Comparison of defense responses of transgenic potato lines expressing three different Rpi genes to specific Phytophthora infestans races based on transcriptome profiling

Potato late blight, one of the most devastating diseases in potato, is caused by the oomycete Phytophthora infestans. Over 20 resistance genes have been cloned including R1, R3a, and R3b. The distinctions between defense response mechanisms mediated by different resistance genes are still unclear. Here we performed transcriptome profiling in three transgenic lines, R1, R3a, and R3b, and wild-type Desiree under inoculation with two P. infestans isolates, 89148 (race 0) and CN152 (super race), using RNA-seq. Compared with wild type, specific differentially expressed genes (DEGs) were identified in the three transgenic lines. The highest number of DEGs occurred in transgenic R3b, with 779 DEGs in response to isolate 89148 and 864 DEGs in response to infection by CN152, followed by transgenic R1 lines with 408 DEGs for isolate 89148 and 267 DEGs for CN152. Based on gene ontology, the most common GO terms (15 for 89148 and 20 for CN152) were enriched in transgenic R3a and R3b lines. This indicates that the defense pathways mediated by R3a and R3b are more similar than those mediated by R1. Further separate GO analysis of up- or down-regulated DEGs showed that the down-regulated DEGs mainly functioned in mediating the resistance of potato to P. infestans 89148 by response to stress biological process and to CN152 by oxidation reduction biological process. KEGG pathways of DNA replication, plant-pathogen interaction and pentose and glucuronate interconversions are unique for transgenic R1, R3a, and R3b lines in incompatible interactions. Quantitative real-time PCR experimental validation confirmed the induced expression of DEGs in the late blight resistance signaling pathway. Our results will lay a solid foundation for further understanding the mechanisms of plant-pathogen interactions, and provide a theoretical reference for durable resistance in potato.

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Plant–Pathogen Molecular Dialogue: Evolution, Mechanisms and Agricultural Implementation

Russian Journal of Plant Physiology ◽

10.1134/s1021443721020072 ◽

2021 ◽

Vol 68 (2) ◽

pp. 197-211

Author(s):

E. E. Khavkin

Keyword(s):

Late Blight ◽

Resistance Genes ◽

Plant Pathogen ◽

Crop Production ◽

Virulence Genes ◽

Durable Resistance ◽

Plant Diseases ◽

Plant Colonization ◽

Molecular Evidence ◽

Pathogen Virulence

Abstract Plant diseases persistently challenge sustainable crop production worldwide. The most economical and eco-friendly way to effectively deal with this problem is to breed new cultivars with stable and durable resistance. Current progress towards this goal has been reinforced by considerable advancements in the molecular studies of pathogens and host plants. These advancements have greatly benefited from recently developed methods to research into gene structure and activity, especially the “omics” technologies. These steps forward are vividly represented by the case of late blight, which is economically the most important disease of potato and tomato (Solanum L.). Late blight became a popular model of multidimensional plant-microbe interactions, and newly obtained molecular evidence has considerably reshaped both our vision of plant–pathogen molecular dialogue and our approach to mitigating this disease. Drawing on recent publications, this review will focus on genome of the causal agent of disease, the oomycete Phytophthora infestans (Mont.) de Bary, and its already characterized genes of virulence, with particular emphasis on their evolution, which underlines the exceptional genetic and phenotypic plasticity of this pathogen. Specially highlighted is the diversity of the immediate tools of virulence—effectors, which interact with potato target molecules, alter host physiology and facilitate plant colonization. Turning to plant defense barriers, the reviewer elaborates on the polymorphism and evolution of Solanum genes providing for plant resistance to P. infestans. The repertoire of P. infestans virulence genes in agrocenoses and the diversity of resistance genes in potato wild relatives are explored as regards the agriculture-oriented implementation of new molecular knowledge. The multifaceted approach to late blight combines the search for new resistance genes in genetic collections, the characterization of their function and stacking these genes in potato cultivars in order to breed new donors of long-lasting and durable resistance together with express assessment of pathogen virulence genes.

