Greenhouse Gas Profile of a Plastic Material Derived from a Genetically Modified Plant

Abstract: This article reports an assessment of the global warming potential associated with the life cycle of a biopolymer (poly(hydroxyalkanoate) or PHA) produced in genetically engineered corn developed by Monsanto. The grain corn is harvested in a conventional manner, and the polymer is extracted from the corn stover (i.e., residues such as stalks, leaves and cobs), which would be otherwise left on the field. While corn farming was assessed based on current practice, four different hypothetical PHA production scenarios were tested for the extraction process. Each scenario differed in the energy source used for polymer extraction and compounding, and the results were compared to polyethylene (PE). The first scenario involved burning of the residual biomass (primarily cellulose) remaining after the polymer was extracted from the stover. In the three other scenarios, the use of conventional energy sources of coal, oil, and natural gas were investigated. This study indicates that an integrated system, wherein biomass energy from corn stover provides energy for polymer processing, would result in a better greenhouse gas profile for PHA than for PE. However, plant-based PHA production using fossil fuel sources provides no greenhouse gas advantage over PE, in fact scoring worse than PE. These results are based on a "cradle-to-pellet" modeling as the PHA end-of-life was not quantitatively studied due to complex issues surrounding the actual fate of postconsumer PHA.     

The Biosynthesis of a Novel Nicotine Alkaloid in the Trichomes of Nicotiana stocktonii

N-Hydroxyacylnornicotine, newly discovered from fresh plant tissue, was found entirely in the trichome exudate produced at the epidermis of the aerial part of Nicotiana stocktonii. Nicotine and nornicotine, but not N-hydroxyacylnornicotine, were present inside of the trichomes as well as other internal parts of the plant. Only nicotine was found in bleeding sap squeezed from cut roots or stems. Feeding of leaves with 2′-¹⁴C-labeled nicotine primarily yielded labeled nicotine, nornicotine, and N-hydroxyacylnornicotine. When similarly labeled nornicotine was fed to leaves as a precursor, a labeled N-hydroxyacylnornicotine was obtained, with a higher specific activity than with the [2′-¹⁴C]nicotine feeding. Based on these results, a synthesis route is suggested where nicotine is converted in the leaf to nornicotine, followed by trichome conversion of nornicotine to N-hydroxyacylnornicotine, and rapid secretion of this product.     

多重PCR技术快速检测烤后烟叶转基因成分

为提高烟叶转基因成分检测效率,建立了烤后烟叶中3种外源基因成分的多重PCR检测技术体系,通过优化该反应体系中的Mg2+、d NTP、r Taq酶、引物浓度及退火温度等参数,实验最适条件为将4对相等摩尔浓度引物预先按1∶1∶1∶1(V/V)混合,在Mg2+为1.5 mmol/L、d NTP为125μmol/L、r Taq酶1 U、混合引物1μmol/L,DNA 100 ng,退火温度65℃,循环数35个的反应条件下,在一次PCR反应中便可同时检测烟草内源基因NR,外源基因35S启动子、NOS终止子、NPT II筛选标记基因。优化后的反应体系能够有效地检测出转基因烤后烟叶百分比含量为0.9%(V/V)的转基因成分。     

The Role of Genetically Modified Mesenchymal Stem Cells in Urinary Bladder Regeneration

