转基因植物的生态风险

转基因植物已在很多国家大规模商业化种植,并且取得了显著的经济效益。同时有关转基因植物潜在的生态风险已引起广泛的关注。本文从转基因植物人侵危害、对非靶标有益生物直接和间接的影响、害虫对抗虫转基因植物产生抗性、抗病毒转基因植物带来的潜在风险等方面论述了转基因植物可能潜在的生态安全性问题。     

Phytoremediation of Metals Using Transgenic Plants

An ideal plant for environmental cleanup can be envisioned as one with high biomass production, combined with superior capacity for pollutant tolerance, accumulation, and/or degradation, depending on the type of pollutant and the phytoremediation technology of choice. With the use of genetic engineering, it is feasible to manipulate a plant's capacity to tolerate, accumulate, and/or metabolize pollutants, and thus to create the ideal plant for environmental cleanup. In this review, we focus on the design and creation of transgenic plants for phytoremediation of metals. Plant properties important for metal phytoremediation are metal tolerance and accumulation, which are determined by metal uptake, root-shoot translocation, intracellular sequestration, chemical modification, and general stress resistance. If we know which molecular mechanisms are involved in these tolerance and accumulation processes, and which genes control these mechanisms, we can manipulate them to our advantage. This review aims to give a succinct overview of plant metal tolerance and accumulation mechanisms, and to identify possible strategies for genetic engineering of plants for metal phytoremediation. An overview is presented of what has been achieved so far regarding the manipulation of plant metal metabolism. In fact, both enhanced metal tolerance and accumulation have been achieved by overproducing metal chelating molecules (citrate, phytochelatins, metallothioneins, phytosiderophores, ferritin) or by the overexpression of metal transporter proteins. Mercury volatilization and tolerance was achieved by introduction of a bacterial pathway. The typical increase in metal accumulation as the result of these genetic engineering approaches is 2-to 3-fold more metal per plant, which could potentially enhance phytoremediation efficiency by the same factor. As for the applicability of these transgenics for environmental cleanup, results from lab and greenhouse studies look promising for several of these transgenics, but field studies will be the ultimate test to establish their phytoremediation potential, their competitiveness, and risks associated with their use.     

Phytoremediation of Phosphorus-Enriched Soils

After numerous years of application of phosphorus (P) fertilizer and/or manures, many soils accumulate high levels of P, increasing the risk of P transfer via surface runoff. While current and future regulations may limit P application to soil, little effort is given to the reduction of soils already enriched with P. To evaluate the potential of phytoremediation for decreasing available P in soil, the P uptake of several crops and vegetables grown on a high P soil was studied. Plant shoot and root P contents, plant-available soil P, and potential P removal under optimum conditions were examined. None of the plants were “true” P hyperaccumulators, although all removed some P from the soil. Whole corn plants (for silage) and Indian mustard removed the highest amount of P; 114 and 108 kg ha^-1, respectively. If only the grain from corn and Indian mustard were harvested, significantly lower amounts of P were removed from the field. These results show that there is extensive variation in the uptake of P by various crop species. For soils highly enriched with P, it may be desirable to grow a plant species capable of removing above average concentrations of P and to harvest the whole plant because a large fraction of the plant P is contained within the leaves and stems.     

转拟南芥ICE1基因增强烟草抗寒性的研究

ICE1是CBF冷响应通道的上游转录调控因子,通过与CBF启动子中MYC顺式作用元件的结合激活CBF3基因表达.采用RT-PCR方法,从拟南芥获得AtICE1基因,将AtICE1导入pCAMBIA1301构建35S:AtICE1植物表达载体.通过根癌农杆菌GV3101,将AtICE1基因导人烟草,T1代植株经潮霉素抗性筛选,PCR、RT-PCR检测,结果表明AtICE1基因已经整合到烟草基因组中,并在转录水平表达;在正常生长条件下,转基因烟草与对照烟草的生长未见明显区别,而在瞬时低温冻害下,转基因烟草存活率明显高于对照烟草植株,说明Atl-CEI基因可以提高低温敏感作物的耐寒性.     

