Interactive Regulation of Hormone and Resistance Gene in Proline Metabolism Is Involved in Effector-Triggered Immunity or Disease Susceptibility in the Xanthomonas campestris pv. campestris–Brassica napus Pathosystem

To characterize cultivar variations in hormonal regulation of the transition between pattern-triggered immunity (PTI) and effector-triggered immunity or susceptibility (ETI or ETS), the responses of resistance (R-) genes, hydrogen peroxide, and proline metabolism in two Brassica napus cultivars to contrasting disease susceptibility (resistant cv. Capitol vs. susceptible cv. Mosa) were interpreted as being linked to those of endogenous hormonal levels and signaling genes based on a time course of disease symptom development. Disease symptoms caused by the Xanthomonas campestris pv. campestris (Xcc) infections were much more developed in cv. Mosa than in cv. Capitol, as shown by an earlier appearance (at 3 days postinoculation [3 DPI]) and larger V-shaped necrosis lesions (at 9–15 DPI) in cv. Mosa. The cultivar variations in the R-genes, hormone status, and proline metabolism were found in two different phases (early [0–3 DPI] and later [9–15 DPI]). In the early phase, Xcc significantly upregulated PTI-related cytoplasmic kinase (Botrytis-induced kinase-1 [BIK1]) expression (+6.3-fold) with salicylic acid (SA) accumulation in cv. Capitol, while relatively less (+2.6-fold) with highly increased jasmonic acid (JA) level in cv. Mosa. The Xcc-responsive proline accumulation in both cultivars was similar to upregulated expression of proline synthesis-related genes (P5CS2 and P5CR). During the later phase in cv. Capitol, Xcc-responsive upregulation of ZAR1 (a coiled-coil-nucleotide binding site-leucine-rich repeat [CC-NB-LRR-type R-gene]) was concomitant with a gradual increase in JA levels without additional proline accumulation. However, in cv. Mosa, upregulation of TAO1 (a toll/interleukin-1 receptor-nucleotide binding site-leucine-rich repeat [TIR-NB-LRR-type R-gene]) was consistent with an increase in SA and abscisic acid (ABA) levels and resulted in an antagonistic depression of JA, which led to a proline accumulation. These results indicate that Xcc-induced BIK1- and ZAR1-mediated JA signaling interactions provide resistance and confirm ETI, whereas BIK1- and TAO1-enhanced SA- and/or ABA-mediated proline accumulation is associated with disease susceptibility (ETS).

Pseudomonas aeruginosa associated pulmonary infections and in vitro amplification virulent rhamnolipid (rhlR) gene

Background Pseudomonas aeruginosa is a common opportunistic pathogenic bacterium with the ability to develop a strong communication pathway by quorum sensing system and different virulent factors. Among the various important secretions of P. aeruginosa rhamnolipid is important biological detergent, believed to be involved in the development of the biofilm and intercellular communication. It readily dissolves the lung surfactants that are then easily catalyzed by the phospholipases and in this way is involved in the acute pulmonary infection. Objective research work was designed to investigate virulence and gene associated with virulence in P. aeruginosa responsible for pulmonary infections. Methods In current study polymerase chain reaction (PCR) was used for the detection of the rhlR (rhamnolipid encoding) gene of isolated strains. A number of assays were performed that ensured its virulent behavior. Disc diffusion method was used to check its antibiotic resistance. Isolated strains were resistant to a number of antibiotics applied. Result It was found that males are more prone to respiratory infections as compared to females. Male members with age of 44-58 and 59-73 are at a higher risk, while females with age of 44-58 are also at a risk of pulmonary infections. Antibiotic resistance was observed by measuring zone of inhibition in strains GCU-SG-M4, GCU-SG-M3, GCU-SG-M5, GCU-SG-M2, GCU-SG-M1 and GCU-SG-M6. GCU-SG-M2 was resistant to fluconazole (FLU), clarithromycin (CLR), cefixime (CFM) and Penicillin (P10). No zone of inhibition was observed. But it showed unusual diffused zone around the Ak and MEM antibiotic discs. rhl R gene and 16s rRNA gene were characterized and analyzed. Conclusion Findings from current study would help in raising awareness about antibiotic resistance of P. aeruginosa, and also the sequence of rhl R gene can be used as the diagnostic marker sequence to identify the virulent rhl R gene sequence from the samples when isolated from sputum of Pneumonia patients.

