Leaf Stripe Research Articles

Acidovorax sacchari sp. nov., a pathogen causing red stripe of sugarcane in Japan.

Phytopathogenic bacteria (MAFF 311311T and MAFF 311313) were isolated from sugarcane plants exhibiting leaf stripe symptoms associated with red stripe disease in Okinawa Prefecture, Japan. The strains were Gram-reaction-negative, aerobic, motile with one polar flagellum, rod-shaped and non-spore-forming. The genomic DNA G+C content was 69.0 mol%, and the major cellular fatty acids (>10 % of the total fatty acids) included summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 0 and summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c). Phylogenomic analyses using whole-genome sequences consistently placed these strains within the genus Acidovorax. However, their phylogenetic positions did not correspond to any known species within this genus. Comparative analyses, including average nucleotide identity and digital DNA-DNA hybridization with closely related species, yielded values below the thresholds for prokaryotic species delineation (95-96 and 70 %, respectively), with the highest values observed for Acidovorax oryzae ATCC 19882T (93.98 and 54.3 %, respectively). Phenotypic characteristics, cellular fatty acid composition and a repertoire of secretion systems and their effectors can differentiate these strains from their closest relatives. The phenotypic, chemotaxonomic and genotypic data obtained in this study indicate that MAFF 311311T and MAFF 311313 constitute a novel species within the genus Acidovorax, for which we propose the name Acidovorax sacchari sp. nov., with MAFF 311311T (=ICMP 25276T) designated as the type strain.

Identification of Leaf Stripe Resistance Genes in Hulless Barley Landrace Teliteqingke from Qinghai-Tibet Plateau

Leaf stripe disease, caused by Pyrenophora graminea, is a seed-borne fungal disease that significantly impacts hulless barley (Hordeum vulgare var. nudum) production on the Qinghai-Tibet Plateau. This study aimed to identify genetic factors conferring resistance to the leaf stripe by analyzing an F3 population derived from a cross between the resistant landrace Teliteqingke and the susceptible landrace Dulihuang. Genetic analysis revealed that resistance in Teliteqingke was governed by two dominant genes. Using bulked segregant analysis combined with an SNP array (BSA-SNP) and RNA-seq, we identified two candidate regions on chromosomes 3H and 7H. Further analysis focused on chromosome 3H, which revealed a candidate genomic region containing seven potential disease-resistance genes. Among these, RT-qPCR experiments demonstrated significant expression induction of HORVU.MOREX.r3.3HG0232110.1 (encoding a RING/U-box superfamily protein) and HORVU.MOREX.r3.3HG0232410.1 (encoding a bZIP transcription factor) showed significant expression induction following inoculation with P. graminea. These genes are strong candidates for the resistance mechanism against leaf stripes in Teliteqingke. These results provide a foundation for functional validation of these genes and offer valuable insights for breeding disease-resistant hulless barley.

Transcriptome, miRNA, and degradome sequencing reveal the leaf stripe (Pyrenophora graminea) resistance genes in Tibetan hulless barley

Barley leaf stripe, a disease mainly caused by Pyrenophora graminea (P. graminea) infection, severely affects barley yield and quality and is one of the most widespread diseases in barley production. However, little is known about the underlying molecular mechanisms of leaf stripe resistance. In this study, the transcript expression profiles of normal and infected leaves of resistant Tibetan hulless barley (Hordeum vulgare L. var. nudum Hook. f.) variety Kunlun 14 and susceptible variety Z1141 were analyzed by RNA sequencing (RNA-seq). The results showed a total of 7,669 and 5,943 differentially expressed genes (DEGs) were found in resistant and susceptible Kunlun 14 and Z1141, respectively, with 8,916 DEGs found between Kunlun 14 and Z1141. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis of the 8,916 DEGs identified many significantly enriched categories and pathways, of which a plant–pathogen interaction pathway, containing a total of 102 genes (100 known genes and two novel genes), was found, that was very important for the study of the leaf stripe resistance mechanism. Using RNA-seq, small RNA sequencing (miRNA-seq) combined with degradome sequencing (degradome-seq), four pairs associated with leaf-stripe miRNAs and target genes were obtained, namely Hvu-miR168-5p and Argonaute1 (HvAGO1), Hvu-novel-52 and growth-regulating factor 6 (HvGRF6), Hvu-miR6195 and chemocyanin-like protein (CLP), and Hvu-miR159b and gibberellin-dependent MYB (GAMYB). Transformation of the important target gene HvAGO1 into Arabidopsis verified that HvAGO1 could against Botrytis cinerea. Then RNA-seq and miRNA-seq of Arabidopsis transformed with overexpressed of HvAGO1 were performed. Based on the above research results, we constructed a Protein-Protein Interaction (PPI) network of barley leaf stripe resistance. This study lays the foundation for the study of the barley leaf stripe resistance mechanism and provides new targets for the genetic improvement of disease-resistant barley varieties.

