Articles
Flowcytomeric analysis shows stable genome size in tissue culture raised plants of saffron (Crocus sativus L.)
Article number
1200_5
Pages
31 – 40
Language
English
Abstract
Saffron is conventionally propagated through corms and multiplication rate is low.
Corms are infested with fungal pathogens.
Therefore, in vitro propagation is an important alternate for fast multiplication of disease free true to type planting material and stability of genome size is an important aspect.
In the present study, directly regenerated shoots, somatic embryo derived plants and mother plants i.e., plants produced under field conditions were assessed by flow cytometery for genome size stability.
Estimation of Nuclear DNA content with emphasis on nuclei preparation and testing of inhibitors for PI binding was carried out. Secale cereale was used as internal reference standard.
Gailbraith buffer was found best in terms of more count of intact nuclei, less debris percent, low coefficient of variation (CV) and reduced chi square (RCS) values among the tested buffers. 3C nuclear DNA of somatic embryo derived and directly formed shoots was 9.74±0.02 and 9.73±0.04 pg and 1C genome size was 4.81±0.01×109 bp and 4.80±.0.01×109 bp, respectively. 3C nuclear DNA and IC genome size of mother plant was 9.77±0.02 and 4.82±0.01×109 bp, respectively indicating thereby that genome size of tissue culture raised plants remained stable.
Corms are infested with fungal pathogens.
Therefore, in vitro propagation is an important alternate for fast multiplication of disease free true to type planting material and stability of genome size is an important aspect.
In the present study, directly regenerated shoots, somatic embryo derived plants and mother plants i.e., plants produced under field conditions were assessed by flow cytometery for genome size stability.
Estimation of Nuclear DNA content with emphasis on nuclei preparation and testing of inhibitors for PI binding was carried out. Secale cereale was used as internal reference standard.
Gailbraith buffer was found best in terms of more count of intact nuclei, less debris percent, low coefficient of variation (CV) and reduced chi square (RCS) values among the tested buffers. 3C nuclear DNA of somatic embryo derived and directly formed shoots was 9.74±0.02 and 9.73±0.04 pg and 1C genome size was 4.81±0.01×109 bp and 4.80±.0.01×109 bp, respectively. 3C nuclear DNA and IC genome size of mother plant was 9.77±0.02 and 4.82±0.01×109 bp, respectively indicating thereby that genome size of tissue culture raised plants remained stable.
Authors
K. Devi, M. Sharma, S. Chanda, P.S. Ahuja
Keywords
internal reference, Secale cereale, 3C DNA, buffer standardization, propidium iodide, somatic embryo
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