Evaluation of Cultivated and Wild Brinjal Germplasm against Bacterial Wilt Disease with Tollinterleukin-1 Receptors (TIR)-NBS-LRR Type R-gene Specific Degenerate Primer

The experiment was conducted at C-Block Farm of Bidhan Chandra Krishi Viswavidyalaya, Kalyani, West Bengal, India during 2017–18 to screen eight brinjal germplasm lines against BW disease using tollinterleukin-1 receptors (TIR)-NBS-LRR type R-gene specific degenerate primer. The study showed that wild genotype S. torvum was highly resistant to bacterial wilt incidence with no wilting symptom whereas two cultivated genotypes (Utkal Anushree and Utkal Madhuri) and one wild genotype S. sisymbriifolium were found to be resistant to BW disease. Out of the 7 germplasm sequences, 2 had no match with R-genes whereas the remaining 5 sequences have 70-93% homology with R-genes of other plant species submitted in Gene Bank sequence database. Nearly 90% sequence identity of brinjal NBS-LRR RGA was found by analyzing through BLASTn with NBS-LRR RGAs of other solanaceous crops. Two cultivated resistant genotypes (Utkal Madhuri and Utkal Tarini) were similar to the wild resistant type S. sisymbriifloium, while cultivable resistant genotype Utkal Anushree was highly different at sequence level. Two cultivable susceptible genotypes (BCB-30 and Garia) showed high level of similarity among them and they were strongly associated with the wild susceptible genotype S. macrocarpum. Two cultivable genotypes Utkal Anushree and Utkal Madhuri could be utilized in future breeding programme and two wild genotypes S. torvum and S. sisymbriifolium could be used as resistant rootstocks in brinjal grafting.

Bioremediation of arsenic contamination from the environment: New approach to sustainable resource management

Present acceleration of Arsenic [As] exposure leads to severe health problems. Modern scientific approaches look towards potent bio-agents for the removal of such types of contaminations in sustainable ways. Microbes can potentially change the redox potential, solubility, pH by different complex reactions during bioremediation. There are many enzymes present in the microbial system which are involved in methylation such as As (V) reductase, monomethyl arsonic acid reductase, As (III) methyltransferase, and MMA (III) methyltransferase. On the other hand, microbes have As transformation ability and changed into different extractable forms with sulfide minerals such as arsenopyrite (FeAsS), enargite (Cu3AsS4) and realgar (As4S4). In some bacteria, the As-operon machinery thiol group bind with As, itdetoxifies its toxicity. Ars R gene and arsenic reductase enzyme (Ars C) play the key role in the reduction of As (V) to As (III) and detoxify by being transported outside of the cell by Ars AB As chemiosmotic efflux system. In fungi, As (V) is reduced to As (III) by the arsenate reductase and GSH glutathione converted into GSSH glutathione disulfide. In plants, As (III) conjugates with phytochelatin (PC) or GSH glutathione and accumulates in the vacuole or is converted into less toxic forms in the presence of arsenic reductase enzyme. This review focused on the potentiality and mechanisms of different microbes for As-detoxification in a sustainable manner.

Phytophthora infestans: An Overview of Methods and Attempts to Combat Late Blight

Phytophthora infestans (Mont.) de Bary is one of the main pathogens in the agricultural sector. The most affected are the Solanaceae species, with the potato (Solanum tuberosum) and the tomato (Solanum lycopersicum) being of great agricultural importance. Ornamental Solanaceae can also host the pests Petunia spp., Calibrachoa spp., as well as the wild species Solanum dulcamara, Solanum sarrachoides, etc. Annual crop losses caused by this pathogen are highly significant. Although the interaction between P. infestans and the potato has been investigated for a long time, further studies are still needed. This review summarises the basic approaches in the fight against the late blight over the past 20 years and includes four sections devoted to methods of control: (1) fungicides; (2) R-gene-based resistance of potato species; (3) RNA interference approaches; (4) other approaches to control P. infestans. Based on the latest advances, we have provided a description of the significant advantages and disadvantages of each approach.