Pseudo-linkage or real-linkage of rust resistance genes in a wheat-Thinopyrum intermedium translocation line.

We analysed the chromosomal structures of two wheat-Thinopyrum intermedium addition lines Z4 and Z5 and resolved the linkage relationship between the leaf rust and stripe rust resistance genes in Z4. Wheat addition lines Z4 and Z5 carrying rust resistance genes from Thinopyrum intermedium (JJJsJsStSt, 2n = 6x = 42) together with three wheat lines involved in the production of these addition lines were analysed by rust response, 90K SNP genotyping, and molecular cytogenetic analysis. Seedling leaf rust (LR) responses to five diverse pathotypes indicated that the LR resistance gene(s) was located in translocation chromosome T3DS-3AS.3AL-7StS common to Z4 and Z5. The stripe rust (YR) resistance gene(s) was located in translocation chromosome T3AL-7StS.7StL, which is unique to Z4, based on the seedling YR responses to four diverse pathotypes. Backcross and selfed populations involving the addition lines and various wheat cultivars were studied to understand the inheritance of the alien resistance genes. Although inheritance studies indicated genetic linkage, the alien genes for resistance to leaf rust (LR) and stripe rust (YR) in Z4 were present in different wheat-Th. intermedium translocation chromosomes. We found that LR and YR were in pseudo-linkage, rather than true linkage.

Genotyping-by-sequencing uncovers a Thinopyrum 4StS·1JvsS Robertsonian translocation linked to multiple stress tolerances in bread wheat

KeymessageGBS read coverage analysis identified a Robertsonian chromosome from twoThinopyrumsubgenomes in wheat, conferring leaf and stripe rust resistance, drought tolerance, and maintaining yield stability.Agropyron glael (GLAEL), a Thinopyrum intermedium × Th. ponticum hybrid, serves as a valuable genetic resource for wheat improvement. Despite its potential, limited knowledge of its chromosome structure and homoeologous relationships with hexaploid wheat (Triticum aestivum) has restricted the full exploitation of GLAEL's genetic diversity in breeding programs. Here, we present the development of a 44-chromosome wheat/GLAEL addition line (GLA7). Multicolor genomic in situ hybridization identified one chromosome arm from the St subgenome of Th. intermedium, while the other arm remained unclassified. Genotyping-by-sequencing (GBS) read coverage analysis revealed a unique Robertsonian translocation between two distinct Thinopyrum subgenomes, identified as 4StS·1JvsS. The GLA7 line demonstrated strong adult plant resistance to both leaf rust and stripe rust under natural and artificial infection conditions. Automated phenotyping of shoot morphological parameters together with leaf relative water content and yield components showed that the GLA7 line exhibited elevated drought tolerance compared to parental wheat genotypes. Three years of field trials showed that GLA7 exhibits similar agronomic performance and yield components to the wheat parents. This unique addition line holds promise for enhancing wheat's tolerance to multiple stresses through the introduction of new resistance genes, as well as its ability to mitigate the effects of temporary water limitation during flowering, all without negatively impacting wheat performance.