Plants, animals and the logic of development

Multicellular plants and animals have evolved independently from a unicellular, last common ancestor. Each lineage started with a common toolkit of functioning genes and evolved to complex, multicellular forms. Comparison of the genes used to serve similar functions shows how organisms can use different genes for similar ends and thereby reveals the principles of development.     

Plants, animals and the logic of development

Multicellular plants and animals have evolved independently from a unicellular, last common ancestor. Each lineage started with a common toolkit of functioning genes and evolved to complex, multicellular forms. Comparison of the genes used to serve similar functions shows how organisms can use different genes for similar ends and thereby reveals the principles of development.     

Plants, animals and the logic of development

Multicellular plants and animals have evolved independently from a unicellular, last common ancestor. Each lineage started with a common toolkit of functioning genes and evolved to complex, multicellular forms. Comparison of the genes used to serve similar functions shows how organisms can use different genes for similar ends and thereby reveals the principles of development.     

Simple Identification of Transgenic Arabidopsis Plants Carrying a Single Copy of the Integrated Gene

Transgenic Arabidopsis thaliana plants carrying a single copy of integrated DNA can be identified by single-step genomic polymerase chain reaction. The reaction employs two sets of primer pairs with the same melting temperature that amplify the amplicons derived from the integrated T-DNA together with those from an endogenous single-copy gene as a reference. When the band intensity ratio is one, this means that the transgenic plants are carrying a single copy of the integrated gene per haploid.     

突变体在植物光破坏防御研究中的应用

用突变体研究植物的光破坏防御机制是一种有效的方法，与传统方法相比，具有专一性强、副效应少、直接在基因水平上起作用、可信度高等优点。本文简要介绍了突变体的获得、筛选、鉴定，并总结了几类与光破坏防御有关的突变体。     

Comparative Evaluation of Metal Phytoremediation Potential of Trees,Grasses, and Flowering Plants from Tannery-Wastewater-Contaminated Soil in Relation with Physicochemical Properties

This study reports a comparative account of metal accumulation in the trees, grasses, and flowering plants from agricultural fields contaminated with tannery wastewater. Soil physico-chemical properties along the pollution gradient and soil depth were analyzed. Monitoring and assessment of the plants growing on contaminated sites revealed that the accumulation of Cr in the aboveground part of the trees ranged from 1.87 to 34.44 μg g^?1 dw with maximum concentration in Dendro-calamus strictus (34.44 μg g^?1 dw). Chrysanthemum coronarium and Tagetes erecta showed better accumulation of Cr than other flowering plants. Separate field experiments were conducted on the contaminated area. The shoots of Vetiveria zizanoides (532 mg 4 m^?2) and Cymbopogan winterianus (535.46 mg 4 m^?2) have shown almost similar removal potential of Cr, with maximum removal potential in the roots of C. winterianus (1206.43 mg 4 m^?2). Seasonal flowering plants (i.e., C. coronarium) have shown better accumulation of Cr than T. erecta. The results indicate that the plants of V. zizanoides, C. coronarium, and C. winterianus are suitable for phytoremediation of contaminated sites and trees can successfully be used for phytostabilization.     

Antisense suppression of l-galactose dehydrogenase in Arabidopsis thaliana provides evidence for its role in ascorbate synthesis and reveals light modulated l-galactose synthesis

l-Galactose dehydrogenase (l-GalDH), a novel enzyme that oxidizes l-Gal to l-galactono-1,4-lactone (l-GalL), has been purified from pea seedlings and cloned from Arabidopsis thaliana. l-GalL is a proposed substrate for ascorbate biosynthesis in plants, therefore the function of l-GalDH in ascorbate biosynthesis was investigated by overexpression in tobacco and antisense suppression in A. thaliana. In tobacco the highest expressing lines had a 3.5-fold increase in extractable activity, but this did not increase leaf ascorbate concentration. Arabidopsis thaliana, transformed with an antisense l-GalDH construct, produced lines with 30% of wild-type activity. These had lower leaf ascorbate concentration when grown under high light conditions. l-Gal pool size increased in antisense transformants with low l-GalDH activity, and l-Gal concentration was negatively correlated with ascorbate. The results provide direct evidence for a role of l-GalDH in ascorbate biosynthesis. Ascorbate pool size in A. thaliana is increased by acclimation to high light, but l-GalDH expression was not affected. l-Gal accumulation was higher in antisense plants acclimated to high light, indicating that the capacity to synthesize l-Gal from GDP-mannose is increased. Because the only known function of l-GalL is ascorbate synthesis, these antisense plants provide an opportunity to investigate ascorbate function with minimal effects on carbohydrate metabolism. Measurements of other antioxidants revealed an increase in ascorbate- and pyrogallol-dependent peroxidase activity in low-ascorbate lines. As ascorbate is the major hydrogen peroxide-scavenging antioxidant in plants, this could indicate a compensatory mechanism for controlling hydrogen peroxide concentration.     