Genotypic and phenotypic dissection of Xanthomonas oryzae pv. oryzae on Indica rice cultivars for bacterial blight resistance

Aim: To evaluate the level of virulence of different Xoo isolates/ pathotypes of Eastern and North-eastern India and to identify the suitable donors in rice cultivars having various R-gene combination against virulent Xoo races of Bacterial Blight disease of rice. Methodology: Thirty six Xoo isolates were collected from different places of Eastern and North-eastern India and genetic diversity/ similarity was examined by genotyping of pathotypes using JEL1/JEL2 markers. The 34 Indica rice cultivars carrying different R-gene combination were selected and grown in net house and inoculated artificially with Xoo inoculants from these races/ isolates bacterial of blight disease. Results: The selected 36 Xoo isolates of Eastern and North-eastern India were grouped into seven different isolates/ races based on their genetic diversity using JEL1/JEL2 markers. Among 34 Indica rice cultivars, three or more R-gene combination (xa5 + xa13 + Xa21 and/or Xa4 + xa5 + xa13 + Xa21) cultivars exhibited highly resistant as compared to cultivars with single and double gene combination cultivars against most of the Xoo isolates/ races. Interpretation: The cultivars may determine different level of resistance due to complementary effect of inheritance of suitable R-gene combination. Identified donors may be used for rice resistance breeding programme for Eastern and North-eastern India.

SNPs, InDels, and Microsatellites within and near to Rice NBS-LRR Resistance Gene Candidates

Plant resistance genes (R-genes) drive the immune responses of crops against specific pathotypes of disease-causing organisms. Over time, genetic diversity in R-genes and R-pseudogenes has arisen among different rice varieties. This bioinformatics study was carried out to (i) predict the full sets of candidate nucleotide-binding site leucine-rich repeat (NLR) R-genes present in six rice genomes; (ii) detect variation within candidate R-genes; (iii) identify potential selectable markers within and near to LRR genes among 75 diverse indica rice genomes. Four high quality indica genomes, plus the standard japonica and indica reference genomes, were analysed with widely available bioinformatic tools to identify candidate R-genes and R-pseudogenes. They were detected in clusters, consistent with previous studies. BLAST analysis of cloned protein sequences of 31 R-gene loci gave confidence in this approach for detection of cloned NLR R-genes. Approximately 10% of candidate R-genes were located within 1 kb of a microsatellite (SSR) marker. Sequence comparisons among indica rice genomes detected SNPs or InDels in 334 candidate rice R-genes. There were significantly more SNPs and InDels within the identified NLR R-gene candidates than in other types of gene. The genome-wide locations of candidate R-genes and their associated markers are presented here for the potential future development of improved disease-resistant varieties. Limitations of in silico approaches used for R-gene discovery are discussed.

Improved isolation and detection of toxigenic Vibrio parahaemolyticus from coastal water in Saudi Arabia using immunomagnetic enrichment

