Alkaloid production and capacity for methyljasmonate induction by hairy roots of two species in Tribe Anthocercideae, family Solanaceae

2015 ◽  
Vol 42 (8) ◽  
pp. 792 ◽  
Author(s):  
Suzanne M. Ryan ◽  
Kathleen D. DeBoer ◽  
John D. Hamill

In addition to producing medicinally important tropane alkaloids, some species in the mainly Australian Solanaceous tribe Anthocercideae, sister to genus Nicotiana, are known to also contain substantial levels of the pyridine alkaloids nicotine and nornicotine. Here, we demonstrate that axenic hairy root cultures of two tribe Anthocercideae species, Cyphanthera tasmanica Miers and Anthocercis ilicifolia ssp. ilicifolia Hook, contain considerable amounts of both nicotine and nornicotine (~0.5–1% DW), together with lower levels of the tropane alkaloid hyoscyamine (<0.2% DW). Treatment of growing hairy roots of both species with micromolar levels of the wound stress hormone methyl-jasmonate (MeJa) led to significant increases (P < 0.05) in pyridine alkaloid concentrations but not of hyoscyamine. Consistent with previous studies involving Nicotiana species, we also observed that transcript levels of key genes required for pyridine alkaloid synthesis increased in hairy roots of both Anthocercideae species following MeJa treatment. We hypothesise that wound-associated induction of pyridine alkaloid synthesis in extant species of tribe Anthocercideae and genus Nicotiana was a feature of common ancestral stock that existed before the separation of both lineages ~15 million years ago.

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Khoa Van Nguyen ◽  
Benyakan Pongkitwitoon ◽  
Thanika Pathomwichaiwat ◽  
Unchera Viboonjun ◽  
Sompop Prathanturarug

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.


Author(s):  
Fumihito Hasebe ◽  
Honoka Yuba ◽  
Takashi Hashimoto ◽  
Kazuki Saito ◽  
Nobutaka Funa ◽  
...  

Abstract Tropane alkaloids, including clinically important hyoscyamine and scopolamine, are produced in the roots of medicinal plant species, such as Atropa belladonna, from the Solanaceae family. Recent molecular and genomic approaches have advanced our understanding of the metabolic enzymes involved in tropane alkaloid biosynthesis. A non-canonical type III polyketide synthase (PKS), pyrrolidine ketide synthase (PYKS), catalyzes a two-step decarboxylative reaction, which involves imine-ketide condensation indispensable to tropane skeleton construction. In this study, we generated pyks mutant A. belladonna hairy roots via CRISPR/Cas9-mediated genome editing and analyzed the metabolic consequences of the loss of PYKS activity on tropane alkaloids, providing insights into a crucial role of the scaffold-forming reaction in the biosynthetic pathway.


2021 ◽  
Author(s):  
jinyu Bao ◽  
Xu Lu ◽  
Lei Ma ◽  
Xiumin Zhang ◽  
Peng Tian ◽  
...  

Abstract Hairy roots obtained by infecting broccoli (Brassica oleracea L. var. Italic Planch) leaves with Agrobacterium rhizogenes (ATCC15834) had the characteristics of phytohormone autonomy, genetic stability and can produce a large amount of anti-cancer substance Sulforaphane (SF) and SF biosynthetic precursors substance Glucoraphanin (GRA). The production of SF in hairy roots of broccoli increased significantly under the induction of exogenous signal molecule Methyl jasmonate (MeJA). However, the molecular mechanism of MeJA treatment hairy roots of broccoli have not been reported. In this study, the optimal concentration of MeJA for having treatment broccoli hairy roots were selected based on the yield of GRA and SF. After grew 18 days, broccoli hairy roots were treated with 10 mmol/L MeJA for 0, 3, 6, 9 and 12 h respectively. Compared with 0 h, the yields of GRA and SF increased under other treatments. The highest yield of GRA and SF was noted at 9 h, which were 2.22-fold and 1.74-fold of 0 h, respectively. Broccoli (Brassica oleracea L. var. botrytis L) as reference genome, and 1195 co-differentially expressed genes (DEGs) at 0, 3, 6, 9 and 12 h was observed under treatment 10 mmol/L MeJA, which there were 3,826 down-regulated and 574 up-regulated genes. The six key genes that regulated Glucosinolates (GLS) synthesis, MAM1, CYP79B1, CYP83B1, UGT74B1, and FMOGS-OX5, were up-regulated at 0 and 3 h, and down-regulated at the rest of the time; BCAT2 was up-regulated at 6, 9, 12 h, and at 0, 3 h expression was down-regulated, transcription factors MYB34 and MYB122 were up-regulated at 3 h, and down-regulated at other time points, MYB51 was up-regulated at 0, 3 h and down-regulated at 6, 9, 12 h. The pathway diagram of GRA biosynthesis and transformation pathway in broccoli hairy roots treatment by MeJA simulated, and the molecular mechanis0m of GRA biosynthesis and SF accumulation in broccoli hairy roots under MeJA treatment were revealed.


