scholarly journals Alleviation of Phytophthora infestans Mediated Necrotic Stress in the Transgenic Potato (Solanum tuberosum L.) with Enhanced Ascorbic acid Accumulation

Plants ◽  
2019 ◽  
Vol 8 (10) ◽  
pp. 365 ◽  
Author(s):  
Ill-Min Chung ◽  
Baskar Venkidasamy ◽  
Chandrama Prakash Upadhyaya ◽  
Gurusaravanan Packiaraj ◽  
Govindasamy Rajakumar ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Transgenic Plants ◽  
Phytophthora Infestans ◽  
Solanum Tuberosum L ◽  
Transgenic Potato ◽  
Pr Genes ◽  
Pathogen Infection ◽  
Untransformed Control ◽  
Expression Of Genes ◽  
Pathogenesis Related

Potato is the most widely cultivated non-cereal crop in the world, and like any other crop, it is susceptible to yield losses because of various factors, including pathogen attacks. Among the various diseases of potato, late blight caused by the oomycete Phytophthora infestans is considered as the most devastating disease worldwide. In this study, transgenic potato plants overexpressing the D-galacturonic acid reductase (GalUR) gene with an enhanced level of cellular L-ascorbate (L-AsA) were challenged with Phytophthora infestans to determine the level of stress tolerance induced in those plants. With the onset of pathogen infection, necrotic lesions progressively expanded and became necrotic in the control plants. The transgenic potato lines with enhanced ascorbic acid showed reduced necrotic lesions. Hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels were relatively lower in transgenic plants compared to the untransformed control (UT) plants. The mRNA expressions of pathogenesis-related (PR) genes, such as pathogenesis related 1 (PR1) and phenylalanine ammonia-lyase (PAL) were slightly higher in GalUR overexpressing transgenic lines as compared to the untransformed control plants. Pathogen infection also altered the mRNA expression of genes associated with gibberellic acid (GA) and abscisic acid (ABA) biosynthesis. Furthermore, the increase in various antioxidant enzymes was also observed in the gene expression analysis with the transgenic plants. The complete loss of the pathogen growth and disease occurrence was not observed in our study; however, the findings indicated that an increase in the level of cellular L-ascorbate in the transgenic potato leads to enhanced cellular antioxidants, PR genes and plant defense hormones, such as GA and ABA resulting in the reduction of the disease symptoms caused by the Phytophthora infestans.

Overexpression of Solanum tuberosum respiratory burst oxidase homolog A (StRbohA) promotes potato tolerance to Phytophthora infestans.

2021 ◽  
Author(s):  
Atta Soliman ◽  
Lorne R. Adam ◽  
Pawanpuneet K. Rehal ◽  
Fouad Daayf
Keyword(s):  
Solanum Tuberosum ◽  
Transgenic Plants ◽  
Phytophthora Infestans ◽  
Respiratory Burst ◽  
Defense Mechanisms ◽  
Transgenic Potato ◽  
Plant Tolerance ◽  
Potato Plants ◽  
Pathogenesis Related ◽  
Respiratory Burst Oxidase

Reactive oxygen species (ROS) represent one of the first lines of plants’ biochemical defense against pathogens. Plants’ respiratory burst oxidase homologs (RBOHs) produce ROS as by-products in several cellular compartments. In potato tubers, Solanum tuberosum respiratory burst oxidase homolog (StRBOHs) are involved in suberization and healing of wounded tissues. StRbohA has been tested in the model plant Arabidopsis thaliana, which led to enhanced plant defense against the soil-borne pathogen Verticillium dahliae. Here, we showed that overexpressing StRbohA in potato plants enhancesd plant tolerance to the oomycete Phytophthora infestans, the causal agent of late blight disease. Transgenic potato plants expressing StRbohA showed reduced disease symptoms (necrosis) compared to the wild type check. The In parallel, the expression of pathogenesis-related genes (PRs), RBOHs, antioxidation-related genes CPRX1, PRX2, APRX1, CAT1, and CAT2, and genes involved in the biosynthesis pathways of jasmonic and salicylic acids (ICS, PAL1, PAL2, LOX1, LOX2, and LOX3) exhibited significant increases in the transgenic plants in response to infection. Following higher expression of RBOHs, ROS accumulated more in inoculation sites of the transgenic plants. ROS act as signals that activate gene expression in the SA biosynthesis pathway, leading to the accumulation of SA and triggering SA-based defense mechanisms. SA-responsive pathogenesis-related genes (PRs) showed higher expression in the transgenic plants, which resulted in the restriction of pathogen growth in plant tissues. These results represent a demonstration of the effective role of StRbohA in enhancing potato defense against P. infestans.

scholarly journals Isolate specific responses of the non-host grass Brachypodium distachyon to the fungal pathogen Zymoseptoria tritici, compared to wheat.

