The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.

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CHAMANI E.; EFTEKHARI Z.; GHANBARI A.R.; HEYDARI H.R.; ARSHAD M.

Title

The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.

Author(s)

Keywords

BulbletQ2

CallusQ1

Essential oilQ1

In Vitro cultureQ1

Plant growth regulatorQ2

Abstract

Introduction: Fritillaria imperialis L. is an ornamental and medicinal plant native to mountainous regions of Iran. This plant genetic resource is in danger of extinction, Because of grazing livestock and pest outbreaks. Therefore, micro propagation of Fritillaria through in vitro regeneration is essential for conservation and commercial production. Thymol and Carvacrol are one of the main Essential oil compounds in family Lamiaceae. Material and Methods: Fritillaria imperialis L. bulbs in dormancy stage obtained from mountainous regions of Lorestan in Iran and were placed in cold room at +4 ° C for 4-6 weeks. Then, Bulbs were surfacesterilized with 70% ethanol for 45 s followed by immersion in 5% (v/v) NaOCl solution for 20 min with gentle agitation, and then rinsed three times in sterile double distilled water. Present study was conducted in two separate experiments. In first experiment, effect of different concentration of Thymol and Carvacrol and in second experiment, different concentration of NAA and BA on in vitro characteristics of Fritillaria was evaluated. Explants (1× 1 cm) prepared from the lower third of scales with basal plate and were placed in MS basal medium supplemented with different concentrations of Thymol (50, 100, 150 and 300 ppm), Carvacrol (10, 100, 500 and 100 ppm), BA (1, 2 and 4 mg/l) and NAA (1, 2 and 4 mg/l). All cultures were incubated in a growth chamber at 24± 2° C, and a photosynthetic photon flux of 40-60 μ mol m– 2 s– 1 was provided by cool white fluorescent lamps with a 16-h photoperiod. This experiment wascarried out in completely randomized designs with fivereplications. Results and Discussion: Analysis of variance showed that Thymol and Carvacrol were not effective on number of new Bulblets but had significant effects on bulb diameter, number and length or roots, number and length leaves and Callus induction and diameter of Callus obtained from scales (P< 0. 05). The highest rate (3 Bulblets) of Bulblets formation was obtained fromMS medium supplemented with 50 ppm Thymol that showed significantly difference from other treatments. Medium containing 10 ppm Carvacrol gave the highest Bulblet formation (2. 5 Bulblets) between Carvacrol treatments. Investigation of rooting was done by assessment of the number and length of roots. Mean comparison of the effect of cultivar type on root number showed that the largest number of roots per explant was obtained fromMS medium containing 50 ppm Thymol. Lowest number of roots observed in mediums supplemented with 300 ppm Thymol and 100 ppm Carvacrol. The best medium for increasing the root length per explant (10. 90 cm) was MS medium supplemented with 100 ppm Carvacrol, while the least increasing in root length per explant observed fromculture mediums contained 300 ppm Thymol and 100 ppm Carvacrol. Also, the largest number of leave formation obtained fromculture medium supplemented with 50 ppm Thymol that significantly higher than other treatments. Statistical analysis (ANOVA) of the data showed that high frequency Callus induction and formation occurred in MS mediums contained 50, 100 and 150 ppm Thymol and 10 ppm Carvacrol and culture mediums supplemented with 300 ppm Thymol and 1000 ppm Carvacrol showed least Callus induction. In contrast, largest Callus diameter observed in culture mediums supplemented with 300 ppm Thymol and 500, 100 ppm Carvacrol. Statistical analysis of results showed that different concentrations of BA and NAA had significant effects on Bulblets number and Bulblets diameter (P<0. 05). High frequency of Bulblets formation obtained fromMS mediums containing 2 mg/l and least Bulblets formed in culture medium supplemented with 4 mg/l BA. Stimart and Ascher (1978) observed highest number of Bulblets formation in culture of LiliumlongiflorumL. Scales on LS medium supplemented with 0. 3 mg/l NAA that correlated with our study. Also, Pierik and Steegmans (1975) explained that auxins versus Gibberellins increased number of bulb formation in Hyacinthus orientalis. Analysis of variance showed that effects of BA and NAA only was significant in root and leaf number and no significant in root and leaf length. The highest and lowest number of roots observed in MS mediums supplemented with 2 mg/l NAA and 4 mg/l BA, respectively. Pierik (1998) demonstrated that Cytokinins in low concentrations induced cell division but in high concentrations inhibited root formation. Nuth (1998) concluded that in LiliumLongiflorum medium containing NAA significantly increased root formation than PGRs free medium. Statistical analysis of results showed that different concentrations of BA and NAA had significant effects on Callus induction and formation in Fritillariaimperialis L. (P<0. 05). High rate of Callus formation was obtained fromMS mediums supplemented with 2 and 4 mg/l NAA and 2 mg/l BA. Conclusion: Thymol and Carvacrol as additives to tissue culture medium can improve some parameters of Fritillariaimperialis L. in vitro propagation like Bulblet diameter, roots and leaves number and size. In this context, Thymol showed better responses than Carvacrol and 50 ppm concentration of Thymol was more effective. Such plant compounds needs to be more considered in tissue culture mediums. Also we concluded that Auxins in comparison to Cytokinins was more effective in propagation of Fritillariaimperialis L. through in vitroBulblet formation.

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APA: Copy

CHAMANI, E., EFTEKHARI, Z., GHANBARI, A.R., HEYDARI, H.R., & ARSHAD, M.. (2018). The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.. JOURNAL OF HORTICULTURE SCIENCE (AGRICULTURAL SCIENCES AND TECHNOLOGY), 32(2 ), 185-197. SID. https://sid.ir/paper/142409/en

Vancouver: Copy

CHAMANI E., EFTEKHARI Z., GHANBARI A.R., HEYDARI H.R., ARSHAD M.. The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.. JOURNAL OF HORTICULTURE SCIENCE (AGRICULTURAL SCIENCES AND TECHNOLOGY)[Internet]. 2018;32(2 ):185-197. Available from: https://sid.ir/paper/142409/en

IEEE: Copy

E. CHAMANI, Z. EFTEKHARI, A.R. GHANBARI, H.R. HEYDARI, and M. ARSHAD, “The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.,” JOURNAL OF HORTICULTURE SCIENCE (AGRICULTURAL SCIENCES AND TECHNOLOGY), vol. 32, no. 2 , pp. 185–197, 2018, [Online]. Available: https://sid.ir/paper/142409/en

Scientific Information Database

ISSN: 2588-4824

Journal Paper

The Effect of Carvacrol, Thymol, BA and NAA on in vitro Regeneration in Fritillaria imperialis L.