Ubiquitination in the rice blast fungus Magnaporthe oryzae: from development and pathogenicity to stress responses

Ubiquitination is a vital protein post-translational modification (PTM) prevalent in eukaryotes. This modification regulates multiple cellular processes through protein degradation mediated by the 26S proteasome or affecting protein–protein interaction and protein localization. Magnaporthe oryzae causes rice blast disease, which is one of the most devastating crop diseases worldwide. In M. oryzae, ubiquitination plays important roles in growth, pathogenicity, stress response and effector-mediated plant-pathogen interaction. In this review, we summarize the roles of ubiquitination components in the above biological processes of M. oryzae, including single- or multi-subunit E3s, E2s, components of 26S proteasome and also deubiquitinating enzymes. The essential function of ubiquitination in plant-fungus interaction is also discussed. Moreover, this review presents several issues related to the ubiquitination system in M. oryzae, which need to be further explored in future researches.

Two genomic regions of a sodium azide induced rice mutant confer broad-spectrum and durable resistance to blast disease

Rice blast, one of the most destructive epidemic diseases, annually causes severe losses in grain yield worldwide. To manage blast disease, breeding resistant varieties is considered a more economic and environment-friendly strategy than chemical control. For breeding new resistant varieties, natural germplasms with broad-spectrum resistance are valuable resistant donors, but the number is limited. Therefore, artificially induced mutants are an important resource for identifying new broad-spectrum resistant (R) genes/loci. To pursue this approach, we focused on a broad-spectrum blast resistant rice mutant line SA0169, which was previously selected from a sodium azide induced mutation pool of TNG67, an elite japonica variety. We found that SA0169 was completely resistant against the 187 recently collected blast isolates and displayed durable resistance for almost 20 years. Linkage mapping and QTL-seq analysis indicated that a 1.16-Mb region on chromosome 6 (Pi169-6(t)) and a 2.37-Mb region on chromosome 11 (Pi169-11(t)) conferred the blast resistance in SA0169. Sequence analysis and genomic editing study revealed 2 and 7 candidate R genes in Pi169-6(t) and Pi169-11(t), respectively. With the assistance of mapping results, six blast and bacterial blight double resistant lines, which carried Pi169-6(t) and/or Pi169-11(t), were established. The complementation of Pi169-6(t) and Pi169-11(t), like SA0169, showed complete resistance to all tested isolates, suggesting that the combined effects of these two genomic regions largely confer the broad-spectrum resistance of SA0169. The sodium azide induced mutant SA0169 showed broad-spectrum and durable blast resistance. The broad resistance spectrum of SA0169 is contributed by the combined effects of two R regions, Pi169-6(t) and Pi169-11(t). Our study increases the understanding of the genetic basis of the broad-spectrum blast resistance induced by sodium azide mutagenesis, and lays a foundation for breeding new rice varieties with durable resistance against the blast pathogen.

Soil supplementation with Si, B and Zn and their synergetic effects in reducing severity of wheat blast (Magnaporthe oryzae Triticum)

Wheat blast (Magnaporthe oryzae Triticum) in Bangladesh and South America is recognized as one major limiting factor of wheat production. Its control using chemical pesticides raises concerns about food safety and pesticide resistance, which have dictated the need for alternative blast management approach, nutrient supplementation could be an ecofriendly alternative. Experiments were carried out under confined net house condition for two consecutive cropping seasons. Single doses of the nutrients (Si, B and Zn) were incorporated during soil preparation. Plants of the wheat blast susceptible variety BARI Gom-26 were inoculated with spores (1 x 107 spores ml-1) of Magnaporthe oryzae Triticum at blast vulnerable pre-heading stage of 52 days age. Typical wheat blast symptoms of spike bleaching from top to downward appeared on sight 14 days after inoculation i.e., 66 days age of the crop. Incidence and severity of blast bleaching of spike were scored for four times starting from 68 days age @ three day’s interval. None of the nutrients could stop the incidence of blast on wheat; however, some nutrients reduced the blast incidence significantly. Solo application of Si, B and Zn or combination of two caused significant reduction of spike bleaching. With the mixed application of Si, B and Zn, > 47% reduction of wheat blast severity was obtained. The results revealed that the soil application of silicon, zinc and boron had a synergistic effect on the intensity of blast disease of wheat. Int. J. Agril. Res. Innov. Tech. 11(2): 76-84, Dec 2021

An Allelic Variant of the Broad-Spectrum Blast Resistance Gene Ptr in Weedy Rice is Associated with Resistance to the Most Virulent Blast Race IB-33

