Morphophysiological and molecular evidence supporting the augmentative role of Piriformospora indica in mitigation of salinity in Cucumis melo L.

Salinity is one of the major limiting factors in plant growth and productivity.Cucumis melo L.is a widely cultivated plant,but its productivity is significantly influenced by the level of salinity in soil.Symbiotic colonization of plants with Piriformospora indica has shown a promotion in plants growth and tolerance against biotic stress.In this study,physiological markers such as ion analysis,antioxidant determination,proline content,electrolyte leakage and chlorophyll measurement were assessed in melon cultivar under two concentrations(100 and 200 mM)of NaCl with and without P.indica inoculation.Results showed that the endophytic inoculation consistently upregulated the level of antioxidants,enhanced plants to antagonize salinity stress.The expression level of an RNA editing factor(SLO2)which is known to participate in mitochondria electron transport chain was analyzed,and its full mRNA sequence was obtained by rapid amplification of cDNA ends(RACE).Under salinity stress,the expression level of SLO2 was increased,enhancing the plant’s capability to adapt to the stress.However,P.indica inoculation further elevated the expression level of SLO2.These findings suggested that the symbiotic association of fungi could help the plants to tolerate the salinity stress.     

Effects of environmental factors on gross caloric values of three life-forms aquatic plants on the Qinghai-Xizang Plateau,China北大核心CSCD

Aims Gross caloric value (GCV) reflected plants' capability of converting solar energy. It provided a reliable indicator of plants' adaptations to environments in perspective of energy conversion and fixation. The aims of this study were (1) to illustrate the characteristics of GCV of aquatic plants on the Qinghai-Xizang Plateau, (2) to explore the geographical and environmental patterns and (3) to discuss the underlying mechanisms in forming the patterns. Methods In July and August 2015, we collected 533 samples of aquatic plants' leaves in 143 field sites on the Qinghai-Xizang Plateau, and measured their GCV using SDACM-4000 oxygen bomb calorimeter. Together with mean annual temperature (MAT) of climatic factor and properties of water body, this study compared the differences of GCV among submerged, emergent and floating-leaved plants by analysis of variance (ANOVA) and Tukey's HSD. We further regressed GCV of submerged and emergent plants against geographical and climatic factors and properties of water body by simple regression to explore the relative effects of environmental factors on GCV. Important findings On the Qinghai-Xizang Plateau, the mean GCV was (15.95 ± 3.90) kJ·g-1. Among the three life forms, the rank of GCV was the emergent plants (18.10 kJ·g-1) > the floating-leaved plants (16.77 kJ·g-1) > the submerged plants (14.31 kJ·g-1). With an increasing latitude, the GCV of emergent and submerged plants increased. Only GCV of emergent plants decreased with increasing altitude and temperature. The GCV of emergent and submerged plants increased with increased water salinity. Dissolved oxygen had significant negative effects on emergent plants, while pH value had no significant effects.     

Increased Vitamin C Content Accompanied by an Enhanced Recycling Pathway Confers Oxidative Stress Tolerance in Arabidopsis

Vitamin C(L-ascorbic acid,AsA) has important antioxidant and metabolic functions in both plants and animals.Once used,ascorbic acid can be regenerated from its oxidized form in a reaction catalyzed by dehydroascorbate reductase(DHAR,EC 1.8.5.1).To analyze the physiological role of DHAR catalyzing the reduction of DHA to ascorbate in environmental stress adaptation,we examined whether increasing the level of AsA through enhanced AsA recycling would limit the deleterious effects of oxidative stress.A chimeric construct consisting of the double CaMV35S promoter fused to the Myc-dhar gene was introduced into Arabidopsis thaliana.Transgenic plants were biochemically characterized and tested for responses to oxidative stress.Western blot indicated that the dhar-transgene was successfully expressed.In homozygous T4 transgenic seedlings,DHAR overexpression was increased up to 1.5 to 5.4 fold,which enhanced foliar ascorbic acid levels 2-to 4.25-fold and ratio of AsA/DHA about 3-to 16-fold relative to wild type.In addition,the level of glutathione,the reductant used by DHAR,also increased as did its redox state.When whole plants were treated with high light and high temperature stress or in vitro leaf discs were subjected to 10 μM paraquat,transgenic plants showed a larger AsA pool size,lower membrane damage,and a higher level of chlorophyll compared with controls.These data suggested that increasing the plant vitamin C content through enhanced ascorbate recycling could limit the deleterious effects of environmental oxidative stress.     

