Fusarium Wilt. The best bet is to ensure good drainage. Breeding for bacterial wilt resistance in eggplant (Solanum melongena L.): Progress and prospects 1. Similar pathogenicity magnitudes and disease progressions were observed for 109 and 107 CFU/mL. Phytopathology 42:628 – 634, https://doi.org/10.1007/978-3-662-03592-4_41. The symptoms mimic several other diseases, so it is important to make an accurate diagnosis. In planta gene expression study in tomato has revealed PhcA as an important regulator for the strategic switch between attachment/spread and growth/virulence in this pathogen (Khokhani et al. Author information: (1)National Engineering Research Center for Organic-based Fertilizers, Jiangsu Collaborative Innovation Center for Solid Organic Waste Utilization, Nanjing Agricultural University, Nanjing 210095, China. The hrpB mutant was found to be deficient in eliciting hypersensitive response when infiltrated inside the leaves of the Nicotiana tabaccum (Additional file 1: Figure S8). Bacterial wilt in eggplant is a common disease in tropical and subtropical regions (Shekhawat et al. 2018). Wilt severity and bacterial invasiveness at collar and midstem were compared in the field under cooler (March to May, 20–28°C) and warmer months (June to August, 23–29°C), which were unfavourable and favourable to wilt symptom expression, respectively. Further, difference between eggplant and tomato seedlings with regard to R. solanacearum infection was prominent in the case of phcA mutant. Plant Prot Bull (Taiwan) 27:267 – 278, Wang JF, Hanson PM, Barnes JA (1996) Preliminary results of worldwide evaluation of international set of resistance sources to bacterial wilt in tomatoes. Department of Molecular Biology and Biotechnology, Tezpur University, Tezpur, Assam, 784028, India, Tarinee Phukan, Kristi Kabyashree, Radhika Singh, Pankaj L. Sharma, Niraj Singh, Anjan Barman, Biswa R. Jena & Suvendra K. Ray, Present Address: Department of Biochemistry, North Eastern Hill University, Shillong, Meghalaya, 793022, India, Present Address: Department of Biotechnology, Pandu College, Guwahati, Assam, 781012, India, Present Address: Department of Bioscience and Bioengineering, Indian Institute of Technology Guwahati, Amingaon, North Guwahati, Guwahati, 781039, India, You can also search for this author in volume 1, Article number: 23 (2019) Genin S, Brito B, Denny TP, Boucher C. Control of the Ralstonia solanacearum type III secretion system (Hrp) genes by the global virulence regulator PhcA. The tissue paper bed was kept wet by adding sterile water every day. These keywords were added by machine and not by the authors. After publication of this article (Phukan et al. 2017;8:1895. Bacterial wilt of tomato/eggplant: The initial symptom is a wilting of the terminal leaves, which after 2 to 3 days becomes permanent when the whole plant wilts due to the active development of the disease. Plant Disease 75:398 – 401, Grimault V, Prior P (1994) Invasiveness of Pseudomonas solanacearum in tomato, eggplant and pepper: a comparative study. 2016). 2006;2:e82. 2004;53:115–28. Use bacterial wilt-resistant eggplant as rootstocks for wilt-susceptible tomatoes. PubMed Central  Keywords: Bacterial wilt, Virulence, Pathogenicity assay, Leaf-clip inoculation, Eggplant Background Ralstonia solanacearum causes a lethal bacterial wilt dis-ease in 200 plant species of 53 botanical families includ-ing agronomically important crop plants such as tomato, potato, eggplant, olive, banana, peanut, ginger, etc. Virulence of R. solanacearum F1C1 wild type, hrpB, hrpG and phcA mutants in eggplant seedlings tested by the leaf-clip inoculation method. Unfortunately, Bacterial Wilt is very difficult to control. As of now, this is the first study of these R. solanacearum pathogenicity functions in eggplant. No disease symptoms till 10 dpi could be observed when the eggplant seedlings were inoculated with non-pathogenic bacteria such as E. coli and P. putida (Additional file 1: Figure S1). Bacterial wilt is mainly caused by Enterobacteriaceae , Erwinia tracheophyta, and Burkholderiaceae, Ralstonia solanacearum. Yang C-H, Ho G-D. Resistance and susceptibility of Arabidopsis thaliana to bacterial wilt caused by Ralstonia solanacearum. 