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Nematodes are among the most important pathogens in agriculture, greatly reducing crop biomass and yield. The direct effects of nematodes on above– and belowground plant parts are well known, but the broad range of indirect effects, especially on carbon (C) and phosphorus (P) cycles underground, remains unknown. For the first time, using soil zymography, we analyzed the indirect effects of Meloidogyne incognita cellobiohydrolase and phosphatase. The rhizosphere of lupine ( Lupinus polyphyllus L.), a species sensitive to pathogens with high P demand, was selected to study the activity, distribution and localization of two enzymes responsible for C and P cycling: The distribution patterns of cellobiohydrolase and phosphatase demonstrated that M. incognita induced the formation of knots as well as cluster roots, which corresponded to hotspot locations on zymogram images for both enzymes. Increased C release by nematode–infected roots into the soil led to a decrease in the overall activity of cellobiohydrolase and especially at hotspots (by ∼ 20 times). In contrast, the increased P demand of infected plants raised the phosphatase activity, leading to an increase in the rhizosphere extent around the roots and especially of the hotspot area (by 6 times). Remarkably, this 1 mm increase of rhizosphere extent in 2D equals a 2-fold increment in soil volume (3D) for nutrient mobilization. We conclude that nematode infection not only has direct effects by changing root morphology, but also induces a number of subsequent biochemical changes (e.g. enzyme activities and consequently nutrient mobilization) in the rhizosphere, affecting C and P cycling.

Mountainous grasslands are typically important habitats both for fauna and flora but increasingly suffer from invasions by neophytes (i.e. Lupinus polyphyllus Lindl.) in most German low mountain areas, which eventually threatens species richness. Regular defoliation is required to eliminate the invasion, however, at present options to handle the harvested biomass are limited. Integrated generation of solid fuel and biogas from biomass (IFBB) and anaerobic digestion (AD) are two possible options to utilise the biomass and convert it into energy. There is substantial environmental impact associated with the energy and resource usage during conversion of the biomass into fuel and during usage of fuels and co-products obtained. This study examines IFBB and AD to identify the best option in terms of environmental impacts and primary energy usage, also looking at alternatives for process parameters along the life cycle that would reduce environmental impacts. It was found that IFBB was a better option compared to AD, as it had higher environmental and primary energy savings across all grassland sites. Higher energy conversion efficiency of IFBB resulted in higher greenhouse gas (GHG) and energy savings, even though the energy usage for the processing steps were higher compared to AD. Biomass yield was positively related to the savings, providing better GHG and energy savings for grasslands containing invasive species. There were no savings in terms of acidification (AP) and eutrophication potential (EP) for both IFBB and AD, however AP and EP was lower using IFBB compared to AD. Hence, biomass originating from mountainous grasslands with lupine invasion could be effectively utilised with IFBB, as this option had lower environmental impacts and higher energy savings compared to AD. Biomass from non-invaded grasslands could also be converted effectively using IFBB, hence IFBB could be used to utilise the harvested biomass in the situation where the invasion is eliminated.

The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.

Lupines (Lupinus spp.) are a common legume found on western U.S. rangelands. Lupinus spp. may contain quinolizidine and/or piperidine alkaloids that can be toxic and/or teratogenic to grazing livestock. Lupinus leucophyllus and Lupinus polyphyllus in particular represent important species in the rangelands of southeast Washington and northeast Oregon. The objectives of this study were to identify the alkaloid profiles of these two species, and to explore the phylogenetic relationship among the different populations using amplified fragment length polymorphism (AFLP) markers. Two chemotypes were found among the 14 accessions of Lupinus leucophyllyus. Chemotype A contained anagyrine, a potent teratogen in cattle, and thus would pose a risk to cause crooked calf syndrome in grazing cattle, while Chemotype B did not contain anagyrine and poses no teratogenic risk. No alkaloids were detected in the L. polyphyllus plants collected in this area. Phylogenetic analysis showed that L. leucophyllus accessions with the same chemotype that were geographically proximal were closely grouped in the cladogram; however, accessions that were geographically proximal that represented different chemotypes did not cluster together. Taken together, these results suggest that for the studied species, chemotype is the principle factor in determining relatedness followed by geography.