Deciphering the Enigma of Annual Recurrence of Wheat Rusts in India

ABSTRACTIn the absence of alternate hosts, the epidemiology of wheat rusts has remained elusive in many countries including India. This paper synthesises salient information from historical records collected at the Indian Institute of Wheat and Barley Research, from surveys of alternate and alternative hosts, and from observations in off‐season wheat disease monitoring nurseries. The primary source of rust inoculum to wheat in the plains of India was previously suggested to originate from off‐season wheat, self‐sown plants and green bridges in the hills. However, nowadays there is negligible cultivation of off‐season wheat in the hills, but still recurrence of wheat rust is a regular phenomenon in India. As observed previously, none of the Berberis spp. or grasses could be related to wheat rusts in this study. Thus, the wheat rust pathogens appear to multiply clonally in India. At present, new virulent pathotypes of Puccinia striiformis f. sp. tritici are identified initially from the plains of Punjab and other areas in the foothills of Himachal Pradesh, and after 6–7 years these pathotypes become predominant in the hills. Moreover, owing to the cultivation of traditional wheat germplasm, less virulent pathotypes of the stripe rust pathogen occur in the higher hills, and most of the present‐day wheat varieties are resistant to these pathotypes. The avirulence/virulence profiles of Indian pathotypes of Puccinia spp. on wheat indicate that our pathotype flora is different from that of adjoining countries and continents, which rules out their incursion from the adjoining countries. Wheat rust infection in the plains appears to be independent of the hills and movement of both leaf rust and stripe rust occurs both ways gradually year after year. It appears that the wheat rusts survive on grasses or other non‐gramineous hosts occurring locally near water bodies, forests, grasslands or damp areas in the hills and plains of India.

Pgmiox mediates stress response and plays a critical role for pathogenicity in Pyrenophora graminea, the agent of barley leaf stripe

Barley leaf stripe is an important disease caused by Pyenophora graminea that affects barley yields in the world. Ascorbic acid (AsA) interacts with key elements of a complex network orchestrating plant defense mechanisms, thereby influencing the outcome of plant-pathogen interaction. Myo-inositol oxygenase (MIOX) is a pivotal enzyme involved in plants development and environmental stimuli. However, MIOX has described functions in plants but has not been characterized in fungi. In this study, we characterized the Pgmiox gene in P. graminea pathogenesis through annotated on the metabolic pathway of ascorbic acid aldehyde. Our analysis suggested that the Pgmiox protein had a typical conserved MIOX domain. Multiple alignment analysis indicated that the P. graminea MIOX orthologue clustered with MIOX proteins of Pyrenophora species. RNA interference successfully reduced transcript abundance of Pgmiox in six transformant lines compared to wild type, and the transformants were further less virulent on the host plant barley. Transformants of Pgmiox had significant reductions in vegetative growth and pathogenicity, which had increased resistance to tebuconazole and carbendazim. In addition, Pgmiox is associated with ionic, drought, osmotic, oxidative, and heavy metal stress tolerance in P. graminea. In conclusion, our findings reveal that Pgmiox may be widely utilized by fungi to enhance pathogenesis and holds significant potential for the development of durable P. graminea resistance through genetic modifications.

Identification of Rust Resistance Genes in Wheat (Triticum aestivum L.) Using Molecular Markers and Host–Pathogen Interaction Tests

ABSTRACTThe leaf rust (Puccinia triticina f. sp. tritici), stripe rust (Puccinia striiformis f. sp. tritici), and stem rust (Puccinia graminis f. sp. tritici) are major fungal constraints affecting wheat production worldwide. Identifying and deploying wheat varieties with diverse resistance are the best ways to manage all the rusts. Therefore, a continuous search goes on to identify diverse germplasm with effective rust resistance that expresses at different stages of plant growth (seedling and adult plant). A set of 22 rust resistant wheat genotypes and 4 checks (controls), viz., Avocet‐Yr10, Avocet ‐Yr15, Agra Local, and respective positive checks were studied for characterising rust resistance genes using host–pathogen interactions complemented by molecular markers. Among 22 elite genotypes, 05 genotypes amplified 191 bp fragment with marker PSY1E1, confirmed the presence of gene Lr19/Sr25. These genotypes also expressed resistance to most virulent leaf rust pathotypes, 77‐5 and 77‐9 in host–pathogen interaction test (HPI). Seven genotypes showed the presence of Lr34/Yr18/Sr57/Pm38/Ltn1 in homozygous state, whereas G4 showed its presence in heterozygous condition. Among 22 genotypes, 16 genotypes possessed Yr10. Five genotypes (22.7%) exhibited two gene combinations, Lr19/Sr25, and Yr10 as revealed through the detection of 191 bp fragment with marker PSY1E1 and 260 bp fragment with co‐dominantly inherited microsatellite marker Xpsp3000, respectively. All five genotypes (G2, G3, G8, G9, and G18) also expressed brown glumes controlled by the gene Rg1 tightly linked to Yr10 on the 1BS chromosome. Broad spectrum rust resistance present in these lines in good agronomic backgrounds could be used as potent genetic donors for diverse and durable rust resistance breeding programmes in wheat.

