Paula S. Duggan

Cyanobacteria–bryophyte symbioses

Cyanobacteria are a large group of photosynthetic prokaryotes of enormous environmental importance, being responsible for a large proportion of global CO(2) and N(2) fixation. They form symbiotic associations with a wide range of eukaryotic hosts including plants, fungi, sponges, and protists. The cyanobacterial symbionts are often filamentous and fix N(2) in specialized cells known as heterocysts, enabling them to provide the host with fixed nitrogen and, in the case of non-photosynthetic hosts, with fixed carbon. The best studied cyanobacterial symbioses are those with plants, in which the cyanobacteria can infect the roots, stems, leaves, and, in the case of the liverworts and hornworts, the subject of this review, the thallus. The symbionts are usually Nostoc spp. that gain entry to the host by means of specialized motile filaments known as hormogonia. The host plant releases chemical signals that stimulate hormogonia formation and, by chemoattraction, guide the hormogonia to the point of entry into the plant tissue. Inside the symbiotic cavity, host signals inhibit further hormogonia formation and stimulate heterocyst development and dinitrogen fixation. The cyanobionts undergo morphological and physiological changes, including reduced growth rate and CO(2) fixation, and enhanced N(2) fixation, and release to the plant much of the dinitrogen fixed. This short review summarizes knowledge of the cyanobacterial symbioses with liverworts and hornworts, with particular emphasis on the importance of pili and gliding motility for the symbiotic competence of hormogonia.

Fate of genetically modified maize DNA in the oral cavity and rumen of sheep

The polymerase chain reaction (PCR) technique was used to investigate the fate of a transgene in the rumen of sheep fed silage and maize grains from an insect-resistant maize line. A 1914-bp DNA fragment containing the entire coding region of the synthetic cryIA(b) gene was still amplifiable from rumen fluid sampled 5 h after feeding maize grains. The same target sequence, however, could not be amplified from rumen fluid sampled from sheep fed silage prepared from the genetically modified maize line. PCR amplification of a shorter (211-bp), yet still highly specific, target sequence was possible with rumen fluid sampled up to 3 and 24 h after feeding silage and maize grains, respectively. These findings indicate that intact transgenes from silage are unlikely to survive significantly in the rumen since a DNA sequence 211-bp long is very unlikely to transmit genetic information. By contrast, DNA in maize grains persists for a significant time and may, therefore, provide a source of transforming DNA in the rumen. In addition, we have examined the biological activity of plasmid DNA that had previously been exposed to the ovine oral cavity. Plasmid extracted from saliva sampled after incubation for 8 min was still capable of transforming competent Escherichia coli to kanamycin resistance, implying that DNA released from the diet within the mouth may retain sufficient biological activity for the transformation of competent oral bacteria.

Molecular Analysis of Genes in Nostoc punctiforme Involved in Pilus Biogenesis and Plant Infection

Hormogonia are the infective agents in many cyanobacterium-plant symbioses. Pilus-like appendages are expressed on the hormogonium surface, and mutations in pil-like genes altered surface piliation and reduced symbiotic competency. This is the first molecular evidence that pilus biogenesis in a filamentous cyanobacterium requires a type IV pilus system.

Signalling in Cyanobacteria–Plant Symbioses

Cyanobacteria are a morphologically diverse and widespread group of phototrophic bacteria, many of which are capable of nitrogen fixation. They form symbioses with a wide range of eukaryotic hosts including fungi (lichens and Geosiphon pyriformis), diatoms, dinoflagellates, sponges, ascidians (sea squirts), corals and plants. The best understood are the plant symbioses, which are the subject of this chapter. In the cyanobacteria–plant associations, the cyanobacteria provide the host with fixed nitrogen and usually adopt a heterotrophic form of nutrition, using fixed carbon supplied by the plant, enabling them to occupy regions of the host, such as the roots, that receive little or no light. Most cyanobacterial symbionts of plants belong to the genus Nostoc, members of which fix nitrogen in specialised cells known as heterocysts, which provide the necessary microoxic environment for the functioning of the oxygen-sensitive enzyme nitrogenase. These cyanobacteria, which are immotile for most of their life cycles, produce specialised motile filaments known as hormogonia, as a means of dispersal and as the infective agents in plant symbioses. Host plants improve their chances of infection by releasing external chemical signals that both stimulate hormogonia formation and serve as chemoattractants. However, within the symbiotic tissue the plant releases hormogonia-repressing factors to ensure the conversion of hormogonia into heterocyst-containing, nitrogen-fixing filaments.

Mutation at Different Sites in the Nostoc punctiforme cyaC Gene, Encoding the Multiple-Domain Enzyme Adenylate Cyclase, Results in Different Levels of Infection of the Host Plant Blasia pusilla

The filamentous cyanobacterium Nostoc punctiforme forms symbioses with plants. Disruption of the catalytic domain of the N. punctiforme adenylate cyclase (CyaC) significantly increased symbiotic competence, whereas reduced infectivity was observed in a mutant with a disruption close to the N terminus of CyaC. The total cellular cyclic AMP levels were significantly reduced in both mutants.

Symbiosis between the cyanobacterium Nostoc and the liverwort Blasia requires a CheR-type MCP methyltransferase

In response to environmental change, the cyanobacterium Nostoc punctiforme ATCC 29133 produces highly adapted filaments known as hormogonia that have gliding motility and serve as the agents of infection in symbioses with plants. Hormogonia sense and respond to unidentified plant-derived chemical signals that attract and guide them towards the symbiotic tissues of the host. There is increasing evidence to suggest that their interaction with host plants is regulated by chemotaxis-related signal transduction systems. The genome of N. punctiforme contains multiple sets of chemotaxis (che)-like genes. In this study we characterize the large che5 locus of N. punctiforme. Disruption of NpR0248, which encodes a putative CheR methyltransferase, results in loss of motility and significantly impairs symbiotic competency with the liverwort Blasia pusilla when compared with the parent strain. Our results suggest that chemotaxis-like elements regulate hormogonia function and hence symbiotic competency in this system.