Bacterial Blight Resistance Research Articles

A Novel Single Base Mutation in OsSPL42 Leads to the Formation of Leaf Lesions in Rice

Rice spotted-leaf mutants serve as valuable resources for studying plant programmed cell death (PCD) and disease resistance mechanisms, making them crucial for research on disease resistance in rice. Map-based cloning was used to identify and clone the spotted-leaf gene OsSPL42. Then, functional complementation and CRISPR/Cas9 techniques were also employed to further validate the function of this gene. By applying leaf clippings for bacterial blight (BB) inoculation, the BB resistance of different rice lines was assessed. The results in this study were as follows: The OsSPL42 behaved as a recessive nuclear gene and was narrowed down to a 111 kb region on chromosome 8. All T0 transgenic rice plants in the complementation experiments exhibited a wild-type phenotype, without any lesion spots on the rice leaves. This suggests that the LOC_Os08g06100 encoding O-methyltransferase is the candidate gene for the mutant spl42. The OsSpl42 is widely expressed and the OsSPL42-GFP protein is mainly localized in the cytoplasm. OsSPL42 overexpression lines are more susceptible to BBs, which indicates that OsSPL42 may act as a negative regulator of rice resistance to BB. In summary, we speculate that OsSPL42 plays an important role in the regulation of pathogen response, providing new insights into plant defense mechanisms.

Marker Assisted Selection in Advanced Backcross Population of Rice Variety, MTU1010 for Bacterial Blight and Blast Resistance

The present study was aimed for identification of the bacterial blight and blast resistant lines in the advanced back cross population of MTU-1010 using marker assisted foreground selection and also for evaluation of agro-morphological characters. The M-16-59 introgressed line (derived from an intercrossing of BC2F1 plants of MTU-1010 x GPP2 and MTU-1010 x NLR 145) is developed under ongoing DBTDBSRR subproject -IV possessing Xa21, xa13, Pi1 and Pi54 resistance genes having broad spectrum resistance to bacterial blight and blast is used as a donor parent and MTU-1010 was used as a recurrent parent for the back cross. Before attempting the backcross both the parents were verified for the target genes along with the original donors GPP2 and NLR 145 using gene specific/linked molecular markers viz., xa13-promo for xa13 gene, pTA248 for Xa21 gene, RM 224 for Pi1 and Pi54 MAS for Pi54. under the F1 generation (equivalent to BC3F1 because two backcrosses were completed earlier), 120 plants were screened, and 16 plants were confirmed for xa13, Xa21, Pi1, and Pi54 genes under heterozygous conditions. These confirmed plants were assessed for agro-morphological characteristics such as yield, grain features, and plant type. The results showed that the confirmed heterozygous plants were comparable to MTU-1010 and passed to the next round of selection and evaluation. The BC1F1-198th plant (96.8%) was selfed to produce BC1F2 population. Twenty five plants were advanced to BC1F3 generation based on BB resistance and above other phenotypic characters. The selected BC1F3 progenies were screened for blast and BB resistance. Four BC1F3 progenies with four target genes (xa13xa13Xa21Xa21Pi54Pi54Pi1Pi1) showed very high level of resistance to both the diseases. BC1F2-198-52nd line found similar to MTU1010 with respect to yield and yield related characters besides showing resistance to both BB and blast.Improved lines of MTU1010 can be advanced for multi-location testing under All India Coordinated Rice Improvement Project (AICRIP) for their evaluation and possible release for the benefit of rice farmers.

Phosphatidylcholine Transfer Protein OsPCTP Interacts with Ascorbate Peroxidase OsAPX8 to Regulate Bacterial Blight Resistance in Rice

Rice phosphatidylcholine transfer protein (PCTP), which contains a steroidogenic acute regulatory protein-related lipid transfer (START) domain, responds to bacterial blight disease. Overexpression of OsPCTP quantitatively enhances resistance to pathogen in rice, whereas depletion of it has the opposite effect. Further analysis showed that OsPCTP physically interacts with OsAPX8, a ROS scavenging enzyme, and influences ascorbate peroxidase enzymatic activity in vivo. In addition, the expression of pathogenesis-related genes PR1a, PR1b and PR10 were significantly induced in OsPCTP-OE plants compared with that in wild-type plants ZH11. Taken together, these results suggested that OsPCTP mediates bacterial blight resistance in rice through regulating the ROS defense pathway.

