Agrobacterium-mediated genetic transformation of ajowan (Trachyspermum ammi (L.) Sprague): an important industrial medicinal plant

Niazian, Mohsen; Sadat-Noori, Seyed Ahmad; Tohidfar, Masoud; Galuszka, Petr; Mortazavian, Seyed Mohammad Mahdi

doi:10.1016/j.indcrop.2019.02.005

Cited by 30 publications

(14 citation statements)

References 60 publications

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“…However, untransformed leaves showed signs of wilting at a drought stress of 100 mM mannitol and exhibited severe chlorosis with increased concentrations of NaCl (above 50 mM). These results are in agreement with previous findings that suggested salt and drought stress hamper plant growth and development (Ali and Ismail, 2014;Niazian et al, 2019). Nevertheless, the transgenic leaves survived extreme stress conditions due to the overexpression of rol genes and the production of stress-tolerant metabolites, as supported by previous reports (Bulgakov et al, 2012;Kayani et al, 2016aKayani et al, , 2017.…”

Section: Abiotic Stress Tolerance and Photosynthetic Indices Of The Tsupporting

confidence: 93%

Enhanced stress tolerance in transformed Ajuga bracteosa Wall. ex Benth. regenerants by upregulated gene expression of metabolic pathways

Rubnawaz¹,

Kayani²,

Mahmood³

et al. 2020

Turk J Bot

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The rol oncogenes of Agrobacterium rhizogenes enhance the production of medicinally important compounds in plants and provide a first barrier against the overproduction of reactive oxygen species during biotic and abiotic stress. This study was designed to evaluate the expression of genes involved in biosynthetic pathways and their impact on metabolic contents and environmental stress tolerance in regenerated Ajuga bracteosa Wall. ex Benth. After successful transformation, real-time quantitative PCR confirmed the increased expression (1.94-6.59-fold) of HMGR, HDS, FDS, PAL, and TTG1 genes in transgenic lines. Furthermore, GC-MS coupled with principal component analysis revealed diverse concentrations of 97 metabolites in A. bracteosa. Transgenic lines showed greater survival under multiple stresses. This was revealed by significant chlorophyll content (8.13-21 µmoles/m 2), higher quantum efficiency of PSII (F v /F m), and the performance index (PI abs) value. Similarly, catalase and peroxidase enzyme activities were enhanced during extreme drought (300-400 mM mannitol) and salinity (150-200 mM NaCl) conditions, compared to untransformed control. Wild type control plant leaves were completely necrotized by Aspergillus fumigatus (FCBP 66) and Fusarium solani (FCBP 0291), whereas transformed leaves had improved antifungal resistance. In conclusion, our data suggest that rolABC genes have a significant impact on the synthesis of metabolites involved in enhancing multistress tolerance in A. bracteosa.

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Section: Abiotic Stress Tolerance and Photosynthetic Indices Of The Tsupporting

confidence: 93%

Enhanced stress tolerance in transformed Ajuga bracteosa Wall. ex Benth. regenerants by upregulated gene expression of metabolic pathways

Rubnawaz¹,

Kayani²,

Mahmood³

et al. 2020

Turk J Bot

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“…Thymol is a valuable bioactive compound with strong germicide, anti-spasmodic and fungicidal effects (Niaizian et al 2018). Different in vitro-based methods, including arti cial tetraploidy induction (Noori et al 2017), genetic engineering (Niazian et al 2019;Nomani et al 2019), and biotic-abiotic elicitation (Razavizadeh et al 2020), have been applied to improve the valuable bioactive compounds of ajowan. The present study was conducted to evaluate the effect of SA elicitation on regeneration e ciency, enzymatic antioxidants activity and secondary metabolite production in shoot cultures of ajowan under the arti cially induced drought stress (PEG 6000).…”

Section: Introductionmentioning

confidence: 99%

Integrative effects of stress- and stress tolerance-inducing elicitors on in vitro bioactive compounds of ajowan [Trachyspermum ammi (L.) Sprague] medicinal plant