2020 ◽  
Author(s):  
Ailsing Reilly ◽  
Sujit Jung Karki ◽  
Anthony Twamley ◽  
Anna M.M Tiley ◽  
Steven Kildea ◽  
...  
Keyword(s):  
Fungal Pathogen ◽  
Plant Immunity ◽  
Brachypodium Distachyon ◽  
Septoria Tritici Blotch ◽  
Septoria Tritici ◽  
Pr Genes ◽  
Zymoseptoria Tritici ◽  
Expression Of Genes ◽  
Pathogenesis Related ◽  
Defence Gene Expression

Septoria tritici blotch (STB) is an important foliar disease of wheat that is caused by the fungal pathogen Zymoseptoria tritici. The grass Brachypodium distachyon has been used previously as a model system for cereal-pathogen interactions. In this study, we examined the non-host resistance (NHR) response of B. distachyon to two different Z. tritici isolates in comparison to wheat. These isolates vary in aggressiveness on wheat cv. Remus displaying significant differences in disease and pycnidia coverage. Using microscopy, we found that similar isolate specific responses were observed for H2O2 accumulation and cell death in both wheat and B. distachyon. Despite this, induction of isolate specific patterns of defence gene expression by Z. tritici did differ between B. distachyon and wheat. Our results suggest that phenylalanine ammonia lyase (PAL) expression may be important for NHR in B. distachyon while pathogenesis-related (PR) genes and expression of genes regulating reactive oxygen species (ROS) may be important to limit disease in wheat. Future studies of the B. distachyon-Z. tritici interaction may allow identification of conserved plant immunity targets which are responsible for the isolate specific responses observed in both plant species.

Transgenic Potato Plants Expressing Soybean ß-1,3-Endoglucanase Gene Exhibit an Increased Resistance to Phytophthora infestans

1998 ◽  
Vol 53 (11-12) ◽  
pp. 1012-1016 ◽  
Author(s):  
Maria Borkowska ◽  
Magdalena Krzymowska ◽  
Andrzej Talarczyk ◽  
Malik F. M. Awan ◽  
Ludmila Yakovleva ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Phytophthora Infestans ◽  
Cell Walls ◽  
Fungal Pathogens ◽  
Pcr Amplification ◽  
Transgenic Potato ◽  
Fungal Cell ◽  
Potato Genome ◽  
Potato Plants ◽  
Transgenic Potato Plants

Abstract Soybean β-1,3-endoglucanase represents a model system for studies on early plant re­sponses to infection by fungal pathogens, and it has been implicated in the release of elicitors from fungal cell walls. In the present study, potato plants were transformed with the soybean β-1,3-endoglucanase cDNA via Agrobacterium delivery system. The transfer of the gene into potato genome was confirmed by (i) PCR amplification, (ii) Northern blot analyses, and (Hi) an increase in the activity of β-1,3-endoglucanase in transgenic plants. The transformation resulted in an increased resistance of selected transgenic plants to infection by Phytophthora infestans, an important pathogen.

scholarly journals Novel Approach to Parasitic Weed Control Based on Inducible Expression of Cecropin in Transgenic Plants

2003 ◽  
Author(s):  
Radi Aly ◽  
James H. Westwood ◽  
Carole L. Cramer
Keyword(s):  
Transgenic Plants ◽  
Weed Control ◽  
Control Strategies ◽  
Solanum Tuberosum L ◽  
Transgenic Potato ◽  
Inducible Expression ◽  
Growth And Yield ◽  
Parasitic Weeds ◽  
Small Proteins ◽  
Enhanced Resistance