A defect in the PINOID serine/threonine kinase affects leaf shape in cucumber

<正>Examining the plants in any forest or meadow reveals a remarkable diversity of leaf shape, suggesting the importance of this trait for adaptation to various environmental conditions (reviewed in Nicotra et al.2011). Indeed, leaf shape may be constrained by biomechanical factors and affects thermoregulation,     

转基因番木瓜的抗病性及分子鉴定

对T1代转番木瓜环斑病毒(PRV)复制酶(RP)突变体基因的两个番木瓜株系,进行了抗病性和分子生物学分析。结果表明,转基因番木瓜对PRV抗性达到高抗或免疫,目的基因RP遗传至转基因后代并在RNA水平表达,PCR可检测到CaMV35S启动子序列、标记基因NPTII。 Abstract:Virus resistance in field and molecular biological characterizations of the transgenes were analyzed for two lines of T1 generation of transgenic papaya with the replicase mutant gene from papaya ringspot virus (PRV).The transgenic plants showed highly resistant or immune against PRV.Results indicated that the transgenes inherited to and expressed at RNA level in the progenies.     

Fading indirect effects in a warming arctic tundra

Indirect interactions in food webs can strongly influence the net effect of global change on ecological communities yet they are rarely quantified and hence remain poorly understood. Using a 22-year time series, we investigated climate-induced and predator-mediated indirect effects on grazing intensity in the tundra food web of Bylot Island, which experienced a warming trend over the last two decades. We evaluated the relative effects of environmental parameters on the proportion of plant biomass grazed by geese in wetlands and examined the temporal changes in the strength of these cascading effects. Migrating geese are the dominant herbivores on Bylot Island and can consume up to 60% of the annual production of wetland graminoids. Spring North Atlantic Oscillation, mid-summer temperatures and summer abundance of lemmings （prey sharing predators with geese） best-explained annual variation in grazing intensity. Goose grazing impact increased in years with high temperatures and high lemming abundance. However, the strength of these indirect effects on plants changed over time. Grazing intensity was weakly explained by environmental factors in recent years, which were marked by a sharp increase in plant primary production and steady decrease in grazing pressure. Indirect effects do not seem to be reversing the direct positive effect of warming on wetland plants. We suggest that cascading effects on plants may lag considerably behind direct effects in vertebrate dominated arctic communities, especially where key herbivore populations are strongly affected by factors outside of the Arctic [Current Zoology 60 （2）： 189-202, 2014].     

植物基因工程表达载体的改进和优化策略

外源基因在转基因植物中表达效率低一直是令相关研究者困扰的一个问题.转基因植物的生物安全性最近在全球范围内开始引起世人的担忧.本文简要介绍了近年来在植物表达载体构建方面所采用的一些新策略,这些策略有助于增强外源基因的表达水平、提高生物工程体的安全性。 Abstract:The low expression level of foreign genes in transgenic plants is a puzzling question for researchers in plant genetic engineering.The biosafety concern is now causing a growing public wariness of transgenic plants around the world.This article reviews the new strategies currently used in construction of plant expression vector for plant genetic engineering.These strategies are important for obtaining better expression of foreign genes and developing safer transgenic plants.     