Recent studies have demonstrated that mesenchymal stem cells (MSCs) combined with CD34⁺ hematopoietic/stem progenitor cells (HSPCs) can function as surrogate urinary bladder cells to synergistically promote multi-faceted bladder tissue regeneration. However, the molecular pathways governing these events are unknown. The pleiotropic effects of Wnt5a and Cyr61 are known to affect aspects of hematopoiesis, angiogenesis, and muscle and nerve regeneration. Within this study, the effects of Cyr61 and Wnt5a on bladder tissue regeneration were evaluated by grafting scaffolds containing modified human bone marrow derived MSCs. These cell lines were engineered to independently over-express Wnt5a or Cyr61, or to exhibit reduced expression of Cyr61 within the context of a nude rat bladder augmentation model. At 4 weeks post-surgery, data demonstrated increased vessel number (~250 vs ~109 vessels/mm²) and bladder smooth muscle content (~42% vs ~36%) in Cyr61OX (over-expressing) vs Cyr61KD (knock-down) groups. Muscle content decreased to ~25% at 10 weeks in Cyr61KD groups. Wnt5aOX resulted in high numbers of vessels and muscle content (~206 vessels/mm² and ~51%, respectively) at 4 weeks. Over-expressing cell constructs resulted in peripheral nerve regeneration while Cyr61KD animals were devoid of peripheral nerve regeneration at 4 weeks. At 10 weeks post-grafting, peripheral nerve regeneration was at a minimal level for both Cyr61OX and Wnt5aOX cell lines. Blood vessel and bladder functionality were evident at both time-points in all animals. Results from this study indicate that MSC-based Cyr61OX and Wnt5aOX cell lines play pivotal roles with regards to increasing the levels of functional vasculature, influencing muscle regeneration, and the regeneration of peripheral nerves in a model of bladder augmentation. Wnt5aOX constructs closely approximated the outcomes previously observed with the co-transplantation of MSCs with CD34⁺ HSPCs and may be specifically targeted as an alternate means to achieve functional bladder regeneration.     

The Association of Methylenetetrahydrofolate Reductase Genotypes with the Risk of Childhood Leukemia in Taiwan

Background

Acute lymphoblastic leukemia (ALL) is the most prevalent type of pediatric cancer, the causes of which are likely to involve an interaction between genetic and environmental factors. To evaluate the effects of the genotypic polymorphisms in methylenetetrahydrofolate reductase (MTHFR) on childhood ALL risk in Taiwan, two well-known polymorphic genotypes of MTHFR, C677T (rs1801133) and A1298C (rs1801131), were analyzed to examine the extent of their associations with childhood ALL susceptibility and to discuss the MTHFR genotypic contribution to childhood ALL risk among different populations.

Methodology/Principal Findings

In total, 266 patients with childhood ALL and an equal number of non-cancer controls recruited were genotyped utilizing PCR-RFLP methodology. The MTHFR C677T genotype, but not the A1298C, was differently distributed between childhood ALL and control groups. The CT and TT of MTHFR C677T genotypes were significantly more frequently found in controls than in childhood ALL patients (odds ratios=0.60 and 0.48, 95% confidence intervals=0.42–0.87 and 0.24–0.97, respectively). As for gender, the boys carrying the MTHFR C677T CT or TT genotype conferred a lower odds ratio of 0.51 (95% confidence interval=0.32–0.81, P=0.0113) for childhood ALL. As for age, those equal to or greater than 3.5 years of age at onset of disease carrying the MTHFR C677T CT or TT genotype were of lower risk (odds ratio= 0.43 and 95% confidence interval=0.26–0.71, P=0.0016).

Conclusions

Our results indicated that the MTHFR C677T T allele was a protective biomarker for childhood ALL in Taiwan, and the association was more significant in male patients and in patients 3.5 years of age or older at onset of disease.     

Protein Crystallography Reveals a Role for the FS0 Cluster of Escherichia coli Nitrate Reductase A (NarGHI) in Enzyme Maturation

