scholarly journals Development of Hairy Root Cultures for Biomass and Triterpenoid Production in Centella asiatica

Plants ◽  
2022 ◽  
Vol 11 (2) ◽  
pp. 148
Author(s):  
Seungeun Baek ◽  
Jong-Eun Han ◽  
Thanh-Tam Ho ◽  
So-Young Park
Keyword(s):  
Hairy Root ◽  
Mass Production ◽  
Adventitious Roots ◽  
Transformation Efficiency ◽  
Growth Index ◽  
Centella Asiatica ◽  
Asiatic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Petiole Explants

Centella asiatica (Apiaceae) is a tropical/subtropical medicinal plant, which contains a variety of triterpenoids, including madecassoside, asiaticoside, madecassic acid, and asiatic acid. In this study, we tested the efficiency of hairy root (HR) induction in C. asiatica from leaf and petiole explants. Leaves and petioles collected from C. asiatica plants were suspended in agro-stock for 30 min and co-cultured with Agrobacterium rhizogenes for 3 days to induce HR formation. The transformation efficiency of leaf and petiole explants was approximately 27% and 12%, respectively. A total of 36 HR lines were identified by PCR-based amplification of rol genes, and eight of these lines were selected for further analysis. Among all eight HR lines, the petiole-derived lines HP4 and HP2 displayed the highest growth index (37.8) and the highest triterpenoids concentration (46.57 mg∙g−1), respectively. Although triterpenoid concentration was >2-fold higher in leaves than in petioles of C. asiatica plants, the accumulation of triterpenoids in petiole-derived HR cultures was 1.4-fold higher than that in leaf-derived HR cultures. Additionally, in both leaf- and petiole-derived HR cultures, terpenoid production was higher in HRs than in adventitious roots. These results demonstrate that the triterpenoid content in the explant does not affect the triterpenoid content in the resultant HRs. The HR culture of C. asiatica could be scaled up to enable the mass production of triterpenoids in bioreactors for the pharmaceutical and cosmetic industries.

scholarly journals Effects of methyl jasmonate on the growth and triterpenoid production of diploid and tetraploid Centella asiatica (L.) Urb. hairy root cultures

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Khoa Van Nguyen ◽  
Benyakan Pongkitwitoon ◽  
Thanika Pathomwichaiwat ◽  
Unchera Viboonjun ◽  
Sompop Prathanturarug
Keyword(s):  
Methyl Jasmonate ◽  
Hairy Root ◽  
Hairy Roots ◽  
Centella Asiatica ◽  
Root Culture ◽  
Hairy Root Culture ◽  
Asiatic Acid ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Meja Treatment

AbstractIn this study, the effects of methyl jasmonate (MeJA) on the phytomass and triterpenoid production of diploid and tetraploid Centella asiatica hairy roots were investigated. Hairy root cultures were obtained from diploid and induced tetraploid plants of C. asiatica infected by Agrobacterium rhizogenes strain ATCC 43057. MeJA triggered triterpenoid production in both ploidy hairy roots, whereas triterpenoids were not produced in the untreated hairy roots. Among the treatments, the 50 µM MeJA treatment yielded the maximum triterpenoid production in diploid hairy roots of 27.25 ± 0.27 µg/mg Dry weight (DW) total triterpenoid at day 21. For the tetraploid hairy root cultures, the 28th-day hairy root culture produced a maximum amount of triterpenoids of 16.29 ± 6.32 µg/mg DW in response to the 50 µM MeJA treatment, whereas the 100 µM MeJA treatment produced a similar triterpenoid amount (16.31 ± 9.24 µg/mg DW) at day 14. Moreover, in response to 50 µM MeJA, we obtained different ratios of aglycone to glycoside, i.e., 1:7 and 1:2, between the diploid and tetraploid hairy root cultures. Asiaticoside was the dominant phytochemical, followed by asiatic acid and madecassic acid. This study provides valuable information for producing triterpenoids for C. asiatica commercial products and preparations by using hairy root cultures.

