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- Frenkel, Martin, et al.
(author)
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An illustrated gardener's guide to transgenic Arabidopsis field experiments
- 2008
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In: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 180:2, s. 545-555
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Journal article (peer-reviewed)abstract
- Field studies with transgenic Arabidopsislines have been performed over 8 yr, to better understand the influence that certain genes have on plant performance. Many (if not most) plant phenotypes cannot be observed under the near constant, low-stress conditions in growth chambers, making field experiments necessary. However, there are challenges in performing such experiments: permission must be obtained and regulations obeyed, the profound influence of uncontrollable biotic and abiotic factors has to be considered, and experimental design has to be strictly controlled. The aim here is to provide inspiration and guidelines for researchers who are not used to setting up such experiments, allowing others to learn from our mistakes. This is believed to be the first example of a ‘manual’ for field experiments with transgenic Arabidopsisplants. Many of the challenges encountered are common for all field experiments, and many researchers from ecological backgrounds are skilled in such methods. There is huge potential in combining the detailed mechanistic understanding of molecular biologists with ecologists’ expertise in examining plant performance under field conditions, and it is suggested that more interdisciplinary collaborations will open up new scientific avenues to aid analyses of the roles of genetic and physiological variation in natural systems.
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- Frenkel, Martin, et al.
(author)
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Improper excess light energy dissipation in Arabidopsis results in a metabolic reprogramming
- 2009
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In: BMC Plant Biology. - : Springer Science and Business Media LLC. - 1471-2229. ; 9:12, s. 1-16
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Journal article (peer-reviewed)abstract
- Background: Plant performance is affected by the level of expression of PsbS, a key photoprotective protein involved in the process of feedback de-excitation (FDE), or the qE component of non-photochemical quenching, NPQ. Results: In studies presented here, under constant laboratory conditions the metabolite profiles of leaves of wild-type Arabidopsis thaliana and plants lacking or overexpressing PsbS were very similar, but under natural conditions their differences in levels of PsbS expression were associated with major changes in metabolite profiles. Some carbohydrates and amino acids differed ten-fold in abundance between PsbS-lacking mutants and over-expressers, with wild-type plants having intermediate amounts, showing that a metabolic shift had occurred. The transcriptomes of the genotypes also varied under field conditions, and the genes induced in plants lacking PsbS were similar to those reportedly induced in plants exposed to ozone stress or treated with methyl jasmonate (MeJA). Genes involved in the biosynthesis of JA were up-regulated, and enzymes involved in this pathway accumulated. JA levels in the undamaged leaves of field-grown plants did not differ between wild-type and PsbS-lacking mutants, but they were higher in the mutants when they were exposed to herbivory. Conclusion: These findings suggest that lack of FDE results in increased photooxidative stress in the chloroplasts of Arabidopsis plants grown in the field, which elicits a response at the transcriptome level, causing a redirection of metabolism from growth towards defence that resembles a MeJA/JA response.
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5. |
- Johansson Jänkänpää, Hanna, 1981-, et al.
(author)
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Arabidopsis thaliana with reduced capacity for non-photochemical quenching show altered herbivore preference
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Other publication (other academic/artistic)abstract
- Under natural conditions, plants have to cope with a multitude of stresses, two of those are light-stress and herbivory. Plants have evolved several mechanisms to avoid the damage done by strong and fluctuating light and one important photoprotection mechanism is the qE-type of non-photochemical quenching (NPQ) where the PsbS protein is involved. We have compared Arabidopsis thaliana wild type and two "photoprotection genotypes", npq4 and oePsbS that, respectively, lack and overexpress PsbS. In dual-choice feeding experiments on field-grown plants with a specialist (Plutella xylostella) and a generalist (Spodoptera littoralis) insect herbivore, both herbivores preferred the plants with higher expression of PsbS. Also both herbivores survived equally well on the different genotypes but for oviposition, female adults of Plutella xylostella preferred plants with lower expression of PsbS. No difference in the amount and composition of the ten most prominent glucosinolates — the most important substances in the Arabidopsis chemical warfare against herbivores — were found between the genotypes. When leaves of the three genotypes, after transfer from a growth chamber to the field, were profiled for changes in composition of metabolites using GC-MS, we found significant differences in metabolite composition. This suggests that differences in herbivore preferences were rather a consequence of changes in the primary metabolism of the plant rather than differences in the composition of typical "defence compounds". In npq4, superoxide accumulated under high light conditions and is likely to directly, or indirectly after dismutation to H2O2, trigger the metabolic change.