OsGRF6-OsYUCCA1/OsWRKY82 Signaling Cascade Upgrade Grain Yield and Bacterial Blight Resistance in Rice.

As a major crop in the world, the sustainable development of rice is often severely restricted by bacterial blight. Breeding crops with resistance is an efficient way to control bacterial blight. However, enhancing resistance often incurs a fitness penalty, making it challenging to simultaneously increase bacterial blight resistance and yield potential. In this study, it is found that OsGRF6, besides being a high-yield gene, can significantly improve rice bacterial blight resistance. Compared with wild-type, the lesion lengths of transgenic material overexpressing OsGRF6 are significantly reduced after inoculation with Xanthomonas oryzae pv. oryzae (Xoo). Furthermore, OsGRF6 can directly bind to the promoters of OsYUCCA1 and OsWRKY82, upregulating their transcription and thereby increasing rice bacterial blight resistance and yield. Haplotypic analysis based on the promoter and genome sequence combined with evolutionary analysis revealed that OsGRF6 is mainly comprised by the OsGRF6XI and OsGRF6GJ subtypes. The superior haplotype OsGRF6Hap4 increased its transcriptional activity and contributed to bacterial blight resistance and rice yield. Together, this study provides theoretical support for further revealing the synergistic regulatory mechanism and genetic improvement of rice high yield and bacterial blight resistance, offering a new strategy for developing disease-resistant cultivars.

Cellulose synthase-like OsCSLD4: a key regulator of agronomic traits, disease resistance, and metabolic indices in rice.

Cellulose synthase-like OsCSLD4 plays a pivotal role in regulating diverse agronomic traits, enhancing resistance against bacterial leaf blight, and modulating metabolite indices based on the multi-omics analysis in rice. To delve deeper into this complex network between agronomic traits and metabolites in rice, we have compiled a dataset encompassing genome, phenome, and metabolome, including 524 diverse accessions, 11 agronomic traits, and 841 metabolites, enabling us to pinpoint eight hotspots through GWAS. We later discovered four distinct metabolite categories, encompassing 15 metabolites that are concurrently present on the QTL qC12.1, associated with leaf angle of flag and spikelet length, and finally focused the cellulose synthase-like OsCSLD4, which was pinpointed through a rigorous process encompassing sequence variation, haplotype, ATAC, and differential expression across diverse tissues. Compared to the wild type, csld4 exhibited significant reductions in the plant height, flag leaf length, leaf width, spikelet length, 1000-grain weight, grain width, grain thickness, fertility, yield per plant, and bacterial blight resistance. However, there were significant increase in tiller numbers, degree of leaf rolling, flowering period, growth period, grain length, and empty kernel rate. Furthermore, the content of four polyphenol metabolites, excluding metabolite N-feruloyltyramine (mr1268), notably rose, whereas the levels of the other three polyphenol metabolites, smiglaside C (mr1498), 4-coumaric acid (mr1622), and smiglaside A (mr1925) decreased significantly in mutant csld4. The content of amino acid L-tyramine (mr1446) exhibited a notable increase, whereas the alkaloid trigonelline (mr1188) displayed a substantial decrease among the mutants. This study offered a comprehensive multi-omics perspective to analyze the genetic mechanism of OsCSLD4, and breeders can potentially enhance rice's yield, bacterial leaf blight resistance, and metabolite content, leading to more sustainable and profitable rice production.

