Effects of Different Hormone Treatments on Nonembryogenic and Embryogenic Callus Induction and Time-Term Enzyme Treatments on Number and Viability of Isolated Protoplasts in Saffron (Crocus Sativus L.)

Darvishi, Elahe; Zarghami, Reza; Mishani, C Abd; Omidi, Mansoor

doi:10.17660/actahortic.2007.739.35

Cited by 13 publications

(8 citation statements)

References 7 publications

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“…They observed the highest frequency of callus induction (44%) on MS medium supplemented with 2.4-D (2 mg L -1 ) and Kinetin (0.5 mg L -1 ), culture from apical meristem. Darvishi et al (2006) also reported that a media containing NAA and BAP induced nonembryogenic calli; in contrast media containing 2.4-D and BAP had the best effect on induction of embryogenic calli in saffron.…”

mentioning

confidence: 93%

In Vitro Indirect Somatic Embryogenesis and Secondary Metabolites Production in the Saffron: Emphasis on Ultrasound and Plant Growth Regulators

Firoozi¹,

Zare²,

Sofalian³

et al. 2019

Tarım Bilimleri Dergisi

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In the present study, the effects of ultrasound and plant growth regulators on in vitro callogenesis and secondary metabolites production in saffron calli were investigated. Accordingly, the saffron corms surface sterilized, sonicated and cultured on different concentrations of plant growth regulators (0.5, 1, 2 and 4 mg L-1 2,4-D or NAA in combination with 0.5 and 1 mg L-1 Kin or BAP). The percentage of callus induction, callus yield (fresh weight) and embryogenic callus formation were recorded and secondary metabolites of calli were measured by UV/VIS spectrophotometer three months after culture. The results indicated that sonication of the saffron corm explants significantly increased the in vitro callus induction and growth. So, the highest callus induction (100%) and yield (4.68 g) was achieved with sonicated explants cultured on MS medium supplemented with 2 mg L-1 1-naphthaleneacetic acid (NAA) and 0.5 mg L-1 kinetin (Kin). Somatic embryogenesis was significantly influenced by plant growth regulator regimes and the MS medium supplemented with 0.5 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) plus 0.5 mg L-1 6-benzylaminopurine (BAP) and 0.5 mg L-1 NAA plus 0.5 mg L-1 Kin exhibited the highest percentage (75 and 72, respectively) of somatic embryogenesis. Secondary metabolite content of the callus cells was significantly different among the plant growth regulator regimes and the highest production of picrocrocin and safranal were occurred on the medium containing 0.5 mg L-1 2,4-D plus 0.5 mg L-1 BAP and 1 mg L-1 NAA plus 1 mg L-1 BAP.

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mentioning

confidence: 93%

In Vitro Indirect Somatic Embryogenesis and Secondary Metabolites Production in the Saffron: Emphasis on Ultrasound and Plant Growth Regulators

Firoozi¹,

Zare²,

Sofalian³

et al. 2019

Tarım Bilimleri Dergisi

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“…The medium was solidified with 0.7% agar and cultures were incubated in a growth chamber under dark conditions at 25ºC for callus induction. Different growth regulator combinations were tested for callus induction (Table 2) as suggested by Sheibani et al (2007), Darvishi et al (2007), Majourhay et al (2007), Sharifi and Ebrahimzade (2010), Blazquez et al (2009).…”

Section: Methodsmentioning

confidence: 99%

Factors Affecting Callus Induction and Organogenesis in Saffron (Crocus sativus L.)

Vahedi

Kalantari

Salami

2014

Plant Tissue Cult. & Biotech.

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Saffron (Crocus sativus L.) belongs to Iridaceae and is known an important native commercial plants in Iran for its high value of saffron. The best growth regulator composition for callus production from corms and sahoot regeneration from callus were determined. Saffron corms harvested from previous crops are generally used for future cropping cycles. However, this practice causes major yield losses due to the heavy attack by different pathogens. Availability of healthy disease free planting materials is of great importance for successful cultivation of saffron. By this investigation the best composition of growth regulators for callus production from corms and shoot regeneration from callus were determined. Callus induction of Crocus sativus L. was investigated by using different combinations NAA, 2, 4-D and TDZ, BA and Kn. The highest frequency of callus induction was observed in medium containing 2 mg/l 2, 4-D + 1 mg/l BA followed by 1 mg/l 2, 4-D + 0.15 mg/l Kn. However, in case of growth parameters such as diameter and the area of calli the best result was obtained in the medium supplemented with 2 mg/l 2, 4-D + 1 mg/l BA. In some treatments, calli were transferred to organogenesis stage after two subcultures. For sprouting of shoots transferred to medium containing 1 mg/l BAP and 1 mg/l NAA. Statistical analyses indicated that the treatment containing 5 mg/l NAA and 5 mg/l TDZ proved to be the best growth regulator treatment for shoot regeneration from the saffron calli.

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“…Studies undertaken for over last four decades have confirmed effective regeneration of callus and intact plantlets from corm explants in media fortified with IAA and/or 2,4-D with the addition of cytokinins [27]. Some of the interesting observations include; induction of embryonic callus using 2,4-D+Kn [28], non-embryonic callus with NAA+BAP [29], multiple shoot generation using NAA+BAP [30] and development of salt tolerant saffron using 2,4-D+BAP [31]. Out of the different combinations of plant growth hormones used in this study, MS medium supplemented with TDZ 0.5 mg/L+IAA 1 mg/L+ activated charcoal 0.1 g/L at 15 ± 2 °C resulted into white, shining, filled somatic embryos in 8 weeks (Table 2), whereas the other concentrations of TDZ and IAA resulted into pale yellow, hard non-embryogenic structure.…”

Section: Cytokinins Auxins and Carbohydrate Source For Cormlet Produmentioning

confidence: 99%

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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Effects of Different Hormone Treatments on Nonembryogenic and Embryogenic Callus Induction and Time-Term Enzyme Treatments on Number and Viability of Isolated Protoplasts in Saffron (Crocus Sativus L.)

Cited by 13 publications

References 7 publications

In Vitro Indirect Somatic Embryogenesis and Secondary Metabolites Production in the Saffron: Emphasis on Ultrasound and Plant Growth Regulators

In Vitro Indirect Somatic Embryogenesis and Secondary Metabolites Production in the Saffron: Emphasis on Ultrasound and Plant Growth Regulators

Factors Affecting Callus Induction and Organogenesis in Saffron (Crocus sativus L.)

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

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