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Production of plants resistant to Alternaria carthami via organogenesis and somatic embryogenesis of safflower cv. NARI-6 treated with fungal culture filtrates

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Abstract

The present study describes a system for efficient plant regeneration via organogenesis and somatic embryogenesis of safflower (Carthamus tinctorius L.) cv. NARI-6 in fungal culture filtrates (FCF)-treated cultures. FCF was prepared by culturing Alternaria carthami fungal mycelia in selection medium for host-specific toxin production. Cotyledon explants cultured on callus induction medium with different levels of FCF (10–50%) produced embryogenic callus. In organogenesis, 42.2% microshoots formed directly from embryogenic callus tissues in plant regeneration medium with 40% FCF. Isolated embryogenic callus cultured on embryo induction medium containing 40% FCF induced 50.2% somatic embryogenesis. Embryo germination percentage was decreased from 64.5 to 28 in embryo maturation medium containing 40% FCF. However, nine plantlets from organogenesis and 24 plantlets from somatic embryogenesis were selected as FCF-tolerant. Alternaria carthami fungal spores (5 × 105 spores/ml) sprayed on the leaves of FCF-tolerant plants showed enhanced survival rate over control plants, which plants were more susceptible to fungal attack. The number of leaf spot lesions per leaf was decreased from 3.4 to 0.9 and their lesion length was also reduced from 2.9 to 0.7 mm in organogenic derived FCF-tolerant plants over control. In somatic embryo derived FCF-tolerant plants, the number of lesions was decreased from 3.1 to 0.4 and the lesion size was also reduced to 2.7–0.5 mm when compared to the control. This study also examined antioxidant enzyme activity in FCF-tolerant plants. Catalase (CAT) activity was slightly decreased whereas peroxidase (POD) activity was increased to a maximum of 42% (0.19 μmol min−1 mg−1 protein) from organogenesis and 47% (0.23 μmol min−1 mg−1 protein) from embryogenesis in FCF-tolerant plants. Superoxide dismutase (SOD) activity was also increased to 17% (149 U mg−1 protein) and 19.5% (145 U mg−1 protein) in FCF-tolerant plants derived from organogenesis and somatic embryogenesis when compared with control plants.

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Abbreviations

PIC:

Picloram

TDZ:

Thidiazuron, N-phenyl-N′-1,2,3-thiadiazol-5-yl-urea

GA3 :

Gibberellic acid

NAA:

α-naphthaleneacetic acid

AgNO3 :

Silver nitrate

PDA:

Potato dextrose agar

PGRs:

Plant growth regulators

T.HCl:

Thiamine hydrochloride

FCF:

Fungal culture filtrate

MSG:

Murashige and Skoog germination medium (Becwar et al. 1990)

CAT:

Catalase

POD:

Peroxidase

SOD:

Superoxide dismutase

NARI:

Nimbkar Agricultural Research Institute

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Acknowledgements

We are grateful to Prasad RD, Directorate of Oilseeds Research, Rajendranagar, Hyderabad, India for providing Alternaria carthami culture. We thank Nimbhkar Agricultural Research Institute (NARI), Maharashtra, India for providing seeds for this study.

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Correspondence to B. D. Ranjitha Kumari.

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Vijaya Kumar, J., Ranjitha Kumari, B.D., Sujatha, G. et al. Production of plants resistant to Alternaria carthami via organogenesis and somatic embryogenesis of safflower cv. NARI-6 treated with fungal culture filtrates. Plant Cell Tiss Organ Cult 93, 85–96 (2008). https://doi.org/10.1007/s11240-008-9346-4

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Keywords

  • Carthamustinctorius L.
  • Medicinal plant
  • Embryogenic calli
  • Organogenesis
  • Somatic embryogenesis
  • Plant regeneration
  • Alternaria leaf spot
  • Disease resistance
  • Antioxidant enzymes