Call number: LD2668 .T4 1962 A48

Call number: LD2668 .T4 1962 W45

The Avena Coleoptile Test indicated that there were no indolyl- 3-acetic acid-like growth substances present in treated municipal wastewater from the Activated Sludge Sewage Treatment Plant in Tucson, Arizona. Using the bioassay of kinetin (6-furfurlyamino purine) as a standard in delaying chlorophyll degradation, wastewater extract was capable of maintaining chlorophyll in wheat (Triticum aestivum L.) leaf tissue; however, toxic contaminants present in municipal wastewater may have prevented a consistent dose response. In the process of repeated chromatography, kinetin broke down into two zones that were capable of retarding chlorophyll degradation. After repeated chromatography, wastewater extract and degradation products of kinetin resulted in similar Rf values. Wastewater extract obtained through repeated chromatography resulted in increased total chlorophyll retention, with increased concentrations; however, the nature of the cytokinin-like growth substances present in wastewater appeared to be unique. The high yields of high protein plant products obtained from plants grown with wastewater may have resulted from cytokinin-like growth substances present in treated municipal wastewater.

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The plant growth-promoting bacterium (PGPB) Pseudomonas putida UW4, previously isolated from the rhizosphere of common reeds growing on the campus of University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentration of salt, cold, heavy metals, drought and phytopathogens. The known mechanisms used by P. putida UW4 to promote plant growth include 1-aminocyclopropane-1-carboxylate (ACC) deaminase, indole-3-acetic acid (IAA) synthesis and siderophore production. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. putida UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,431 predicted protein-coding sequences that occupy 87.4% of the genome. Nineteen genomic islands were predicted and thirty one complete putative insertion sequences were identified. Genome analyses were conducted in order to better characterize the general features of the UW4 genome. Genes potentially involved in plant growth promotion such as IAA biosynthesis, trehalose production, siderophore production, and acetoin synthesis were identified, which will facilitate a better understanding of the mechanisms of plant-microbe interactions. Moreover, genes that contribute to the environmental fitness of UW4 were also determined including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Central metabolic pathways helped elucidate the physiological roles of diverse metabolites of UW4. Unexpectedly, whole-genome comparison with other completely sequenced Pseudomonas sp. revealed that UW4 is more similar to the fluorescens group rather than to the putida group. More surprisingly, a putative type III secretion system (T3SS) was found in the UW4 genome, and T3SS was thought to be essential for bacterial pathogenesis. Although putative T3SS was observed in other non-pathogenic Pseudomonas spp. previously, this is the first report indicating that a T3SS in a Pseudomonas sp. is highly similar to the one from Salmonella spp.

Bacteria that provide benefit to plants are considered to be plant growth-promoting bacteria (PGPB) and can facilitate plant growth by a number of different mechanisms. Plant growth-promoting bacteria that are able to utilize the plant compound 1-aminocyclopropane-1-carboxylate (ACC) as a sole source of nitrogen, as a consequence of possessing the enzyme ACC deaminase, can protect host plants from a number of environmental stresses. In addition to ACC deaminase, PGPB may utilize other mechanisms to facilitate plant growth including IAA synthesis, siderophore production, phosphate solubilization activity, ammonia production, and antibiotic production. Plant growth-promoting bacterial endophytes employ similar plant growth promotion mechanisms to those used by rhizospheric PGPB. In fact, bacterial endophytes are PGPB that go one step further and colonize the inside of the plant tissues and provide more efficient and prompted protection to their hosts compared to those that bind exclusively to the plant’s rhizosphere. Therefore, it is likely that endophytic plant growth-promoting bacteria will be superior to similar non-endophytic bacterial strains in promoting plant growth under a wide range of environmental conditions. In the work reported here, new bacterial endophytes were isolated and characterized. Among twenty-five ACC deaminase positive strains, two best strains were selected and ACC deaminase deficient mutants were constructed. The ability of two newly isolated 1-aminocyclopropane-1-carboxylate (ACC) deaminase-containing plant growth-promoting bacterial endophytes Pseudomonas fluorescens YsS6, Pseudomonas migulae 8R6 and their ACC deaminase deficient mutants was shown to 1) delay the senescence of mini carnation cut flowers and 2) to facilitate tomato plant growth under salinity stress. In the mini carnation flower senescence evaluation, the only difference between wild-type and mutant bacterial endophytes was ACC deaminase activity, our results demonstrate that this enzyme is directly responsible for a significant delay in flower senescence. Despite containing ACC deaminase activity, the rhizosphere-binding PGPB Pseudomonas putida UW4 was not taken up by the cut flowers and therefore had no effect on prolonging flower shelf life. In evaluating the effect of bacterial endophytes under salt stress, tomato plants treated with either of the wild-type strains of the two selected bacterial endophytes demonstrated early flowering and fruiting and had significantly greater numbers of flowers, buds, and fruits than either the corresponding ACC deaminase mutant strain-treated plants or the control plants. Although both bacterial endophytes P. fluorescens YsS6 and P. migulae 8R6 showed significant plant growth-promotion capabilities, P. migulae 8R6 demonstrated better plant growth facilitation under salt stress than did P. fluorescens YsS6. P. migulae 8R6 treated tomato plants demonstrated the least sodium uptake, the highest chlorophyll content, and highest fresh and dry biomass. The results of the work presented here suggest that ACC deaminase containing selected bacterial endophytes could be employed as environmentally friendly adjuncts to agricultural and horticultural practice.

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