Abstract
An efficient plant regeneration protocol was developed for Basilicum polystachyon (L.) Moench using shoot tip from in vitro germinated plant. Both shoot multiplication and root induction were initiated from shoot tip explants in Murashige and Skoog’s (MS) basal medium supplemented with N6-benzylaminopurine (BAP) and 6-furfurylaminopurine (Kin) combination with 1-naphthaleneacetic acid (NAA) and without any plant growth regulator. Among the different concentrations and combinations of growth regulators, the highest number of shoots per explants was induced on 13.32 μM BAP with 0.53 μM NAA. It was also found that the multiplication of shoots along with roots induced in MS medium without any plant growth regulators. The in vitro grown plants were successfully hardened and acclimatized in the field with a 99% survival rate. The results obtained from HPLC analysis established the presence of a significant amount of endogenous auxin, viz. indole-3-acetic acid acid and indole-3-butyric acid in the shoot and root tips of B. polystachyon. This is the first report of a successful multiplication of B. polystachyon in absence of plant growth regulators and the presence of an abundant quantity of endogenous auxin in root and shoot tips using Reversed-Phase High-Performance Liquid Chromatography (RP-HPLC) coupled with ultraviolet–visible (UV–Vis) detector.
Key message
This is the first report on identification of endogenous indole-3-acetic acid and indole-3-butyric acid from the shoot and root tips of B. polystachyon (L.) Moench.
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Abbreviations
- ANOVA:
-
Analysis of variance
- BAP:
-
N6-Benzylaminopurine
- D:
-
Day
- GA:
-
Gibberellic acid
- h:
-
Hour
- IAA:
-
Indole-3-acetic acid
- IBA:
-
Indole-3-butyric acid
- Kin:
-
6-Furfurylaminopurine
- lb:
-
Pound
- m:
-
Meter
- MDR:
-
Multidrug-resistant
- mg:
-
Milligram
- MGT:
-
Mean germination time
- min:
-
Minute
- mm:
-
Millimeter
- MS:
-
Murashige and Skoog
- NAA:
-
1-Naphthaleneacetic acid
- PGR:
-
Plant growth regulator
- RP-HPLC:
-
Reverse Phase- High-Performance Liquid Chromatography
- Rt :
-
Retention time
- SE:
-
Standard error
- Sec:
-
Second
- UV–Vis:
-
Ultraviolet–visible
- μl:
-
Microliter
- μM:
-
Micromole
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Acknowledgements
We sincerely acknowledge the Department of Biotechnology, The University of Burdwan, Burdwan 713104, WB, India for giving the necessary laboratory and instrumentation facilities (HPLC under BOOST, Department of Science & Technology, and Biotechnology, Government of West Bengal, India). We also convey our sincere thanks to Mr. Kaushik Sarkar, Technical Assistant, Grade-II, Department of Biotechnology, The University of Burdwan, for his kind help in HPLC analysis.
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Government of West Bengal, India for financial assistance through State-Funded Fellowship.
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The authors have made the following declarations regarding their contributions: SD and IC conceived the design of the experiments. SD and IC monitored the research work. KWS collected and analyzes sample data. SD contributed to writing the manuscript. All the authors read and approved the final manuscript. This is a part thesis work of SD.
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Communicated by Francisco de Assis Alves Mourão Filho.
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Das, S., Sultana, K.W. & Chandra, I. In vitro micropropagation of Basilicum polystachyon (L.) Moench and identification of endogenous auxin through HPLC. Plant Cell Tiss Organ Cult 141, 633–641 (2020). https://doi.org/10.1007/s11240-020-01824-3
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DOI: https://doi.org/10.1007/s11240-020-01824-3