Background Vibrio parahaemolyticus is recognized globally as a cause of foodborne gastroenteritis and its widely disseminated in marine and coastal environment throughout the world. The main aim of this study was conducted to investigate the presence of toxigenic V. parahaemolyticus in costal water in the Eastern Province of Saudi Arabia by using immunomagnetic separation (IMS) in combination with chromogenic Vibrio agar medium and PCR targeting toxR gene of species level and virulence genes. Methods A total of 192 seawater samples were collected from five locations and enriched in alkaline peptone water (APW) broth. One-milliliter portion from enriched samples in APW were mixed with an immunomagnetic beads (IMB) coated with specific antibodies against V. parahaemolyticus polyvalent K antisera and separated beads with captured bacteria streaked on thiosulfate citrate bile salts sucrose (TCBS) agar and CHROMagar Vibrio (CaV) medium. Results Of the 192 examined seawater samples, 38 (19.8%) and 44 (22.9%) were positive for V. parahaemolyticus, producing green and mauve colonies on TCBS agar and CaV medium, respectively. Among 120 isolates of V. parahaemolyticus isolated in this study, 3 (2.5%) and 26 (21.7%) isolates of V. parahaemolyticus isolated without and with IMB treatment tested positive for the toxin regulatory (toxR) gene, respectively. Screening of the confirmed toxR gene-positive isolates revealed that 21 (17.5%) and 3 (2.5%) were positive for the thermostable direct hemolysin (tdh) encoding gene in strains isolated with IMB and without IMB treatment, respectively. None of the V. parahaemolyticus strains tested positive for the thermostable related hemolysin (trh) gene. In this study, we found that the CaV medium has no advantage over TCBS agar if IMB concentration treatment is used during secondary enrichment steps of environmental samples. The enterobacterial repetitive intergenic consensus (ERIC)-PCR DNA fingerprinting analysis revealed high genomic diversity, and 18 strains of V. parahaemolyticus were grouped and identified into four identical ERIC clonal group patterns. Conclusions The presented study reports the first detection of tdh producing V. parahaemolyticus in coastal water in the Eastern Province of Saudi Arabia.

Breeding for Resistance to Fusarium Wilt of Tomato: A Review

For over a century, breeders have worked to develop tomato (Solanum lycopersicum) cultivars with resistance to Fusarium wilt (Fol) caused by the soilborne fungus Fusarium oxysporum f. sp. lycopersici. Host resistance is the most effective strategy for the management of this disease. For each of the three Fol races, resistance has been introgressed from wild tomato species, predominately in the form of R genes. The I, I-2, I-3, and I-7 R genes have each been identified, as well as the corresponding Avr effectors in the fungus with the exception of Avr7. The mechanisms by which the R gene protein products recognize these effectors, however, has not been elucidated. Extensive genetic mapping, gene cloning, and genome sequencing efforts support the development of tightly-linked molecular markers, which greatly expedite tomato breeding and the development of elite, Fol resistant cultivars. These resources also provide important tools for pyramiding resistance genes and should support the durability of host resistance.

Resilience response to yellow leaf curl disease and identification of resistance gene analogs (RGA) in pepper (Capsicum annuum)

Ayu DK, Maharijaya A, Syukur M, Hidayat SH. 2021. Resilience response to yellow leaf curl disease and identification of resistance gene analogs (RGA) in pepper (Capsicum annuum). Biodiversitas 22: 4731-4739. Pepper yellow leaf curl disease (PYLCD) caused by infection of Begomovirus is a serious threat to pepper production worldwide. Identification of the resistance gene analogs (RGA) and resilience response of pepper against PYLCD is needed especially for selection resistance genotype. Evaluation of resistance response involving 28 pepper genotypes was carried out through transmission of Begomovirus using whitefly (Bemisia tabaci) as vector. The result showed that IPB C12 and F4-012328-6-3 were potential resistance genotypes. A total of 15 R gene analogs (CaRGA) containing NBS motif, namely CARGA1 to CARGA15, were identified by degenerated PCR amplification and database mining. The alignment of deduced amino acid sequence revealed conservation of subdomains Ploop (GKTT), kinase2 (LVVLDDV), RNBSB/kinase3 (IILTTR) and GLPL. BLASTp analysis indicated that 15 RGA showed high homology at deduced amino acid level with R gene identified such as whitefly resistance protein Mi-1.2, Pvr 9 gene for potyvirus, Begomovirus resistance protein, TRGA15 and RGA 13 for putative late blight resistance. Phylogenetic analysis exhibited that isolated sequences distinguished into CNL-NBS groups. These pepper RGA could be considered as candidate sequences of resistance genes.