Improvement and prediction of secondary metabolites production under yeast extract elicitation of Azadirachta indica cell suspension culture using response surface methodology

Abstract

Neem is a medicinal plant used as antimalarial, antibacterial, antiviral, insecticide, and antimicrobial drug. This study aimed to investigate and predict the effect of yeast extract and sampling time on cell growth, secondary metabolites synthesis, SQS1 and MOF1 genes expression by response surface methodology. The highest fresh and dry cell weights were 580.25 g/L and 21.01 g/L, respectively obtained 6 days after using 100 mg/L yeast extract. The highest azadirachtin accumulation and production were 16.08 mg/g DW and 219.78 mg/L obtained 2 and 4 days, respectively after using 25 mg/L yeast extract. Maximum mevalonic acid accumulation (1.75 mg/g DW) and production (23.77 mg/L) were observed 2 days after application of 50 mg/L yeast extract. The highest amount of squalene accumulation (0.22 mg/g DW) and production (4.53 mg/L) were achieved 4 days after using 50 mg/L yeast extract. Prediction results exhibited the highest azadirachtin accumulation (13.61 mg/g DW) and production (190.50 mg/L), mevalonic acid accumulation (0.50 mg/g DW) and production (5.57 mg/L), and squalene accumulation (0.30 mg/g DW) by using 245 mg/L yeast extract for 2 days, 71 mg/L yeast extract for 2 days, 200 mg/L yeast extract for 4.96 days, without yeast extract for 6.54 days and 4 days, respectively. Also, it was predicted that the highest squalene production is achieved by long-term exposure to high concentrations of yeast extract. The qRT-PCR analysis displayed the maximum relative gene expression of SQS1 and MOF1 by using 150 and 25 mg/L yeast extract for 4 and 2 days treatment.

Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

“…These decreases were varied depending on the auxin type, so that, the amount of decline of rutin, quercetin and kaempferol in the treatments containing NAA were significantly higher than those of the containing 2,4-D. On the other hands, the calli grown on the medium containing Kin produced the higher amount of rutin, quercetin and kaempferol as compared to those on the medium containing BAP, especially at 1 mg/L level. Farjaminezhad and Garoosi ( 2021 ) reported the higher azadirachtin production in Azadirachta indica cell suspension culture in the medium containing 2 mg/L Kin. In Lian et al ( 2002 ) study on Panax ging , saponin productivity increased when the medium supplemented with a low concentration of cytokinins, but the growth of the cells was not affected.…”

“…Biotechnological techniques, especially plant cell and tissue culture, play an essential role in exploring alternative approaches for the production of desirable therapeutic compounds from plants (Farjaminezhad et al 2013 ) and environmental adaptation (Farjaminezhad and Garoosi 2021 ). The callus induction and establishment of suspension cultures are two basic techniques, which have been used for production of wide range of phytochemicals, including therapeutic and antioxidant compounds (Colgecen et al 2011 ).…”

Enhancement of shoot proliferation and evaluation of biotic elicitation effects on anatomical changes of pseudo stem and anti-lipid peroxidation activity of Curcuma mangga Val.

“…The small number and fragile xylem cells could greatly reduce the absorption of nutrients, and this was linked directly with the severe growth retardation and gross morphological abnormality of the plantlets cultured in proliferation medium supplemented with yeast extract and chitosan as reported in Abraham et al (2011). Yeast extract was reported to induce a complex stress response resulted in activation of secondary metabolites production (Farjaminezhad and Garoosi, 2021;Kochan et al, 2017) but resulted in slow cell growth (Hedayati et al, 2021) and cell damage at high concentration (Sánchez-Sampedro et al, 2005).…”

Covid-19 ile mücadelede bitkilerin rolü

“…Bitkisel sekonder metabolitler, bitkinin doğal yaşam döngüsünün devamı için gerekli olmamakla birlikte, bitkilerin çevre ile olan adaptasyonlarında büyük önem taşıyan metabolitlerdir [50]. Bitkilerde bulunan sekonder metabolitlerin çeşitli biyolojik ve ekolojik özellikleri mevcuttur.…”

“…Bitkilerden sekonder metabolitlerin üretimi; bitki genetiği, çevresel faktörler, iklim, mevsim, büyüme periyodu, bitki kısmı, hasat öncesi ve sonrasındaki işlemlerin yanı sıra, ekstraksiyon tekniklerine de bağlı olarak değişiklik gösterebilmektedir. Tüm bu faktörler, klasik yöntemlerle sekonder metabolit üretimini kısıtlamakta, bu noktada ise bitki doku kültürü uygulamaları devreye girmektedir [50]. Bitki doku kültürü, farklı bitki hücreleri, organları veya dokularının totipotensi özelliğinden faydalanmak adına aksenik koşullarda kültüre alınması işlemidir [72].…”