Pyramiding of multiple resistant genes of blast and bacterial blight diseases in the background of rice (Oryza sativa) mega variety BRRI dhan29

Blast and bacterial blight (BB) are the two major rice diseases in the world including Bangladesh. In this study, BB resistance genes (Xa21 and xa13) and blast resistance (Pi9 and Pb1) genes were pyramided into a mega variety, BRRI dhan29 through marker-assisted backcross breeding. IRBB58 was used as a BB-resistant donor and Pi9-US2, and Pb1-US2 were used as blast-resistant donors. Backcross was done between BRRI dhan29 and donor parents to develop BC3F1 population and then selfing was done to develop BC3F6 population. BC3F2 population was genotyped and phenotyped for segregation analysis and BC3F6 was evaluated for genotyping, phenotyping and morphological traits and yields. Chi-square analysis of BC3F2 data revealed that blast and BB resistance followed the single gene mendelian fashion (1:2:1 and 3:1). Two to four gene combinations were found in the advanced lines of the BC3F6 population. The yield of the advanced lines ranged from 6.42 (t ha-1) to 9.5 (t ha-1) and they showed resistant against blast and BB with mean disease scores ranging from 0.67 to 2.33 and 0.33 to 2.33, respectively. Finally, eight lines having four genes (xa13, Xa21, Pi9 and Pb1) were selected for multilocational (five locations) trials for yield performance and disease reaction. Mean yield data of eight advanced lines of all locations were varied from 6.48±0.15 to 8.38±0.11 t ha-1 and all the lines showed resistant reactions against blast (score 0.53 to 1) and BB (score 0.6 to 0.87) disease. The highest yield was found in BR (Path) 13800-BC3-224-12 (G28, 8.38±0.11 t ha-1) followed by BR (Path) 13800-BC3-134-252 (G26, 8.28±0.08 t ha-1) and BR (Path) 13800-BC3-136-115 (G12, 8.24±0.07 t ha-1). Pyramided advanced lines of this study could be released as BB and blast-resistant varieties or could be utilized as donor parents in resistant breeding.

First Comprehensive Evaluation of Genetic Variability for Bacterial Blight Resistance in Wild Punica granatum L. Populations from the North-Western Himalayas

Identifying and selecting disease-resistant sources from wild germplasm pools is pivotal due to their ecological benefits and cost-effectiveness. This study encompassed the survey and collection of wild pomegranate genotypes alongside commercial cultivars, followed by screening against native pathogens. Additionally, biochemical analyses were conducted to elucidate defense mechanisms in both resistant and susceptible genotypes. Cuttings from selected germplasm across four districts were obtained, grown for one year, and assessed for pathogenicity and biochemical responses. Microscopic and molecular analyses confirmed the pathogen as Xanthomonas axonopodis pv. punicae (Xap), identified by its distinctive morphology and a 1500 bp amplicon along with phylogeny analysis. Germplasm screening via detached and attached leaf assays identified genotypes with minimal disease severity, highlighting SH-14 and SH-16 as highly resistant in the attached leaf assay. Principal component analysis categorized genotypes based on disease parameters, with PC1 explaining approximately 75% of the variability. Agglomerative hierarchical clustering further grouped genotypes into ten clusters, emphasizing clusters IX and X for their significant resistance traits. Biochemical studies revealed that the resistant genotype SH-16 exhibited higher levels of constitutive defense components like chlorophyll and vitamin C, as well as induced defense components such as phenols, flavonoids, and antioxidants, compared to the susceptible cultivar Bhagwa. These findings underscore the critical roles of both constitutive and induced defense mechanisms in conferring pomegranate resistance to Xap. The study's comprehensive approach in evaluating genetic and biochemical resistance provides valuable insights for breeding disease-resistant pomegranate varieties, marking a significant advancement in exploring pomegranate wild relatives from the Himalayan region for their resistance to bacterial blight.

Development and Validation of an SNP Marker for Identifying Xanthomonas oryzae pv. oryzae Thai Isolates That Break xa5-Mediated Bacterial Blight Resistance in Rice.