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Resistance to Phytophthora infestans in Lycopersicon pennellii

Plant Disease ◽

10.1094/pdis-91-8-1045 ◽

2007 ◽

Vol 91 (8) ◽

pp. 1045-1049 ◽

Cited By ~ 34

Author(s):

Christine D. Smart ◽

Steven D. Tanksley ◽

Hilary Mayton ◽

William E. Fry

Keyword(s):

Late Blight ◽

Phytophthora Infestans ◽

Mapping Population ◽

Durable Resistance ◽

Introgression Lines ◽

Phenotypic Variance ◽

Plant Populations ◽

Lycopersicon Pennellii ◽

Trait Locus ◽

Different Strains

To determine if the desert tomato, Lycopersicon pennellii, possesses resistance to late blight, caused by Phytophthora infestans, two plant populations were analyzed. Resistance was identified through assessments of disease progress in an F2 mapping population (L. esculentum × L. pennellii) and in a series of introgression lines (L. pennellii into L. esculentum). Levels of resistance varied widely among individuals within each population. However, the response of individuals to different strains of P. infestans was consistent. In the mapping population, a quantitative trait locus (QTL) was detected near marker T1556 on chromosome 6. This QTL accounted for 25% of the phenotypic variance in the population. The occurrence of this QTL was confirmed from analysis of the introgression lines (ILs), where IL 6-2 (containing marker T1556) was the most resistant IL in 2002 and the second most resistant IL in 2001. The identification of an additional QTL for resistance to late blight in tomato will aid in the development of durable resistance to this devastating disease.

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The Effectiveness of Induced Defense Responses in a Susceptible Potato Genotype Depends on the Growth Rate of Phytophthora infestans

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi-03-18-0064-r ◽

2019 ◽

Vol 32 (1) ◽

pp. 76-85 ◽

Cited By ~ 1

Author(s):

Cécile Thomas ◽

Romain Mabon ◽

Didier Andrivon ◽

Florence Val

Keyword(s):

Growth Rate ◽

Late Blight ◽

Phytophthora Infestans ◽

Induced Defense ◽

Plant Immunity ◽

Defense Responses ◽

Alternative Methods ◽

Gene Expressions

Phytophthora infestans causes the devastating potato late blight disease, which is widely controlled with fungicides. However, the debate about chemical control is fueling a promotion toward alternative methods. In this context, the enhancement of natural plant immunity could be a strategy for more sustainable protection. We previously demonstrated that a concentrated culture filtrate (CCF) of P. infestans primes defense reactions in potato. They are genotype-dependent and metabolites produced decrease pathogen growth in vitro but not in vivo on tubers. Induced potato defenses are assumed to affect P. infestans life history traits depending on strains. This assumption was studied in vivo through induced leaflets on a susceptible genotype inoculated with four P. infestans strains differing for lesion growth rate. This study combines both defenses mechanistic analysis and ecological observations. Defense-gene expressions were thus assessed by quantitative reverse transcription-polymerase chain reaction; pathogen development was simultaneously evaluated by measuring necrosis, quantifying mycelial DNA, and counting sporangia. The results showed that CCF pretreatment reduced the pathogenicity differences between slow- and fast-growing strains. Moreover, after elicitation, PR-1, PR-4, PAL, POX, and THT induction was strain-dependent. These results suggest that P. infestans could develop different strategies to overcome plant defenses and should be considered in biocontrol and epidemic management of late blight.

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Pengujian Ketahanan Klon-klon Hasil Silangan Tanaman Kentang Transgenik dengan Nontransgenik terhadap Penyakit Hawar Daun Phytophthora infestans di Lapangan Uji Terbatas

Jurnal Hortikultura ◽

10.21082/jhort.v22n2.2012.p187-196 ◽

2013 ◽

Vol 22 (2) ◽

pp. 187 ◽

Cited By ~ 1

Author(s):

Alberta Dinar Ambarwati ◽

Muhamad Herman ◽

Edi Lisanto ◽

Euis Suryaningsih ◽

Eri Sofiari

Keyword(s):