Seasonal Variation of δ13C of Four Tree Species: A Biological Integrator of Environmental Variables

Foliar δ13C values, an indicator of long-term intercellular carbon dioxide concentration and, thus,of long-term water use efficiency (WUE) in plants, were measured for Pinus massoniana Lamb., P. elliottii Engelm., Cunninghamia laceolata (Lamb.) Hook., and Schima superba Gardn. et Champ. in a restored forest ecosystem in the Jiazhu River Basin. Seasonal variation and the relationship between the foliar δ13Cvalues of the four species and environmental factors (monthly total precipitation, monthly average air temperature, relative humidity, atmospheric pressure, and monthly total solar radiation and evaporation)were investigated. The monthly δ13C values and WUE of the four species increased with increasing precipitation, air temperature, solar radiation, and evaporation, whereas δ13C values of the four species decreased with increasing relative humidity and atmospheric pressure. Despite significant differences in δ13C seasonal means for the four species, our results demonstrate a significant convergence in the responses of δ13C values and WUE to seasonal variations in environmental factors among the species investigated and that the δ13C signature for each species gives a strong indication of environmental variables.     

Genotoxic evaluation of an industrial effluent from an oil refinery using plant and animal bioassays

Polycyclic aromatic hydrocarbons (PAHs) are genotoxic chemicals commonly found in effluents from oil refineries. Bioassays using plants and cells cultures can be employed for assessing environmental safety and potential genotoxicity. In this study, the genotoxic potential of an oil refinery effluent was analyzed by means of micronucleus (MN) testing of Alium cepa, which revealed no effect after 24 h of treatment. On the other hand, primary lesions in the DNA of rat (Rattus norvegicus) hepatoma cells (HTC) were observed through comet assaying after only 2 h of exposure. On considering the capacity to detect DNA damage of a different nature and of these cells to metabolize xenobiotics, we suggest the association of the two bioassays with these cell types, plant (Allium cepa) and mammal (HTC) cells, for more accurately assessing genotoxicity in environmental samples.     

Evaluation of genotoxicity of Pseudevernia furfuracea (L.) Zopf by RAPD analysis

We investigated the suitability and applicability of Pseudevernia furfuracea (L.) Zopf for environmental genotoxicity assessment. P. furfuracea lichen specimens were collected from 10 different Pinus species, in every 5 km, starting from around an iron-steel factory located in the central area of Karabük Province up to Yenice Forest. The impact of the pollution sources such as iron-steel factory, roads and railroads, industry, heavy traffic, and waste treatment plants on the heavy metal accumulation in lichens is known. DNA changes in P. furfuracea samples exposed naturally to various polluted sites were analyzed by RAPD to know the influence of the environmental pollution on the hereditary material of the organisms. Twenty-five different primers were tested and 10 yielded clear and reproducible bands. The present study shows the suitability of the lichen samples for the detection of genotoxicity and also provides information about the level of potential genotoxic agents around a steel mill.     

鱼微核试验筛检水体诱变物的应用与研究

对五种试验污染物的遗传毒性采用鱼微核试验技术进行了筛检,试验结果表明鱼微核试验用作筛检诱变物的遗传毒性有效的,对监测水体的质量也具有潜在的价值;同时,对国内外学将各种各样的技术应用到鱼微核式试验进行了总结,概述了微核形成的机理;在此基础上,提出应用鱼微核试验筛检水体诱变物时应注意的问题。     

Genotoxicity of 2,4-D and dicamba revealed by transgenic Arabidopsis thaliana plants harboring recombination and point mutation markers

The phenoxy herbicides 2,4-D and dicamba are released daily into the environment in large amount. The mechanisms of genotoxicity and mutagenicity of these herbicides are poorly understood, and the available genotoxicity data is controversial. There is a cogent need for a novel genotoxicity monitoring system that could provide both reliable information at the molecular level, and complement existing systems.We employed the transgenic Arabidopsis thaliana 'point mutation' and 'recombination' plants to monitor the genetic effects of the herbicides 2,4-D and dicamba. We found that both herbicides had a significant effect on the frequency of homologous recombination A-->G mutation. Neither herbicides affected the T-->G mutation frequency. Interestingly, these transgenic biomonitoring plants were able to detect the presence of phenoxy herbicides at concentrations that were lower than the guideline levels for Drinking Water Quality. The results of our studies suggest that our transgenic system may be ideal for the evaluation of the genotoxicity of herbicide-contaminated water. Moreover, the unique ability of the plants to detect both double-strand breaks (homologous recombination) and point mutations provides tremendous potential in the study of molecular mechanisms of genotoxicity and mutagenicity of phenoxy herbicides.     

