Tutkimus kevätrypsin (Brassica campestris L. var. annua) rasvahappojen analytiikasta ja jalostuksesta

The fatty acid composition of the seed oil of summer turnip rape (Brassica campestris L. var. annua) was investigated by gas liquid chromatography (GLC). The reliability of conventional sampling methods in capillary GC was compared with that of the new on column and PTV (programmed temperature vaporizer) techniques, with particular reference to the determination of fatty acid variation. In order to develop new, well-adapted turnip rape strains with improved oil quality, a breeding programme for a higher linoleic acid content, based on individual plant selection, was performed in 1978—85. The results showed that the conventional sampling techniques involving sample transfer to a hot injector were very unreliable as regards precision and accuracy. This was especially the case in the determination of trace fatty acid levels. The PTV methods with splitless and solvent split mode were as precise as cold on-column injection. The PTV sampling modifications, which are all superior to classical techniques, were even more suitable for routine analyses than on-column injection, where several restrictions are met. The analytical error with PTV for most of the compounds represented less than 1 % of the variation found for fatty acids within a turnip rape variety. The breeding experiments indicated that the level of linoleic acid can be increased under open-pollinated conditions in the field without affecting the α-linolenic acid content. The greenhouse conditions, on the other hand, were found to have a considerable environmental influence on the variation of these compounds, resulting in no response to linoleic-acid selection. In field trials, several strains with yields comparable to the varieties commonly cultivated in Finland were selected with a higher linoleic acid content (up to25 %). Most of them also contained no erucic acid. The new evidence concerning its beneficial physiological effects indicate that rapeseed oil should be considered as a serious alternative among sources of essential fatty acids. Such aspects should also be taken into account in future breeding of rapeseed fatty acids.Kotimainen rypsiöljy on viimeisen kymmenen vuoden aikana saanut yhä tärkeämmän aseman elintarviketeollisuuden raaka-aineena. Nykyisistä rypsilajikkeista saatava öljy on varsin kilpailukykyistä, sillä erityisesti erukahappopitoisuuden suhteen laatuvaatimukset meillä ovat varsin tiukat. Viljelyä ei voida kuitenkaan suuressa määrin lisätä, sillä jalostavan teollisuuden mahdollisuudet korvata tuontiöljyjä kotimaisella raaka-aineella ovat suhteellisen rajoitetut johtuen ensisijaisesti rypsiöljyn alhaisesta linolihappopitoisuudesta. Tämän tutkimuksen tarkoituksena oli yksilövalintaa apuna käyttäen kohottaa kevätrypsin siemenöljyn linolihappopitoisuutta sekä lutkia valinnanvaikutuksia rasvahappokoostumukseen ja satoisuuteen. Tutkimuksessa verrattiin edelleen perinteisten ja uusimpien kaasukromatografisten menetelmien luotettavuutta rasvahappoanalytiikassa ja pyrittiin optimoimaan jalostustyöhön parhaiten soveltuva analyysitekniikka. Jalostuskokeet tehtiin Hankkijan kasvinjalostuslaitoksella Hyrylässä vuosina 1978—85 kanadalaista alkuperää olevasta kevätrypsiaineistosta. Linolihappolinjojen lopullinen karsinta suoritettiin satoisuuden perusteella. Vapaapölytteisissä pelto-oloissa tehdyt jalostuskokeet osoittivat, että linolihappotasoa on mahdollista kohottaa yksilövalinnalla vaikuttamatta kuitenkaan toisen monityydyttämättömän komponentin, α-linoleenihapon, määrään. Parhaat valintalinjat sisälsivät linolihappoa n. 25 %, minkä lisäksi useat olivat erukahapottomia ja satoisuudeltaan nykyisten lajikkeiden veroisia. Kasvihuoneolosuhteiden todettiin sitä vastoin aiheuttavan huomattavaa lisävaihtelua, eikä valinnan vaikutusta linolihappotasoon voitu osoittaa. Pyrittäessä edelleen kohottamaan rypsiöljyn linolihappopitoisuutta saattaisi alhaisen α-linoleenihappotason huomioiminen yksilövalinnassa samanaikaisesti olla merkityksellistä linolihappovalinnan tehostamiseksi. Alhaisesta monityydyttämättömien rasvahappojen määrästä huolimalta rypsiöljyllä on todettu olevan varsin edulliset fysiologiset vaikutukset eritoten kolesterolitasoon. Tässä valossa on rypsiöljyä pidettävä vakavasti otettavana vaihtoehtona välttämättömien rasvahappojen lähteenä, mikä myös tulevaisuudessa on huomioitava rasvahappokoostumusta jalostettaessa