Xanthomonas oryzae pv. oryzae (Xoo) is a pathogenic bacterium responsible for bacterial blight (BB) disease in rice, primarily mediated by the interaction between the plant and pathogen. The virulence mechanism involves the activation of the Sugars Will Eventually be Exported Transporter (SWEET) gene family in rice by transcription activator-like effectors derived from Xoo. The BB resistance gene xa5 has been identified as one of the most effective genes against Thai Xoo isolates, but xa5-mediated resistance-breaking Xoo strains have emerged. This study aimed to develop a single nucleotide polymorphism (SNP) marker for precise identification of xa5-mediated resistance-breaking Xoo. Comparative genomics of Thai Xoo isolates Xoo16PK001 and Xoo16PK002, which were incompatible and compatible with rice variety IRBB5 carrying xa5, respectively, identified eight SNP positions for the development of an SNP marker. The SNP marker XooE6 yields a specific 1,143 bp PCR product unique to Xoo16PK002. Screening 61 Thai isolates using XooE6 identified two positives: Xoo20PL010 and Xoo20UT002. Inoculation tests on rice varieties IRBB5 and IRBB13 demonstrated compatibility with IRBB5 and incompatibility with IRBB13, which bears Xa5 and xa13. Xoo16PK001 (XooE6-negative) showed different virulence. Inoculation on IRBB21 harboring Xa5, Xa13, and Xa21 resulted in partial resistance to both XooE6-positive and -negative strains. XooE6-positive strains up-regulated SWEET11 and suppressed SWEET14 in IRBB5, while Xoo16PK001 slightly induced SWEET11 but activated SWEET14 in IRBB13. This highlights the potential of XooE6 to identify xa5-mediated resistance-breaking Xoo strains and elucidate their pathogenic mechanisms through the upregulation of SWEET11.

Genome-wide association study of bacterial blight resistance in soybean.

Bacterial blight caused by Pseudomonas syringae pv. glycines (Psg) is a widespread foliar disease. Although four Resistance to Pseudomonas syringae pv. glycinea (Rpg) 1 ~ 4 (Rpg1~4) genes that have been observed to segregate in a Mendelian pattern have been reported to confer resistance to Psg in soybean, the genetic basis of quantitative resistance to bacterial blight in soybean remains unclear. In the present study, the Psg resistance of two soybean association panels consisting of 573 and 213 lines, respectively, were phenotyped in multiple environments in 2014 - 2016. Genome-wide association study (GWAS) were performed using 2 models FarmCPU and BLINK to identify Psg resistance loci. A total of 40 soybean varieties with high level of Psg resistance were identified, and 14 quantitative trait loci (QTLs) were detected on 12 soybean chromosomes. These QTLs were identified for the first time. The majority of the QTLs were only detected in one or the other association panels, while qRPG-18-1 was detected in both association panels for at least one growing season. A total of 46 candidate Psg resistance genes were identified from the qRpg_13_1, qRPG-15-1, and qRPG-18-1 loci based on gene function annotation. In addition, we found the genomic region covering rpg1-b and rpg1-r harbored the synteny with a genomic region on chromosome 15, and identified 16 nucleotide binding site - leucine-rich repeat (NBS-LRR) genes as the candidate Psg resistance genes from the synteny blocks. This study provides new information for dissecting the genetic control of Psg resistance in soybean.

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Genomic and transcriptomic analyses of the elite rice variety Huizhan provide insight into disease resistance and heat tolerance

The indica rice variety Huizhan shows elite traits of disease resistance and heat tolerance. However, the underlying genetic basis of these traits is not fully understood due to limited genomic resources. Here, we used Nanopore long-read and next-generation sequencing technologies to generate a chromosome-scale genome assembly of Huizhan. Comparative genomics analysis uncovered a large chromosomal inversion and expanded gene families that are associated with plant growth, development and stress responses. Functional rice blast resistance genes, including Pi2, Pib and Ptr, and bacterial blight resistance gene Xa27, contribute to disease resistance of Huizhan. Furthermore, integrated genomics and transcriptomics analyses showed that OsHIRP1, OsbZIP60, the SOD gene family, and various transcription factors are involved in heat tolerance of Huizhan. The high-quality genome assembly and comparative genomics results presented in this study facilitate the use of Huizhan as an elite parental line in developing rice varieties adapted to disease pressure and climate challenges.

