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136 In vitro Production of Indian citrus ringspot virus-free Plants of Kinnow Mandarin (Citrus nobilis LourxC. deliciosaTenora) by Nucellar Embryo Culture

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Title 136
In vitro Production of Indian citrus ringspot virus-free Plants of Kinnow Mandarin (Citrus nobilis LourxC. deliciosaTenora) by Nucellar Embryo Culture
 
Creator Singh , Bikram
Sharma , S
Rani, G
Zaidi, A A
Hallan, Vipin
Nagpal, A
Virk , G S
 
Subject Plant viruses
 
Description Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lourx C. deliciosa Tenora) were produced from a virus-infected plant using nucellar embryo culture. The parent kinnow plant was tested by indirect ELISA and RT-PCR before using its explants. An amplified product of 539 bp (partial cp gene) was obtained by RT-PCR in ICRSV infected plants. The nucellar embryos obtained from seeds collected from immature fruits of the infected plant were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of 2,4-D and malt extract (ME) alone or in various combinations. Maximum embryogenic callus induction (33.33%) was observed on MS medium supplemented with 2,4-D (9.02 μM) in combination with malt extract (400 mg L-1). Transfer of embryogenic calli to MS medium containing different concentrations of malt extract alone or in combination with ABA resulted in somatic embryogenesis with a maximum of 56.94% cultures in MS medium supplemented with malt extract (500 mg L-1) and ABA (7.56 μM). Cotyledonary shaped embryos when transferred to different strengths of MS medium supplemented with NAA (10.74 μM) developed into complete plantlets in maximum of 72.22% cultures on ½ MS medium. The plantlets were successfully acclimatizated, transferred to screen house and indexed for ICRSV employing indirect ELISA and RT-PCR and all were found negative of virus. A distinct feature of this study is the induction of somatic embryogenesis from nucellar embryos to produce virus-free plants.
 
Date 2006
 
Type Article
PeerReviewed
 
Identifier Singh , Bikram and Sharma , S and Rani, G and Zaidi, A A and Hallan, Vipin and Nagpal, A and Virk , G S (2006) 136 In vitro Production of Indian citrus ringspot virus-free Plants of Kinnow Mandarin (Citrus nobilis LourxC. deliciosaTenora) by Nucellar Embryo Culture. Plant Pathology Journal, 5. pp. 274-282.
 
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