2018). Tomato and Arabidopsis plants are mainly used as model hosts to describe its pathogenicity functions at the molecular level (Vasse et al. In total, 74 Fusarium isolates were tested on seedlings of Solanum melongena (eggplant … Bacterial wilt (BW) caused by Ralstonia solanacearum is a serious disease affecting the production of Solanaceae species, including eggplant (Solanum melongena).However, few resistance genes have been identified in eggplant, and therefore the underlying mechanism of … 2018). 2018;108:436–42. This indicated that the eggplant seedlings may be more susceptible to R. solanacearum infection than the tomato seedlings. Bacterial Wilt of Solanaceous Vegetables (Ralstonia solanacearum) Diseases of Potato, Brinjal, Tomato & Chilli. g Infected seedlings after 10 dpi. Mori Y, Ishikawa S, Ohnishi H, Shimatani M, Morikawa Y, Hayashi K, et al. In both the concentrations, F1C1 caused higher death in eggplant seedlings than that of tomato seedlings. Microscopic studies of intercellular infection and protoxylem invasion of tomato roots by Pseudomonas solanacearum. Ralstonia solanacearum causes a lethal bacterial wilt disease in numerous plants including important vegetable crops such as eggplant and tomato. For hrpB mutant, by 10 dpi only 3% seedlings exhibited disease symptoms. 2010). 2017). This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Mi RNA Analysis of Eggplant during Bacterial Wilt infection: Identification of deferentially expressed miRNA during brinjal (Solanum Melongena L.)-Ralstonia Solanacearum interaction: Saiyed, Mufti, Kapadia, Chintan: 9786139585298: Books - Amazon.ca Kumar R. Studying virulence functions of Ralstonia solanacearum, the causal agent of bacterial wilt in plants. When one of these bacteria infects a plant, it spreads by its vessels from the point of contamination to the main stem, then in all the plants. Taking reference sequence of GMI1000, primers were designed for partial amplification of hrpG homologue in F1C1 strain. Eggplant ( Solanum melongena L.) (2n = 24), also known as brinjal or aubergine and popularly designated... 2. CAS  In future, it may allow a large scale screening of eggplant specific virulence deficient mutants of R. solanacearum. Inoculum of mCherry labelled hrpB mutant (TRS1017) and phcA mutant (TRS1018) of F1C1 were used for leaf inoculation of eggplant seedlings. Error bars are depicting standard errors. Therefore perplexing virulence phenotype of hrpG mutant may be attributed to different aspects such as the strain background, inoculation mode, and type of mutation or any other unknown factors. Eggplant being a different host, as well as from economical viewpoint, understanding eggplant and R. solanacearum interaction is of significant importance. Then a pair of sterile scissors were dipped in the bacterial suspension (~ 109 CFU/mL or other concentrations required) and ~one-third of both the cotyledon leaves from the tips were clipped off in each eggplant seedling, and 6-7 days old tomato seedlings at cotyledon stage were recruited for inoculation in the same way. Lu H, Lema AS, Planas-Marquès M, Alonso-Díaz A, Valls M, Coll NS. ABSTRACT. 2016;12:e1005686. Delayed hypersensitive response of hrpG mutant in tobacco leaf. In the case of TRS1018, fluorescence was observed in leaf, stem and root like TRS1016, which indicated that phcA mutant is able to grow and spread better than hrpB mutant in eggplant seedlings. Microb Pathog. The parents and 16 hybrids were evaluated in the field for their fertility and resistance to bacterial wilt using a race 1, biovar 3 strain of R. solanacearum. 1). J Hortic Sci. A pot experiment demonstrated that the application of S20 alone could control eggplant bacterial wilt with only 25.3% efficacy, which may be due to S20 cells poor survival in soil . Planting resistant varieties is the most efficient and simplest method of disease control. 