Association between molecular markers and resistance to bacterial blight using binary logistic analysis

The most effective strategy for managing wheat bacterial blight caused by Pseudomonas syringae pv. syringae is believed to be the use of resistant cultivars. Researching the correlation between molecular markers and stress resistance can expedite the plant breeding process. The current study aims to evaluate the response of 27 bread wheat cultivars to bacterial blight disease in order to identify resistant and susceptible cultivars and to pinpoint ISSR molecular markers associated with bacterial blight resistance genes. ISSR markers are recommended for assessing a plant's disease resistance. This experiment is focused on identifying ISSR molecular markers linked to bacterial blight resistance. After applying the bacterial solution to the leaves, we performed sampling to determine the infection percentage in the leaves at different intervals (7, 14, and 18 days after spraying). In most cultivars, the average leaf infection percentage decreased 18 days after spraying on young leaves. However, in some cultivars such as Niknegad, Darab2, and Zarin, leaf infection increased in older leaves and reached up to 100% necrosis. In our study, 12 ISSR primers generated a total of 170 bands, with 156 being polymorphic. The primers F10 and F5 showed the highest polymorphism, while the F7 primer exhibited the lowest polymorphism. Cluster analysis grouped these cultivars into four categories. The resistant group included Qods, Omid, and Atrak cultivars, while the semi-resistant and susceptible groups comprised the rest of the cultivars. Through binary logistic analysis, we identified three Super oxide dismutase-related genes that contribute to plant resistance to bacterial blight. These genes were linked to the F3, F5, and F12 primers in regions I (1500 bp), T (1000 bp), and G (850 bp), respectively. We also identified seven susceptibility-associated genes. Atrak, Omid, and Qods cultivars exhibited resistance against bacterial blight, and three genes associated with this resistance were linked to the F3, F5, and F12 primers. These markers can be used for screening or transferring tolerance to other wheat cultivars in breeding programs.

Salicylic acid application against bacterial blight resistance in Xa21-introgression Thai rice cultivar ‘Phitsanulok 2’

Rice cultivar ‘Phitsanulok 2’ (‘PSL2’) is popularly grown in lower-northern Thailand, because it has good agricultural traits with high-quality and high-yield grains. However, one significant limitation of rice ‘PSL2’ is its susceptibility to bacterial blight (BB) disease caused by bacteria <i>Xanthomonas oryzae</i> pv. <i>oryzae (Xoo)</i>, leading to considerable yield losses. Effort approaches have been used to enhance the BB resistance of rice ‘PSL2’ by introducing the resistance gene <i>Xa21</i> and using salicylic acid (SA). Thus, the objective of this study was to determine the effects of SA application on BB disease reduction in rice cultivars (‘PSL2’, ‘IRBB21’, and backcrossed lines ‘PSL2-<i>Xa21’</i> in BC<sub>4</sub>F<sub>6</sub>) after<i> Xoo</i>16PK002 inoculation, compared to a mock treatment control. The results showed that the SA-pretreated rice (2mM) exhibited significantly induced upregulation of the <i>Xa21</i> gene expression before the <i>Xoo</i>16PK002-infected rice cultivars ‘IRBB21’ and ‘PSL2-<i>Xa21</i>-BC<sub>4</sub>F<sub>6</sub>’. Additionally, the SA-pretreated rice ‘PSL2-<i>Xa21</i>-BC<sub>4</sub>F<sub>6</sub>’ demonstrated significant suppression of <i>Xoo</i>16PK002 proliferation (from 1.4 to 2.4-fold reduction), compared to the H<sub>2</sub>O pretreatment. Similarly, the SA-pretreated rice ‘PSL2-<i>Xa21</i>-BC<sub>4</sub>F<sub>6</sub>’ exhibited increased resistance against BB disease, i.e. reduced lesion lengths (10.0-10.9 cm at 14 DAI and 24.6-28.0 cm at 21 DAI), compared to the water pretreatment (16.8-18.0 cm at 14 DAI and 34.9-35.1 cm at 21 DAI). In summary, the application of the SA-pretreatment in the rice introgression lines ‘PSL2-<i>Xa21</i>-BC<sub>4</sub>F<sub>6</sub>’ resulted in improved defense mechanisms against bacteria <i>Xoo</i>16PK002 through reduced appearance of symptoms and inhibited <i>Xoo</i> multiplication.

