scholarly journals The Arabidopsis expansin gene (AtEXPA18) is capable to ameliorate drought stress tolerance in transgenic tobacco plants

Author(s):  
Alireza Abbasi ◽  
Meysam Malekpour ◽  
Sajjad Sobhanverdi

Abstract Expansins are cell wall proteins that, due to changes in pH, causing the expansion of the cell walls. In this study, a previously gene construct designed based on a root-specific gene, AtEXPA18, was utilized to assess its potential roles on different morphological, physiological, and cellular levels of generated transgenic tobacco plants in response to moderate and severe drought stress. AtEXPA18 gene was successfully transferred to the tobacco plants through an agrobacterium-mediate transformation system. Upon obtaining the second generation, tobacco transgenic plants were confirmed by conventional polymerase chain reaction (PCR) technique alongside reverse transcription PCR (RT-PCR) using specific primers. Under drought stress, the transgenic lines showed remarkable growth and significantly improved based on morphological traits such as height and stem diameter, leaf area, leaf number, root dry weight, and Abscisic acid (ABA) levels of leaves compared control plants. As a result, the Cytokinin content of transgenic plants has increased under severe stress levels. Notably, the area's expansion for abaxial epidermal cells under the microscope confirmed in transgene cells compared with the -transgene cells. These results, altogether, could support the AtEXPA18 gene implication in cell expansion and improving tolerance capacity of transgenic crops under drought stress.

Author(s):  
Ai-Hua Wang ◽  
Lan Yang ◽  
Xin-Zhuan Yao ◽  
Xiao-Peng Wen

AbstractPhosphoethanolamine N-methyltransferase (PEAMTase) catalyzes the methylation of phosphoethanolamine to produce phosphocholine and plays an important role in the abiotic stress response. Although the PEAMT genes has been isolated from many species other than pitaya, its role in the drought stress response has not yet been fully elucidated. In the present study, we isolated a 1485 bp cDNA fragment of HpPEAMT from pitaya (Hylocereus polyrhizus). Phylogenetic analysis showed that, during its evolution, HpPEAMT has shown a high degree of amino acid sequence similarity with the orthologous genes in Chenopodiaceae species. To further investigate the function of HpPEAMT, we generated transgenic tobacco plants overexpressing HpPEAMT, and the transgenic plants accumulated significantly more glycine betaine (GB) than did the wild type (WT). Drought tolerance trials indicated that, compared with those of the wild-type (WT) plants, the roots of the transgenic plants showed higher drought tolerance ability and exhibited improved drought tolerance. Further analysis revealed that overexpression of HpPEAM in Nicotiana tabacum resulted in upregulation of transcript levels of GB biosynthesis-related genes (NiBADH, NiCMO and NiSDC) in the leaves. Furthermore, compared with the wild-type plants, the transgenic tobacco plants displayed a significantly lower malondialdehyde (MDA) accumulation and higher activities of the superoxide dismutase (SOD) and peroxidase (POD) antioxidant enzymes under drought stress. Taken together, our results suggested that HpPEAMT enhanced the drought tolerance of transgenic tobacco.


2004 ◽  
Vol 84 (3) ◽  
pp. 785-790 ◽  
Author(s):  
Jianhong Yao, Xiuyun Zhao ◽  
Huaxiong Qi, Bingliang Wan ◽  
Fei Chen, Xiaofen Sun ◽  
Shanqian Yu ◽  
Kexuan Tang

Tobacco leaf discs were transformed with a plasmid, pBIAHA, containing the selectable marker neomycin phosphotransferase gene (nptII) and an Arisaema heterophyllum agglutinin gene (aha) via Agrobacterium tumefaciens-mediated transformation. Thirty-two independent transgenic tobacco plants were regenerated. PCR and Southern blot analyses confirmed that multiple copies of the aha gene had integrated into the plant genome. Northern blot analysis revealed that the aha gene was expressed at various levels in the transgenic plants. Insect bioassay test showed that transgenic plants expressing multiple copies of the aha gene reduced the rate of population increase of the peach potato aphid (Myzus persicae Sulzer). This is the first report that transgenic tobacco plants expressing the aha gene display enhanced resistance to aphids. Key words: Insect bioassay, Arisaema heterophyllum agglutinin, transformation, transgenic tobacco, peach potato aphid (Myzus persicae Sulzer)