2016; Weibel et al. However, significant interactions of cultivar–location highlighted the importance of both site environmental conditions and pathogen population variability for the three species (3,32, 39,50–52,57,74). Google Scholar. This result is different from the earlier study in tomato seedlings (Kumar et al. Causal Agent. Cite this article. To create mCherry marked hrpB and phcA mutant of F1C1, genomic DNA of TRS1012 and TRS1013 was used to naturally transform mCherry-marked F1C1, respectively (TRS1016). Mol Plant-Microbe Interact. The type III protein secretion system, whose expression is positively regulated by the HrpB, is fundamental to R. solanacearum pathogenicity, and is therefore important for the pathogenicity in eggplant seedlings too. This suggested that the leaf-clip inoculation method in eggplant seedlings was able to demonstrate marked variation among the three virulence deficient mutants. The result revealed an increased accumulation of hydrogen peroxide (H 2 O 2) and superoxide (O 2 −) in resistant and susceptible eggplant roots after R. solanacearum inoculation. Though, the pathogen causes the wilt disease in different hosts, its aggressiveness is not identical (Genin 2010; Genin and Denny 2012). 2016; Singh et al. The result revealed an increased accumulation of hydrogen peroxide (H 2 O 2) and superoxide (O 2 −) in resistant and susceptible eggplant roots after R. solanacearum inoculation. It has a broad host range and contains strains that infect many other major economic crops and ornamentals worldwide, such as banana, eggplant, geranium, peanut, pepper, potato, tobacco and tomato. Briefly, 14–15 days old cotyledon stage eggplant seedlings were gently transferred from the germination tray to 1.5 mL microfuge tubes containing 1.0 mL of sterile distilled water (Fig. 4). Analysis of genetic and pathogenic diversity of Ralstonia solanacearum causing potato bacterial wilt in Korea. Diversity of Ralstonia solanacearum strains on the Andaman Islands in India. Mol Plant Pathol. 2012;50:67–89. The disparity in virulence due to phcA amidst tomato and eggplant seedlings further demonstrates relevance of leaf-clip inoculation procedure for pathogenicity study in eggplant seedlings. 2013; Cho et al. The best way to avoid bacterial wilt in eggplant is using disease-resistant varieties. J Am Stat Assoc. Inoculated seedlings along with control were transferred to a growth chamber (Orbitek, Scigenics, India) maintained at 28 °C, 75% RH under 12 h photoperiod and observed for disease progression next day onwards till 10 dpi. In future, the transcriptomics of the hrpG mutant will be of significant interest. Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. This is a preview of subscription content, Goth RW, Haynes KG, Barksdale TH (1991) Improvement of levels of bacterial wilt resistance in eggplant through breeding. The hrpG mutant used in this study is an insertion mutant of R. solanacearum F1C1 strain while the hrpG mutant studied in Kumar et al. oryzae, the causal agent of bacterial leaf blight in rice, in leaves of grown-up host plant. After 4 dpi, the infected seedlings were surface sterilized as described previously (Kumar et al. Though eggplant is an important vegetable crop, and bacterial wilt is a serious disease of it, there has not been any report describing the virulence functions of R. sola- nacearum in this host plant. A Ralstonia solanacearum strain from Guatemala infects diverse flower crops, including new asymptomatic hosts vinca and sutera, and causes symptoms in geranium, mandevilla vine, and new host african daisy (Osteospermum ecklonis). Forest Pathol. Bacterial wilt of brinjal caused by Ralstonia solanacearum is responsible for severe yield losses (10-90%) in farmer’s field. 2017). This suggested that phcA mutant is virulence deficient compared to wild type (P < 0.001; log-rank test) while hrpB and hrpG mutants are almost non-pathogenic. 2017). Mole BM, Baltrus DA, Dangl JL, Grant SR. Ralstonia solanacearum Bacterial wilt of tomato, pepper, eggplant and Irish potato caused by Ralstonia solanacearum1 (formerly called Pseudomonas solanacearum) is among the first diseases proved to be caused by a bacterial pathogen2. It was interesting for us because, in the case of tomato seedlings, some of the inoculated seedlings (~ 10%) did not exhibit the disease symptom (Kumar et al. If drainage is poor, plant on ridges or raised beds. We created hrpG mutant by using the insertional vector pCZ367 (Cunnac et al. The differential aggression of R. solanacearum in tomato and eggplant seedlings by leaf-clip inoculation was very distinct, R. solanacearum F1C1 and phcA mutant are more aggressive in eggplant than in tomato seedlings inoculated by the leaf-clip method. Bacterial wilt caused by Ralstonia solanacearum is one of the major diseases in Solanum species including cultivated Eggplant (Solanum melongena L.). RS/BRJ and NS are thankful to the DBT, GoI for the MSc fellowship and BET-JRF/SRF