Sheath Blight and Bacterial Blight Resistance in Rice: Mechanisms, Progress and Future Perspectives for Sustainable Rice Production

Rice (Oryza sativa L.) is a staple food for nearly half of the world's population, including Pakistan. But several diseases are always posing a threat to rice farming, with bacterial blight and sheath blight being the two most common causes. R. solani is the main source of rice sheath blight and stood crop’s most destructive diseases. Reviewing published data on pathogenicity and disease management is crucial to developing effective crop protection against sheath blight. It also helps identify research gaps that need more in-depth investigation. Although there has been progress in identifying rice and pathogen-related genes linked to pathogenesis, the underlying processes are still unknown. Research on the applications of; agronomic techniques, chemical control, biological control, and genetic improvement in disease management strategies has been conducted. Optimizing the application of nitrogen fertilizer in combination to plant-plant spacing can minimize the transmission of infection, whereas SMART agricultural technologies; like crop monitoring using Unmanned Aerial Systems help detect and treat sheath blight disease early on. Biological agents and natural fungicides can be used to effectively prevent sheath blight while reducing the negative effects on the environment. Genetic strategies that hold potential to control sheath blight include the use of exogenous dsRNA to suppress pathogen gene expression, genome editing to create rice lines less susceptible to sheath blight, and the development of transgenic rice lines that over express or silence genes related to pathogenesis. The pathogen’s flexibility, the absence of resistant rice types, the lack of single resistance genes for breeding, and farmers' restricted access to informative programs about optimum management methods all work against effective crop protection against sheath blight.

Pyramiding of bacterial blight resistance genes into promising restorer BRRI31R line through marker-assisted backcross breeding and evaluation of agro-morphological and physiochemical characteristics of developed resistant restorer lines.

BRRI31R is one of the Bangladesh's most promising restorer lines due to its abundant pollen producing capacity, strong restoring ability, good combining ability, high outcrossing rate and genetically diverse from cytoplasmic male sterile (CMS) line. But the drawback of this line is that it is highly susceptible to bacterial blight (BB) disease of rice caused by Xanthomonas oryzae pv. oryzae. The present study highlighted the pyramiding of effective BB resistance genes (xa5, xa13 and Xa21) into the background of BRRI31R, through marker-assisted backcrossing (MABC). Backcross progenies were confirmed and advanced based on the foreground selection of target genes. Pyramided lines were used for pathogenicity test against five Bangladeshi Xanthomonas oryzae (BXo) races (BXo93, BXo220, BXo822, BXo826, BXo887) and confirmed the dominant fertility restore genes, Rf3 and Rf4 and further validated against SNP markers for more confirmation of target resistance genes. All pyramided restorer lines consisted of Xa4 (in built), xa5, xa13, Xa21, and Chalk5 with two fertility restorer genes, Rf3, Rf4. and these restorer lines showed intermediate amylose content (<25%). Restorer lines BRRI31R-MASP3 and BRRI31R-MASP4 showed high levels of resistance against five virulent BXo races and SNP genotyping revealed that these lines also contained a blast resistance gene Pita races. Gene pyramided restorer lines, BRRI31R-MASP3 and BRRI31R-MASP4 can directly be used as a male parent for the development of new BB resistant hybrid rice variety or could be used as a replacement of restorer line of BRRI hybrid dhan5 and 7 to enhance the quality of hybrid seeds as well as rice production in Bangladesh.