Enhanced production of asiaticoside from hairy root cultures of Centella asiatica (L.) Urban elicited by methyl jasmonate

2007 ◽  
Vol 26 (11) ◽  
pp. 1941-1949 ◽  
Author(s):  
Ok-Tae Kim ◽  
Kyong-Hwan Bang ◽  
Yu-Su Shin ◽  
Min-Jeong Lee ◽  
Su-Jin Jung ◽  
...  
Keyword(s):  
Methyl Jasmonate ◽  
Hairy Root ◽  
Centella Asiatica ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Enhanced Production

scholarly journals SYNTHESIS OF BIOLOGICALLY ACTIVE COMPOUNDS IN HAIRY ROOTS OF ASTRAGALUS PENDULIFLORUS LAM.

2020 ◽  
pp. 209-221
Author(s):  
Elena Valerievna Ambros ◽  
Ol'ga Viktorovna Kotsupiy ◽  
Tat'yana Abdulkhailovna Kukushkina ◽  
Tat'yana Vital'yevna Zheleznichenko ◽  
Tat'yana Ivanovna Novikova
Keyword(s):  
Hairy Root ◽  
Hairy Roots ◽  
Virulent Strain ◽  
High Growth ◽  
Growth Index ◽  
Biologically Active ◽  
Pcr Analysis ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Active Metabolites

Agrobacterium rhizogenes – mediated genetic transformation of medicinal plant A. penduliflorus Lam. using A4-RT, R-1601, 15834 SWISS strains was performed. The competences for transformation of three types of explants: hypocotyls, cotyledons, and primary shoots were tested. The virulent strain (15834 SWISS) and types of explants for transformation (primary shoots and cotyledons) with high growth index (I) were determined. The frequency of transformation of cotyledons by strain 15834 SWISS after 4 weeks of cultivation was 15.4% (I = 59.6), hypocotyls – 9.1% (I = 7.3) and primary shoots – 37.5% (I = 21.0). After 8 weeks of cultivation I increased 4.5 times for primary shoots (I = 94.5 ± 0.20) and cotyledons (I = 265.8 ± 0.35), for cultures from hypocotyls – 5.97 times (I = 43.6 ± 0.30). The roots’ transgenic status and the absence of agrobacterium contamination were confirmed by PCR analysis using rolB-, virC-specific primers. The lines of hairy roots characterized by active increases of biomass with high content of biologically active metabolites were selected, moreover, the content of metabolites in hairy root cultures exceeded their content in the roots of introduced plants. The maximum accumulation of compounds was found in hairy roots obtained from primary shoots (pectins – up 7.8%, protopectins – up 15.3%) and cotyledons (tannins – up 16.1%, triterpenic saponins – up 30.5%) after 8 weeks of cultivation. High perfor-mance liquid chromatography (HPLC) analysis demonstrated that hydrolysates of extracts of hairy roots from primary shoots contained 2 flavonol aglycones – quercetin and isorhamnetin whereas ethanol extracts were characterized by presence of quercetin and 4 flavonoid components. In hairy roots from cotyledons the maximum of phenolic compounds (PСs) content did not differ significantly at 8 and 12 weeks of cultivation (1.38 ± 0.01 and 1.49 ± 0.06% of dry weight, respectively). The content of PCs in hairy roots from primary shoots increased two-fold from 4 to 12 weeks of cultivation (up 1.24 ± 0.18%). To the best of our knowledge, this is the first efficient protocol reported for the establishment of hairy root cultures in A. penduliflorus using A. rhizogenes.

scholarly journals Variability of Bioactive Glucosinolates, Isothiocyanates and Enzyme Patterns in Horseradish Hairy Root Cultures Initiated from Different Organs

Molecules ◽  
2019 ◽  
Vol 24 (15) ◽  
pp. 2828 ◽  
Author(s):  
Bertóti ◽  
Böszörményi ◽  
Alberti ◽  
Béni ◽  
M-Hamvas ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Horseradish Peroxidase ◽  
Hairy Root ◽  
Growth Index ◽  
Biologically Active ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Daily Growth ◽  
Hairy Root Clones ◽  
Enzyme Patterns