Activation of the NaCl- and drought-induced RD29A and RD29B promoters by constitutively active Arabidopsis MAPKK or MAPK proteins

Mitogen-activated protein (MAP) kinases mediate cellular responses to a wide variety of stimuli. Activation of a MAP kinase (MAPK) occurs after phosphorylation by an upstream MAP kinase kinase (MAPKK). The Arabidopsis thaliana genome encodes 10 MKKs, but few of these have been shown directly to activate any of the 20 Arabidopsis MAPKs (AtMPKs) and NaCl-, drought- or abscisic acid (ABA)-induced genes RD29A or RD29B. We have constructed the constitutively activated form for nine of the 10 AtMKK proteins, and tested their ability to activate the RD29A and RD29B promoters and also checked the ability of the nine activated AtMKK proteins to phosphorylate 11 of the AtMPK proteins in transient assays. The results show that three proteins, AtMKK1, AtMKK2 and AtMKK3, could activate the RD29A promoter, while these three and two additional AtMKK6/8 proteins could activate the RD29B promoter. Four other proteins, AtMKK7/AtMKK9 and AtMKK4/AtMKK5, can cause hypersensitive response (HR) in tobacco leaves using transient analysis. The activation of the RD29A promoter correlated with four uniquely activated AtMPK proteins. A novel method of activating AtMPK proteins by fusion to a cis-acting mutant of a human MAPK kinase MEK1 was used to confirm that specific members of the AtMPK gene family can activate the RD29A stress pathway.     

Construction of Synthetic Genes for Analogs of Spider Silk Spidroin 1 and Their Expression in Tobacco Plants

To obtain transgenic tobacco plants expressing recombinant analogs of spider dragline silk spidroin 1, artificial 1f5 and 1f9 coding for spidroin 1 analogs were 3"-fused in-frame with the reporter lichenase gene. The Tr2" weak constitutive promoter of Agrobacterium tumefaciens T-DNA and the strong constitutive promoter of the cauliflower mosaic virus 35S RNA gene were used as regulatory elements. The expression cassettes were used to transform agrobacteria and then introduced in tobacco leaf disks. On evidence of Southern hybridization, transgenic plants each carried a single copy of a hybrid gene, which corresponded in size to the constructed one. Zymography and Western blotting revealed full-length hybrid proteins in leaf extracts of transgenic plants. The results testified that plants can maintain and express synthetic genes for spider silks and, consequently, may be used as a convenient producer of recombinant silk analogs.     

用绿色荧光蛋白监测转基因植物中选择标记基因的消除

绿色荧光蛋白(GFP)可直接进行活体观察,它的这个优点可被用于监测转基因植物中选择标记基因的消除。为此,构建了植物表达载体pGNG,将绿色荧光蛋白基因(gfp)和卡那霉素抗性基因表达盒(NosP-nptll-NosT)一起克隆在两个同向的lox位点间,在第一个lox位点上游置有CaMV 35S启动子以驱动GFP表达,第二个lox位点下游置有不含启动子的大肠杆菌β-葡萄糖醛酸酶(GUS)基因。首先在含卡那霉素(Kan)的培养基上筛选出转pGNG的烟草,借助绿色荧光可容易地检出表达GFP的转化体。然后用另一转化载体pCambia1300Cre二次转化表达GFP的转基因植物,利用另一选择标记基因潮霉素抗性基因(hpt)进行筛选,在获得的再生植株中,Cre重组酶的表达消除了转化体中两lox位点间的gfp和nptll。实验结果表明可借助GFP荧光的消失,快速选出nptII被消除的二次转化体,同时GUS(作为目的蛋白) 在CaMV 35S启动子驱动下获得表达。最后利用后代的分离将hpt和cre除去。     