Evaluation Methods, Resistant Germplasm, and Breeding for Resistance to Bacterial Blight in Cotton: A Review

Bacterial blight (BB) caused by Xanthomonas citri pv. malvacearum (Xcm) is an important cotton (Gossypium spp.) production problem. In the U.S., BB has been controlled effectively using resistant cultivars and acid-delinted seed since the 1970s; however, resurgence of BB occurred in the early 2010s because of negligence in breeding. This review provides an up-to-date account on the pathogen, resistance evaluation methods, resistant germplasm lines, and breeding methods. Twenty-two Xcm races have been reported worldwide, and race 18 is currently the only one found in production fields in the U.S. To evaluate cotton for BB resistance, a pressure-sprayer-based method with surfactant in the field and a cotyledon-scratching-based method in the greenhouse are most often used. Breeding for BB resistance was highly successful in Sudan between the late 1930s and 1960s, when many resistance genes were transferred to G. barbadense from G. arboreum, G. herbaceum, G. anomalum, and G. hirsutum. Breeding for BB resistance commenced in the U.S. in the 1940s, leading to development of numerous resistant Upland cultivars. Although backcrossing was often used to transfer resistance genes in early years, forward breeding has been the breeding method of choice. Currently, some and possibly all resistant cultivars in the U.S. possess the resistance gene B12, which confers immunity with no water-soaked symptoms. Although B12-based resistance has held for a long time, identification of new resistant sources is needed to prevent an epidemic of BB due to evolution or introduction of possible new virulent Xcm races.

KIN10-mediated HB16 protein phosphorylation and self-association improve cassava disease resistance by transcriptional activation of lignin biosynthesis genes.

Cassava bacterial blight significantly affects cassava yield worldwide, while major cassava cultivars are susceptible to this disease. Therefore, it is crucial to identify cassava disease resistance gene networks and defence molecules for the genetic improvement of cassava cultivars. In this study, we found that MeHB16 transcription factor as a differentially expressed gene in cassava cultivars with contrasting disease resistance, positively modulated disease resistance by modulating defence molecule lignin accumulation. Further investigation showed that MeHB16 physically interacted with itself via the leucine-Zippe domain (L-Zip), which was necessary for the transcriptional activation of downstream lignin biosynthesis genes. In addition, protein kinase MeKIN10 directly interacted with MeHB16 to promote its phosphorylation at Ser6, which in turn enhanced MeHB16 self-association and downstream lignin biosynthesis. In summary, this study revealed the molecular network of MeKIN10-mediated MeHB16 protein phosphorylation improved cassava bacterial blight resistance by fine-tuning lignin biosynthesis and provides candidate genes and the defence molecule for improving cassava disease resistance.

Precision mapping and expression analysis of recessive bacterial blight resistance gene xa-45(t) from Oryza glaberrima.

Bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo), is one of the most devastating diseases of rice leading to huge yield losses in Southeast Asia. The recessive resistance gene xa-45(t) from Oryza glaberrima IRGC102600B, mapped on rice chromosome 8, spans 80Kb with 9 candidate genes on Nipponbare reference genome IRGSP-1.0. The xa-45(t) gene provides durable resistance against all the ten Xanthomonas pathotypes of Northern India, thus aiding in the expansion of recessive bacterial blight resistance gene pool. Punjab Rice PR127, carrying xa-45(t), was released for wider use in breeding programs. This study aims to precisely locate the target gene among the 9 candidates conferring resistance to bacterial blight disease. Sanger sequencing of all nine candidate genes revealed seven SNPs and an Indel between the susceptible parent Pusa 44 and the resistant introgression line IL274. The genotyping with polymorphic markers identified three recombinant breakpoints for LOC_Os08g42370, and LOC_Os08g42400, 15 recombinants for LOC_Os08g423420 and 26 for LOC_Os08g42440 out of 190 individuals. Relative expression analysis across six time intervals (0, 8, 24, 48, 72, and 96h) after bacterial blight infection showed over expression of LOC_Os08g42410-specific transcripts in IL274 compared to Pusa 44, with a significant 4.46-fold increase observed at 72h post-inoculation. The Indel marker at the locus LOC_Os08g42410 was found co-segregating with the phenotype, suggesting its candidacy towards xa-45(t). The transcript abundance assay provides strong evidence for the involvement of LOC_Os08g42410 in the resistance conferred by the bacterial blight gene xa-45(t).

