In vitro selection of gamma irradiated shoots of ginger ( Zingiber officinale Rosc.) against Fusarium oxysporum f.sp. zi
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ORIGINAL ARTICLE
In vitro selection of gamma irradiated shoots of ginger (Zingiber officinale Rosc.) against Fusarium oxysporum f.sp. zingiberi and molecular analysis of the resistant plants Vishal Sharma1 · Manisha Thakur1 · Manica Tomar2 Received: 30 December 2019 / Accepted: 19 August 2020 © Springer Nature B.V. 2020
Abstract In the present investigation, an attempt was made to develop Fusarium yellows resistant plants of ginger (Zingiber officinale Rosc.) var. Himgiri using in vitro mutagenesis and selection technique. Axenic in vitro cultures were subjected to gamma irradiation (10–100 Gy) for mutation induction. L D50 was calculated after every 4 weeks, and was observed to be 15 Gy after 16 weeks of irradiation. Surviving 10 Gy irradiated shoots were cultured on selective medium containing different concentrations (0–20%) of fungal culture filtrate (FCF) obtained from Fusarium oxysporum f.sp. zingiberi for in vitro selection. FCF concentration of 17.5% in the selective medium was found to be the highest on which 5% shoots survived after first selection cycle, which further reduced to 1.1% after third continuous cycle of selection. It was followed by 4.3% shoot survival after third selection cycle on 15% FCF concentration. Surviving, selected shoots from 15 and 17.5% FCF were multiplied on multiplication medium and rooted plantlets were hardened with 100% survival. On in vivo evaluation 46.4 and 52% plants selected at 15 and 17.5% FCF were found to be highly resistant. Molecular analysis with disease specific SSR primers differentiated between FCF selected, tissue culture propagated and gamma irradiated plants. Two unique bands were obtained in 15 and 17.5% FCF selected plantlets with GIN 6 and GIN 9 primers, sequencing, of which showed 98 and 97% homology with disease resistance protein-like gene CC-NBS-LRR from clones ZwP627 and ZoP620 of Z. officinale. After in vivo bioassay, SSR analysis of selected highly resistant plants again confirmed the unique bands with both the primers. The DNA sequences obtained from these primers have been published in GenBank under accession number MN497252 and MN497253. Key message A combination of in vitro mutagenesis using gamma irradiations followed by selection mediated by fungal culture filtrate produced ginger plants resistant to Fusarium yellows disease. Keywords Fungal culture filtrate · Gamma rays · Ginger · In vitro selection · Mutations · Resistance
Introduction Communicated by Amita Bhattacharya. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11240-020-01919-x) contains supplementary material, which is available to authorized users. * Manisha Thakur [email protected] 1
Department of Biotechnology, Dr YS Parmar University of Horticulture and Forestry, Nauni‑Solan, HP 173230, India
Department of Plant Pathology, Dr YS Parmar University of Horticulture and Forestry, Nauni‑Solan, HP 173230, India
2
Ginger (Zingiber officinale Rosc.) a monocotyledonous perennial herb belongs to family Zingiberacea
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