Horseradish hairy root cultures are suitable plant tissue organs to study the glucosinolate–myrosinase–isothiocyanate system and also to produce the biologically active isothiocyanates and horseradish peroxidase, widely used in molecular biology. Fifty hairy root clones were isolated after Agrobacterium rhizogenes infection of surface sterilized Armoracia rusticana petioles and leaf blades, from which 21 were viable after antibiotic treatment. Biomass properties (e.g. dry weight %, daily growth index), glucosinolate content (analyzed by liquid chromatography-electronspray ionization-mass spectrometry (LC-ESI-MS/MS)), isothiocyanate and nitrile content (analyzed by gas chromatography-mass spectrometry (GC-MS)), myrosinase (on-gel detection) and horseradish peroxidase enzyme patterns (on-gel detection and spectrophotometry), and morphological features were examined with multi-variable statistical analysis. In addition to the several positive and negative correlations, the most outstanding phenomenon was many parameters of the hairy root clones showed dependence on the organ of origin. Among others, the daily growth index, sinigrin, glucobrassicin, 3-phenylpropionitrile, indole-3-acetonitrile and horseradish peroxidase values showed significantly higher levels in horseradish hairy root cultures initiated from leaf blades.

scholarly journals A protocol for sonication-assisted Agrobacterium rhizogenes-mediated transformation of haploid and diploid sugar beet (Beta vulgaris L.) explants.

2014 ◽  
Vol 61 (1) ◽  
Author(s):  
Magdalena Klimek-Chodacka ◽  
Rafal Baranski
Keyword(s):  
Sugar Beet ◽  
Agrobacterium Rhizogenes ◽  
Hairy Root ◽  
Hairy Roots ◽  
Transformation Efficiency ◽  
Binary Vector ◽  
Root Cultures ◽  
Treatment Combination ◽  
Petiole Explants ◽  
Diploid Genotype

Hairy root cultures obtained after Agrobacterium rhizogenes-mediated genetic transformation can serve as a model system for studying plant metabolism and physiology, or can be utilized for the production of secondary metabolites. So far no efficient protocol of hairy root development in sugar beet has been publically released. In this work, two A. rhizogenes strains (A4T and LBA1334) carrying a binary vector pBIN-m-gfp5-ER or pCAMBIA1301 possessing gfp and uidA reporter genes were used to transform petiole explants of haploid and diploid sugar beet genotypes. Five treatment combinations of sonicated-assisted Agrobacterium-mediated transformation were compared. Hairy roots appeared on 0% to 54% of explants depending on the treatment combination used. The highest frequency was achieved when explants of a diploid genotype were sonicated for 15 s in the inoculum containing A. rhizogenes of OD600=0.5 and then co-cultured for three days. Using the same treatment combinations the explants of haploid genotypes developed hairy roots with the frequency ranging from 10% to 36%. Transformation efficiency was independent on the bacterial strain used. The results indicate that haploid sugar beet explants are amenable to transformation using A. rhizogenes, and that the efficiency of that process can be increased by applying short ultrasound treatment.

Elicitor-enhanced Silymarin production in hairy root cultures of Silybum marianum L. Gaertn

Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
T Hasanloo ◽  
H Rahnama ◽  
R Sepehrifar ◽  
MR Shams
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Silybum Marianum

Clone selection of in vitro horseradish hairy root cultures

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
R Bertóti ◽  
Á Alberti ◽  
A Böszörményi ◽  
R Könye ◽  
T Horváth ◽  
...  
Keyword(s):  
Hairy Root ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Clone Selection ◽  
Selection Of

scholarly journals Carrot hairy roots: factories for secondary metabolite production

2020 ◽  
Vol 71 (22) ◽  
pp. 6861-6864
Author(s):  
María A Pedreño ◽  
Lorena Almagro
Keyword(s):  
Phenolic Compounds ◽  
Secondary Metabolite ◽  
Hairy Root ◽  
Hairy Roots ◽  
Hairy Root Cultures ◽  
Root Cultures ◽  
Black Carrot ◽  
Metabolite Production ◽  
Experimental Botany ◽  
Induced Changes

This article comments on: Barba-Espín G, Chen S-T, Agnolet S, Hegelund JN, Stanstrup J, Christensen JH, Müller R, Lütken H. 2020. Ethephon-induced changes in antioxidants and phenolic compounds in anthocyanin-producing black carrot hairy root cultures. Journal of Experimental Botany 71, 7030–7045.

Sign in / Sign up

Export Citation Format

Share Document