Understanding the nature of blast resistance in combined bacterial leaf blight and blast gene pyramided lines of rice variety tellahamsa.

Rice blast and bacterial leaf blight (BLB) are the most limiting factors for rice production in the world which cause yield losses typically ranging from 20 to 30% and can be as high as 50% in some areas of Asia especially India under severe infection conditions. An improved line of Tellahamsa, TH-625-491 having two BLB resistance genes (xa13 and Xa21) and two blast resistance genes (Pi54 and Pi1) with 95% Tellahamsa genome was used in the present study. TH-625-491 was validated for all four target genes and was used for backcrossing with Tellahamsa. Seventeen IBC1F1 plants heterozygous for all four target genes, 19 IBC1F2 plants homozygous for four, three and two gene combinations and 19 IBC1F2:3 plants also homozygous for four, three and two gene combinations were observed. Among seventeen IBC1F1 plants, IBC1F1-62 plant recorded highest recurrent parent genome (97.5%) covering 75 polymorphic markers. Out of the total of 920 IBC1F2 plants screened, 19 homozygous plants were homozygous for four, three and two target genes along with bacterial blight resistance. Background analysis was done in all 19 homozygous IBC1F2 plants possessing BLB resistance (possessing xa13, Xa21, Pi54 and Pi1 in different combinations) with five parental polymorphic SSR markers. IBC1F2-62-515 recovered 98.5% recurrent parent genome. The four, three and two gene pyramided lines of Tellahamsa exhibited varying resistance to blast. Results show that there might be presence of antagonistic effect between bacterial blight and blast resistance genes since the lines with Pi54 and Pi1 combination are showing better resistance than the combinations with both bacterial blight and blast resistance genes.

Genetic Enhancement of Blast and Bacterial Leaf Blight Resistance in Rice Variety CO 51 through Marker-Assisted Selection

The increased use of chemicals in rice farming poses significant issues regarding the emergence of pesticide/fungicide resistance and environmental sustainability concerns. This study was aimed at the genetic improvement of blast, bacterial leaf blight (BB) and gall midge resistance in a popular rice variety CO 51 which already harbours a blast resistance gene Pi54. Efforts were made to pyramid an additional blast resistance gene Pi9 along with two BB resistance genes (xa13 and Xa21) and two gall midge resistance genes (Gm1 and Gm4) into an elite rice variety CO 51 to enhance the resistance level to biotic stresses. The superior lines were selected using functional markers conferring resistance to blast (NBS4 and Pi54MAS linked to Pi9 and Pi54 genes, respectively) and BB [(xa13Prom (xa13) and pTA248 (Xa21)] and SSR markers linked to Gm1 (RM1328) and Gm4 (RM22550) for phenotypic screening and agronomic evaluation. The genotyping and phenotyping of F6 and BC2F6 progenies of CO 51 X 562-4, for agronomic traits and resistance to BB and blast, identified ten superior progenies in F6 and five superior progenies in BC2F6. The breeding lines harbouring both xa13+Xa21 exhibited high levels of resistance to BB (score ≤ 1 cm) and Pi9+Pi54 exhibited strong resistance to blast (score ≤ 2). Identified lines can be evaluated further for varietal improvement or utilised as genetic stocks in breeding programs.