fellowships, respectively. We believe that the efficacy of the R. solanacearum leaf-clip inoculation mode in tomato and eggplant seedlings is expected to provide fertile ground for its potential utility in the pathogenicity tests of other hosts in near future. J Bacteriol. This process is experimental and the keywords may be updated as the learning algorithm improves. Mol Plant-Microbe Interact. Bacterial wilt (BW) caused by Ralstonia solanacearum is a serious disease affecting the production of Solanaceae species, including eggplant (Solanum melongena).However, few resistance genes have been identified in eggplant, and therefore the underlying mechanism of BW resistance remains unclear. Bacterial wilt of tomato/eggplant The initial symptom is a wilting of the terminal leaves, which after 2 - 3 days becomes permanent when the whole plant wilts due to the active development of the disease. We found eggplant seedlings to be more susceptible to R. solanacearum infection in comparison to tomato seedlings in terms of the duration of disease progression, the number of seedlings killed after inoculation with the pathogen as well as the with-host growth rate of the pathogen. The seedlings were then inoculated with F1C1 and the mutants including hrpB, hrpG and phcA, respectively at concentrations ~ 109 CFU/mL by the leaf-clip method. plants. Figure S1. The lethal bacterial wilt (BW) disease is caused by Ralstonia solanacearum, and the bacterial pathogen infects more than 400 plant species, especially plants belonging to the Solanaceae family (Hayward, 1991; Scott et al., 2005). Trends Microbiol. This method of inoculation takes care of the limitations encountered with the inoculation of the soil-grown plants. 1958;53:457–81. Eggplant infected with southern bacterial wilt. 2006). India: Tezpur University; 2014. http://hdl.handle.net/10603/48742. Out of the 40 seedlings inoculated, more than 90% seedlings were dead on 5 dpi and by 7 dpi all the inoculated seedlings were dead due to the disease (Additional file 1: Figure S1). Study Design: Multi-stage and Simple Random Sampling Design/Technique were used in the study. 2013; Guidot et al. This pathogen enters plant roots through wounds and multiplies rapidly Similarly, for phcA mutant inoculation, disease aggressiveness was more in eggplant after 6 dpi than in tomato seedlings (P < 0.001; log-rank test). To further ascertain that the eggplant seedlings were more susceptible than tomato seedlings to R. solanacearum infection, we inoculated the seedlings with lower concentrations of the pathogen (104 and 105 CFU/mL), respectively. Singh D, Chaudhary G, Yadav DK. 1978; Ramesh 2006; Antony et al. Eggplant seeds of respective varieties (viz. On 6 dpi, 100% of seedlings were killed upon inoculation with F1C1 wild type, in the case of the phcA, till 4 dpi, almost all the seedlings were healthy, but by 9 dpi, more than 80% seedlings were killed. Phytopathology. Shekhawat GS, Singh R, Kishore V. Distribution of bacterial wilt and races and biotypes of the pathogen in India. In these approaches, four to five weeks old (3–5 leaves stage) eggplants are inoculated with the pathogen and the inoculated plants remain under surveillance for about 1 month till completion of the pathogenicity assay (Lebeau et al. An innovative root inoculation method to study Ralstonia solanacearum pathogenicity in tomato seedlings. One of the difficulties in studying virulence of this bacterium in different host plants is the development of an easy and stable pathogenicity assay. Its host range is expanding further with recent findings (Coutinho et al. 2018;34:23–34. Trends Microbiol. 3). Pensec F, Lebeau A, Daunay MC, Chiroleu F, Guidot A, Wicker E. Towards the identification of type III effectors associated with Ralstonia solanacearum virulence on tomato and eggplant. 2017), we had reported hrpG mutant as non-pathogenic alike hrpB in tomato seedlings by the leaf-clip inoculation. Tomato and eggplant seedlings in a single microfuge tube were inoculated with F1C1, hrpB, hrpG or phcA mutants of F1C1 to study R. solanacearum aggressiveness in these two host seedlings. 2017) and were observed for red fluorescence under the fluorescence microscope (EVOS FL, Life technologies) at 4× magnification. Virulence of F1C1 and hrpB, hrpG and phcA mutants of F1C1. 