selection of high producing cell lines, media optimization, elicitation, culturing of differentiated cells (organ cultures), immobilization. In recent years metabolic engineering has opened a new promising perspectives for improved production in a plant or plant cell culture.     

Preliminary observations on the role of Acacia gum chemistry in Acacia utilization by Galago senegalensis in Kenya

A census and preliminary observations on the ranging and feeding behavior of Galago senegalensis were made in a savannah-woodland site in Kenya. Population densities in three slightly different habitats were all approximately 1.5 animals/ha. Ranging patterns suggested the existence of territorial behavior. The diet of the galagos was exclusively insects and gums of two species of Acacia. The gum of A. drepanolobium appeared to be preferred to that of A. xanthophloea. The chemistry of the gums suggests that this preference is not due simply to the total level of phenolics or to avoidance of condensed tannins but may relate to the presence of compounds (e. g., flavonoids) having nutritional or hormonelike action.     

Biotechnological interventions for harnessing podophyllotoxin from plant and fungal species: current status,challenges, and opportunities for its commercialization

Podophyllotoxin is an aryltetralin lignan synthesized in several plant species, which is used in chemotherapies for cancers and tumor treatment. More potent semisynthetic derivatives of podophyllotoxin such as etoposide and teniposide are being developed and evaluated for their efficacy. To meet the ever increasing pharmaceutical needs, species having podophyllotoxin are uprooted extensively leading to the endangered status of selective species mainly Sinopodophyllum hexandrum. This has necessitated bioprospection of podophyllotoxin from different plant species to escalate the strain on this endangered species. The conventional and non-conventional mode of propagation and bioprospection with the integration of biotechnological interventions could contribute to sustainable supply of podophyllotoxin from the available plant resources. This review article is focused on the understanding of different means of propagation, development of genomic information, and its implications for elucidating podophyllotoxin biosynthesis and metabolic engineering of pathways. In addition, various strategies for sustainable production of this valuable metabolite are also discussed, besides a critical evaluation of future challenges and opportunities for the commercialization of podophyllotoxin.     

四种植物次生物质对橘小实蝇抗敌百虫品系的抗药性影响

为明确各种植物次生物质对橘小实蝇Bactrocera dorsalis （Hendel）抗药性的影响,本研究以室内选育的橘小实蝇对敌百虫高抗品系为研究对象,使用不同剂量的芸香苷、槲皮素、2-十三烷酮、单宁酸等4种植物次生物质分别连续饲喂3代该品系成虫和幼虫,采用药膜法测定毒力,获得抗性倍数和抗性增长率,分析明确植物次生物质种类、剂量与橘小实蝇抗药性发展之间关系。研究结果表明4种物质中较高浓度芸香苷处理橘小实蝇对敌百虫的抗性明显增强,总体呈现处理代次越多抗药性增强程度越大的趋势。F1代0.05%、0.25%芸香苷剂量处理抗性倍数分别为198.05、172.13,明显高于对照（110.61）,抗性增长率分别为1.79、1.56;F2代0.01%、0.05%、0.25%剂量处理抗性倍数由对照的89.49分别增大至137.20、146.38、171.48,其抗性增长率分别为1.53、1.64、1.92;F3代0.01%、0.05%、0.25%剂量下抗性倍数由对照的61.33分别增大至128.12、137.23、211.36,其抗性增长率明显增大（2.09、2.24、3.45）。部分浓度单宁酸处理增强或者降低了橘小实蝇对敌百虫的抗性,F1代0.002%浓度处理时抗性增长率为1.60,而F2代0.25%、F3代0.05%、0.25%浓度处理时抗性增长率降低至0.68、0.70和0.67。槲皮素、2-十三烷酮多个浓度处理多代均未对橘小实蝇抗敌百虫品系的抗药性产生影响。本研究显示一些植物次生物质可对橘小实蝇抗药性产生正面或者负面的影响,其作用与物质种类、浓度、接触时间等有关。