Expression and regulation of theRSI-1 gene during lateral root initiation

The tomato geneRSI-1 was previously identified as a molecular marker for auxin-induced lateral root initiation. We have further characterized the expression mode of theRSI-1 gene in tomato andArabidopsis thaliana. Northern blot analyses revealed that the gene was induced specifically by auxin in tomato roots and hypocotyls. For experiments with transgenic plants, the 5′ flanking region of theRSI-1 gene was linked to a GUS reporter gene, then transformed into tomato andArabidopsis. In these transgenic tomato plants, GUS activity was detected at the sites of initiation for lateral and adventitious roots. Expression of the fusion gene was auxin-dependent and tissue-specific. This was consistent with results from the northern blot analyses. In transgenicArabidopsis, the overall expression pattern of theRSI-GUS gene, including tissue specificity and auxin inducibility, was comparable to that in transgenic tomato seedlings. These results indicate that an identical regulatory mechanism for lateral root initiation might be conserved in both plants. Thus, the expression mode of theRSI-CUS gene inArabidopsis mutants defective in lateral root development should be investigated to provide details of this process.     

拟南芥盐胁迫应答相关基因AtGRP9的克隆及表达

利用拟南芥cDNA文库在裂殖酵母中的功能性表达 ,从拟南芥中分离了一个编码富甘氨酸蛋白 (glycine richprotein ,GRP)的cDNA克隆 (其基因被命名为AtGRP9)。在裂殖酵母中大量表达AtGRP9cDNA能显著提高细胞的耐盐性。在拟南芥中 ,AtGRP9基因的表达受盐胁迫诱导 ,其表达具有根器官特异性 ;而且 35S启动子组成形成超量表达植株的耐盐性明显提高。这些实验结果表明拟南芥AtGRP9可能是一盐胁迫应答相关基因。     

强化植物修复重金属污染土壤的策略及其机制

重金属对生态环境、农业生产、人类健康等诸多方面造成重要危害。植物修复因其具有经济有效、绿色生态等优点,已经成为土壤重金属污染修复研究领域的热点。由于植物重金属毒害、修复耗时过长等因素致使植物修复技术受限于研究阶段而不能广泛应用于实践。采用科学合理的强化措施提高植物修复的效率可能是解决该矛盾的关键之一。讨论了根瘤菌、丛枝菌根真菌、溶磷微生物和内生真菌构建的微生物-植物共生系统在强化植物修复过程中的具体应用;概述了EDTA、EDDS等螯合剂在改变土壤中重金属可溶态,促进重金属从土壤向植株转运的重要作用;介绍了植物中编码金属转运蛋白、金属硫蛋白、植物螯合肽等与重金属转运和代谢相关的基因在植物修复领域的实际应用;归纳了上述强化策略主要机制为微生物促进植物生长、缓解重金属植物毒性以及提高了土壤中重金属生物利用度,从而促进重金属在富集植物中积累和植物生物量的增加;最后总结并展望了植物修复强化技术在今后研究的重点及存在的问题。综述植物修复技术采用的主要强化策略及其机制,旨在为利用植物修复技术治理土壤重金属污染提供重要参考。     

The Phytoremediation Potential of Thallium-Contaminated Soils Using Iberis and Biscutella Species

Biscutella laevigata and Iberis intermedia were sampled from sites near St Laurent le Minier, Southern France, and B. laevigata was also sampled from Rocca San Silvestro, Tuscany, Italy. Soils associated with the rhizosphere of each plant were also sampled. Both Biscutella laevigata and Iberis intermedia accumulate inordinately high concentrations of thallium (1.94 and 0.4%, respectively) in their above-ground dry tissue. The levels of thallium accumulated by both species were strongly correlated with both the total and extractable concentrations of thallium in the soils. Concentrations of zinc, cadmium, and lead were below the threshold for hyperaccumulation. It is proposed that B. laevigata and/or I. intermedia could be used for phytoremediation or phytomining of thallium-contaminated soils. Such an operation would involve the repeated cropping of either species, until an acceptable level of thallium in the soils was reached. Additionally, the harvested plant material could be burnt and the resulting ash smelted to produce an economically viable ‘crop’ of thallium.