Induction of Cell Death in Transgenic Plants Expressing a Fungal Glucose Oxidase

Hydrogen peroxide (H2O2) has been implicated in the induction of plant defense genes and programmed cell death. Expression of a chimeric fungal glucose oxidase (GO) gene driven by a pathogen- and wound-inducible promoter was evaluated in transgenic tobacco and canola as a possible tool for engineering plant cell death and defense gene induction. Expression of this gene under the control of a peroxidase gene promoter resulted in the accumulation of relatively low levels of H2O2 in the young leaves of transgenic tobacco plants and this was not sufficient to cause any visible cell death and defense gene induction as measured by PR-1a mRNA induction. Older leaves of transgenic tobacco plants, however, exhibited visible necrotic lesions and constitutively expressed PR-1a mRNA when grown under high light conditions. Inoculation of cotyledons of control and transgenic canola with Leptosphaeria maculans resulted in rapid cotyledon senescence in the transgenic plants. Strong activators of the peroxidase promoter, i.e., wounding and inoculation of transgenic plants with Cercospora nicotianae, were not sufficient to trigger any additional visible cell death in transgenic tobacco plants, compared with controls. However, when exogenous glucose was supplied to transgenic tissue, massive cell death and PR-1a gene induction were observed in tobacco. Exogenously applied salicylic acid further increased the rate and extent of cell death. Our results suggest that efficacy of GO expression for the induction of cell death is restricted by glucose supply in the plants and are consistent with a role for salicylic acid in the potentiation of plant cell death by H2O2.

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Overexpression of the pitaya phosphoethanolamine N-methyltransferase gene (HpPEAMT) enhanced simulated drought stress in tobacco

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-021-02040-3 ◽

2021 ◽

Author(s):

Ai-Hua Wang ◽

Lan Yang ◽

Xin-Zhuan Yao ◽

Xiao-Peng Wen

Keyword(s):

Drought Stress ◽

Stress Response ◽

Transgenic Plants ◽

Drought Tolerance ◽

Transgenic Tobacco ◽

Sequence Similarity ◽

Amino Acid Sequence Similarity ◽

Transgenic Tobacco Plants ◽

Wild Type ◽

Tobacco Plants

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.

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Transgenic tobacco expressing an Arisaema heterophyllum agglutinin gene displays enhanced resistance to aphids

Canadian Journal of Plant Science ◽

10.4141/p03-036 ◽

2004 ◽

Vol 84 (3) ◽

pp. 785-790 ◽

Cited By ~ 6

Author(s):

Jianhong Yao, Xiuyun Zhao ◽

Huaxiong Qi, Bingliang Wan ◽

Fei Chen, Xiaofen Sun ◽

Shanqian Yu ◽

Kexuan Tang

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Myzus Persicae ◽

Transgenic Tobacco Plants ◽

Insect Bioassay ◽

Tobacco Plants ◽

Potato Aphid ◽

Enhanced Resistance ◽

Multiple Copies ◽

Peach Potato Aphid

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)

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

10.21203/rs.3.rs-204710/v1 ◽

2021 ◽

Author(s):

Alireza Abbasi ◽

Meysam Malekpour ◽

Sajjad Sobhanverdi

Keyword(s):

Drought Stress ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Dry Weight ◽

Conventional Polymerase Chain Reaction ◽

Specific Gene ◽

Severe Drought ◽

Transgenic Tobacco Plants ◽

Tobacco Plants ◽

Reverse Transcription Pcr

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.

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Overexpression of Grapevine VvIAA18 Gene Enhanced Salt Tolerance in Tobacco

International Journal of Molecular Sciences ◽

10.3390/ijms21041323 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1323 ◽

Cited By ~ 1

Author(s):

Wei Li ◽

Changxi Dang ◽

Yuxiu Ye ◽

Zunxin Wang ◽

Laibao Hu ◽

...

Keyword(s):

Salt Stress ◽

Salt Tolerance ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Transcriptional Activation ◽

Late Embryogenesis Abundant Protein ◽

Pcr Analysis ◽

Vitis Vinifera L ◽

Transgenic Tobacco Plants ◽

Tobacco Plants

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.

