The Physiological Mechanism of Improved Formaldehyde Resistance in Petunia hybrida Harboring a Mammalian cyp2e1 Gene

AbstractCytochrome P450 CYP2E1 is mainly present in hepatocytes in the livers of mammals, where it plays an important role in the metabolism of xenobiotic organic substances. Previous studies showed that transgenic petunia (Petunia hybrid) plants harboring a mammalian cyp2e1 gene (designated cyp2e1-transgenic petunia) exhibited increased resistance to formaldehyde stress. In this study, we used cyp2e1-transgenic petunia plants to analyze physiological indexes related to formaldehyde stress responses. The results indicated that under formaldehyde stress, the malondialdehyde content in cyp2e1-transgenic petunia plants was lower than in β-glucuronidase gene (gus)-transgenic and wild-type petunia plants. The activities of both superoxide dismutase and peroxidase in the cyp2e1-transgenic plants were higher than in gus-transgenic and wild-type plants. The alcohol dehydrogenase activity was slightly increased and more glutathione was consumed. Additionally, under formaldehyde stress, the levels of plant hormones including indole-3-acetic acid, zeatin and abscisic acid in cyp2e1-transgenic petunia plants displayed decreasing trends, whereas the level of gibberellic acid displayed an increasing trend. In contrast, the indole-3-acetic acid, zeatin and abscisic acid levels in gus-transgenic and wild-type petunia plants displayed increasing trends, whereas the gibberellic acid level displayed a decreasing trend. At 72h after incubation of 0.5g of cyp2e1-transgenic petunia plants in 40mL of treatment solution containing formaldehyde at 50mg · L−1, the formaldehyde content remaining in the treatment solution was close to zero while approximately half of original formaldehyde remained in the treatment solutions containing gus-transgenic and wild-type petunia plants

Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde

The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants

Transgenic plants of Petunia hybrida harboring the CYP2E1 gene efficiently remove benzene and toluene pollutants and improve resistance to formaldehyde

The CYP2E1 protein belongs to the P450 enzymes family and plays an important role in the metabolism of small molecular and organic pollutants. In this study we generated CYP2E1 transgenic plants of Petunia using Agrobacterium rhizogenes K599. PCR analysis confirmed that the regenerated plants contained the CYP2E1 transgene and the rolB gene of the Ri plasmid. Southern blotting revealed the presence of multiple copies of CYP2E1 in the genome of transgenic plants. Fluorescent quantitative PCR revealed exogenous CYP2E1 gene expression in CYP2E1 transgenic plants at various levels, whereas no like expression was detected in either GUS transgenic plants or wild-types. The absorption of benzene and toluene by transgenic plants was analyzed through quantitative gas chromatography. Transgenic plants with high CYP2E1 expression showed a significant increase in absorption capacity of environmental benzene and toluene, compared to control GUS transgenic and wild type plants. Furthermore, these plants also presented obvious improved resistance to formaldehyde. This study, besides being the first to reveal that the CYP2E1 gene enhances plant resistance to formaldehyde, also furnishes a new method for reducing pollutants, such as benzene, toluene and formaldehyde, by using transgenic flowering horticultural plants

Leakage Model of Tubing and Casing Premium Connection Based on Sinusoidal Contact Simulation between Rough Surfaces

This paper proposed a semi-theoretical model to quantitatively predict leakage rate of tubing and casing premium connections. The geometric parameters of the sealing surface profile approximated by a sinusoidal micro-convex surface were first obtained based on the random normal distribution sampling method. With the actual area prediction formula for elastic–plastic contact of an axisymmetric sinusoidal micro-convex body based on the equivalent simulation principle, the circumferential leakage width and radial average leakage height of the micro-leakage channel between sealing surfaces were then acquired with the surface roughness and geometric mean contact pressure. At last, the actual micro-leakage rate of the premium connection was derived by considering the non-uniform contact pressure distribution between sealing surfaces. An example was investigated to validate the model and reveal the sealing and leakage characteristics, and anti-leakage measures were proposed. The results show that average contact pressure, circumferential leakage width, and radial average leakage height between sealing surfaces were non-uniformly distributed. The leakage rate of a premium connection decreases exponentially with an increase in radial interference between sealing surfaces. In order to reduce leakage rate, it is beneficial to increase radial interference and lower sealing surface roughness

Transcriptional and protein structural characterization of homogentisate phytyltransferase genes in barley, wheat, and oat

Abstract Background Homogentisate phytyltransferase (HPT) is the critical enzyme for the biosynthesis of tocopherols (vitamin E), which are the major lipid-soluble antioxidants and help plants adapt to various stress conditions. HPT is generally strictly conserved in various plant genomes; however, a divergent lineage HPT2 was identified recently in some Triticeae species. The molecular function and transcriptional profiles of HPT2 remain to be characterized. Results In this study, we performed comprehensive transcriptome data mining of HPT1 and HPT2 in different tissues and stages of barley (Hordeum vulgare), wheat (Triticum aestivum), and oat (Avena sativa), followed by qRT-PCR experiments on HPT1 and HPT2 in different tissues of barley and wheat. We found that the common HPT1 genes (HvHPT1, TaHPT1s, and AsHPT1s) displayed a conserved transcriptional pattern in the three target species and were universally transcribed in various tissues, with a notable preference in leaf. In contrast, HPT2 genes (HvHPT2, TaHPT2, and AsHPT2) were specifically transcribed in spike (developmentally up-regulated) and shoot apex tissues, displaying a divergent tissue-specific pattern. Cis-regulatory elements prediction in the promoter region identified common factors related to light-, plant hormone-, low temperature-, drought- and defense- responses in both HPT1s and HPT2s. We observed the transcriptional up-regulation of HvHPT1 and HvHPT2 under various stress conditions, supporting their conserved function in environmental adaption. We detected a clear, relaxed selection pressure in the HPT2 lineage, consistent with the predicted evolution pattern following gene duplication. Protein structural modelling and substrate docking analyses identified putative catalytic amino acid residues for HvHPT1 and HvHPT2, which are strictly conserved and consistent with their function in vitamin E biosynthesis. Conclusions We confirmed the presence of two lineages of HPT in Triticeae and Aveninae, including hexaploid oat, and characterized their transcriptional profiles based on transcriptome and qRT-PCR data. HPT1s were ubiquitously transcribed in various tissues, whilst HPT2s were highly expressed in specific stages and tissue. The active transcription of HPT2s, together with its conserved cis-elements and protein structural features, support HPT2s’ role in tocopherol production in Triticeae. This study is the first protein structural analysis on the membrane-bound plant HPTs and provides valuable insights into its catalytic mechanism

Coherent extreme ultraviolet free-electron laser with echo-enabled harmonic generation

The echo-enabled harmonic generation (EEHG) scheme holds promising prospects for efficiently generating intense coherent radiation at very high harmonics of a conventional ultraviolet seed laser. We report the lasing of the EEHG free-electron laser (FEL) at an extreme ultraviolet (EUV) wavelength with a seeded FEL facility, the Shanghai soft x-ray FEL. For the first time, we have benchmarked the basic theory of EEHG by measuring the bunching factor distributions over one octave down to the EUV region. Our results demonstrated the key advantages of the EEHG FEL, i.e., generation of very high harmonics with a small laser-induced energy spread and insensitivity to beam imperfections, and marks a great step towards fully coherent x rays with the EEHG scheme