Late Blight ◽

Phytophthora Infestans ◽

Field Trial ◽

Durable Resistance ◽

Transgenic Potato ◽

Resistance Testing ◽

Solanum Bulbocastanum ◽

Phenotypic Resistance ◽

Rb Gene ◽

Confined Field Trial

STRAK. Tanaman kentang transgenik Katahdin event SP904 dan SP951 mengandung gen RB, yang diisolasi dari spesies liar kentang diploid Solanum bulbocastanum. Gen RB mempunyai ketahanan yang bersifat durable dengan spektrum yang luas terhadap ras-ras Phytophthora infestans di Amerika Serikat. Dalam perakitan tanaman kentang tahan penyakit hawar daun P. infestans di Indonesia, transgenik Katahdin dijadikan sebagai donor tahan dalam persilangan dengan varietas rentan Atlantik dan Granola. Klon-klon hasil silangan dianalisis secara molekuler mengandung gen RB. Penelitian dilakukan untuk menguji ketahanan klon-klon hasil silangan tanaman kentang transgenik dengan nontransgenik terhadap isolat P. infestans di lapangan uji terbatas (LUT) yang berlokasi di Balai Penelitian Tanaman Sayuran (Balitsa), Lembang dari bulan Oktober 2009 sampai dengan Maret 2010. Klon-klon yang diuji ialah 12 klon hasil silangan Atlantik x transgenik Katahdin SP904 (A); 15 klon hasil silangan Atlantik x transgenik Katahdin SP951 (B); 17 klon hasil silangan Granola x transgenik Katahdin SP904 (C); dan 20 klon hasil silangan Granola x transgenik Katahdin SP951 (D). Atlantik dan Granola digunakan sebagai kontrol rentan, sedangkan transgenik Katahdin sebagai kontrol tahan. Pengamatan dimulai ketika muncul gejala awal, yaitu pada 26, 32, 39, 46, dan 53 hari setelah tanam. Ketahanan tanaman semakin menurun dengan bertambahnya periode pengamatan, diikuti meningkatnya intensitas penyakit dan AUDPC. Semua klon yang diuji menunjukkan keragaman dalam ketahanan fenotipik terhadap hawar daun P. infestans. Klon-klon hasil silangan Atlantik x transgenik Katahdin SP951 mempunyai nilai AUDPC 697, yang hampir sama dengan transgenik Katahdin SP904 yaitu 698,5. Klon-klon Granola x transgenik Katahdin SP951 mempunyai nilai AUDPC 687,5 lebih kecil dibandingkan transgenik Katahdin SP904. Hasil penelitian ini juga menunjukkan bahwa klon-klon tersebut mempunyai ketahanan yang lebih tinggi dibandingkan transgenik Katahdin SP904. Pada pengamatan 46 hari setelah tanam atau 20 hari setelah infeksi diperoleh tiga klon tahan yaitu B49 (skor 7,5), C111 (skor 7,1), dan D26 (skor 7,3). Ketahanan ini lebih tinggi daripada transgenik Katahdin SP904 (skor 5,1) dan transgenik Katahdin SP951 (skor 6,4). <br /><br />ABSTRACT. Ambarwati, AD, Herman, M, Listanto, E, Suryaningsih, E and Sofiari, E 2012. Resistance Testing on Transgenic and Nontransgenic Potato Clones Against Late Blight Phytophthora infestans in Confined Field Trial. Transgenic potato Katahdin event SP904 and SP951 containing RB gene, which were isolated from a wild diploid potato species, Solanum bulbocastanum. RB gene showed durable resistance with broad spectrum to all known races of P. infestans in the USA. In development of potato resistant to late blight P. infestans in Indonesia, Katahdin transgenic were used as a resistant donor and crossed with susceptible varieties i.e. Atlantic and Granola. Clones derived from the crossing were molecularly analyzed and had RB gene contain. Experiment was conducted to assess the resistance of the clones derived from crossing of Katahdin transgenic and nontransgenic to P. infestans in confined field trial (CFT), located at the Indonesian Vegetable Research Institute (IVEGRI), Lembang from October 2009 to March 2010. Several clones tested were 12 clones of Atlantic x Katahdin transgenic SP904 (A); 15 clones of Atlantic x Katahdin transgenic SP951 (B); 17 clones of Granola x Katahdin transgenic SP904 (C); and 20 clones of Granola x Katahdin transgenic SP951 (D). Atlantic and Granola were used as susceptible control whereas Katahdin transgenic as resistant control. Observation was started as late blight symptoms and detected at 26, 32, 39, 46, and 53 days after planting. Plant resistance decreases with increasing period of observation, followed by increasing disease intensity and AUDPC. All clones tested showed variation in phenotypic resistance to late blight P. infestans. Clones derived from crossing of Atlantic x Katahdin transgenic SP951 had AUDPC score 697 and almost similar to Katahdin transgenic SP904 (698.5). Clones derived from crossing of Granola x Katahdin transgenic SP951 had AUDPC score 687.5 and smaller than Katahdin transgenic SP904. The results also indicated that these clones had higher resistance than Katahdin transgenic SP904. Observation at 46 days after planting or 20 days after infection resulted three resistant clones i.e. B49 (score 7.5), C111 (score 7.1); and D26 (score 7.3). This resistance was higher than Katahdin transgenic SP904 (score 5.1) and Katahdin transgenic SP951 (score 6.4). <br />

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

International Journal of Molecular Sciences ◽

10.3390/ijms222111423 ◽

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.