Our overall goal was to engineer crop plants with enhanced resistance to Orobanche (broomrape) based on the inducible expression of sarcotoxin-like peptide (SLP). A secondary objective was to localize small proteins such as SLP in the host-parasite union in order to begin characterizing the mechanism of SLP toxicity to Orobanche. We have successfully accomplished both of these objectives and have demonstrated that transgenic tobacco plants expressing SLP under control of the HMG2 promoter show enhanced resistance to O. aegyptiaca and O. ramosa . Furthermore, we have shown that proteins much larger than the SLP move into Orobanche tubercles from the host root via either symplastic or apoplastic routes. This project was initiated with the finding that enhanced resistance to Orobanche could be conferred on tobacco, potato, and tomato by expression of SLP (Sarcotoxin IA is a 40-residue peptide produced as an antibiotic by the flesh fly, Sarcophaga peregrina ) under the control of a low-level, root-specific promoter. To improve the level of resistance, we linked the SLP gene to the promoter from HMG2, which is strongly inducible by Orobanche as it parasitizes the host. The resulting transgenic plants express SLP and show increased resistance to Orobanche. Resistance in this case is manifested by increased growth and yield of the host in the presence of the parasite as compared to non-transgenic plants, and decreased parasite growth. The mechanism of resistance appears to operate post-attachment as the parasite tubercles attached to the transgenic root plants turned necrotic and failed to develop normally. Studies examining the movement of GFP (approximately 6X the size of SLP) produced in tobacco roots showed accumulation of green fluorescence in tubercles growing on transformed plants but not in those growing on wild-type plants. This accumulation occurs regardless of whether the GFP is targeted to the cytoplasm (translocated symplastically) or the apoplastic space (translocated in xylem). Plants expressing SLP appear normal as compared to non-transgenic plants in the absence of Orobanche, so there is no obvious unintended impact on the host plant from SLP expression. This project required the creation of several gene constructs and generation of many transformed plant lines in order to address the research questions. The specific objectives of the project were to: 1. Make gene constructs fusing Orobanche-inducible promoter sequences to either the sarcotoxin-like peptide (SLP) gene or the GFP reporter gene. 2. Create transgenic plants containing gene constructs. 3. Characterize patterns of transgene expression and host-to-parasite movement of gene products in tobacco ( Nicotiana tabacum L.) and Arabidopsis thaliana (L.). 4. Characterize response of transgenic potato ( Solanum tuberosum L.) and tomato ( Lycopersicon esculentum Mill .) to Orobanche in lab, greenhouse, and field. Objectives 1 and 2 were largely accomplished during the first year during Dr. Aly's sabbatical visit to Virginia Tech. Transforming and analyzing plants with all the constructs has taken longer than expected, so efforts have concentrated on the most important constructs. Work on objective 4 has been delayed pending the final results of analysis on tobacco and Arabidopsis transgenic plants. The implications of this work are profound, because the Orobanche spp. is an extremely destructive weed that is not controlled effectively by traditional cultural or herbicidal weed control strategies. This is the first example of engineering resistance to parasitic weeds and represents a unique mode of action for selective control of these weeds. This research highlights the possibility of using this technique for resistance to other parasitic species and demonstrates the feasibility of developing other novel strategies for engineering resistance to parasitic weeds.

scholarly journals Phenotypic performance of transgenic potato (Solanum tuberosum L.) plants with pyramided rice cystatin genes (OCI and OCII)

2015 ◽  
Vol 67 (3) ◽  
pp. 957-964 ◽  
Author(s):  
Aleksandar Cingel ◽  
Jelena Savic ◽  
Tatjana Cosic ◽  
Martin Raspor ◽  
Nabil Ghalawenji ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Solanum Tuberosum L ◽  
Transgenic Potato ◽  
Field Conditions ◽  
Plant Performance ◽  
Field Environment ◽  
Transgenic Lines ◽  
Agronomical Traits ◽  
Wild Type Control

