نوع مقاله : مقاله پژوهشی
نویسندگان
1 گروه پژوهشی بیوتکنولوژی گیاهان باغبانی، پژوهشکده بیوتکنولوژی صنعتی، سازمان جهاد دانشگاهی خراسان رضوی، مشهد، ایران.
2 گروه بیوتکنولوژی و به‌نژادی گیاهی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران.
3 گروه بیوتکنولوژی و به‌نژادی گیاهی، دانشکده کشاورزی، دانشگاه فردوسی مشهد، مشهد، ایران
4 گروه پژوهشی بیوتکنولوژی گیاهان باغبانی، پژوهشکده بیوتکنولوژی صنعتی، سازمان جهاد دانشگاهی خراسان رضوی، مشهد، ایران
چکیده
کلیدواژه‌ها
موضوعات
عنوان مقاله [English]
نویسندگان [English]
Hairy root culture of medicinal plants is a promising method for producing herbal medicinal compounds. This study focused on Datura innoxia, a plant known for its use in treating various diseases, with the goal of cultivating its hairy roots to produce the alkaloids hyoscyamine and scopolamine. In addition, the research aimed to explore the potential for enhancing the production of these medicinal compounds using biotic elicitors, including methyl jasmonate, yeast extract, and salicylic acid. The investigation into the effects of these different elicitors on the growth rate and alkaloid production in Datura hairy roots yielded significant findings. Treatment with methyl jasmonate did not alter the growth rate of the hairy roots but resulted in a 30% increase in alkaloid production compared to the control. On the other hand, yeast extract treatment did not change alkaloid production after 48 hours. However, the application of salicylic acid led to a notable increase in alkaloid levels. Overall, the results demonstrated that the use of elicitors is a crucial approach to enhance alkaloid production in hairy root cultures. These findings suggest that application of various elicitors in hairy root culture can be an effective strategy for maximizing the production of valuable medicinal compounds. Thus, using biotic elicitors like methyl jasmonate and salicylic acid offers a viable method to boost the yield of important alkaloids in Datura innoxia, making hairy root culture a highly effective tool in the production of medicinal plant compounds.
کلیدواژه‌ها [English]
Extended Abstract
Introduction
Hairy root culture, a promising technique in plant biotechnology, has garnered significant interest in recent years for producing herbal medicinal compounds. Datura, known for its therapeutic potential in treating various diseases, particularly cancer and digestive system issues, was the focus of this research. This study aimed to optimize the cultivation conditions of Datura innoxia hairy roots to enhance the production of the alkaloids hyoscyamine and scopolamine.
Material and Methods
To conduct this experiment, Datura innoxia seeds were first surface-sterilized and cultivated in MS medium. Leaf explants from sterile seedlings were then co-cultivated with Agrobacterium rhizogenes strains A4, 15834, and MSU in LB liquid medium. Induced roots, approximately 2 to 3 cm in length, were excised and transferred to B5 liquid medium on a shaker at 90 rpm in the dark. After two months, the rate and stability of root growth were assessed. The bacterial species were confirmed by amplifying the rolC gene, located in the T-DNA region of the hairy root-inducing plasmid, using PCR with specific primers. The effects of different elicitors—methyl jasmonate, salicylic acid, and yeast extract—on the growth and alkaloid production were investigated. Preliminary identification of tropane alkaloids in the samples was done using Dragendorff reagent. The alkaloid content in each sample was carefully measured using spectrophotometry. To enhance accuracy in quantifying alkaloids across different treatments, HPLC analysis was also performed on the samples with the highest alkaloid content.
Result and Discussion
The study on the effect of methyl jasmonate on the growth rate and alkaloid production revealed that methyl jasmonate effectively enhanced alkaloid production, increasing it by 30% compared to the control. Salicylic acid and yeast extract did not significantly affect the growth rate of hairy roots, as there was no substantial difference in the dry weight of hairy roots across treatments. However, alkaloid production varied significantly between treatments. Yeast extract did not change alkaloid content after 48 hours compared to the control. In contrast, salicylic acid treatment resulted in a significant increase in alkaloid content after 48 hours. These findings indicate that elicitors can influence tropane alkaloid production in Datura hairy roots, but their effectiveness depends on optimization alongside other environmental factors. Additionally, HPLC analysis showed a significant increase in hyoscyamine content with methyl jasmonate treatment, whereas the increase in scopolamine was not significant.
Conclusion
The results of this experiment demonstrated that adding an elicitor as a stimulating factor enhances tropane alkaloid production in Datura hairy roots. Specifically, methyl jasmonate increased alkaloid production by 30% compared to the control treatment.
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