Mapping of Aegilops speltoides derived leaf rust and stripe rust resistance genes using 35K SNP array

Wheat is an essential food commodity cultivated throughout the world. However, this crop faces continuous threats from fungal pathogens, leaf rust (LR) and stripe rust (YR). To continue feeding the growing population, these major destructors of wheat must be effectively countered by enhancing the genetic diversity of cultivated germplasm. In this study, an introgression line with hexaploid background (ILsp3603) carrying resistance against Pt pathotypes 77−5 (121R63-1), 77−9 (121R60-1) and Pst pathotypes 46S119 (46E159), 110S119 (110E159), 238S119 (238E159) was developed from donor wheat wild progenitor, Aegilops speltoides acc pau 3603. To understand the genetic basis of resistance and map these genes (named Lrsp3603 and Yrsp3603), inheritance studies were carried out in F6 and F7 mapping population, developed by crossing ILsp3603 with LR and YR susceptible cultivar WL711, which revealed a monogenic (single gene) inheritance pattern for each of these traits. Bulk segregant analysis combined with 35 K Axiom SNP array genotyping mapped both genes as separate entities on the short arm of chromosome 6B. A genetic linkage map, comprising five markers, 1 SNP, 1 PLUG and three gene based SSRs, covered a genetic distance of 12.65 cM. Lrsp3603 was flanked by markers Tag-SSR14 (located proximally at 2.42 cM) and SNP AX-94542331 (at 3.28 cM) while Yrsp3603 was mapped at one end closest to AX-94542331 at 6.62 cM distance. Functional annotation of Lrsp3603 target region (∼ 1 Mbp) revealed 10 gene IDs associated with disease resistance mechanisms including three encoding typical R gene domains.

Characterization and identification of Burkholderia gladioli causing stalk rot of sweet corn in Thailand

AbstractStalk rot disease is becoming an important disease in sweet corn crops because it causes significant yield losses of the crop. In July 2022, typical symptoms of stem rot were observed in fields of Mae Hia Agricultural Research, Demonstration and Training Center in Chiang Mai province of Thailand. The symptoms exhibited by affected crop included circular water‐soaked lesions, rot and necrotic tissue at the base of the stalk. Koch's postulates were fulfilled with reisolation of the inoculated strain from the inoculated tissues, and confirmation was obtained by phenotypic characters and 16S rRNA sequences. The isolated colony on nutrient agar was cream‐coloured, circular, with a convex elevation, entire margin and smooth surface; the optical character was translucent and exuded a yellowish pigment that diffused in the nutrient agar, gram‐negative, lacked arginine dihydrolase, occurrence of the oxidase and catalase reaction. The strains were able to utilize d‐galactose and glucose but were unable to utilize d‐maltose, d‐cellobiose, d‐fructose, d‐mannose, mannitol, lactose and sucrose. The strains showed positive results for gelatine liquefaction test, citrate utilization and casein hydrolysis, but starch hydrolysis and indole production were negative result. These characteristics are the same as those described previously for Burkholderia gladioli. Disease severity in sweet corn was 43.8%, with infections showing spots with dark margins and soft rot on mature leaves and flag leaves. Disease severity in waxy corn and maize were 21.9% and 15.6%, respectively, showing a few light or dark brown spots at the infection sites. Therefore, this represents the first report of leaf stripe and stalk rot disease in sweet corn in Thailand.