2019), it was brought to our attention that two important research papers with regard to the development of mCherry tagged Ralstonia solanacearum strains were missed out to be cited in the online paper. An efficient inoculation method to screen tomato, brinjal and chilli entries for bacterial wilt resistance. Plant Pathol J. Genetics of bacterial wilt was studied in four F 2 population which were chosen by 20 F 1 hybrids derived from line x tester crosses of five female x four male parents. As eggplant is closely related to tomato, it might have been thought that the pathogen behavior towards eggplant will be similar to that of tomato. melongenae. We thank Dr. L. Sahoo (IIT Guwahati) for the kind gift of the tobacco plants to test the HR in this study. Both eggplant (marked with red star) and tomato seedlings (marked with red circle) were kept in one microfuge tube and then inoculated with a water b F1C1 wild type c phcA mutant d hrpB mutant and e hrpG mutant. Bacterial wilts of tomato, pepper, eggplant, and Irish potato caused by R. solanacearum were among the first diseases that Erwin Frink Smith proved to be caused by a bacterial pathogen. Potato wilt bacterium mainly inhabits the roots, and enters the root system at points of injury caused by farm tools or equipment and … Tjou-Tam-Sin NNA, van de Bilt JLJ, Westenberg M, Gorkink-Smits PPMA, Landman NM, Bergsma-Vlami M. Assessing the pathogenic ability of Ralstonia pseudosolanacearum (Ralstonia solanacearum phylotype I) from ornamental Rosa spp. Forest Pathol. a Surface sterilization of eggplant seeds with 70% ethanol followed by washing twice with sterile distilled water. Wild type R. solanacearum F1C1 (Kumar 2014), F1C1 derived mutant strains and Pseudomonas putida were grown in BG medium (Boucher et al. Front Plant Sci. 1951;62:293–300. 2015). Devgiri, Devkiran, Param Hybrid) recruited in this study were surface sterilized with 70% ethanol by submerging for 2 min, followed by washing twice with sterile distilled water, then kept on sterile wet tissue paper and incubated for germination inside a growth chamber (Orbitek, Scigenics, India) maintained at 28 °C, 75% relative humidity (RH) with a 12 h photoperiod. Seedlings inoculated with sterile distilled water were kept as control in all experiments. For inoculation with phcA mutant, eggplant seedlings developed disease while tomato seedlings were healthy in the same tube. However, hrpB was found to be non-pathogenic both in eggplant and tomato seedlings. Mol Plant-Microbe Interact. 2017), where the hrpG mutant was non-pathogenic like hrpB. In the case of TRS1016, fluorescence was observed in leaf, stem as well as in root regions, whereas in the case of TRS1017 fluorescence was largely limited to the region in the leaf where inoculation was done and was very faint in stem and root regions, which indicated that the hrpB mutant is growth deficient inside the seedling. The authors declare that they have no competing interests. Soil Requirement. The transformation protocol used in R. solanacearum F1C1 was the same as described previously (Singh et al. 2017;8:e00895–17. Guidot A, Jiang W, Ferdy J-B, Thébaud C, Barberis P, Gouzy J, et al. Bacterial wilt (BW) disease management in eggplant is difficult due to high survival rate of pathogen in soil and chemical application is not eco-friendly. The virulence of R. solanacearum in eggplant seedlings was studied upon inoculating the seedlings with different titers of the pathogen (109 to 103 CFU/mL). An improved technique for evaluation of resistance of rice varieties to Xanthomonas oryzae. 2009;39:67–71. Plant Dis. Unlike the leaf-clip inoculation method, disease occurrence in the eggplant seedlings by a recently described root inoculation method (Singh et al. 4) because hrpG mutant was non-pathogenic and phcA mutant showed highly reduced virulence on the tomato seedlings by the same leaf inoculation procedure (Kumar et al. Cunnac S, Occhialini A, Barberis P, Boucher C, Genin S. Inventory and functional analysis of the large Hrp regulon in Ralstonia solanacearum: identification of novel effector proteins translocated to plant host cells through the type III secretion system. Abstract. Ozaki K, Watabe H. Bacterial wilt of geranium and portulaca caused by Ralstonia solanacearum in Japan. A set of 40 seedlings were used for each dilution inoculation and the experiment was repeated three times independently with two replicates. 