2020 ◽  
Vol 21 (4) ◽  
pp. 1323 ◽  
Author(s):  
Wei Li ◽  
Changxi Dang ◽  
Yuxiu Ye ◽  
Zunxin Wang ◽  
Laibao Hu ◽  
...  

In plants, auxin/indoleacetic acid (Aux/IAA) proteins are transcriptional regulators that regulate developmental process and responses to phytohormones and stress treatments. However, the regulatory functions of the Vitis vinifera L. (grapevine) Aux/IAA transcription factor gene VvIAA18 have not been reported. In this study, the VvIAA18 gene was successfully cloned from grapevine. Subcellular localization analysis in onion epidermal cells indicated that VvIAA18 was localized to the nucleus. Expression analysis in yeast showed that the full length of VvIAA18 exhibited transcriptional activation. Salt tolerance in transgenic tobacco plants and Escherichia. coli was significantly enhanced by VvIAA18 overexpression. Real-time quantitative PCR analysis showed that overexpression of VvIAA18 up-regulated the salt stress-responsive genes, including pyrroline-5-carboxylate synthase (NtP5CS), late embryogenesis abundant protein (NtLEA5), superoxide dismutase (NtSOD), and peroxidase (NtPOD) genes, under salt stress. Enzymatic analyses found that the transgenic plants had higher SOD and POD activities under salt stress. Meanwhile, component analysis showed that the content of proline in transgenic plants increased significantly, while the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) decreased significantly. Based on the above results, the VvIAA18 gene is related to improving the salt tolerance of transgenic tobacco plants. The VvIAA18 gene has the potential to be applied to enhance plant tolerance to abiotic stress.


2009 ◽  
Vol 46 (2) ◽  
pp. 63-75 ◽  
Author(s):  
Roya Razavizadeh ◽  
Ali Ehsanpour

Effects of salt stress on proline content, expression of delta-1-pyrroline-5-carboxylate synthetase, and activities of catalase and ascorbate peroxidase in transgenic tobacco plantsIn arid and semiarid regions, soil salinity limits crop production. Proline accumulation in transgenic plants results in increased stress tolerance, but the underlying mechanism was unclear. To elucidate it, effects of salt stress on the expression pattern of Δ1-pyrroline-5-carboxylate synthetase (P5CS), proline content, catalase (CAT), and ascorbate peroxidase (APX) activities were analyzed in transgenic tobacco (Nicotiana tabacumcv. Wisconsin). Transgenic tobacco plants containing CaMV 35S promoter and theP5CSgene from moth bean (Vigna aconitifolia), linked to theNPTIIgene, were culturedin vitrowith or without 300 mM NaCl. The expression pattern ofP5CSwas evaluated using semiquantitative RT-PCR (reverse transcription-polymerase chain reaction). Time-course experiments showed an increase in proline content after 4 h of the treatment. The level ofP5CStranscripts was increased significantly in leaves and roots of transgenic plants after 24 and 48 h of treatment. This rise in transcripts was concomitant with the highest increase in proline content. In addition, CAT and APX activities increased under salt stress, and their highest activities were observed after 24 and 48 h of NaCl treatment. These results suggest thatP5CSis an inducible gene regulating the activities of CAT and APX and the accumulation of proline in plants subjected to salt stress.


Author(s):  
Coenraad R. van Beek ◽  
Tapiwa Guzha ◽  
Nolusindiso Kopana ◽  
Cornelius S. van der Westhuizen ◽  
Sanjib K. Panda ◽  
...  