Rice resistance (R) genes have been effectively deployed to prevent blast disease caused by the pathogen Magnaporthe oryzae, one of the most serious threats for stable rice production worldwide. Weedy rice competing with cultivated rice may carry novel or lost R genes. The QTL qBR12.3b was previously mapped between two single nucleotide polymorphism (SNP) markers 10,633,942 bp and 10,820,033 bp in a black hull awned (BHA) weed strain using a weed-crop mapping population under greenhouse conditions. In the present study, we found a portion of the known resistance gene Ptr encoding a protein with 4 armadillo repeats and confers a broad spectrum of blast resistance. We then analyzed the sequences of the Ptr gene from weedy rice, PtrBHA, identified a unique amino acid glutamine (Gln) at protein position 874. Minor changes of protein conformation of the PtrBHA gene were predicted through structural analysis of PtrBHA suggesting the product of PtrBHA is involved in disease resistance. A gene-specific codominant marker HJ17-13 from PtrBHA was then developed to distinguish alleles in weed and crop. The existence of the PtrBHA gene in 207 individuals of the same mapping population where qBR12.3b was mapped using this gene-specific marker. Disease reactions of 207 individuals and their parents to IB-33 were evaluated. The resistant individuals had the PtrBHA whereas the susceptible individuals did not suggest HJ17-13 is reliable to predict qBR12.3b. Taken together, this newly developed marker and weedy rice genotypes carrying qBR12.3b are useful for blast improvement using marker assisted selection.

Identifying mutations in sd1, Pi54 and Pi-ta, and positively selected genes of TN1, the first semidwarf rice in Green Revolution

Background: Taichung Native 1 (TN1) is the first semidwarf rice cultivar that initiated the Green Revolution. As TN1 is a direct descendant of the Dee-geo-woo-gen cultivar, the source of the sd1 semidwarf gene, the sd1 gene can be defined through TN1. Also, TN1 is susceptible to the blast disease and is described as being drought-tolerant. However, genes related to these characteristics of TN1 are unknown. Our aim was to identify and characterize TN1 genes related to these traits. Results: Aligning the sd1 of TN1 to Nipponbare sd1, we found a 382-bp deletion including a frameshift mutation. Sanger sequencing validated this deleted region in sd1, and we proposed a model of the sd1 gene that corrects errors in the literature. We also predicted the blast disease resistant (R) genes of TN1. Orthologues of the R genes in Tetep, a well-known resistant cultivar that is commonly used as a donor for breeding new blast resistant cultivars, were then sought in TN1, and if they were present, we looked for mutations. The absence of Pi54, a well-known R gene, in TN1 partially explains why TN1 is more susceptible to blast than Tetep. We also scanned the TN1 genome using the PosiGene software and identified 11 genes deemed to have undergone positive selection. Some of them are associated with drought-resistance and stress response. Conclusions: We have redefined the deletion of the sd1 gene in TN1, a direct descendant of the Dee-geo-woo-gen cultivar, and have corrected some literature errors. Moreover, we have identified blast resistant genes and positively selected genes, including genes that characterize TN1's blast susceptibility and abiotic stress response. These new findings increase the potential of using TN1 to breed new rice cultivars.

Biologicals and New Generation Fungicides in the Management of Blast Disease in Rice

Background: Blast is a devastating disease in rice production. The current research tested the efficacy of biologicals and fungicides against blast. Aqueous extracts of Azadirachta indica, Ocimum basalicum, Cymbopogan flexousus, Thymus vulgaris, Aloe vera, Tagetes patula, Cordia curassavica, Aegle marmelos, Allium fistulosum, Syzygium aromaticum, and Calotropis gigantea were tested in vitro at 5, 10, and 15% concentrations against Magnaporthe oryzae. Also, two isolates, Bacillus cereus OG2L and B. subtilis OG2A, and one isolate, Azotobacter SAG19, and, similarly, five new generation fungicides were evaluated at three different doses against blast pathogen M. oryzae in vitro. The treatments that exhibited the best performance in vitro were further evaluated against blast disease under field conditions.Results: Extracts of T. patula (5%), C. gigantea (5%), C. curassavica (10%), A. fistulosum (10%), and A. marmelos (15%) showed greater than 81% inhibition to M. oryzae in vitro. Likewise, all three biocontrol agents, viz. B. cereus OG2L, B. subtilis OG2A, and Azotobacter SAG19, demonstrated more than 50% inhibition of mycelial growth of pathogen in vitro, and the commercial formulations of fungicides Propineb, Trifloxystrobin+Tebuconazole, Tebuconazol+Triadimenol, Bacillus subtilis Strain QST 713, and Cinnamon Oil 8%+Clove Oil 2% also significantly inhibited M. oryzae. Under field conditions, C. curassavica (10%), A. marmelos (15%), C. gigantea (5%); B.cereus OG2L; B.subtilis OG2A; Proineb and Trifloxystrobin+Tebuconazole were effective in reducing disease in addition to yield enhancement.Conclusion: Overall, the plant extracts (C. curassavica at 10%, A. marmelos at 15%, C. gigantea at 5%); bio-agents (B. cereus OG2L, B. subtilis OG2A both at 2g/l), and new generation fungicides Antracol 70WP (Proineb) at 2.5 g/l and Nativo 75 WG (Trifloxystrobin+Tebuconazole) at 0.5 g/l provided effective control against blast disease and superior plant growth and yield compared with other treatments and untreated control.