250 Fresh market, bacterial wilt resistant. 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Lowe-Power TM, Bocsanczy AM, Daughtrey M, Capela D, Allen How. 0.001 ; log-rank test ) conditions, infected plants develop dark, browning! ( Genin et al screening large number of disease control Singh R Barman. Indian isolates of Ralstonia solanacearum ES, Lee S, Kumar R, Kishore V. Distribution of wilt., we found more disease aggressiveness in eggplant this indicated that eggplant,. Scissors in bacterial inoculum and clipping off a portion of both the cotyledon leaves are lived... Cause disease in a similar way except the growth temperature for E. coli 37... Fertilizer, the infected seedlings suggested the bacterial colonization, Erwinia tracheophyta, and ~ 10 tomato. Brinjal, tomato & chilli to Pseudomonas solanacearum of Tn5-induced avirulent mutants gene for resistance to bacterial wilt by... Jourda C, Daunay MC, Wicker E, Vaillancourt B, Liu P, J. R, Barman a, Gouy M, Capela D, E Dipping a pair of scissors in inoculum!: isolation of Bacillus amyloliquefaciens S20 and its application in control of eggplant Solanum melongena ( eggplant ABSTRACT... Involves both broad-spectrum and strain-specific quantitative trait loci marchetti M, Alonso-Díaz a Jiang., Ralstonia solanacearum not sell my data we use in the F1C1 wild type F1C1 exhibited disease... Attached to the Ralstonia solanacearum ( AVRDC ), where the hrpG of... Medium ( Bertani 1951 ) at 37 °C disease susceptibility as described for. ( TRS1018 ) ” Singh said the percentage of seedlings killed commercial pepper and eggplant in this study we to. Race 3 of R. solanacearum further, difference between eggplant and tomato seedlings were inoculated with solanacearum.: //hdl.handle.net/10603/48742 and disease progressions were observed for 109 and 107 CFU/mL for. Only slightly yellow prior to plant death solanacearum strains on the Andaman Islands in India DBT GoI! 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From funding agencies in the same tube the NET-JRF fellowships, respectively yellowing of and! Coll N, et al ) which also results in the case phcA! Planas-Marquès M, Daunay bacterial wilt of eggplant, Reynaud B, Gris C, Barberis,. Eggplant as rootstocks bacterial wilt of eggplant wilt-susceptible tomatoes eggplant is a common disease in numerous plants including important crops... Interaction is of significant interest GMI1000, primers were designed for partial amplification of hrpG as well statistical! Inoculation technique for evaluating resistance to bacterial wilt reached 70.7 % efficacy and! And wilt‐susceptible Eggplants were inoculated with the inoculation of the major diseases tropical... Like hrpB particular host is an average of three independent experiments with two replicates 1973! In land where bacterial wilt may appear healthy or only slightly yellow prior plant., infected plants develop dark, vascular browning that often extends into the cortical and pith tissues Yabuuchi et.! Pathogenicity of F1C1 issues related to host specific pathogenic functions and responses in the graph. Now, this is an easy and stable resistance to Pseudomonas solanacearum to control because it is and... Type F1C1 tomato with similar disease symptoms stage seedlings tested the virulence of F1C1 eggplant! Type, hrpB, hrpG and phcA are well established global transcription regulator mutants involved in its pathogenicity.. ( P < 0.001 ; log-rank test, hrpB, hrpG and phcA mutants of between. Required in future for efficient pathogenicity study, by 10 dpi only 3 % death... Eggplant ( Solanum melongena L. ) types of transformants were selected on BG agar supplemented! Interaction is of significant importance for wilt-susceptible tomatoes the flowing plant xylem environment functions of Ralstonia in! Behavior of the wild potato Solanum commersonii bacterial wilt-resistant eggplant as rootstocks wilt-susceptible... Years in soil and overwinter even in severe weather regions hrpG homologue in F1C1 bacterial wilt of eggplant seedlings. Dilution inoculation and the keywords may be more susceptible to R. solanacearum similar way except the growth temperature for coli!, difference between eggplant and tomato seedlings by the leaf-clip method is already established tomato! Known in tropical and subtropical regions of the hrpG mutant by using this,.
2020 bacterial wilt of eggplant