The evaluation of transgenic plants which is usually carried out under controlled conditions in culture rooms and greenhouses can yield valuable information about the influence of introduced genes on a transgenic plant phenotype. However, an overall assessment of plant performance can only be made by testing transgenic plants in the field environment. Thus, the effects of pyramided rice cystatin genes OCI and OCII on morphological parameters of transgenic potato cv. Desiree, Dragacevka and Jelica lines were compared under in vitro, greenhouse, and field conditions. All analyzed OC co-expressing transgenic lines exhibited normal phenotype, both in vitro and in greenhouse conditions. In the field environment, eight of nine OCI/OCII lines were similar to the wild-type control plants in their general phenotypic appearance. Yield parameters, such as tuber number and tuber weight for these phenotypically normal OCI/OCII lines, were also comparable to the controls. Only transgenic cv. Jelica line 4 plants exhibited slightly reduced growth, atypical leaf morphology and, contrary to the plants of other transgenic lines and untransformed controls, failed to flower. However, despite the phenotypic and developmental changes under field conditions, the OCI/OCII Jelica line 4 did not exhibit a significant decrease in tuber yield. Stacking of OCI and OCII genes preserves important attributes of the parental lines, confirming that this approach could be suitable for improving agronomical traits in potato.

scholarly journals CRISPR/Cas9-Mediated SlMYBS2 Mutagenesis Reduces Tomato Resistance to Phytophthora infestans

2021 ◽  
Vol 22 (21) ◽  
pp. 11423
Author(s):  
Chunxin Liu ◽  
Yiyao Zhang ◽  
Yinxiao Tan ◽  
Tingting Zhao ◽  
Xiangyang Xu ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Late Blight ◽  
Phytophthora Infestans ◽  
Gene Knockout ◽  
Disease Index ◽  
Oxygen Species ◽  
Wild Type ◽  
Pr Genes ◽  
Knock Out ◽  
Pathogenesis Related

Phytophthora infestans (P. infestans) recently caused epidemics of tomato late blight. Our study aimed to identify the function of the SlMYBS2 gene in response to tomato late blight. To further investigate the function of SlMYBS2 in tomato resistance to P. infestans, we studied the effects of SlMYBS2 gene knock out. The SlMYBS2 gene was knocked out by CRISPR-Cas9, and the resulting plants (SlMYBS2 gene knockout, slmybs2-c) showed reduced resistance to P. infestans, accompanied by increases in the number of necrotic cells, lesion sizes, and disease index. Furthermore, after P. infestans infection, the expression levels of pathogenesis-related (PR) genes in slmybs2-c plants were significantly lower than those in wild-type (AC) plants, while the number of necrotic cells and the accumulation of reactive oxygen species (ROS) were higher than those in wild-type plants. Taken together, these results indicate that SlMYBS2 acts as a positive regulator of tomato resistance to P. infestans infection by regulating the ROS level and the expression level of PR genes.

scholarly journals Obtaining of transgenic potato plants (Solanum tuberosum L.) that contain antisense sequence of prolindehydrogenase gene

2018 ◽  
Vol 22 ◽  
pp. 299-304
Author(s):  
O. O. Ovcharenko ◽  
V. A. Rudas ◽  
N. L. Shcherbak ◽  
M. V. Kuchuk
Keyword(s):  
Solanum Tuberosum ◽  
Transgenic Plants ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
Practical Interest ◽  
Transgenic Potato ◽  
Proline Dehydrogenase ◽  
Potato Plants ◽  
Antisense Sequence ◽  
Transgenic Potato Plants

Aim. The aim of our work was to obtain transgenic potato plants of Ukrainian varieties with the expression of a double-stranded RNA suppressor of proline dehydrogenase gene. We propose the decrease of proline degradation level and increase of overall proline concentration in obtained transgenic plants. Methods. The Agrobasterium tumefaciens-mediated method of genetic transformation to obtain transgenic plants of potato was used. Internodes of aseptic potato plants were transformed with a binary vector pBi2E containing an inverted repeats of two copies of proline dehydrogenase gene’s first exon and the gene of neomycin phosphotransferase II (nptII). Results. As a result of experiments kanamycin resistant transgenic potato lines of Deseiree, Belarusian 12 and Slavianka varieties were obtained. The transgenic nature of the obtained plants was confirmed by PCR with primers specific to the first exon of proline dehydrogenase and to nptII genes. Conclusions. The optimized conditions of genetic transformation and used agrobacterial strain allow to obtain the transgenic plants of a model potato variety Désirée, as well as varieties Belorussian 12 and Slovyanka which are of practical interest for cultivation in Ukraine. Keywords: transgenic plants, potatoes (Solanum tuberosum L.), stress resistance, proline.

