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This account presents information on all aspects of the biology of Robinia pseudoacacia L. that are relevant to understanding its ecological characteristics and behaviour. The main topics are presented within the standard framework of the Biological Flora of the British Isles: distribution, habitat, communities, responses to biotic factors, responses to environment, structure and physiology, phenology, floral and seed characters, herbivores and disease, and history and conservation.Robinia pseudoacacia, false acacia or black locust, is a deciduous, broad-leaved tree native to North America. The medium-sized, fast-growing tree is armed with spines, and extensively suckering. It has become naturalized in grassland, semi-natural woodlands and urban habitats. The tree is common in the south of the British Isles and in many other regions of Europe.Robinia pseudoacacia is a light-demanding pioneer species, which occurs primarily in disturbed sites on fertile to poor soils. The tree does not tolerate wet or compacted soils. In contrast to its native range, where it rapidly colonizes forest gaps and is replaced after 15-30years by more competitive tree species, populations in the secondary range can persist for a longer time, probably due to release from natural enemies.Robinia pseudoacacia reproduces sexually, and asexually by underground runners. Disturbance favours clonal growth and leads to an increase in the number of ramets. Mechanical stem damage and fires also lead to increased clonal recruitment. The tree benefits from di-nitrogen fixation associated with symbiotic rhizobia in root nodules. Estimated symbiotic nitrogen fixation rates range widely from 23 to 300kgha(-1)year(-1). The nitrogen becomes available to other plants mainly by the rapid decay of nitrogen-rich leaves.Robinia pseudoacacia is host to a wide range of fungi both in the native and introduced ranges. Megaherbivores are of minor significance in Europe but browsing by ungulates occurs in the native range. Among insects, the North American black locust gall midge (Obolodiplosis robiniae) is specific to Robinia and is spreading rapidly throughout Europe. In parts of Europe, Robinia pseudoacacia is considered an invasive non-indigenous plant and the tree is controlled. Negative impacts include shading and changes of soil conditions as a result of nitrogen fixation.
During this PhD project three technical platforms were either improved or newly established in order to identify interesting genes involved in SNF, validate their expression and functionally characterise them. An existing 5.6K cDNA array (Colebatch et al., 2004) was extended to produce the 9.6K LjNEST array, while a second array, the 11.6K LjKDRI array, was also produced. Furthermore, the protocol for array hybridisation was substantially improved (Ott et al., in press). After functional classification of all clones according to the MIPS database and annotation of their corresponding tentative consensus sequence (TIGR) these cDNA arrays were used by several international collaborators and by our group (Krusell et al., 2005; in press). To confirm results obtained from the cDNA array analysis different sets of cDNA pools were generated that facilitate rapid qRT-PCR analysis of candidate gene expression. As stable transformation of Lotus japonicus takes several months, an Agrobacterium rhizogenes transformation system was established in the lab and growth conditions for screening transformants for symbiotic phenotypes were improved. These platforms enable us to identify genes, validate their expression and functionally characterise them in the minimum of time. The resources that I helped to establish, were used in collaboration with other people to characterise several genes like the potassium transporter LjKup and the sulphate transporter LjSst1, that were transcriptionally induced in nodules compared to uninfected roots, in more detail (Desbrosses et al., 2004; Krusell et al., 2005). Another gene that was studied in detail was LjAox1. This gene was identified during cDNA array experiments and detailed expression analysis revealed a strong and early induction of the gene during nodulation with high expression in young nodules which declines with the age of the nodule. Therefore, LjAox1 is an early nodulin. Promoter:gus fusions revealed an LjAox1 expression around the nodule endodermis. The physiological role of LjAox1 is currently being persued via RNAi. Using RNA interference, the synthesis of all symbiotic leghemoglobins was silenced simultaneously in Lotus japonicus. As a result, growth of LbRNAi lines was severely inhibited compared to wild-type plants when plants were grown under symbiotic conditions in the absence of mineral nitrogen. The nodules of these plants were arrested in growth 14 post inoculation and lacked the characteristic pinkish colour. Growing these transgenic plants in conditions where reduced nitrogen is available for the plant led to normal plant growth and development. This demonstrates that leghemoglobins are not required for plant development per se, and proves for the first time that leghemoglobins are indispensable for symbiotic nitrogen fixation. Absence of leghemoglobins in LbRNAi nodules led to significant increases in free-oxygen concentrations throughout the nodules, a decrease in energy status as reflected by the ATP/ADP ratio, and an absence of the bacterial nitrogenase protein. The bacterial population within nodules of LbRNAi plants was slightly reduced. Alterations of plant nitrogen and carbon metabolism in LbRNAi nodules was reflected in changes in amino acid composition and starch deposition (Ott et al., 2005). These data provide strong evidence that nodule leghemoglobins function as oxygen transporters that facilitate high flux rates of oxygen to the sites of respiration at low free oxygen concentrations within the infected cells.