We have used site-directed mutagenesis, EPR spectroscopy, redox potentiometry, and protein crystallography to monitor assembly of the FS0 [4Fe-4S] cluster and molybdo-bis(pyranopterin guanine dinucleotide) cofactor (Mo-bisPGD) of the Escherichia coli nitrate reductase A (NarGHI) catalytic subunit (NarG). Cys and Ser mutants of NarG-His⁴⁹ both lack catalytic activity, with only the former assembling FS0 and Mo-bisPGD. Importantly, both prosthetic groups are absent in the NarG-H49S mutant. EPR spectroscopy of the Cys mutant reveals that the E_m value of the FS0 cluster is decreased by at least 500 mV, preventing its participation in electron transfer to the Mo-bisPGD cofactor. To demonstrate that decreasing the FS0 cluster E_m results in decreased enzyme activity, we mutated a critical Arg residue (NarG-Arg⁹⁴) in the vicinity of FS0 to a Ser residue. In this case, the E_m of FS0 is decreased by 115 mV, with a concomitant decrease in enzyme turnover to ∼30% of the wild type. Analysis of the structure of the NarG-H49S mutant reveals two important aspects of NarGHI maturation: (i) apomolybdo-NarGHI is able to bind GDP moieties at their respective P and Q sites in the absence of the Mo-bisPGD cofactor, and (ii) a critical segment of residues in NarG, ⁴⁹HGVNCTG⁵⁵, must be correctly positioned to ensure holoenzyme maturation.     

Persistence of Genetically Modified Potatoes in the Field

Volunteers from genetically modified (GM) potatoes may pose an environmental problem if allowed to grow in the field after the annual crop is harvested. We tested whether they are more likely to produce volunteers than non-GM potatoes. Specifically, we compared the number of volunteers, number of tubers per plant, tuber size, and their vertical distribution in the soil. More volunteer plants came from non-GM potatoes than from GM potatoes, but the number and size of tubers were similar between the two. Vertical distribution of the tubers differed significantly, with most non-GM tubers being found in shallower soil (<2 cm deep). Our results suggest that spontaneous GM volunteers may emerge and produce tubers to a degree similar to that of the non-GM plants. No viable volunteers emerged from GM tubers in the next growing season, probably deterred by winter frost and a period of low soil temperatures (below −2°C) at our study site. However, in regions with warmer climates, such GM volunteers may survive Winter and produce more plants the following year.     

植物中硝酸还原酶和亚硝酸还原酶的作用

植物通过硝酸盐同化途径以硝酸盐和氨的形式吸收氮元素。硝酸盐的同化是一个受到严格控制的过程,其中两个先后参加反应的酶——硝酸还原酶(NR)和亚硝酸还原酶(NiR)对初级氮的同化起主要调控。在高等植物中,NR和NiR基因的转录及转录后加工受到各种内在和外在因素的影响,翻译后调控是消除亚硝酸盐积累的重要机制。随着分子生物学技术的发展,可以更容易地通过突变体和转基因方式来研究NR和NiR基因的调控。     

Rooting and the Metabolism of Nicotine in Tobacco Callus Cultures

The usefulness of exogenous nicotine as a factor in the induction of morphogenesis in a tobacco tissue culture medium has been demonstrated. Nicotiana rustica callus cell cultures were grown on a modified Murashige and Skoog medium with 2 mg/l indoleacetic acid (IAA) and 0.2 mg/l kinetin (MMS). Root morphogenesis was induced in roller tube callus cell cultures and solid callus cell cultures grown on MMS without kinetin supplemented with 10–100 mg/l nicotine. Optimal nicotine concentration for root induction was 50 mg/l. Other tests using varying combinations of IAA, kinetin and nicotine produced no obvious morphogenesis, although some changes in the amount of callus growth and endogenous protein concentration did correlate with nicotine concentration relative to the presence of IAA and/or kinetin. In liquid MMS medium, ¹⁴C-nicotine was primarily incorporated into the protein fraction of cultured cells while primarily incorporated into the cell wall and/or cell membrane fraction of cells cultured on MMS without kinetin in the medium. In MMS without IAA and MMS without both IAA and kinetin, there was incorporation, but to a lesser extent in both the protein and the cell wall and/or cell membrane fractions.     