scholarly journals Ascorbic Acid Deficiency in Arabidopsis Induces Constitutive Priming That is Dependent on Hydrogen Peroxide, Salicylic Acid, and the NPR1 Gene

2010 ◽  
Vol 23 (3) ◽  
pp. 340-351 ◽  
Author(s):  
Madhumati Mukherjee ◽  
Katherine E. Larrimore ◽  
Naushin J. Ahmed ◽  
Tyler S. Bedick ◽  
Nadia T. Barghouthi ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Salicylic Acid ◽  
Disease Resistance ◽  
Pseudomonas Syringae ◽  
Messenger Rna ◽  
Bacterial Pathogen ◽  
Pathogen Resistance ◽  
Wild Type ◽  
Pr Genes ◽  
Pathogenesis Related

The ascorbic acid (AA)-deficient Arabidopsis thaliana vtc1-1 mutant exhibits increased resistance to the virulent bacterial pathogen Pseudomonas syringae. This response correlates with heightened levels of salicylic acid (SA), which induces antimicrobial pathogenesis-related (PR) proteins. To determine if SA-mediated, enhanced disease resistance is a general phenomenon of AA deficiency, to elucidate the signal that stimulates SA synthesis, and to identify the biosynthetic pathway through which SA accumulates, we studied the four AA-deficient vtc1-1, vtc2-1, vtc3-1, and vtc4-1 mutants. We also studied double mutants defective in the AA-biosynthetic gene VTC1 and the SA signaling pathway genes PAD4, EDS5, and NPR1, respectively. All vtc mutants were more resistant to P. syringae than the wild type. With the exception of vtc4-1, this correlated with constitutively upregulated H2O2, SA, and messenger RNA levels of PR genes. Double mutants exhibited decreased SA levels and enhanced susceptibility to P. syringae compared with the wild type, suggesting that vtc1-1 requires functional PAD4, EDS5, and NPR1 for SA biosynthesis and pathogen resistance. We suggest that AA deficiency causes constitutive priming through a buildup of H2O2 that stimulates SA accumulation, conferring enhanced disease resistance in vtc1-1, vtc2-1, and vtc3-1, whereas vtc4-1 might be sensitized to H2O2 and SA production after infection.

scholarly journals LncRNA39026 Enhances Tomato Resistance to Phytophthora infestans by Decoying miR168a and Inducing PR Gene Expression

2020 ◽  
Vol 110 (4) ◽  
pp. 873-880 ◽  
Author(s):  
Xinxin Hou ◽  
Jun Cui ◽  
Weiwei Liu ◽  
Ning Jiang ◽  
Xiaoxu Zhou ◽  
...  
Keyword(s):  
Phytophthora Infestans ◽  
Noncoding Rna ◽  
Target Gene ◽  
Expression Level ◽  
Regulation Mechanism ◽  
Pr Genes ◽  
Tomato Plants ◽  
Expression Levels ◽  
Pathogenesis Related ◽  
Pr Gene Expression

Our previous study has indicated that a long noncoding RNA (lncRNA), lncRNA39026, can be responsive to Phytophthora infestans infection. However, the function and regulation mechanism of lncRNA39026 during tomato resistance to P. infestans are unknown. In this study, an lncRNA39026 sequence was cloned from tomato Zaofen No. 2, and this sequence contained an endogenous target mimicry for miR168a, which might suppress the expression of miR168a. LncRNA39026 was strongly downregulated at 3 h in the tomato plants infected with P. infestans, and its expression level displayed a negative correlation with the expression level of miR168a and a positive correlation with the expression levels of SlAGO1 genes (target gene of miR168a) upon P. infestans infection. Tomato plants in which lncRNA39026 was overexpressed displayed enhanced resistance to P. infestans, decreased level of miR168a, and increased level of SlAGO1, whereas the resistance was impaired, level of miR168a was increased, and level of SlAGO1 was decreased after lncRNA39026 silencing. In addition, lncRNA39026 could also induce the expression of pathogenesis-related (PR) genes, as shown by increased and decreased expression levels of PR genes in tomato plants with overexpressed and silenced lncRNA39026, respectively. The result demonstrated that lncRNA39026 might function to decoy miR168a and affect the expression of PR genes in tomato plants, increasing resistance to disease.

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