A comprehensive understanding of the effects of invasive plants on native species requires identification of both the mechanisms of interaction and the spatial scale over which they act. Indirect interactions involving mobile organisms such as pollinators are likely to be scale-dependent, yet most studies examining effects of invasive species on pollination of native plants have considered effects across a single distance between interacting species. We examined the effects of the invasive herb Lupinus polyphyllus on pollination of two native herbs using multiple distances between the invasive and the natives. We recorded pollinator visitation and seed production in the native herbs Lotus corniculatus and Lychnis viscaria at 0, 5 m or 200 m away from L. polyphyllus. To reduce the influence of confounding factors, we used experimentally established populations of the invasive and potted individuals of the natives. In the immediate vicinity to L. polyphyllus, visitation to L. corniculatus was higher than 200 m away, and seed production per flower was higher than 5 m and 200 m away. In L. viscaria, bumblebee visitation was higher adjacent to L. polyphyllus than 5 m and 200 m away, but total pollinator visitation and reproductive success did not vary with distance. The results indicate that L. polyphyllus facilitates pollination of the native plants, and that this occurs at a very local spatial scale as effects dropped off already at a distance of 5 m. Presence of L. polyphyllus could benefit both pollinators and pollination of native herbs, and these positive effects should be considered along with likely negative effects due to resource competition. Moreover, the results illustrate the necessity to consider scale-dependent effects when assessing the impact of invasive flowering plants on native pollination interactions.

The alkaloid profile of white lupine Lupinus albus L. cv. ‘Boros’ and ‘Butan’, narrow-leaf lupine L. angustifolius L. cv. ‘Bojar’, ‘Graf’, ‘Karo’, ‘Mirela’, and ‘Sonet’, yellow lupine L. luteus L. cv. ‘Dukat’, ‘Parys’, ‘Perkoz’, and ‘Talar’, and wild species big-leaf lupine L. polyphyllus Lindl. was studied. The pea aphid Acyrthosiphon pisum Harris probing behaviour was monitored using the electrical penetration graph (EPG) technique. Four types of feeding behaviour were recognized. First, on highly acceptable L. luteus cv. ‘Dukat’, the total and mean duration of probing, time to reach phloem phase, and the duration of the first phloem phase were comparable to those in aphids on control plant Pisum sativum. Second, on partially acceptable L. luteus cv. ‘Talar’, pathway activities were slightly impeded, and the probes were more numerous and slightly shorter than on control plants. The phloem phase occurred sporadically, and feeding was terminated early after a brief period of ingestion. Third, on unpalatable but acceptable L. angustifolius cv. ‘Bojar’ and ‘Sonet’, and L. luteus cv. ‘Parys’ and ‘Perkoz’, the probes were numerous and usually epidermal. The phloem phase occurred rarely, and when it did, it was short and consisted mainly of watery salivation. Finally, on unacceptable L. albus cv. ‘Boros’ and ‘Butan’, L. angustifolius cv. ‘Graf’, ‘Karo’, ‘Mirela’, and L. polyphyllus, total probing time and individual probes were short and phloem phase did not occur. Eighteen alkaloids were identified: one piperidine alkaloid (ammodendrine), one indole (gramine), ten quinolizidine alkaloids (one tricyclic and nine tetracyclic compounds), and six esters. All lupine varieties that contained lupanine, its derivatives and especially their esters appeared to be unacceptable to the pea aphid, independent of the total concentration of any specific lupanine alkaloid. In contrast, sparteine and its derivatives did not seem to affect aphid probing significantly.

Biotin is an essential micronutrient needed but not synthesized by animals. We performed a field experiment where we measured biotin levels and the community composition of nematodes in the rhizosphere of two legumes, the native Lotus corniculatus and an invasive alien, Lupinus polyphyllus. The relative abundance of root-feeding nematodes and the biotin concentration in nematodes were lower in the rhizosphere of the alien. We are the first to show that biotin levels in indigenous nematode populations are associated with the host plant species, and that the difference may affect nematode community.