Genotoxicity risk assessment: a proposed classification strategy

Recent advances in genetic toxicity (mutagenicity) testing methods and in approaches to performing risk assessment are prompting a renewed effort to harmonize genotoxicity risk assessment across the world. The US Environmental Protection Agency (EPA) first published Guidelines for Mutagenicity Risk Assessment in 1986 that focused mainly on transmissible germ cell genetic risk. Somatic cell genetic risk has also been a risk consideration, usually in support of carcinogenicity assessments. EPA and other international regulatory bodies have published mutagenicity testing requirements for agents (pesticides, pharmaceuticals, etc.) to generate data for use in genotoxicity risk assessments. The scheme that follows provides a proposed harmonization approach in which genotoxicity assessments are fully developed within the risk assessment paradigm used by EPA, and sets out a process that integrates newer thinking in testing battery design with the risk assessment process. A classification strategy for agents based on inherent genotoxicity, dose-responses observed in the data, and an exposure analysis is proposed. The classification leads to an initial level of concern for genotoxic risk to humans. A total risk characterization is performed using all relevant toxicity data and a comprehensive exposure evaluation in association with the genotoxicity data. The result of this characterization is ultimately used to generate a final level of concern for genotoxic risk to humans. The final level of concern and characterized genotoxicity risk assessment are communicated to decision makers for possible regulatory action(s) and to the public.     

Genotoxicity risk assessment: a proposed classification strategy

Recent advances in genetic toxicity (mutagenicity) testing methods and in approaches to performing risk assessment are prompting a renewed effort to harmonize genotoxicity risk assessment across the world. The US Environmental Protection Agency (EPA) first published Guidelines for Mutagenicity Risk Assessment in 1986 that focused mainly on transmissible germ cell genetic risk. Somatic cell genetic risk has also been a risk consideration, usually in support of carcinogenicity assessments. EPA and other international regulatory bodies have published mutagenicity testing requirements for agents (pesticides, pharmaceuticals, etc.) to generate data for use in genotoxicity risk assessments. The scheme that follows provides a proposed harmonization approach in which genotoxicity assessments are fully developed within the risk assessment paradigm used by EPA, and sets out a process that integrates newer thinking in testing battery design with the risk assessment process. A classification strategy for agents based on inherent genotoxicity, dose-responses observed in the data, and an exposure analysis is proposed. The classification leads to an initial level of concern for genotoxic risk to humans. A total risk characterization is performed using all relevant toxicity data and a comprehensive exposure evaluation in association with the genotoxicity data. The result of this characterization is ultimately used to generate a final level of concern for genotoxic risk to humans. The final level of concern and characterized genotoxicity risk assessment are communicated to decision makers for possible regulatory action(s) and to the public.     

Development of genotoxicity test procedures with Episkin, a reconstructed human skin model: towards new tools for in vitro risk assessment of dermally applied compounds?

Today reconstructed skin models that simulate human skin, such as Episkin, are widely used for safety or efficacy pre-screening. Moreover, they are of growing interest for regulatory purposes in the framework of alternatives to animal testing. In order to reduce and eventually replace results of in vivo genotoxicity testing with in vitro data, there is a need to develop new complementary biological models and methods with improved ability to predict genotoxic risk. This can be achieved if these new assays do take into account exposure conditions that are more relevant than in the current test systems. In an attempt to meet this challenge, two new applications using a human reconstructed skin model for in vitro genotoxicity assessment are proposed. The skin is the target organ for dermally exposed compounds or environmental stress. Although attempts have been made to develop genotoxicity test procedures in vivo on mouse skin, human reconstructed skin models have not been used for in vitro genotoxicity testing so far, although they present clear advantages over mouse skin for human risk prediction. This paper presents the results of the development of a specific protocol allowing to perform the comet assay, a genotoxicity test procedure, on reconstructed skin. The comet assay was conducted after treatment of Episkin with UV, Lomefloxacin and UV or 4-nitroquinoline-N-oxide (4NQO). Treatment with the sunscreen Mexoryl was able to reduce the extent of comet signal. A second approach to use reconstructed epidermis in genotoxicity assays is also proposed. Indeed, the skin is a biologically active barrier driving the response to exposure to chemical agents and their possible metabolites. A specific co-culture system (Figure 1) using Episkin to perform the regular micronucleus assay is presented. Micronucleus induction in L5178Y cells cultured underneath Episkin was assessed after treatment of the reconstructed epidermis with mitomycin C, cyclophosphamide or apigenin. This second way of using human reconstructed skin for genotoxicity testing aims at improving the relevance of exposure conditions in in vitro genotoxicity assays for dermally applied compounds.     