1999 ◽  
Vol 80 (11) ◽  
pp. 2813-2822 ◽  
Author(s):  
Liliana F. Franco-Lara ◽  
Kara D. McGeachy ◽  
Uli Commandeur ◽  
Robert R. Martin ◽  
Mike A. Mayo ◽  
...  

A full-length cDNA copy of the genome of Potato leafroll virus (PLRV) was introduced into the genome of tobacco and potato plants by Agrobacterium tumefaciens-mediated transformation. Transgenic lines were obtained in which the transgene was readily detected by PCR with DNA extracted from T1 tobacco seedlings and clonally multiplied potato plants. PLRV-specific genomic and sub- genomic RNAs, coat protein antigen and virus particles were detected in transgenic plants. Aphids fed on the transgenic tobacco plants readily transmitted PLRV to test plants. Infected transgenic tobacco plants, like non-transgenic (WT) PLRV-infected plants, displayed no symptoms of the infection but transgenic plants of potato were severely stunted. In parallel tests, the mean PLRV titres in WT tobacco plants and transgenic tobacco plants were 600 and 630 ng virus/g leaf, respectively, although differences in PLRV titres among transgenic plants were much greater than those among infected WT plants. In similar tests with potato, the mean PLRV titre of WT plants was 50 ng virus/g leaf whereas higher concentrations (up to 3400 ng virus/g leaf) accumulated in transgenic potato plants. In tissue prints of stems, PLRV was detected in similar proportions of phloem cells in transgenic and infected WT plants. In transgenic tobacco and potato plants, but not in infected WT plants, a few stem epidermal cells also contained virus. From tissue prints of transgenic tobacco leaves, it was estimated that about one in 40000 mesophyll cells contained virus, but in transgenic potato, a greater proportion of mesophyll cells was infected.


1994 ◽  
Vol 3 (3) ◽  
pp. 159-166 ◽  
Author(s):  
Cornelia van der Hoeven ◽  
Antje Dietz ◽  
Jorg Landsmann

2006 ◽  
Vol 398 (3) ◽  
pp. 577-583 ◽  
Author(s):  
Meng Lin ◽  
Stefan Rose-John ◽  
Joachim Grötzinger ◽  
Udo Conrad ◽  
Jürgen Scheller

In murine models of Crohn's disease, rheumatoid arthritis and colon cancer, IL-6 (interleukin-6) signalling via the sIL-6R (soluble IL-6 receptor; termed IL-6 trans-signalling) has been shown to promote the pathology associated with these conditions. These detrimental activities can, however, be selectively blocked by soluble forms of the gp130 (glycoprotein 130) receptor. Although sgp130 (soluble gp130) therefore represents a viable therapeutic modality for the treatment of these conditions, the mass manufacture of such biologics is often expensive. The advent of molecular farming has, however, provided an extremely cost-effective strategy for the engineering of recombinant proteins. Here, we describe the expression and production of a biologically active sgp130 variant that is expressed in transgenic tobacco plants as an ELP (elastin-like peptide)-fusion protein (mini-gp130–ELP). Mini-gp130–ELP consists of the first three domains of gp130 (Ig-like domain and cytokine binding module) fused to 100 repeats of ELP. Expression of mini-gp130–ELP did not affect the growth rate or morphology of the transgenic plants, and purification was achieved using inverse transition cycling. This approach led to an overall yield of 141 μg of purified protein per g of fresh leaf weight. The purified mini-gp130–ELP specifically inhibited sIL-6R-mediated trans-signalling as measured by binding to the IL-6–sIL-6R complex and through its ability to block sIL-6R-mediated activation of STAT3 (signal transducer and activator of transcription 3) phosphorylation and proliferation in human hepatoma cells and murine pre-B-cells. Consequently, the present study validates the potential application of molecular farming in transgenic tobacco plants as a strategy for the expression and purification of therapeutically advantageous biologics such as sgp130.


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