An increasing number of invasive species are changing ecosystems around the world. Road verges have commonly become the first footholds of non-native species in the new environments. Regularly mown road verges also offer habitats for meadow flora and fauna, which in Europe have suffered from the radical decline of semi-natural biotopes due to the agricultural modernization. We studied impacts of an invasive plant Lupinus polyphyllus on the plant and Lepidoptera species composition along road verges. The plant species composition was studied on 15 sites (with 1 m 2 quadrats) and butterflies and diurnal moths along 15 transects (with weekly censuses) in SE Finland, each site and transect representing equally lupine invaded verge and an adjacent non-lupine verge. The species richness and diversity of flora and the cover and species richness of low growing (

Potassium independent asparaginase has been purified from the testa of maturing seeds of Lupinus polyphyllus. The enzyme has a Mr of 71 × 103 and is composed of two subunits of Mr 35–36 × 103. Antisera raised against the purified asparaginase cross-reacted with a protein of similar characteristics isolated from the cotyledons and leaves of L. polyphyllus but no reaction was detected with extracts from the roots and nodules following Western blot analysis. Potassium dependent asparaginase was purified to a lesser extent from the testa of pea seeds and was shown to have a Mr of 61 × 103. The potassium independent and dependent enzymes appear to be distinct proteins with different immunological characteristics. The proportion of the two enzymes varies with plant species, organ and developmental age.

In the cell-free extracts of 10-day-old seedlings of L. hirsutus , the acyltransferase activities were detected for tigloyl-CoA: 13α-hydroxymultiflorine O -tigloyltransferase (HMTase) (EC 2.3.1.-), p -coumaroyl-CoA: epilupinine O - coumaroyltransferase (ECTase) (EC 2.3.1.-) and feruloyl-CoA: epilupinine O -feruloyltransferase (EFTase) (EC 2.3.1.-). The HMTase activity was predominantly distributed in hypocotyls and roots of the seedlings. These acyltransferases were partially purified by fractional precipitation with ammonium sulphate and by DEAE-Sepharose chromatography. There was a single HMTase, whose K m values were 37 μM for 13α-hydroxymultiflorine and 80 μM for tigloyl-CoA. HMTase also showed tigloyl-CoA: 13α-hydroxylupanine O -tigloyltransferase (HLTase) (EC 2.3.1.-) activity, although L. hirsutus contains no 13-hydroxylupanine-derived esters. There were two isoenzymes, on the contrary, showing both activities of ECTase and EFTase separated by DEAE-Sepharose chromatography. The activities of HMTase and HLTase were detected in L. hirsutus , L. termis and L. polyphyllus x L. arboreus . However, the activities of ECTase and EFTase were detected only in L. hirsutus .

Summary We isolated and sequenced a cDNA clone (pPLZ17) from a cDNA library of a cell suspension culture of Lupinus polyphyllus . Its nucleotide sequence of 1005 by shows homologies up to 73.9 % to «70 kDa heat shock protein» (HSP70) genes of unrelated organisms. The deduced amino acid sequence is 84 % and 79 % homologous to HSP70 of maize and petunia, respectively. mRNA analysis revealed that pPLZ17 not only encodes the lupin HSP70, but also has a high homology to a low molecular weight HSP transcript of approximately 1000 bp. Apparently, these genes are expressed constitutively in cell suspension culture and their expression is not enhanced by heat shock. In contrast, the HSP70 gene is thermally inducible in the intact lupin plant and callus culture.

Summary We isolated and sequenced a cDNA clone from a cell suspension culture of Lupinus polyphyllus, which encodes about 54 % (701 bp) of a basic peroxidase isoenzyme. The nucleotide sequence is 51.8 % homologous to a peroxidase gene of tobacco and the predicted amino acid sequence shows up to 80 % homology to peroxidase-specific domains of the already known isoenzymes from turnip, horseradish and tobacco. Expression studies indicate that the mature lupin peroxidase mRNA is approximately 1.3 kb in size and is expressed in suspension-cultured cells and in leaf and root tissue of intact lupin plants, but not in stem tissue.

