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

HomePlant DiseaseVol. 107, No. 7First Report of Klebsiella Leaf Streak on Sorghum Caused by Klebsiella variicola in Haryana, India PreviousNext DISEASE NOTE OPENOpen Access licenseFirst Report of Klebsiella Leaf Streak on Sorghum Caused by Klebsiella variicola in Haryana, IndiaVinod Kumar Malik, Manjeet Singh, Pooja Sangwan, Pummy Kumari, Bajrang Lal Sharma, Pavitra Kumari, Preety Verma, Pankaj Yadav, Niharika Sheoran, Ajit Singh, Dalvinder Pal Singh, Satyawan Arya, Navjeet Ahalawat, and Janvi MalikVinod Kumar Malik†Corresponding author: V. K. Malik; E-mail Address: [email protected] and E-mail Address: [email protected]https://orcid.org/0000-0003-1178-5790Department of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Manjeet SinghDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Pooja SangwanDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Pummy KumariForage Section, Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Bajrang Lal SharmaForage Section, Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Pavitra KumariDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Preety VermaDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Pankaj YadavDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Niharika SheoranDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Ajit SinghDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Dalvinder Pal SinghForage Section, Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Satyawan AryaForage Section, Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, Navjeet AhalawatDepartment of Bioinformatics and Computational Biology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this author, and Janvi MalikDepartment of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, IndiaSearch for more papers by this authorAffiliationsAuthors and Affiliations Vinod Kumar Malik1 † Manjeet Singh1 Pooja Sangwan1 Pummy Kumari2 Bajrang Lal Sharma2 Pavitra Kumari1 Preety Verma1 Pankaj Yadav1 Niharika Sheoran1 Ajit Singh1 Dalvinder Pal Singh2 Satyawan Arya2 Navjeet Ahalawat3 Janvi Malik1 1Department of Plant Pathology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, India 2Forage Section, Department of Genetics and Plant Breeding, Chaudhary Charan Singh Haryana Agricultural University, Hisar, India 3Department of Bioinformatics and Computational Biology, Chaudhary Charan Singh Haryana Agricultural University, Hisar, India Published Online:9 Jul 2023https://doi.org/10.1094/PDIS-09-22-2200-PDNAboutSectionsView articlePDFSupplemental ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinked InRedditEmailWechat View articleSorghum (Sorghum bicolor [L.] Moench), one of the top 10 cereal crops in the world, is grown for fodder and seed. In 2019 to 2022, a disease causing short to long leaf streaks was seen in sorghum fields in Hisar (29° 9′ 6.70″N, 75° 43′ 16.04″E), Rohtak (28° 53′ 43.85″N, 76° 36′ 23.81″E), and Mohindergarh (28° 16′ 6.05″N, 76° 9′ 3.36″E), Haryana, between July and October. Reddish-brown streaks were in the interveinal spaces of upper and lower leaves. Disease incidence reached 20 to 30% of plants in affected fields. Diseased leaf tissues were disinfected with 70% alcohol and put in a tube of sterile water. After 30 min, 100 μl of the suspension was inoculated onto nutrient agar (NA) and incubated at 28 ± 2°C for 3 days, and a pure culture was obtained by restreaking on NA (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. 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 confirmed with PCR amplification of Klebsiella specific primers (F: 5ʹ-CGCGTACTATACGCCATGAACGTA-3ʹ; R: 5ʹ-ACCGTTGATCACTTCGGTCAGG-3ʹ) for gyrA gene (Brisse and Verhoef 2001). Discrete 1,500 (16S rDNA) and 300 bp (gyrA) PCR amplicons were observed in a 1% (w/v) agarose gel. Forward and reverse DNA sequencing of both amplicons of the Hisar isolate (VMKV101) was done with a BDT v3.1 Cycle sequencing kit; consensus sequences were generated using SeqMan Ultra (DNASTAR Lasergene). PCR product sequences were deposited in GenBank (MZ569433, 16S rDNA; 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 was 99.66% similar to K. variicola strain FH-1 (CP054254; 297/298 bp). A 16S RNA-based maximum-likelihood phylogenetic tree in MEGA11 (Tamura et al. 2021) found that VMKV101 clustered with K. variicola type strain F2R9. The complete genome (GCA025629215), sequenced by the Ion GeneStudio S5 system using Ion 530 chips (Thermo Fisher Scientific), was 99.03% identical by average nucleotide identity to K. variicola type genome CP045783, with 87.8% genome coverage. To test pathogenicity, a bacterial suspension (10 ml, 1 × 107 CFU/ml) was injected into whole whorls after mechanical injury on 15- to 20-day-old plants of susceptible genotype HC-171, then plants were incubated at 35 ± 2°C and >80% relative humidity. Control plants were injected with sterile distilled water. Initial symptoms on inoculated leaves after 5 to 7 days were narrow, small, longitudinal, reddish-brown streaks. As the disease progressed, streaks increased in number and size, staying reddish brown. The streaks had slightly wavy margins and 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 sides of the leaves. Neighboring streaks coalesced to form large necrotic areas. All inoculated plants had identical symptoms. Controls were symptomless. The reisolated bacterium from diseased leaves had 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 spp. primarily cause diseases in humans and animals, but K. variicola has been found to incite banana soft rot (Fan et al. 2016) 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. This study will inform studies of epidemiology and management of K. variicola on sorghum.The author(s) declare no conflict of interest.References:Brisse, S., and Verhoef, J. 2001. Int. J. Syst. Evol. Microbiol. 51:915. https://doi.org/10.1099/00207713-51-3-915 Crossref, ISI, Google ScholarFan, H. C., et al. 2016. Plant Dis. 100:517. https://doi.org/10.1094/pdis-05-15-0586-pdn Link, ISI, Google ScholarJanse, J. D. 2005. Page 366 in: Phytobacteriology: Principles and Practices. CABI Publishing, Wallingford, U.K. Crossref, Google ScholarMalik, V. K., et al. 2022. Plant Dis. 106:754. https://doi.org/10.1094/pdis-05-21-1019-pdn Link, ISI, Google ScholarMarchesi, J. R., et al. 1998. Appl. Environ. Microbiol. 64:795. https://doi.org/10.1128/AEM.64.2.795-799.1998 Crossref, ISI, Google ScholarTamura, K., et al. 2021. Mol. Biol. Evol. 38:3022. https://doi.org/10.1093/molbev/msab120 Crossref, ISI, Google ScholarThe author(s) declare no conflict of interest.DetailsFiguresLiterature CitedRelated Vol. 107, No. 7 July 2023SubscribeISSN:0191-2917e-ISSN:1943-7692 Download Metrics Article History Issue Date: 26 Jul 2023Published: 9 Jul 2023First Look: 2 Jan 2023Accepted: 30 Dec 2022 Page: 2215 Information© 2023 The American Phytopathological SocietyKeywordsbacteriumdiseaseKlebsiellaleaf streakSorghumThe author(s) declare no conflict of interest.PDF download