Characterization and fine mapping of a white stripe leaf mutant in rice

Characterization and fine mapping of a white stripe leaf mutant in rice

Comprehensive insights into sorghum (Sorghum bicolor) defense mechanisms unveiled: Plant growth-promoting rhizobacteria in combating Burkholderia-induced bacterial leaf stripe disease

Sorghum, a vital cereal crop with significant importance in traditional human health systems, faces severe challenges from biotic stresses. This study aimed to assess the morpho-physiological variations and plant growth promoting rhizobacteria (PGPR)-induced defense mechanisms in sorghum against Burkholderia sp. infection, a well-known cause of bacterial leaf stripe disease. Also, the alterations in the leaf protein band profile of sorghum exposed to Burkholderia infection were assessed via 2-D gel electrophoresis wherein the disappearance of protein bands in diseased leaf tissues was recorded. The selected PGPR, A. chroococcum Beijerinck 1901 (MCC 2351), B. megaterium (MCC 2336), and P. fluorescens (NAIMCC B-00,340), secreted hydrogen cyanide and ammonia, and produced lytic enzymes, cellulase and amylase. When tested in-planta, Burkholderia sp., applied as a foliar spray, more profoundly impacted the growth as compared to seed coat application, resulting in significant reductions in root dry biomass, whole plant length, and total chlorophyll content by 93.9, 73.4 and 53.8 %, respectively, compared to disease-free control. Visible symptoms of bacterial leaf stripe disease like brown discoloration and necrotic lesions were observed in infected sorghum plants. Furthermore, the level of phytochemicals, proline, and malondialdehyde and antioxidants, CAT and SOD, in foliage declined significantly by 46.4, 70.2, 20.7, and 16.9 %, respectively, following inoculation with Bacillus applied as a seed coat. Additionally, Burkholderia-infected plants exhibited cellular damage, mainly in leaf tissues, as revealed by SEM and CLSM. Interestingly, the PGPR inoculation substantially enhanced the measured growth parameters of sorghum while reducing the levels of phytochemicals and antioxidants. These findings suggest that A. chroococcum, B. megaterium, and P. fluorescens, possessing biocontrol activity, can be employed as a profitable, environment and pocket-friendly approach for managing plant diseases and promoting the production of sorghum under biotic stress, thereby sufficing the human food demands.

Exopolysaccharide Production and Precipitation Method as a Tool to Study Virulence Factors.

Acidovorax avenae subsp. avenae (Aaa) is the causal agent of red stripe in sugarcane, a disease characterized by two forms: leaf stripe and top rot. Despite the importance of this disease, little is known about Aaa virulence factors (VFs) and their function in the infection process. Among the different array of VFs exerted by phytopathogenic bacteria, exopolysaccharides (EPSs) often confer a survival advantage by protecting the cell against abiotic and biotic stresses, including host defensive factors. They are also main components of the extracellular matrix involved in cell-cell recognition, surface adhesion, and biofilm formation. EPS composition and properties have been well studied for some plant pathogenic bacteria; nevertheless, there is no knowledge about Aaa-EPS. In this work, we describe a simple and reliable method for EPS production, precipitation, and quantification based on cold precipitation after ethanol addition, which will allow to study EPS characteristics of different Aaa strains and to evaluate the association among EPS (e.g., amount, composition, viscosity) and Aaa pathogenicity.

QTL Mapping for Adult-Plant Resistance to Leaf Rust in Italian Wheat Cultivar Libellula.