An analytical and breeding study on fatty acids in summer turnip rape (brassica campestris L. var. annua)

The fatty acid composition of the seed oil of summer turnip rape (Brassica campestris L. var. annua) was investigated by gas liquid chromatography (GLC). The reliability of conventional sampling methods in capillary GC was compared with that of the new on column and PTV (programmed temperature vaporizer) techniques, with particular reference to the determination of fatty acid variation. In order to develop new, well-adapted turnip rape strains with improved oil quality, a breeding programme for a higher linoleic acid content, based on individual plant selection, was performed in 1978—85. The results showed that the conventional sampling techniques involving sample transfer to a hot injector were very unreliable as regards precision and accuracy. This was especially the case in the determination of trace fatty acid levels. The PTV methods with splitless and solvent split mode were as precise as cold on-column injection. The PTV sampling modifications, which are all superior to classical techniques, were even more suitable for routine analyses than on-column injection, where several restrictions are met. The analytical error with PTV for most of the compounds represented less than 1 % of the variation found for fatty acids within a turnip rape variety. The breeding experiments indicated that the level of linoleic acid can be increased under open-pollinated conditions in the field without affecting the α-linolenic acid content. The greenhouse conditions, on the other hand, were found to have a considerable environmental influence on the variation of these compounds, resulting in no response to linoleic-acid selection. In field trials, several strains with yields comparable to the varieties commonly cultivated in Finland were selected with a higher linoleic acid content (up to25 %). Most of them also contained no erucic acid. The new evidence concerning its beneficial physiological effects indicate that rapeseed oil should be considered as a serious alternative among sources of essential fatty acids. Such aspects should also be taken into account in future breeding of rapeseed fatty acids

Genetic variation and factors affecting the genetic structure of the lichenicolous fungus Heterocephalacria bachmannii (Filobasidiales, Basidiomycota)

Heterocephalacria bachmannii is a lichenicolous fungus that takes as hosts numerous lichen species of the genus Cladonia. In the present study we analyze whether the geographical distance, the host species or the host secondary metabolites determine the genetic structure of this parasite. To address the question, populations mainly from the Southern Europe, Southern Finland and the Azores were sampled. The specimens were collected from 20 different host species representing ten chemotypes. Three loci, ITS rDNA, LSU rDNA and mtSSU, were sequenced. The genetic structure was assessed by AMOVA, redundance analyses and Bayesian clustering methods. The results indicated that the host species and the host secondary metabolites are the most influential factors over the genetic structure of this lichenicolous fungus. In addition, the genetic structure of H. bachmannii was compared with that of one of its hosts, Cladonia rangiformis. The population structure of parasite and host were discordant. The contents in phenolic compounds and fatty acids of C. rangiformis were quantified in order to test whether it had some influence on the genetic structure of the species. But no correlation was found with the genetic clusters of H. bachmannii.Peer reviewe

Methyljasmonate Elicitation Increases Terpenoid Indole Alkaloid Accumulation in Rhazya stricta Hairy Root Cultures

Methyl jasmonate is capable of initiating or improving the biosynthesis of secondary metabolites in plants and therefore has opened up a concept for the biosynthesis of valuable constituents. In this study, the effect of different doses of methyl jasmonate (MeJA) elicitation on the accumulation of terpenoid indole alkaloids (TIAs) in the hairy root cultures of the medicinal plant, Rhazya stricta throughout a time course (one-seven days) was investigated. Gas chromatography-mass spectrometry (GC-MS) analyses were carried out for targeted ten major non-polar alkaloids. Furthermore, overall alterations in metabolite contents in elicited and control cultures were investigated applying proton nuclear magnetic resonance (1H NMR) spectroscopy. Methyl jasmonate caused dosage- and time course-dependent significant rise in the accumulation of TIAs as determined by GC-MS. The contents of seven alkaloids including eburenine, quebrachamine, fluorocarpamine, pleiocarpamine, tubotaiwine, tetrahydroalstonine, and ajmalicine increased compared to non-elicited cultures. However, MeJA-elicitation did not induce the accumulation of vincanine, yohimbine (isomer II), and vallesiachotamine. Furthermore, principal component analysis (PCA) of 1H NMR metabolic profiles revealed a discrimination between elicited hairy roots and control cultures with significant increase in total vindoline-type alkaloid content and elevated levels of organic and amino acids. In addition, elicited and control samples had different sugar and fatty acid profiles, suggesting that MeJA also influences the primary metabolism of R. stricta hairy roots. It is evident that methyl jasmonate is applicable for elevating alkaloid accumulation in “hairy root” organ cultures of R. strica