Niazian

Howyzeh

Sadat-Noori

2021

Plant Cell Tiss Organ Cult

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Bioactive compounds of medicinal plants have a wide range of applications in pharmaceutical, food and other industries. In vitro culture systems have great potential for sustainable production of bioactive compounds of medicinal plants. In the present study, the individual and combined effects of a stress tolerance-inducing (salicylic acid) and a stress-inducing elicitor (polyethylene glycol) were evaluated on regeneration e ciency, antioxidants activity and phytochemical pro le of in vitro shoot cultures of ajowan. Different concentrations of salicylic acid (SA) (0, 10, 20, 40, 80 µM) and polyethylene glycol (PEG 6000) (0, 1, 2, 5%) were added to the shoot regeneration Murashige and Skoog medium containing Kin (1.5 mg/L) and NAA (0.25 mg/L) plant growth regulators. Salicylic acid reduced the adverse effect of PEG treatment on number of regenerated shoots and in vitro rooting. The activities of catalase, superoxide dismutase, and peroxidase enzymatic antioxidants were signi cantly increased in SA + PEG treated plants. The gas chromatography-mass spectrometry (GC-MS)-pro ling revealed quantitative and qualitative phytochemical differences between control and SA + PEG treated plants. The greatest means of p-cymene and thymol bioactive compounds were obtained from in vitro shoots treated with 5% PEG + 40 µM SA. The inter-simple sequence repeats (ISSR) markers proved the genetic stability of in vitro regenerated plants. The presented protocol is useful for large-scale sustainable production of secondary metabolites (SMs) of medicinal plants. The same strategy (stress tolerance-inducing elicitor + stress-inducing elicitor) is applicable to increase valuable SMs in other production systems such as hydroponic, greenhouse and eld conditions.

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“…In another study, to enhance drought and salinity tolerance in T. ammi , betaine aldehyde dehydrogenase gene and hypocotyl explants were used. The results showed enhancement thymol level in both wild type and transformed plants of ajowan (39.2 and 55.07%) by the drought stress [ 57 ]. In another study conducted by Sharma et al [ 58 ], it was found that overexpression of Tryptophan decarboxylase and strictosidine synthase in Catharanthus roseus increased vindoline, catharanthine, and vinblastine content.…”

Section: Discussionmentioning

confidence: 99%

Plant regeneration and transformation of Trachyspermum ammi using Agrobacterium tumefaciens and zygotic embryos

Nomani

Tohidfar

2021

Journal of Genetic Engineering and Biotechnology

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Background Trachyspermum ammi is one of the key medicinal plant species with many beneficial properties. Thymol is the most important substance in the essential oil of this plant. Thymol is a natural monoterpene phenol with high anti-microbial, anti-bacterial, and anti-oxidant properties. Thymol in the latest research has a significant impact on slowing the progression of cancer cells in human. In this research, embryos were employed as convenient explants for the fast and effectual regeneration and transformation of T. ammi. To regenerate this plant, Murashige and Skoog (MS) and Gamborg's B5 (B5) media were supplemented with diverse concentrations of plant growth regulators, such as 6-benzyladenine (BA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D), and kinetin (kin). Transgenic Trachyspermum ammi plants were also obtained using Agrobacterium-mediated transformation and zygotic embryos explants. Moreover, two Agrobacterium tumefaciens strains (EHA101 and LBA4404) harboring pBI121-TPS2 were utilized for genetic transformation to Trachyspermum ammi. Results According to the obtained results, the highest plant-regeneration frequency was obtained with B5 medium supplemented with 0.5 mg/l BA and 1 mg/l NAA. The integrated gene was also approved using the PCR reaction and the Southern blot method. Results also showed that the EHA101 strain outperformed another strain in inoculation time (30 s) and co-cultivation period (1 day) (transformation efficiency 19.29%). Furthermore, HPLC method demonstrated that the transformed plants contained a higher thymol level than non-transformed plants. Conclusions In this research, a fast protocol was introduced for the regeneration and transformation of Trachyspermum ammi, using zygotic embryo explants in 25–35 days. Our findings confirmed the increase in the thymol in the aerial part of Trachyspermum ammi. We further presented an efficacious technique for enhancing thymol content in Trachyspermum ammi using Agrobacterium-mediated plant transformation system that can be beneficial in genetic transformation and other plant biotechnology techniques.

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Agrobacterium-mediated genetic transformation of ajowan (Trachyspermum ammi (L.) Sprague): an important industrial medicinal plant

Cited by 30 publications

References 60 publications

Enhanced stress tolerance in transformed Ajuga bracteosa Wall. ex Benth. regenerants by upregulated gene expression of metabolic pathways

Enhanced stress tolerance in transformed Ajuga bracteosa Wall. ex Benth. regenerants by upregulated gene expression of metabolic pathways

Integrative effects of stress- and stress tolerance-inducing elicitors on in vitro bioactive compounds of ajowan [Trachyspermum ammi (L.) Sprague] medicinal plant

Plant regeneration and transformation of Trachyspermum ammi using Agrobacterium tumefaciens and zygotic embryos

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