Twenty-nine quinolizidine alkaloids have been found and characterized from the combined leaf/hypocotyl extracts of Lupinus polyphyllus. Twelve have not been found previously in this plant and the structure of 10 has been unambiguously assigned from a combination of mass spectroscopy and NMR data, and by direct comparison with synthetic material for the N-carboxymethyl ester, N-carboxyethyl ester and N-formyl derivatives of angustifoline. The co-occurrence of 13α-hydroxylupanine and its 13β-isomer is reported for the first time in a quinolizidine alkaloid-accumulating species.

Protocols for whole plant regeneration of two large seed legumes ( Lupinus angustifolius, L. polyphyllus ) have been developed. Plant regeneration was achieved from callus cultures via organogenic pathway. It was necessary to use 0.54 μM α-naphthalene acetic acid (NAA) and 4.40 μM 6-benzylaminopurine (BA) as well as basal constituents of the II-medium for multiple shoot-bud induction. Only NAA was needed in the rooting medium.

Rabbit antiserum was raised against potassium-independent asparaginase purified from Lupinus polyphyllus . A survey of 54 lines of Lupinus showed that only 11 contained the enzyme in maturing cotyledons with activities > 0.5 μmol/hr per g fr. wt. Potassium-dependent asparaginase activity was detected in a number of the remaining varieties.

Summary Cell suspension cultures of Lupinus polyphyllus form about 100 mmol/1(0.4 %) ethanol when entering the stationary growth phase. 50–70 % of the ethanol produced is released into the culture medium which occupies ca. 50 % of the total volume. The culture medium also contains various hydrolytic and oxidative enzymes. The activities of extracellular peroxidase, «sparteine oxidase», acid phosphatase, esterase, and protease increase during the active growth phase and decrease during stationary growth, when ethanol formation starts.

In leaves of alkaloid producing Lupinus polyphyllus lysine decarboxylase activity is positively correlated with the chlorophyll content during leaf regreening. A similar positive correlation was found between the leaf alkaloid content and lysine decarboxylase activity in L. albus and L. luteus . These results indicate that in lupin leaves lysine decarboxylase is an integrated part of the alkaloid specific biosynthetic sequence. Lysine decarboxylase could also be demonstrated in 46 alkaloidal and non-alkaloidal species out of 17 families of higher plants, including cell cultures of seven species.

Summary The distribution of the first two enzymes of the biosynthetic pathway of quinolizidine alkaloid biosynthesis, lysine decarboxylase and 17-oxosparteine synthase, was studied in the various organs of a flowering Lupinus polyphyllus plant. Lysine decarboxylase activity is present in all plant parts, whereas the activiy of the key enzyme 17-oxosparteine synthase is only expressed in leaf extracts. This indicates that alkaloid synthesis is bound to the leaves, where both enzyme had been localized in chloroplasts previously. The alkaloid content, alkaloid concentration and the alkaloid composition was quantitatively evaluated for the various organs of two flowering L. polyphyllus plants. The highest alkaloid content was found in the roots (38-45% of total alkaloids) and leaves (27-31%), but alkaloids are virtually present in all parts of a plant. Relatively high alkaloid concentrations were found in leaves, in younger parts of the shoot axis and in mature seeds. The alkaloid content of leaves is reduced during senescence by 90% as compared with young leaves. The alkaloid pattern evaluated quantitatively by capillary gas chromatography for the organs of two plants and seeds includes the following compounds: lupanine, sparteine, angustifoline, N-methylangustifoline, dehydrolupanine, 17-oxolupanine, 13-hydroxylupanine and its esters (13-tigloyl-, 13-angeloyl-, 13-benzoyl- and 13-cis and 13-transcinnamoyl-oxylupanine). A high percentage of ester alkaloids and a low content of 13-hydroxylupanine was found in leaves. Roots and seeds, the main sites of alkaloid accumulation, displayed the opposite pattern, i.e. high concentration of 13-hydroxylupanine and low concentration of ester alkaloids.