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Disease Resistance Conferred by Constitutive Expression of a Fungal Glucose Oxidase Gene in Transgenic Tobacco Plants

Asian Journal of Plant Sciences ◽

10.3923/ajps.2013.128.136 ◽

2013 ◽

Vol 12 (3) ◽

pp. 128-136 ◽

Cited By ~ 2

Author(s):

Palaniappan Selvakumar ◽

Singaram Arvinth ◽

Subbiyan Maruthasal ◽

Chin Ho Lin

Keyword(s):

Disease Resistance ◽

Glucose Oxidase ◽

Transgenic Tobacco ◽

Constitutive Expression ◽

Transgenic Tobacco Plants ◽

Tobacco Plants ◽

Oxidase Gene ◽

Glucose Oxidase Gene

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Over-expression of SlJA2 decreased heat tolerance of transgenic tobacco plants via salicylic acid pathway

Plant Cell Reports ◽

10.1007/s00299-017-2100-9 ◽

2017 ◽

Vol 36 (4) ◽

pp. 529-542 ◽

Cited By ~ 10

Author(s):

Zhong-Ming Liu ◽

Meng-Meng Yue ◽

Dong-Yue Yang ◽

Shao-Bo Zhu ◽

Na-Na Ma ◽

...

Keyword(s):

Salicylic Acid ◽

Transgenic Tobacco ◽

Heat Tolerance ◽

Transgenic Tobacco Plants ◽

Tobacco Plants ◽

Over Expression ◽

Salicylic Acid Pathway ◽

Acid Pathway

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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 plants

Biological Letters ◽

10.2478/v10120-009-0002-4 ◽

2009 ◽

Vol 46 (2) ◽

pp. 63-75 ◽

Cited By ~ 12

Author(s):

Roya Razavizadeh ◽

Ali Ehsanpour

Keyword(s):

Salt Stress ◽

Transgenic Plants ◽

Transgenic Tobacco ◽

Expression Pattern ◽

Ascorbate Peroxidase ◽

Proline Content ◽

35S Promoter ◽

Transgenic Tobacco Plants ◽

Tobacco Plants ◽

Moth Bean

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.

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Transformation of tobacco and potato with cDNA encoding the full-length genome of Potato leafroll virus: evidence for a novel virus distribution and host effects on virus multiplication

Journal of General Virology ◽

10.1099/0022-1317-80-11-2813 ◽

1999 ◽

Vol 80 (11) ◽

pp. 2813-2822 ◽

Cited By ~ 34

Author(s):

Liliana F. Franco-Lara ◽

Kara D. McGeachy ◽

Uli Commandeur ◽

Robert R. Martin ◽

Mike A. Mayo ◽

...

Keyword(s):

Transgenic Plants ◽

Transgenic Tobacco ◽

Transgenic Potato ◽

Transgenic Tobacco Plants ◽

Mesophyll Cells ◽

Tobacco Plants ◽

Potato Plants ◽

The Mean ◽

Leafroll Virus ◽

Tissue Prints

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.

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Role for Salicylic Acid in the Activation of Defense Responses in Catalase-Deficient Transgenic Tobacco

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1997.10.7.922 ◽

1997 ◽

Vol 10 (7) ◽

pp. 922-925 ◽

Cited By ~ 20

Author(s):

He Du ◽

Daniel F. Klessig

Keyword(s):

Salicylic Acid ◽

Transgenic Tobacco ◽

Catalase Activity ◽

Defense Responses ◽

Transgenic Tobacco Plants ◽

Strong Light ◽

Tobacco Plants ◽

Enhanced Resistance ◽

The Cross

Transgenic tobacco plants with severely reduced catalase activity were crossed with NahG plants that do not accumulate salicylic acid. Some of the progeny from the cross spontaneously developed necrosis when grown under strong light, similar to that observed on the parental severely catalase-deficient plants. However, in contrast to the parental catalase-deficient plants, these progeny plants did not constitutively express PR-1 genes or develop enhanced resistance. Thus, salicylic acid appears to be required for the induction of these defense responses in catalase-deficient tobacco plants.

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