Methyljasmonate Elicitation Increases Terpenoid Indole Alkaloid Accumulation in Rhazya stricta Hairy Root Cultures

Methyl jasmonate is capable of initiating or improving the biosynthesis of secondary metabolites in plants and therefore has opened up a concept for the biosynthesis of valuable constituents. In this study, the effect of different doses of methyl jasmonate (MeJA) elicitation on the accumulation of terpenoid indole alkaloids (TIAs) in the hairy root cultures of the medicinal plant, Rhazya stricta throughout a time course (one-seven days) was investigated. Gas chromatography-mass spectrometry (GC-MS) analyses were carried out for targeted ten major non-polar alkaloids. Furthermore, overall alterations in metabolite contents in elicited and control cultures were investigated applying proton nuclear magnetic resonance (1H NMR) spectroscopy. Methyl jasmonate caused dosage- and time course-dependent significant rise in the accumulation of TIAs as determined by GC-MS. The contents of seven alkaloids including eburenine, quebrachamine, fluorocarpamine, pleiocarpamine, tubotaiwine, tetrahydroalstonine, and ajmalicine increased compared to non-elicited cultures. However, MeJA-elicitation did not induce the accumulation of vincanine, yohimbine (isomer II), and vallesiachotamine. Furthermore, principal component analysis (PCA) of 1H NMR metabolic profiles revealed a discrimination between elicited hairy roots and control cultures with significant increase in total vindoline-type alkaloid content and elevated levels of organic and amino acids. In addition, elicited and control samples had different sugar and fatty acid profiles, suggesting that MeJA also influences the primary metabolism of R. stricta hairy roots. It is evident that methyl jasmonate is applicable for elevating alkaloid accumulation in “hairy root” organ cultures of R. strica

Methyljasmonate Elicitation Increases Terpenoid Indole Alkaloid Accumulation in Rhazya stricta Hairy Root Cultures

Methyl jasmonate is capable of initiating or improving the biosynthesis of secondary metabolites in plants and therefore has opened up a concept for the biosynthesis of valuable constituents. In this study, the effect of different doses of methyl jasmonate (MeJA) elicitation on the accumulation of terpenoid indole alkaloids (TIAs) in the hairy root cultures of the medicinal plant, Rhazya stricta throughout a time course (one-seven days) was investigated. Gas chromatography-mass spectrometry (GC-MS) analyses were carried out for targeted ten major non-polar alkaloids. Furthermore, overall alterations in metabolite contents in elicited and control cultures were investigated applying proton nuclear magnetic resonance (H-1 NMR) spectroscopy. Methyl jasmonate caused dosage- and time course-dependent significant rise in the accumulation of TIAs as determined by GC-MS. The contents of seven alkaloids including eburenine, quebrachamine, fluorocarpamine, pleiocarpamine, tubotaiwine, tetrahydroalstonine, and ajmalicine increased compared to non-elicited cultures. However, MeJA-elicitation did not induce the accumulation of vincanine, yohimbine (isomer II), and vallesiachotamine. Furthermore, principal component analysis (PCA) of H-1 NMR metabolic profiles revealed a discrimination between elicited hairy roots and control cultures with significant increase in total vindoline-type alkaloid content and elevated levels of organic and amino acids. In addition, elicited and control samples had different sugar and fatty acid profiles, suggesting that MeJA also influences the primary metabolism of R. stricta hairy roots. It is evident that methyl jasmonate is applicable for elevating alkaloid accumulation in "hairy root" organ cultures of R. strica.Peer reviewe

A simplified procedure for indole alkaloid extraction from Catharanthus roseus combined with a semi-synthetic production process for vinblastine