Wheat leaf rust (Lr), which is caused by Puccinia triticina Eriks. (Pt), is one of the most important wheat diseases affecting wheat production globally. Using resistant wheat cultivars is the most economical and environmentally friendly way to control leaf rust. The Italian wheat cultivar Libellula has demonstrated good resistance to Lr in field studies. To identify the genetic basis of Lr resistance in 'Libellula', 248 F6 recombinant inbred lines from the cross 'Libellula'/'Huixianhong' was phenotyped for Lr severity in seven environments: the 2014/2015, 2016/2017, 2017/2018, and 2018/2019 cropping seasons at Baoding, Hebei Province, and the 2016/2017, 2017/2018, and 2018/2019 crop seasons at Zhoukou, Henan Province. Bulked segregant analysis and simple sequence repeat markers were then used to identify the quantitative trait loci (QTLs) for Lr adult-plant resistance in the population. Six QTLs were consequently detected and designated as QLr.hebau-1AL and QLr.hebau-1AS that were presumed to be new and QLr.hebau-1BL, QLr.hebau-3AL, QLr.hebau-4BL, and QLr.hebau-7DS that were identified at similar physical positions as previously reported QTLs. Based on chromosome positions and molecular marker tests, QLr.hebau-1BL and QLr.hebau-7DS share similar flanking markers with Lr46 and Lr34, respectively. Lr46 and Lr34 are race nonspecific adult plant resistance (APR) genes for leaf rust and stripe rust and powdery mildew. QLr.hebau-4BL showed multiple disease resistance to leaf rust, stripe rust, Fusarium head blight, and powdery mildew. The QTL identified in this study, as well as their closely linked markers, may potentially be used in marker-assisted selection in wheat breeding.

Comparative study of virulence factors, pathogenicity and genetic diversity of Acidovorax avenae subsp. avenae, the causal agent of red stripe disease in sugarcane

AbstractAcidovorax avenae subsp. avenae (Aaa) is the causal agent of red stripe in sugarcane, a disease characterized by two forms: leaf stripe and top rot. Despite the importance of this disease, little is known about Aaa infection mechanisms, and studies of virulence factors (VFs) have not been reported. Previous analysis of Aaa isolates obtained from sugarcane fields in Tucumán, Argentina, suggested that pathogenicity was not related to genetic groups determined by rep‐PCR analysis. Therefore, the aims of this work were to analyse VFs produced by Aaa strains from different genetic groups and to evaluate the association among VFs, between VFs and Aaa pathogenicity and between VFs and Aaa genetic diversity. Bacterial motility (sliding, swimming and twitching), extracellular enzyme activities (proteases, amylases and endoglucanases), H2O2‐mediated oxidative stress, cell adhesion and exopolysaccharide (EPS) production were assayed in 14 Aaa isolates from different genetic groups. Pearson's correlation analysis and principal component analysis (PCA) were performed. For each VF analysed, values were variable presenting quantitative traits among the strains. Significant correlations were observed among the VFs, though no clear trend was identified between them and the pathogenicity or genetic group. Results reinforce the idea that a single VF does not account for the infection outcome, but a more complex process occurs where the interplay among VFs must be considered. The results of this study are crucial because they provide insight into the infection mechanism of this bacterium.

Introgression of the bread wheat D genome encoded Lr34/Yr18/Sr57/Pm38/Ltn1 adult plant resistance gene into Triticum turgidum (durum wheat)

Key MessageLack of function of a D-genome adult plant resistance gene upon introgression into durum wheat.The wheat Lr34/Yr18/Sr57/Pm38/Ltn1 adult plant resistance gene (Lr34), located on chromosome arm 7DS, provides broad spectrum, partial, adult plant resistance to leaf rust, stripe rust, stem rust and powdery mildew. It has been used extensively in hexaploid bread wheat (AABBDD) and conferred durable resistance for many decades. These same diseases also occur on cultivated tetraploid durum wheat and emmer wheat but transfer of D genome sequences to those subspecies is restricted due to very limited intergenomic recombination. Herein we have introgressed the Lr34 gene into chromosome 7A of durum wheat. Durum chromosome substitution line Langdon 7D(7A) was crossed to Cappelli ph1c, a mutant derivative of durum cultivar Cappelli homozygous for a deletion of the chromosome pairing locus Ph1. Screening of BC1F2 plants and their progeny by KASP and PCR markers, 90 K SNP genotyping and cytology identified 7A chromosomes containing small chromosome 7D fragments encoding Lr34. However, in contrast to previous transgenesis experiments in durum wheat, resistance to wheat stripe rust was not observed in either Cappelli/Langdon 7D(7A) or Bansi durum plants carrying this Lr34 encoding segment due to low levels of Lr34 gene expression. Key message