Metabolite Profiling Reveals a Role for Atypical Cinnamyl Alcohol Dehydrogenase CAD1 in the Synthesis of Coniferyl Alcohol in Tobacco Xylem

In angiosperms, lignin is built from two main monomers, coniferyl and sinapyl alcohol, which are incorporated respectively as G and S units in the polymer. The last step of their synthesis has so far been considered to be performed by a family of dimeric cinnamyl alcohol dehydrogenases (CAD2). However, previous studies on Eucalyptus gunnii xylem showed the presence of an additional, structurally unrelated, monomeric CAD form named CAD1. This form reduces coniferaldehyde to coniferyl alcohol, but is inactive on sinapaldehyde. In this paper, we report the functional characterization of CAD1 in tobacco (Nicotiana tabacum L.). Transgenic tobacco plants with reduced CAD1 expression were obtained through an RNAi strategy. These plants displayed normal growth and development, and detailed biochemical studies were needed to reveal a role for CAD1. Lignin analyses showed that CAD1 down-regulation does not affect Klason lignin content, and has a moderate impact on G unit content of the non-condensed lignin fraction. However, comparative metabolic profiling of the methanol-soluble phenolic fraction from basal xylem revealed significant differences between CAD1 down-regulated and wild-type plants. Eight compounds were less abundant in CAD1 down-regulated lines, five of which were identified as dimers or trimers of monolignols, each containing at least one moiety derived from coniferyl alcohol. In addition, 3-trans-caffeoyl quinic acid accumulated in the transgenic plants. Together, our results support a significant contribution of CAD1 to the synthesis of coniferyl alcohol in planta, along with the previously characterized CAD2 enzymes. Sequences of NtCAD1-1 and NtCAD1-7 were deposited in GenBank under accession numbers AY911854 and AY911855, respectively.     

烟草去甲基尼古丁产生的机理

烟叶的采收后处理和加工过程中,大量的尼古丁经去甲基化作用生成了去甲基尼古丁,后者是烟草特有的亚硝胺类（tobacco-specific nitrosamines,TSNAs）致癌物——亚硝基去甲基尼古丁（nitrosonomicoline,NNN）的前体,与人类健康息息相关。由于尼古丁去甲基化反应在基础理论研究和商业上的重要价值,长期以来关于这个反应的机制研究一直是学术界的热点。本文讨论了尼古丁去甲基化反应研究的历史概况、反应机制假说的演变及影响该反应的因素,期望通过对烟草尼古丁去甲基化反应研究的总结,为烟草品质提高和低毒害烟草制品的研究与开发提供一定的参考。     

Studies on the Chemical Regulation of Alkaloid Biosynthesis in Tobacco Plants

Effect of auxin treatment upon the alkaloid content of Nicotiana was examined using hydroponically grown plants. Nicotine content of Nicotiana tabacum was reduced by the addition of 3-indoleacetic acid or 2, 4-dichlorophenoxyacetic acid to nutrient solution, an the incorporation of nitrate-¹⁵N into nicotine was decreased by the addition of 2, 4-dichloro-phenoxyacetic acid. Contents of nicotine, nornicotine, anabasine and anatabine in Nicotiana glutinosa were decreased by 2, 4-dichlorophenoxyacetic acid added to the nutrient solution. Foliar applications of eleven kinds of synthetic auxins were effective in reducing nicotine content of Nicotiana tabacum and the effectiveness of the compounds seemed to be concerned with their auxin activities. Foliar application of auxin tended to increase the dry weight of stems whereas decreased that of roots, and, moreover, higher content of soluble sugars and lower content of starch in leaves were observed by the treatment. A possibility of a practical application of auxin to reduce the nicotine content of tobacco plants were drawn from this work.     