A novel and simple method of screening compounds for interaction with DNA: A validation study

We report the development of a simple, cost-effective assay for detecting compounds that have the ability to interact with and modify DNA. Potential uses for the assay lie in the areas of early genotoxicity testing of drug candidates, anticancer and antibiotic drug discovery, environmental monitoring and testing in the food, beverage and cosmetics industries. At present the assay has been used to assess direct-acting compounds only and it is yet to be established whether the assay is compatible with bio-activation. The methodology is based on the oxidative reaction of potassium permanganate with pyrimidine bases, which have become perturbed and more reactive by the agent under test. Results are recorded by use of UV/vis spectroscopy. The adaptation to a multi-well plate format provides the capacity for high throughput utilizing small amounts of compounds. Over 100 compounds, comprising different classes of DNA-binding chemicals as well as non-binding controls, have been put through the assay and the results compared with existing genotoxicity testing data from other methods. The assay has shown to be predictive of the results of other genotoxicity testing methods. We have found that the method is overall predictive of 71% of Ames bacterial reverse-mutation test results (where data are given) encompassing both negative and positive results.     

The ability of plant genotoxicity assays to predict carcinogenicity

A number of assays have been developed which use higher plants for measuring mutagenic or cytogenetic effects of chemicals, as an indication of carcinogenicity. Plant assays require less extensive equipment, materials and personnel than most other genotoxicity tests, which is a potential advantage, particularly in less developed parts of the world. We have analyzed data on 9 plant genotoxicity assays evaluated by the Gene-Tox program of the U.S. Environmental Protection Agency, using methodologies we have recently developed to assess the capability of assays to predict carcinogenicity and carcinogenic potency. All 9 of the plant assays appear to have high sensitivity (few false negatives). Specificity (rate of true negatives) was more difficult to evaluate because of limited testing on non-carcinogens; however, available data indicate that only the Arabidopsis mutagenicity (ArM) test appears to have high specificity. Based upon their high sensitivity, plant genotoxicity tests are most appropriate for a risk-averse testing program, because although many false positives will be generated, the relatively few negative results will be quite reliable.     

Genotoxicity of 2,4-D and dicamba revealed by transgenic Arabidopsis thaliana plants harboring recombination and point mutation markers

The phenoxy herbicides 2,4-D and dicamba are released daily into the environment in large amount. The mechanisms of genotoxicity and mutagenicity of these herbicides are poorly understood, and the available genotoxicity data is controversial. There is a cogent need for a novel genotoxicity monitoring system that could provide both reliable information at the molecular level, and complement existing systems.We employed the transgenic Arabidopsis thaliana ‘point mutation’ and ‘recombination’ plants to monitor the genetic effects of the herbicides 2,4-D and dicamba. We found that both herbicides had a significant effect on the frequency of homologous recombination A→G mutation. Neither herbicides affected the T→G mutation frequency. Interestingly, these transgenic biomonitoring plants were able to detect the presence of phenoxy herbicides at concentrations that were lower than the guideline levels for Drinking Water Quality. The results of our studies suggest that our transgenic system may be ideal for the evaluation of the genotoxicity of herbicide-contaminated water. Moreover, the unique ability of the plants to detect both double-strand breaks (homologous recombination) and point mutations provides tremendous potential in the study of molecular mechanisms of genotoxicity and mutagenicity of phenoxy herbicides.