Detection of some registered barley varieties reactions to barley leaf stripe disease

Cereals are a significant agricultural product group with the highest cultivation area and production in Turkey. Among these cereals, barley is an important cereal used in human and animal nutrition, and the most widelycultivated plant after wheat. There are biotic and abiotic factors which affect the yield and quality parameters of barley. Of the biotic factors, barley leaf stripe (agent: Pyrenophora graminea (anamorph: Drechslera graminea) is an important fungal disease. Infections can occur in diseased seeds and in the presence of suitable environmental conditions. Hence, the use of disease-free seeds and resistant cultivars (cvs.) against this pathogen are crucial. With this in mind, this study was carried out with a highly virulent isolate of Pyrenophora graminea, obtained from the Disease and Pest Resistance Unit culture collection in 57 registered barley cultivars in the greenhouses of the Central Research Institute of Field Crops (TARM) in 2021. The experiments were conducted in 3 replications in a randomized block design. Of 57 registered barley cultivars 62%, 15% and 36% of these barley cultivars expressed resistance, moderate resistance, and susceptible reactions, respectively. As a result of the analysis of variance performed on the results obtained, it was concluded that the difference between the mean values of 57 varieties was statistically significant at 1% level (P

First report of Klebsiella Leaf Streak on Sorghum Caused by Klebsiella variicola in Haryana, India.

Sorghum (Sorghum bicolor [L.] Moench) is one of the top ten cereal crops in the world and is grown for fodder and seed purposes. During the fall of 2019 to 2022, a disease causing small to long streaks on leaves was observed in sorghum fields of Hisar (29° 9' 6.6996'' N, 75° 43' 16.0428'' E), Rohtak (28° 53' 43.8540'' N, 76° 36' 23.8068'' E) and Mohindergarh (28° 16' 6.0492'' N, 76° 9' 3.3552'' E) regions of Haryana between July and October. The reddish brown streaks were observed in the interveinal spaces of upper and lower leaves. The disease incidence reached 20-30% of plants in affected fields. The diseased leaf tissues were disinfected with 70% alcohol and placed in a tube with sterile water. After 30 minutes, 100 µl of the suspension was inoculated onto nutrient agar medium, incubated at 28 ± 2°C for three days, and a pure culture was obtained by restreaking on nutrient agar (Janse, 2005). The rod-shaped gram-negative bacterium with round, cream to white colonies was positive for methyl red, citrate utilization, urease activity, and glucose, lactose, sorbitol, rhamnose and sucrose fermentation tests. The genomic DNA of the bacterial suspension was extracted and 16S rDNA was amplified using universal 27F/1492R primers (Marchesi et al. 1998), resulting in tentative identification as Klebsiella sp. It was further confirmed with PCR amplification of Klebsiella specific primers (F:5'-CGCGTACTATACGCCATGAACGTA-3'; R:5'-ACCGTTGATCACTTCGGTCAGG-3') for gyrA gene (Brisse and Verhoef 2001). Discrete PCR amplicons of 1,500 (16S rDNA) and 300 bp (gyrA) were observed in a 1% (w/v) agarose gel. Forward and reverse DNA sequencing of both amplicons of the Hisar isolate (VMKV101) was carried out using a BDT v3.1 Cycle sequencing kit and consensus sequences were generated by using the program SeqMan Ultra (DNASTAR Lasergene). Sequences of the PCR products were deposited in GenBank with accession numbers MZ569433 (16S rDNA) and OP390080 (gyrA). The 16S rDNA sequence was 97.32% similar to K. variicola strain 13450 (CP026013; 1,450/1,490 bp) and the gyrA sequence had 99.66% similarity to K. variicola strain FH-1 (CP054254; 297/298 bp). A 16S RNA-based phylogenetic tree done by MEGA11 (Tamura et al. 2021) using the Maximum Likelihood method showed that strain VMKV101 clustered with K. variicola type strain F2R9. The complete bacterial genome (GCA025629215), sequenced by the Ion GeneStudio S5 system using Ion 530 chips (Thermo Fisher Scientific), was 99.03% identical by average nucleotide identity (ANI) to the type genome (CP045783) of Klebsiella variicola, with 87.8% genome coverage. For pathogenicity testing, a bacterial suspension (10 ml, 1×107 colony forming units/ml) was injected into the whole whorl after mechanical injury on 15-20 days old seedlings of the susceptible genotype HC-171, then plants were incubated at 35 ± 2°C, >80% relative humidity. Control plants were injected with sterile distilled water. Initial symptoms were observed on leaves of inoculated plants after 5 to 7 days as narrow, small longitudinal reddish brown streaks. As the disease progressed, the streaks on the leaf blade increased in number and size maintaining the reddish brown color. These streaks had slightly wavy margins and were surrounded by bright yellow halos. After 15 to 20 days, the streaks were 0.5 to 2.0 mm wide and 1.0 to 5.0 cm long, occasionally up to 10.0 cm long on both side of the leaves. Over time, neighboring streaks coalesced to form large necrotic areas. All inoculated plants exhibited identical symptoms. No symptoms were observed on control leaves. The reisolated bacterium from diseased sorghum leaves showed exactly the same morphological, biochemical and 16S RNA and gyrA molecular characteristics. To our knowledge, this is the first report of K. variicola causing a leaf streak disease on sorghum. Klebsiella species primarily cause diseases in humans and animals, but K. variicola has been found to incite banana soft rot (Fan et al. 2015) and K. aerogenes to cause stem rot in pearl millet (Malik et al. 2022). Differences of prevalence, spread and control between K. variicola and two other bacteria (Xanthomonas vasicola pv. holcicola causing Bacterial leaf streak; Paraburkholderia andropogonis causing Bacterial leaf stripe) causing leaf streak diseases on sorghum need to be determined. The identification of Klebsiella leaf streak disease lays the groundwork for future investigations into epidemiology and management of K. variicola on sorghum.