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6. |
- Johansson Jänkänpää, Hanna, et al.
(author)
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Metabolic profiling reveals metabolic shifts in Arabidopsis plants grown under different light conditions
- 2012
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In: Plant, Cell and Environment. - : John Wiley & Sons. - 0140-7791 .- 1365-3040. ; 35:10, s. 1824-1836
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Journal article (peer-reviewed)abstract
- Plants have tremendous capacity to adjust their morphology, physiology and metabolism in response to changes in growing conditions. Thus, analysis solely of plants grown under constant conditions may give partial or misleading indications of their responses to the fluctuating natural conditions in which they evolved. To obtain data on growth-condition dependent differences in metabolite levels we compared leaf metabolite profiles of Arabidopsis thaliana growing under three constant laboratory light conditions: 30 (LL), 300 (NL) and 600 (HL) µmol photons m(-2) s(-1) . We also shifted plants to the field and followed their metabolite composition for three days. Numerous compounds showed light-intensity dependent accumulation, including: many sugars and sugar derivatives (fructose, sucrose, glucose, galactose and raffinose); tricarboxylic acid (TCA) cycle intermediates and amino acids (ca. 30% of which were more abundant under HL and 60% under LL). However, the patterns differed after shifting NL plants to field conditions. Levels of most identified metabolites (mainly amino acids, sugars and TCA cycle intermediates) rose after 2 h and peaked after 73 h, indicative of a "biphasic response" and "circadian" effects. The results provide new insight into metabolomic level mechanisms of plant acclimation, and highlight the role of known protectants under natural conditions.
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7. |
- Johansson Jänkänpää, Hanna, et al.
(author)
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Non-photochemical quenching capacity in arabidopsis thaliana affects herbivore behaviour
- 2013
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In: PLOS ONE. - : Public Library of Science (PLoS). - 1932-6203. ; 8:1, s. e53232-
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Journal article (peer-reviewed)abstract
- Under natural conditions, plants have to cope with numerous stresses, including light-stress and herbivory. This raises intriguing questions regarding possible trade-offs between stress defences and growth. As part of a program designed to address these questions we have compared herbivory defences and damage in wild type Arabidopsis thaliana and two "photoprotection genotypes", npq4 and oePsbS, which respectively lack and overexpress PsbS (a protein that plays a key role in qE-type non-photochemical quenching). In dual-choice feeding experiments both a specialist (Plutella xylostella) and a generalist (Spodoptera littoralis) insect herbivore preferred plants that expressed PsbS most strongly. In contrast, although both herbivores survived equally well on each of the genotypes, for oviposition female P. xylostella adults preferred plants that expressed PsbS least strongly. However, there were no significant differences between the genotypes in levels of the 10 most prominent glucosinolates; key substances in the Arabidopsis anti-herbivore chemical defence arsenal. After transfer from a growth chamber to the field we detected significant differences in the genotypes' metabolomic profiles at all tested time points, using GC-MS, but no consistent "metabolic signature'' for the lack of PsbS. These findings suggest that the observed differences in herbivore preferences were due to differences in the primary metabolism of the plants rather than their contents of typical "defence compounds". A potentially significant factor is that superoxide accumulated most rapidly and to the highest levels under high light conditions in npq4 mutants. This could trigger changes in planta that are sensed by herbivores either directly or indirectly, following its dismutation to H2O2.