烟草去甲基尼古丁产生的机理

烟叶的采收后处理和加工过程中, 大量的尼古丁经去甲基化作用生成了去甲基尼古丁, 后者是烟草特有的亚硝胺类(tobacco-specific nitrosamines, TSNAs)致癌物— — 亚硝基去甲基尼古丁(nitrosonornicotine, NNN)的前体, 与人类健康息息相关。由于尼古丁去甲基化反应在基础理论研究和商业上的重要价值, 长期以来关于这个反应的机制研究一直是学术界的热点。本文讨论了尼古丁去甲基化反应研究的历史概况、反应机制假说的演变及影响该反应的因素, 期望通过对烟草尼古丁去甲基化反应研究的总结, 为烟草品质提高和低毒害烟草制品的研究与开发提供一定的参考。     

Population Study of Frequency of Methylenetetrahydrofolate Reductase C677T Gene Polymorphism in Yakutia

The enzyme methylenetetrahydrofolate reductase (MTHFR) catalyzes synthesis of 5-methylenehydrofolate, which is the methyl donor for the conversion of homocysteine to methionine. According to the numerous literature data, polymorphic variant of the MTHFR-encoding gene, C677T, is associated with hyperhomocysteinemia, vascular pathologies, neural tube defects, dementia, perinatal mortality, mental disorders, long-term neurodegenerative disorders, lens displacement, arachnodactyly, and venous thromboses. The present study was focused on the analysis of the C677T polymorphism (missence mutation leading to the replacement of cytosine by thymine at position 677) of the MTHFR gene in three indigenous populations of the Republic of Sakha (Yakutia), living in the settlements of Cheriktei, Byadi, and Dyupsya. Comparison of the genotype and allele frequencies revealed no substantial differences between the three Yakut populations, as well as between Yakuts and other Mongoloid ethnic groups.     

The Growth Response of the Stems of Genetically Modified Tobacco Plants (Nicotiana tabacum'Samsun') to Flexural Stimulation

Genetically modified tobacco plants (Nicotiana tabacum‘Samsun’)with antisense cinnamyl alcohol dehydrogenase DNA, produce secondaryxylem of a reduced tensile stiffness. These plants were grownalongside control plants. The stems of the plants were flexedor protected from flexing over a period of several weeks. Thetensile moduli and second moments of areas of the differenttissues inside the stems were measured and used to calculatethe bending stiffness of the plants. In tobacco, the cylinderof xylem was found to be the most important tissue in determiningthe bending stiffness of the plants. The thickness of the xylemtissue cylinder increased when plants were subjected to flexuralstimulation. This increased the bending stiffness of the stems.The response to mechanical stimulation was found to be correlatedwith tissue strain and the genetically modified plants wereable to exactly compensate for the reduced modulus of theirxylem tissue by increasing the thickness of the xylem tissuecylinder more than in control plants.Copyright 1999 Annals ofBotany Company. Tobacco plants, stem bending, xylem tissue, second moment of area, thigmomorphogenesis, mechanical strain.     

Studies on the Chemical Regulation of Alkaloid Biosynthesis in Tobacco Plants

Effects of helminthosporol and helminthosporic acid (H-acid) on nicotine biosynthesis in Nicotiana were examined. By the addition of these compounds into nutrient solution, nicotine formation in decapitated and intact plants of Nicotiana tabacum was decreased. In sterile culture of excised roots of Nicotiana rustica, H-acid reduced the yield of nicotine exerting no effect on growth rate. Incorporation of l-glutamic acid-U-¹⁴C and dl-ornithine-2-¹⁴C into nicotine in the roots was decreased by the addition of H-acid, and that of nicotinic acid-³H(G) was not influenced. Since H-acid did not enhance destruction of nicotine-³H exogenously added, it is probable that the decrease of nicotine yield by the acid in the root culture is due to reduction in the production rate of the alkaloid.     

转基因食品安全性研究进展

转基因食品的安全性在世界范围内备受关注,近年来,已经成为公众争论的焦点。本文简要综述了转基因食品的现状和安全性,并介绍了我国政府对转基因食品的态度。此外,还对转基因食品的发展前景作了简要展望。