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      An Assessment of the Environmental Impacts of Transgenic Triploid Populus tomentosa in Field Condition

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      Forests

      MDPI AG

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          Abstract

          Populus tomentosa grow rapidly, but are salt susceptible. To quickly and efficiently gain new poplar breeds with better salt resistance, a DREB transcription factor derived from Atriplex hortensis was transformed into triploid Populus tomentosa by our lab, which significantly improved the salt tolerance of host plants. However, environmental impacts of transgenic plants must be assessed before large-scale cultivation in China. Here, we conducted a field trial of AhDREB1 transgenic and non-transgenic triploid Populus tomentosa to assess the impact of transgenic trees on rhizospheric soil microbial communities and allelopathic activity of leaves. No significant differences in the number of soil microbes present were detected between the transgenic lines and the non-transgenic controls. The allelopathic activity of leaves from both the transgenic and non-transgenic lines varied with sampling time, but did not differ significantly between the transgenic and non-transgenic lines. These results indicate that the impact on the environment of AhDREB1 transgenic P. tomentosa did not differ significantly from that of the non-transformed controls for the variables observed in this field trial. We also investigated the persistence of AhDREB1 genes in decomposing transgenic poplar leaf on the soil under natural conditions for five months, and our data indicated that fragments of the genetically modified DNA were not detectable in the field after more than two months. We used a triphenyl tetrazolium chloride test (TTC) (or pollen germination method) and hybridization to test the pollen viability and fertility, respectively, of the transgenic and non-transgenic trees and the results showed that the pollen viability of both the transgenic and non-transgenic trees was extremely low in 2016; the receptor plant may have been sterile.

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          Most cited references 29

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          Gene flow from cultivated rice (Oryza sativa) to its weedy and wild relatives.

           Jung Suh,  Z. Song,  Bao Lu (2003)
          Transgene escape through gene flow from genetically modified (GM) crops to their wild relative species may potentially cause environmental biosafety problems. The aim of this study was to assess the extent of gene flow between cultivated rice and two of its close relatives under field conditions. Experiments were conducted at two sites in Korea and China to determine gene flow from cultivated rice (Oryza sativa L.) to weedy rice (O. sativa f. spontanea) and common wild rice (O. rufipogon Griff.), respectively, under special field conditions mimicking the natural occurrence of the wild relatives in Asia. Herbicide resistance (bar) and SSR molecular finger printing were used as markers to accurately determine gene flow frequencies from cultivated rice varieties to their wild relatives. Gene flow frequency from cultivated rice was detected as between approx. 0.011 and 0.046 % to weedy rice and between approx. 1.21 and 2.19 % to wild rice under the field conditions. Gene flow occurs with a noticeable frequency from cultivated rice to its weedy and wild relatives, and this might cause potential ecological consequences. It is recommended that isolation zones should be established with sufficient distances between GM rice varieties and wild rice populations to avoid potential outcrosses. Also, GM rice should not be released when it has inserted genes that can significantly enhance the ecological fitness of weedy rice in regions where weedy rice is already abundant and causing great problems.
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            Overexpression of mtlD gene in transgenic Populus tomentosa improves salt tolerance through accumulation of mannitol.

            The mtlD gene encoding mannitol-1-phosphate dehydrogenase, which catalyzes the biosynthesis of mannitol from fructose, was cloned from Escherichia coli and transferred to poplar (Populus tomentosa Carr.) through Agrobacterium-mediated transformation. The transgenic plants were screened and selected on Murashige and Skoog (MS) medium containing 30-50 mg l(-1) kanamycin and verified by polymerase chain reaction (PCR) and Southern blotting. Expression of the gene led to synthesis and accumulation of mannitol in the transgenic plants. Gas chromatography and mass spectrometry (GC/MS) and capillary gas chromatography (GC) showed that transgenic plants accumulated much more mannitol in their tissues than the wild-type plants, whether cultured in vitro, or grown hydroponically or in the field. Increased salt tolerance of transgenic plants was observed both in vitro and in hydroponic culture. The transgenic buds rooted normally on MS medium containing 50 mM NaCl, whereas wild-type buds did not. In the 40-day hydroponic experiments, transgenic poplar plants survived in a 75-mM NaCl treatment, whereas the wild-type poplar plants tolerated only 25 mM NaCl. Under the same NaCl stress, stomatal conductance, transpiration rates and photosynthetic rates were all higher in transgenic plants than in wild-type plants, whereas cellular relative conductivity was lower. We demonstrated that the mtlD gene was expressed in transgenic poplar plants, resulting either directly or indirectly in mannitol accumulation and improved salt tolerance. The constant mannitol concentrations in transgenic plants during the NaCl treatments indicated that mannitol accumulation caused by the mtlD gene was not a consequence of NaCl stress. Height growth was reduced by about 50% in the transgenic plants compared with the wild-type plants in the absence of salt; however, relative growth rate was much less influenced by salt stress in transgenic plants than in wild-type plants. The stunted growth of the transgenic plants may in part explain their improved salt tolerance.
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              Characterization of a DRE-binding transcription factor from a halophyte Atriplex hortensis.

              Environmental stresses, such as salinity, drought and cold, can induce the expression of a large amount of genes. Among these are many transcription factors that regulate the expression of downstream genes by specifically binding to cis-elements or forming transcriptional complexes with other proteins. In the present study, a DREB-like transcription factor gene, named AhDREB1, was isolated from a halophyte Atriplex hortensis. AhDREB1 encoded a protein containing a conserved EREBP/AP2 domain featuring the DREB family. In yeast one-hybrid analysis AhDREB1 protein was specifically bound to DRE elements and activated the expression of the reporter genes of HIS3 and LacZ. The AhDREB1 gene was expressed in roots, stems and leaves of A. hortensis. Salinity induced its expression in roots, but not in other organs. Overexpression of AhDREB1 in transgenic tobacco led to the accumulation of its putative downstream genes. The performance of the transgenic lines was also tested under stressed conditions and two lines were found to be stress-tolerant. These results suggest that the AhDREB1 protein functions as a DRE-binding transcription factor and play roles in the stress-tolerant response of A. hortensis.
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                Author and article information

                Journal
                Forests
                Forests
                MDPI AG
                1999-4907
                August 2018
                August 08 2018
                : 9
                : 8
                : 482
                10.3390/f9080482
                © 2018

                https://creativecommons.org/licenses/by/4.0/

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                Self URI (article page): http://www.mdpi.com/1999-4907/9/8/482

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