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<p>This study employed an in vitro selection technique to develop a ginger tetraploid line tolerant to rhizome rot caused by <i>Fusarium oxysporum</i> f. sp. <i>zingiberi</i> and <i>Pythium ultimum</i>. The ploidy level of axenic in vitro tetraploid shoots was confirmed using flow cytometry and cytological approaches and subjected to in vitro selection on selective medium containing different concentrations of both <i>F</i>. <i>oxysporum</i> (12.5–20%) and <i>P. ultimum</i> (7.5–11%) culture filtrates (FPCF). Fusarium culture filtrate (FCF) at 15% and Pythium culture filtrate (PCF) at 7.5% concentration were observed to be the highest dose at which only 6.67% of shoots survived the first cycle, dropping to 3.33% by the third cycle. The selected shoots were multiplied on multiplication medium, followed by hardening on a potting mixture comprising sand, soil, and FYM (1:1:1; v/v/v), resulting in 100% survival. In vivo evaluation of FPCF-selected tetraploid (FPST) plants separated them into three groups, with six plants falling into group 0 signifying the ‘highly tolerant’ category, and 7 and 2 falling into groups 1 &amp; 2 signifying ‘tolerant’ and ‘moderately tolerant’ status. At the morphological level, the selected plants demonstrated superiority over the tetraploid control (TC) and diploid counterparts in parameters related to shoot, leaf, and rhizome characteristics. Biochemical analysis revealed higher carbohydrate, protein, oleoresin, oil, and 6-gingerol content in selected plants, but with less crude fibre than diploid plants. Out of ten R gene-specific SSR markers, nine showed amplification, generating 87 amplicons with 4 polymorphic and 8 monomorphic bands ranging in size between 150 and 500 base pairs. An additional unique band was observed in FPST plants upon amplification with GIN1, which, upon sequencing, showed 97.16% homology with the <i>Z</i>. <i>officinale</i> retrotransposon putative reverse transcriptase gene associated with disease tolerance in ginger. The sequence, with accession number PV135018, has been published in NCBI GenBank.</p>

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In vitro selection and evaluation of colchicine-induced ginger (Zingiber officinale Rosc.) tetraploids against rhizome rot tolerance

  • 期刊论文

  • 发表日期：

    2025年9月18日

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  • Lokesh Thakur,
  • Manisha Thakur,
  • Keshav Kumar,
  • Meenu Gupta,
  • Salej Sood

摘要

This study employed an in vitro selection technique to develop a ginger tetraploid line tolerant to rhizome rot caused by Fusarium oxysporum f. sp. zingiberi and Pythium ultimum. The ploidy level of axenic in vitro tetraploid shoots was confirmed using flow cytometry and cytological approaches and subjected to in vitro selection on selective medium containing different concentrations of both F. oxysporum (12.5–20%) and P. ultimum (7.5–11%) culture filtrates (FPCF). Fusarium culture filtrate (FCF) at 15% and Pythium culture filtrate (PCF) at 7.5% concentration were observed to be the highest dose at which only 6.67% of shoots survived the first cycle, dropping to 3.33% by the third cycle. The selected shoots were multiplied on multiplication medium, followed by hardening on a potting mixture comprising sand, soil, and FYM (1:1:1; v/v/v), resulting in 100% survival. In vivo evaluation of FPCF-selected tetraploid (FPST) plants separated them into three groups, with six plants falling into group 0 signifying the ‘highly tolerant’ category, and 7 and 2 falling into groups 1 & 2 signifying ‘tolerant’ and ‘moderately tolerant’ status. At the morphological level, the selected plants demonstrated superiority over the tetraploid control (TC) and diploid counterparts in parameters related to shoot, leaf, and rhizome characteristics. Biochemical analysis revealed higher carbohydrate, protein, oleoresin, oil, and 6-gingerol content in selected plants, but with less crude fibre than diploid plants. Out of ten R gene-specific SSR markers, nine showed amplification, generating 87 amplicons with 4 polymorphic and 8 monomorphic bands ranging in size between 150 and 500 base pairs. An additional unique band was observed in FPST plants upon amplification with GIN1, which, upon sequencing, showed 97.16% homology with the Z. officinale retrotransposon putative reverse transcriptase gene associated with disease tolerance in ginger. The sequence, with accession number PV135018, has been published in NCBI GenBank.