Mining the Australian Grains Gene Bank for Rust Resistance in Barley.

Global barley production is threatened by plant pathogens, especially the rusts. In this study we used a targeted genotype-by-sequencing (GBS) assisted GWAS approach to identify rust resistance alleles in a collection of 287 genetically distinct diverse barley landraces and historical cultivars available in the Australian Grains Genebank (AGG) and originally sourced from Eastern Europe. The accessions were challenged with seven US-derived cereal rust pathogen races including Puccinia hordei (Ph-leaf rust) race 17VA12C, P. coronata var. hordei (Pch-crown rust) race 91NE9305 and five pathogenically diverse races of P. striiformis f. sp. hordei (Psh-stripe rust) (PSH-33, PSH-48, PSH-54, PSH-72 and PSH-100) and phenotyped quantitatively at the seedling stage. Novel resistance factors were identified on chromosomes 1H, 2H, 4H and 5H in response to Pch, whereas a race-specific QTL on 7HS was identified that was effective only to Psh isolates PSH-72 and PSH-100. A major effect QTL on chromosome 5HL conferred resistance to all Psh races including PSH-72, which is virulent on all 12 stripe rust differential tester lines. The same major effect QTL was also identified in response to leaf rust (17VA12C) suggesting this locus contains several pathogen specific rust resistance genes or the same gene is responsible for both leaf rust and stripe rust resistance. Twelve accessions were highly resistant to both leaf and stripe rust diseases and also carried the 5HL QTL. We subsequently surveyed the physical region at the 5HL locus for across the barley pan genome variation in the presence of known resistance gene candidates and identified a rich source of high confidence protein kinase and antifungal genes in the QTL region.

Identification and functional analysis of the HvWRKY1 gene associated with Qingke (Hordeum vulgare L. var. nudum Hook. f.) leaf stripe disease

Identification and functional analysis of the HvWRKY1 gene associated with Qingke (Hordeum vulgare L. var. nudum Hook. f.) leaf stripe disease