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UJI KETAHANAN GALUR-GALUR KENTANG TRANSGENIK HASIL TRANSFORMASI DENGAN GEN RB TERHADAP PENYAKIT HAWAR DAUN (PHYTOPHTHORA INFESTANS) DI KP PASIRSARONGGE, CIANJUR

JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ◽

10.23960/j.hptt.213141-150 ◽

2013 ◽

Vol 13 (2) ◽

pp. 141-150

Author(s):

Edy Listanto ◽

M. Herman ◽

Eri Sofiari

Keyword(s):

Late Blight ◽

Phytophthora Infestans ◽

First Generation ◽

Block Design ◽

Transgenic Potato ◽

Resistant Plant ◽

Randomized Complete Block Design ◽

Transgenic Lines ◽

Transgenic Potatoes ◽

Rb Gene

Resistance test strains of transgenic potatoes transformed with RB gene to late blight (Phytophthora infestan) in KP Pasirsarongge, Cianjur. Potato late blight caused by Phytophthora infestans (P. infestans) (Mönt.) de Barry continues to be one of the most important crop diseases of all time. Genetic engineering of potato using RB gene for resistant plant to this disease is the most effective and environmental friendly to prevent widespread of late blight. This research aims to perform resistance of transgenic potato lines containing RB gene to lateblight (P. infestans) in Pasirsarongge, Cianjur field trial station. The first generation of transgenic lines were planted on polybag containing soil:manures using randomized complete block design. Tested plant inoculation was done naturaly from inoculum source from border row (Granola) that has been planted at one month before. The symptom was observed at one month after planting and damage scoring was done every three days for five times. Twenty two transgenic lines of tested plant showed various resistance respond to late blight (P. infestans) attack. Three transgenic lines showed highly resistance to late blight (P. infestans) were lines 11, 24, and 25, one transgenic line has resistant level was line 6.

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Phenazine-1-Carboxylic Acid Production by Pseudomonas fluorescens LBUM636 Alters Phytophthora infestans Growth and Late Blight Development

Phytopathology ◽

10.1094/phyto-06-16-0247-r ◽

2017 ◽

Vol 107 (3) ◽

pp. 273-279 ◽

Cited By ~ 9

Author(s):

Christopher K. Morrison ◽

Tanya Arseneault ◽

Amy Novinscak ◽

Martin Filion

Keyword(s):

Carboxylic Acid ◽

Late Blight ◽

Plant Defense ◽

Phytophthora Infestans ◽

Pseudomonas Fluorescens ◽

Quantitative Polymerase Chain Reaction ◽

Direct Interaction ◽

Wild Type ◽

In Planta

Phytophthora infestans causes late blight of potato, one of the most devastating diseases affecting potato production. Alternative approaches for controlling late blight are being increasingly sought due to increasing environmental concerns over the use of chemical pesticides and the increasing resistance of P. infestans to fungicides. Our research group has isolated a new strain of Pseudomonas fluorescens (LBUM636) of biocontrol interest producing the antibiotic phenazine-1-carboxylic acid (PCA). Wild-type LBUM636 was shown to significantly inhibit the growth of Phytophthora infestans in in vitro confrontational assays whereas its isogenic mutant (phzC−; not producing PCA) only slightly altered the pathogen’s growth. Wild-type LBUM636 but not the phzC− mutant also completely repressed disease symptom development on tubers. A pot experiment revealed that wild-type LBUM636 can significantly reduce P. infestans populations in the rhizosphere and in the roots of potato plants, as well as reduce in planta disease symptoms due to PCA production. The expression of eight common plant defense-related genes (ChtA, PR-1b, PR-2, PR-5, LOX, PIN2, PAL-2, and ERF3) was quantified in tubers, roots, and leaves by reverse-transcription quantitative polymerase chain reaction and revealed that the biocontrol observed was not associated with the induction of a plant defense response by LBUM636. Instead, a direct interaction between P. infestans and LBUM636 is required and PCA production appears to be a key factor for LBUM636’s biocontrol ability.

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Comparative Transcriptome Profiling Reveals Compatible and Incompatible Patterns of Potato Toward Phytophthora infestans

G3 Genes|Genome|Genetics ◽

10.1534/g3.119.400818 ◽

2019 ◽

Vol 10 (2) ◽

pp. 623-634 ◽

Cited By ~ 2

Author(s):

Yanfeng Duan ◽

Shaoguang Duan ◽

Miles R. Armstrong ◽

Jianfei Xu ◽

Jiayi Zheng ◽

...

