<i>In vitro</i>regeneration and<i>Agrobacterium</i>mediated transformation in gladiolus

Babu, Punuri Jayasekhar; Chawla, H. S.

doi:10.1080/14620316.2000.11511258

Cited by 16 publications

(8 citation statements)

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“…Therefore, pre-determination of this period helps overcome less frequency of transformation or overgrowth of Agrobacterium. Previous studies of geophytes have established a co-cultivation period of 3 days as best for Agapanthus species [39] whereas a 3-day period resulted in highest GUS expression levels in Typha latifolia [40]. Interestingly, our study demonstrated 2-day period of co-cultivation as optimum for Cas9 expression in saffron.…”

Section: Agrobacterium Tumefaciens Mediated Genetic Transformation Insupporting

confidence: 47%

Development of a system for efficient callus production, somatic embryogenesis and gene editing using CRISPR/Cas9 in Saffron (Crocus sativus L.)

et al. 2020

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Background: Crocus sativus is a recalcitrant plant for genetic transformation and genetic improvement, largely due to difficulties in Agrobacterium mediated transformation and vegetative reproduction. Effective genome editing requires proficient callus production and an efficient method to deliver Cas9 and sgRNAs into the plant. Here, we demonstrate Agrobacterium-mediated transformation of saffron. Further, we developed a CRISPR-Cas9 based system in this plant, for efficient gene knockout or edits in future. Results: Efficient callus production and regeneration confers important benefits in developing competent transformation system in plants. More than 70% multiplication rate of callus initiation was achieved from corm slices of saffron subjected to a two-step sterilization procedure and grown on complete MS medium supplemented with 2,4-D (0.5 mg/L), BAP (1 mg/L), IAA (1 mg/L), photoperiod of 16/8 h and 45% relative humidity at 20 ± 2 °C. In vitro cormlet generation was accomplished in 8 weeks by using mature somatic embryos on MS medium supplemented with TDZ (0.5 mg/L) + IAA (1 mg/L) + Activated charcoal (0.1 g/L) at 15 ± 2 °C. The attempt of using Agrobacterium-mediated transformation resulted in successful integration of the binary vector into the somatic embryos of saffron with a transformation efficiency of 4%. PCR and Southern blot analysis confirmed the integration of Cas9 into saffron. Conclusion: The protocol for callus production, somatic embryogenesis and regeneration was standardised. Successful demonstration of integrated Cas9 in this study constitutes first step in developing strategies for genetic manipulation of saffron, which has so far been considered recalcitrant. Furthering the development of this technology holds significant potential for advancing genetic research in saffron by integrating multigene targeting and/or use of recyclable cassettes.

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Section: Agrobacterium Tumefaciens Mediated Genetic Transformation Insupporting

confidence: 47%

Development of a system for efficient callus production, somatic embryogenesis and gene editing using CRISPR/Cas9 in Saffron (Crocus sativus L.)

et al. 2020

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“…Agrobacterium tumefaciens-mediated transformation of in vitro plants, cormels and seedling leaves lead to tumour formation on 12% of explants, with only 1% of these tumours capable of opine synthesis (Kamo 1997a). Wounding explants prior to co-cultivation improved transformation efficiency (Babu and Chawla 2000). Analysis of four different promoters (mannopine synthase 2, translation elongation factor 1 subunit a, rolD and the cauliflower mosaic virus 35S) within transformed Gladiolus plants revealed that rolD and cauliflower mosaic virus 35S promoters had better patterns (tissue specificity) and levels of expression over the entire plant compared to other promoters (Kamo and Blowers 1999).…”

Section: Transformationmentioning

confidence: 98%

Micropropagation of iridaceae—a review

Ascough

Erwin

Staden

2009

Plant Cell Tiss Organ Cult

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The Iridaceae contains many hundreds of attractive species that have been overlooked for horticultural development. The incredible variation in flower and leaf shape, size and colour suggest this little-tapped resource could offer great opportunities for developing new commercial ornamental products. Micropropagation has increasingly become a valuable tool assisting breeders to release new species and cultivars into the market more rapidly. Here we review the progress made in Iridaceae micropropagation genus by genus, and highlight the potential for future expansion in this field.

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“…Better shoot induction (upto 89%) was observed with top slice of cormel (dia. 1.0 to 1.5cm) in response to MS medium containing 4 mg/l KIN rather than BAP (Babu and Chawla, 2000).…”

Section: Shoot Organogenesismentioning

confidence: 98%