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8. |
- Johansson Jänkänpää, Hanna, 1981-
(author)
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Stress responses of Arabidopsis plants with a varying level of non-photochemical quenching
- 2011
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Doctoral thesis (other academic/artistic)abstract
- When light energy input exceeds the capacity for photosynthesis the plant need to dissipate the excess energy and this is done through non-photo-chemical quenching (NPQ). Photochemical quenching (photosynthesis), NPQ and fluorescence are three alternative faiths of excited chlorophylls. PsbS associates to photosystem II and is involved in NPQ.The results presented in this thesis were generated on Arabidopsis plants and mainly based on wildtype Col-0 together with a mutant deficient in PsbS (npq4) and a transgene overexpressing PsbS (oePsbS). We connect light and herbivore stress and show that the level of PsbS influences the food preference of both a specialist (Plutella) and a generalist (Spodoptera) herbivore as well as oviposition of Plutella. Level of PsbS also affects both metabolomics and transcriptomics of the plant; up-regulation of genes in the jasmonic acid (JA) -pathway and amount of JA has been found in the npq4 plants after herbivory.Since many experiments were performed in field we have also characterized the field plant and how it differs from the commonly used lab plant. We have also studied the natural variation of NPQ in Arabidopsis plants both in the field and the lab. The results show surprisingly no correlation.
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9. |
- Misra, Yogesh, et al.
(author)
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Arabidopsis Plants Grown in the Field and Climate Chambers Significantly Differ in Leaf Morphology and Photosystem Components
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Other publication (other academic/artistic)abstract
- Background: Plants exhibit phenotypic plasticity and respond to differences in environmental conditions by acclimation. We have systematically compared leaves of Arabidopsis thaliana plants grown in the field and under controlled low, normal and high light conditions in the laboratory to determine their most prominent phenotypic differences. Results: Compared to plants grown under field conditions, the ―indoor plants‖ had larger leaves, modified leaf shapes and longer petioles. Their pigment composition also significantly differed; indoor plants had reduced levels of xanthophyll pigments. In addition, Lhcb1 and Lhcb2 levels were up to three times higher in the indoor plants, but differences in the PSI antenna were much smaller, with only the low-abundance Lhca5 protein showing altered levels. Both isoforms of early-light-induced protein (ELIP) were absent in the indoor plants, and they had less non-photochemical quenching (NPQ). The field-grown plants had a high capacity to perform state transitions. Plants lacking ELIPs did not have reduced growth or seed set rates, but their mortality rates were sometimes higher. NPQ levels between natural ecotypes grown under different conditions were not correlated. Conclusion: Our results indicate that comparative analysis of field-grown plants with those grown under artificial conditions is important for a full understanding of plant plasticity and adaptation.
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10. |
- Wagner, Raik, 1976-, et al.
(author)
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Fitness analyses of Arabidopsis thaliana mutants depleted of FtsH metalloproteases and characterization of three FtsH6 deletion mutants exposed to high light stress, senescence and chilling
- 2011
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In: New Phytologist. - : Wiley. - 0028-646X .- 1469-8137. ; 191:2, s. 449-458
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Journal article (peer-reviewed)abstract
- Darwinian fitness analyses were performed, comparing single ftsh mutants with wild-type Arabidopsis thaliana plants grown under controlled laboratory conditions and in the field, by measuring plant size, survival rate, and silique and seed production. Additionally, three genotypes of ΔFtsH6 were analysed, under controlled growth conditions, with respect to both their ability to degrade the light-harvesting complex of photosystem II during senescence and light acclimation. In the field, substantial increases in variegation and reductions in growth were observed in the ΔFtsH2, ΔFtsH5 and ΔFtsH10 mutants; FtsH2 seemed particularly important for plant survival. Despite being grown in relatively cold weather, the ΔFtsH11 mutant displayed strong phenotypic deviations from wild type. Both ΔFtsH10 and ΔFtsH3 mutants exhibited less severe phenotypic changes, but were different from wild-type plants when placed in the field as young plants. When older ΔFtsH3 or ΔFtsH10 mutants were placed outdoors, no phenotypic differences from wild type were observed. Three genotypes of ΔFtsH6 displayed no phenotypic deviations from wild-type plants. Under controlled growth conditions, during senescence and light acclimation, no differences in the amount of chlorophyll or Photosystem II light-harvesting complex b3 (Lhcb3) were detected in ΔFtsH6 mutants compared with the wild type. Therefore, FtsH6 seems to be unimportant for LHCII degradation in vivo.
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