Keyword(s):

Phytophthora Infestans ◽

Clustering Analysis ◽

Transcriptome Profiling ◽

Defense Responses ◽

Differentially Expressed ◽

Incompatible Interaction ◽

Time Points ◽

Gene Enrichment ◽

Profiling Analysis ◽

Insight Into

Late blight, caused by Phytophthora infestans (P. infestans), is a devastating disease in potato worldwide. Our previous study revealed that the Solanum andigena genotype 03112-233 is resistant to P. infestans isolate 90128, but susceptible to the super race isolate, CN152. In this study, we confirmed by diagnostic resistance gene enrichment sequencing (dRenSeq) that the resistance of 03112-233 toward 90128 is most likely based on a distinct new R gene(s). To gain an insight into the mechanism that governs resistance or susceptibility in 03112-223, comparative transcriptomic profiling analysis based on RNAseq was initiated. Changes in transcription at two time points (24 h and 72 h) after inoculation with isolates 90128 or CN152 were analyzed. A total of 8,881 and 7,209 genes were differentially expressed in response to 90128 and CN152, respectively, and 1,083 differentially expressed genes (DEGs) were common to both time points and isolates. A substantial number of genes were differentially expressed in an isolate-specific manner with 3,837 genes showing induction or suppression following infection with 90128 and 2,165 genes induced or suppressed after colonization by CN152. Hierarchical clustering analysis suggested that isolates with different virulence profiles can induce different defense responses at different time points. Further analysis revealed that the compatible interaction caused higher induction of susceptibility genes such as SWEET compared with the incompatible interaction. The salicylic acid, jasmonic acid, and abscisic acid mediated signaling pathways were involved in the response against both isolates, while ethylene and brassinosteroids mediated defense pathways were suppressed. Our results provide a valuable resource for understanding the interactions between P. infestans and potato.

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Enhanced defense responses of tomato plants against late blight pathogen Phytophthora infestans by pre-inoculation with rhizobacteria

Crop Protection ◽

10.1016/j.cropro.2010.07.023 ◽

2010 ◽

Vol 29 (12) ◽

pp. 1406-1412 ◽

Cited By ~ 12

Author(s):

Yongjun An ◽

Seogchan Kang ◽

Ki-Deok Kim ◽

Byung KooK Hwang ◽

Yongchull Jeun

Keyword(s):

Late Blight ◽

Phytophthora Infestans ◽

Defense Responses ◽

Tomato Plants ◽

Late Blight Pathogen

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New Findings on the Resistance Mechanism of an Elite Diploid Wild Potato Species JAM1-4 in Response to a Super Race Strain of Phytophthora infestans

Phytopathology ◽

10.1094/phyto-09-19-0331-r ◽

2020 ◽

Vol 110 (8) ◽

pp. 1375-1387

Author(s):

Jiayi Zheng ◽

Shaoguang Duan ◽

Miles R. Armstrong ◽

Yanfeng Duan ◽

Jianfei Xu ◽

...

Keyword(s):

Disease Resistance ◽

Late Blight ◽

Phytophthora Infestans ◽

Resistance Genes ◽

Resistance Mechanism ◽

Late Blight Resistance ◽

Blight Resistance ◽

Resistance Response ◽

Potato Disease ◽

New Findings

Late blight is a devastating potato disease worldwide, caused by Phytophthora infestans. The P. infestans strain 2013-18-306 from Yunnan is a “supervirulent race” that overcomes all 11 known late blight resistance genes (R1 to R11) from Solanum demissum. In a previous study, we identified a diploid wild-type potato JAM1-4 (S. jamesii) with high resistance to 2013-18-306. dRenSeq analysis indicated the presence of novel R genes in JAM1-4. RNA-Seq was used to analyze the late blight resistance response genes and defense regulatory mechanisms of JAM1-4 against 2013-18-306. Gene ontology enrichment and KEGG pathway analysis showed that many disease-resistant pathways were significantly enriched. Analysis of differentially expressed genes (DEGs) revealed an active disease resistance mechanism of JAM1-4, and the essential role of multiple signal transduction pathways and secondary metabolic pathways comprised of SA-JA-ET in plant immunity. We also found that photosynthesis in JAM1-4 was inhibited to promote the immune response. Our study reveals the pattern of resistance-related gene expression in response to a super race strain of potato late blight and provides a theoretical basis for further exploration of potato disease resistance mechanisms, discovery of new late blight resistance genes, and disease resistance breeding.

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