2015 ◽  
Vol 10 (11) ◽  
pp. 1934578X1501001
Author(s):  
Yeon Bok Kim ◽  
Darwin W. Reed ◽  
Patrick S. Covello

Silene vulgaris (Moench) Garcke (Caryophyllaceae) is widely distributed in North America and contains bioactive oleanane-type saponins. In order to investigate in vitro production of triterpenoid saponins, hairy root cultures of S. vulgaris were established by infecting leaf explants with five strains of Agrobacterium rhizogenes (LBA9402, R1000, A4, 13333, and 15834). The A. rhizogenes strain LBA9402 had an infection of 100% frequency and induced the most hairy roots per plant. Methyl jasmonate (MeJA)-induced changes in triterpenoid saponins in S. vulgaris hairy roots were analyzed. Accumulation of segetalic acid and gypsogenic acid after MeJA treatment was 5-and 2-fold higher, respectively, than that of control root. We suggest that hairy root cultures of S. vulgaris could be an important alternative approach to the production of saponins.


2004 ◽  
Vol 59 (11-12) ◽  
pp. 863-867 ◽  
Author(s):  
Rawia Zayed ◽  
Michael Wink

Hairy root cultures of Brugmansia suaveolens were set up by infection of root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analysing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of tropane alkaloids, such as hyoscyamine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid in order to stimulate the biosynthesis of tropane alkaloids. Profile and amounts of tropane alkaloids were analysed using capillary GLC-MS. At least 18 different tropane alkaloids could be identified. Treatment of the cultures with 200 μᴍ methyljasmonate increased the alkaloid accumulation 25-fold up to a level of 1 mg/g fresh weight as compared to untreated controls. Quercetin enhanced the alkaloid production 10 fold (0.4 mg/g fresh weight) within 24 h. In contrast 100 μᴍ salicylic acid decreased alkaloids to a level of 1 μg/g fresh weight.


1991 ◽  
Vol 46 (7-8) ◽  
pp. 519-521 ◽  
Author(s):  
Katja Doerk ◽  
Ludger Witte ◽  
A. Wilhelm Alfermann

Abstract Tropane alkaloids are typical natural products of solanaceous plants. The patterns of these alkaloids from hairy roots of Hyoscyamus albus were determined by GC-MS analysis. 18 alka-loidal compounds were detected, six of them only in trace amounts. Some of these alkaloids, namely hygrine, 3a-acetoxytropane, 3 β-acetoxytropane and 6-hydroxylittorine as well as the trace compounds N-methylpyrrolidinylhygrine, phenylacetoxytropane and two isomers of feruloyloxytropane were not previously reported to be components of this species. The major compounds formed were found to be hyoscyamine and littorine.


2020 ◽  
Vol 71 (22) ◽  
pp. 6861-6864
Author(s):  
María A Pedreño ◽  
Lorena Almagro

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.


Hairy Roots ◽  
2018 ◽  
pp. 71-93
Author(s):  
Guoyin Kai ◽  
Weiwei Zhao ◽  
Min Shi ◽  
Yao Wang
Keyword(s):  

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