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Author:Böhling, Linda
Title:Development of plumbagin-producing plant cell and tissue cultures
Plumbagiinia tuottavien kasvisolu- ja kudosviljelmien kehittäminen
Publication type:Master's thesis
Publication year:2013
Pages:viii + 89 s. + liitt. 9      Language:   eng
Department/School:Biotekniikan ja kemian tekniikan laitos
Main subject:Biokemia ja mikrobiologia   (Kem-30)
Supervisor:Nordström, Katrina
Instructor:Häkkinen, Suvi ; Rischer, Heiko
OEVS:
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Location:P1 Ark Aalto  2056   | Archive
Keywords:plumbagin
dionaea muscipula
nepenthes maxima
plumbago auriculata
callus culture
suspension culture
hairy root culture
nicotiana tabacum
polyketide synthase
plumbagiini
kallusviljelmä
suspensioviljelmä
karvajuuriviljelmä
polyketidisyntaasi
Abstract (eng): The aim of this work was to develop plumbagin-producing plant cell and tissue cultures and to investigate plumbagin biosynthesis.
The targets of the study were plumbagin-producing plants Dionaea muscipula, Nepenthes maxima and Plumbago auriculata, from which callus, suspension and hairy root cultures were established, and polyketide synthase PinPKS, the function of which was investigated in transgenic Nicotiana tabacum SRI hairy root cultures.

Several callus and suspension cultures were successfully established with all the investigated species.
Callus induction was highest in P. auriculata explants, whereas tissue growth was fastest in N. maxima callus cultures.
Relatively high plumbagin contents were observed in D. muscipula and N. maxima suspensions.
Establishment of hairy root cultures did not succeed, but a few possibly transformed P. auriculata roots were formed.

No plumbagin or related polyketide structures were detected in N. tabacum SRI PinPKS hairy root cultures, but their fatty acid and phytosterol profiles were prominently different compared to control cultures.
It is suggested that PinPKS suppresses the mevalonate pathway, and thus the synthesis of campesterol and stigma sterol, and interferes with fatty acid synthesis resulting in a higher content of oleic acid amide at the expense of palmitic acid content.

As a result of the work, plenty of valuable information was obtained about cell and tissue culture of the investigated species.
The high plumbagin contents observed in D. muscipula and N. maxima suspension cultures prompt extensive studies.
The observed changes in the metabolite contents of N. tabacum SRI PinPKS hairy root cultures were significant and require thorough scrutiny.
Abstract (fin): Tämän työn tarkoituksena oli kehittää plumbagiinia tuottavia kasvisolu- ja kudosviljelmiä sekä tutkia plumbagiinin biosynteesiä.
Tutkimuksen kohteena olivat plumbagiinia tuottavat Dionaea muscipula -, Nepenthes maxima - ja Plumbago auriculata -kasvit, joista perustettiin kallus-, suspensio- ja karvajuuriviljelmiä, sekä PinPKS-polyketidisyntaasi, jonka toimintaa tutkittiin siirtogeenisissä Nicotiana tabacum SRI -karvajuuriviljelmissä.

Useita kallus- ja suspensioviljelmiä perustettiin menestyksekkäästi kaikilla tutkituilla lajeilla.
Kallusinduktio oli korkeinta P. auriculata -eksplanteissa, ja solukon kasvu puolestaan nopeinta N. maxima -kallusviljelmissä.
Suhteellisen korkeita plumbagiinipitoisuuksia havaittiin D. muscipula - ja N. maxima -suspensioissa.
Karvajuuriviljelmien perustaminen ei onnistunut, mutta muutamia mahdollisesti transformoituneita P. auriculata -juuria muodostui.

N. tabacum SRI PinPKS -karvajuuriviljelmissä ei havaittu plumbagiinia tai samankaltaisia polyketidirakenteita, mutta rasvahappo- ja fytosteroliprofiilit olivat niissä selkeästi erilaiset verrattuna kontrolliviljelmiin.
PinPKS:n ehdotetaan supressoivan mevalonaattireittiä, ja siten kampesterolin ja stigmasterolin synteesiä, sekä häiritsevän rasvahapposynteesiä johtaen korkeampaan oleiinihappoamidipitoisuuteen palmitiinihappopitoisuuden kustannuksella.

Työn tuloksena saatiin runsaasti arvokasta tietoa tutkittujen lajien solu- ja kudosviljelystä.
D. muscipula -ja N. maxima -suspensioviljelmissä havaitut korkeat plumbagiinipitoisuudet puoltavat laajamittaisempia tutkimuksia.
N. tabacum SRI PinPKS -karvajuuriviljelmissä havaitut metaboliittipitoisuuksien muutokset olivat merkittäviä ja vaativat perusteellista tarkastelua.
ED:2013-03-01
INSSI record number: 45907
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