Peer reviewe

HPLC-DAD and UHPLC/QTOF-MS Analysis of Polyphenols in Extracts of the African Species Combretum padoides, C. zeyheri and C. psidioides Related to Their Antimycobacterial Activity

Combretum padoides Engl. & Diels, C. psidioides Welv. and C. zeyheri Sond. are used for the treatment of infections and tuberculosis related symptoms in African traditional medicine. In order to verify these uses, extracts were screened for their growth inhibitory effects against M. smegmatis ATCC 14468. Ultra-high pressure liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (UHPLC/QTOF-MS) and GC-MS were used to investigate the polyphenolic composition in the active extracts. The lowest minimum inhibitory concentration (MIC), 625 µg/mL, was shown by a methanol extract of the stem bark of C. psidioides. A butanol extract of C. psidioides gave large inhibition zone diameters (IZD 21 mm) and inhibited 84% of the mycobacterial growth at 312 µg/mL. Combretastatin B-2 and dihydrostilbene derivatives were present in the methanol extract of C. psidioides, whereas the butanol extract of this species contained punicalagin, corilagin, and sanguiin H-4. Methanol and butanol extracts of the stem bark of C. padoides gave large inhibition zone diameters (IZD 26.5 mm) and MIC values of 1250 and 2500 µg/mL, respectively. C. padoides contained an ellagitannin with a mass identical to punicalagin ([M-H]− 1083.0587) and a corilagin like derivative ([M-H]− 633.0750) as well as ellagic acid arabinoside and methyl ellagic acid xyloside. A butanol extract of the roots of C. zeyheri showed mild antimycobacterial activity and contained a gallotannin at m/z [M-H]− 647.0894 as the main compound along with punicalagin and three unknown ellagitannins at m/z [M-H]− 763.0788, 765.0566, and 817.4212. Our results indicate that the studied species of Combretum contain phenolic and polyphenolic compounds with possible potential as leads for antimycobacterial drugs or as adjuvants for conventional anti-TB drugs

Chemical Profiling and Bioactivity of Body Wall Lipids from Strongylocentrotus droebachiensis

The lipids from gonads and polyhydroxynaphthoquinone pigments from body walls of sea urchins are intensively studied. However, little is known about the body wall (BW) lipids. Ethanol extract (55 degrees C) contained about equal amounts of saturated (SaFA) and monounsaturated fatty acids (MUFA) representing 60% of total fatty acids, with myristic, palmitic and eicosenoic acids as major SaFAs and MUFAs, respectively. Non-methylene-interrupted dienes (13%) were composed of eicosadienoic and docosadienoic acids. Long-chain polyunsaturated fatty acids (LC-PUFA) included two main components, n6 arachidonic and n3 eicosapentaenoic acids, even with equal concentrations (15 mu g/mg) and a balanced n6/n3 PUFA ratio (0.86). The UPLC-ELSD analysis showed that a great majority of the lipids (80%) in the ethanolic extract were phosphatidylcholine (60 mu g/mg) and phosphatidylethanolamine (40 mu g/mg), while the proportion of neutral lipids remained lower than 20%. In addition, alkoxyglycerol derivativeschimyl, selachyl, and batyl alcoholswere quantified. We have assumed that the mechanism of action of body wall lipids in the present study is via the inhibition of MAPK p38, COX-1, and COX-2. Our findings open the prospective to utilize this lipid fraction as a source for the development of drugs with anti-inflammatory activity.Peer reviewe

Analysis of indole alkaloids from Rhazya stricta hairy roots by ultra-performance liquid chromatography-mass spectrometry

Rhazya stricta Decne. (Apocynaceae) contains a large number of terpenoid indole alkaloids (TIAs). This study focused on the composition of alkaloids obtained from transformed hairy root cultures of R. stricta employing ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). In the UPLC-MS analyses, a total of 20 TIAs were identified from crude extracts. Eburenine and vincanine were the main alkaloids followed by polar glucoalkaloids, strictosidine lactam and strictosidine. Secodine-type alkaloids, tetrahydrosecodinol, tetrahydro- and dihydrosecodine were detected too. The occurrence of tetrahydrosecodinol was confirmed for the first time for R. stricta. Furthermore, two isomers of yohimbine, serpentine and vallesiachotamine were identified. The study shows that a characteristic pattern of biosynthetically related TIAs can be monitored in Rhazya hairy root crude extract by this chromatographic method.Peer reviewe