Development of activation-tagged gain-of-functional mutants in indica rice line (BPT 5204) for sheath blight resistance

Mahendranath Gandikota; T Krishnakanth Yadav; Raghurami Reddy Maram; Sudhamani Kalluru; M Balachandran Sena; E A Siddiq; Yamini Kalinati Narasimhan; Lakshminarayana R Vemireddy; Anuradha Ghanta

doi:10.1007/s11033-023-09194-7

Development of activation-tagged gain-of-functional mutants in indica rice line (BPT 5204) for sheath blight resistance

Mol Biol Rep. 2024 Mar 2;51(1):381. doi: 10.1007/s11033-023-09194-7.

Authors

Mahendranath Gandikota^{1

2}, T Krishnakanth Yadav¹, Raghurami Reddy Maram², Sudhamani Kalluru³, M Balachandran Sena², E A Siddiq¹, Yamini Kalinati Narasimhan¹, Lakshminarayana R Vemireddy⁴, Anuradha Ghanta⁵

Affiliations

¹ Institute of Biotechnology, Professor Jayashankar Telangana State Agricultural University (PJTSAU), Rajendranagar, Hyderabad, 500030, India.
² ICAR-Indian Institute of Rice Research, Rajendranagar, Hyderabad, 500030, India.
³ Department of Genetics and Plant Breeding, S.V. Agricultural College, Acharya N.G. Ranaga Agricultural University (ANGRAU), Tirupati, 517502, India.
⁴ Department of Molecular Biology and Biotechnology, S.V. Agricultural College, Acharya N.G. Ranaga Agricultural University (ANGRAU), Tirupati, 517502, India. [email protected].
⁵ Institute of Biotechnology, Professor Jayashankar Telangana State Agricultural University (PJTSAU), Rajendranagar, Hyderabad, 500030, India. [email protected].

PMID: 38430361
DOI: 10.1007/s11033-023-09194-7

Abstract

Background: The development of sheath blight (ShB) resistance varieties has been a challenge for scientists for long time in rice. Activation tagging is an efficient gain-of-function mutation approach to create novel phenotypes and to identify their underlying genes. In this study, a mutant population was developed employing activation tagging in the recalcitrant indica rice (Oryza sativa L.) cv. BPT 5204 (Samba Mahsuri) through activation tagging.

Methods and results: In this study, we have generated more than 1000 activation tagged lines in indica rice, from these mutant population 38 (GFP^- RFP⁺) stable Ds plants were generated through germinal transposition at T₂ generation based on molecular analysis and seeds selected on hygromycin (50 mg/L) containing medium segregation analyses confirmed that the transgene inherited as mendelian segregation ratio of 3:1 (3 resistant: 1 susceptible). Of them, five stable activation tagged Ds lines (M-Ds-1, M-Ds-2, M-Ds-3, M-Ds-4 and M-Ds-5) were selected based on phenotypic observation through screening for sheath blight (ShB) resistance caused by fungal pathogen Rhizoctonia solani (R. solani),. Among them, M-Ds-3 and M-Ds-5 lines showed significant resistance for ShB over other tagged lines and wild type (WT) plants. Furthermore, analysed for launch pad insertion through TAIL-PCR results and mapped on corresponding rice chromosomes. Flanking sequence and gene expression analysis revealed that the upregulation of glycoside hydrolase-OsGH or similar to Class III chitinase homologue (LOC_Os08g40680) in M-Ds-3 and a hypothetical protein gene (LOC_Os01g55000) in M-Ds-5 are potential candidate genes for sheath blight resistance in rice.

Conclusion: In the present study, we developed Ac-Ds based ShB resistance gain-of-functional mutants through activation tagging in rice. These activation tagged mutant lines can be excellent sources for the development of ShB resistant cultivars in rice.

Keywords: Ac-Ds transposable elements; Agrobacterium mediated transformation; Activation tagging; BPT 5204 (Samba Mahsuri); Glycoside hydrolase; Rice; Sheath blight.

MeSH terms

Gene Expression Profiling
Oryza* / genetics
Oryza* / microbiology

Grants and funding

BT/PR-13105/AGR/02/684/2009/Department of Biotechnology, Government of India