No description available.

No description available.

E151 (sym15) is characterized as a pleiotropic pea (Pisum sativum L.) mutant. It has been described as having short lateral roots, a short primary root, and a shorter epicotyl than that of the wild type Sparkle. Furthermore, after 4 weeks of growth it was described as a low nodulator since nodulation was rare (Kneen et al., 1994). My main objective is to further characterize the nodulation phenotype of this mutant. Inoculated of the mutant with two separate strains of Rhizobium leguminosarum biovar viciae caused E151 to develop more nodules when infected with 8401 (lacZ) than with 128C53K. These results suggest that E151 exhibits different levels of susceptibility to infection with different strains. Nodule organogenesis was studied by inoculating E151 with a rhizobial strain that constitutively produces ß-galactosidase and then staining cleared whole root sections with the X-Gal substrate. The substrate produced a blue colour and allowed the rhizobial path to be visualized. It was found that nodule organogenesis in the mutant line is delayed at stage C (i.e., IT associated with cortical cell division) and eventually mature nodules form. The rates of N2 fixation (µmol N2/hr)/nodule dry weight (g) were found to be similar between Sparkle and E151 at 14, 21, and 28 days after inoculation. When comparing the nodulation defect of E151 to other pea mutants summarized by Guinel and Geil (2002), it appears that the nodulation phenotype of E151 is unique.

E151 (sym15) is characterized as a pleiotropic pea (Pisum sativum L.) mutant. It has been described as having short lateral roots, a short primary root, and a shorter epicotyl than that of the wild type Sparkle. Furthermore, after 4 weeks of growth it was described as a low nodulator since nodulation was rare (Kneen et al., 1994). My main objective is to further characterize the nodulation phenotype of this mutant. Inoculated of the mutant with two separate strains of Rhizobium leguminosarum biovar viciae caused E151 to develop more nodules when infected with 8401 (lacZ) than with 128C53K. These results suggest that E151 exhibits different levels of susceptibility to infection with different strains. Nodule organogenesis was studied by inoculating E151 with a rhizobial strain that constitutively produces ß-galactosidase and then staining cleared whole root sections with the X-Gal substrate. The substrate produced a blue colour and allowed the rhizobial path to be visualized. It was found that nodule organogenesis in the mutant line is delayed at stage C (i.e., IT associated with cortical cell division) and eventually mature nodules form. The rates of N2 fixation (µmol N2/hr)/nodule dry weight (g) were found to be similar between Sparkle and E151 at 14, 21, and 28 days after inoculation. When comparing the nodulation defect of E151 to other pea mutants summarized by Guinel and Geil (2002), it appears that the nodulation phenotype of E151 is unique.

Contemporary proximity extension assays (PEAs) are used for qualitative proteinquantifications in serological samples, with possibilities for scaling assays in multiplex. Medical research can however benefit from robust immunoassays functional for assessingprotein levels in other types of biospecimens. Formalin-fixed paraffin embedded (FFPE)tissues have long been used for morphological studies. The proteome encapsulated byextensive cross-linking from formalin fixation has however impeded the development ofproteomic analysis from the vast biorepositories FFPE-tissues constitute. In this study, Ipresent a proof of concept for assessing FFPE-samples in multiplex format through PEA.Furthermore, a homogenization and protein extraction protocol for assessing fresh-frozentissue with PEA is presented, together with a novel sample buffer for which remarkable risesin protein detection can be seen in several protein assays. Together, these findings extend theapplication area of PEA to tissues together with improved quantification characteristics.

Genes encoding representatives of two subfamilies from the vicilin storage protein family in pea (Pisum sativum L.) have been sequenced and characterised. One, encoding convicilin, shows that this protein differs from vicilin by the insertion of a hydrophilic region near the N-terminus. The transcription start point has been determinedand the pattern of expression in developing seeds elucidated. By theexpression of this gene in tobacco, the specific polypeptide product ofthe gene was identified as a minor component of convicilin , with a lowerMr than the major species . The other gene subfamily investigated wasThat encoding the vicilin 47,000 Mr polypeptide. A gene and a cDNA weresequenced, and the gene found to diverge from the cDNA in the 3" regionof the coding sequence. No product from this divergent gene could beidentified.A member of the legumin gene family (legK) was sequenced and found to be inactive due to a mutation of the start codon. The region of DNAencoding the start codon of this gene was amplified by polymerase chainreaction from a pea line in which the gene was known to be active . Thesequence of this revealed the presence of a normal start codon. Two dimensional protein gels were run with seed extracts from these twolines , and the product of (legK) demonstrated by its occurrence in theline with the functional gene. A method for the extraction and purification of the major pea root protein was established. The protein was shown to have a Mr of 16,000 and not to be susceptible to cleavage by cyanogen bromide. Partial amino acid sequence data was obtained from the purified protein .A differential screen of cDNA from purple and green poddedvarieties of pea was conducted, and differentially expressed cDNAsisolated . The nature of the expression of these cDNAs was studied in thetwo lines and the cause of instability in the purple podded phenotypeinvestigated . A genomic library was constructed from the purple poddedline . Two genes were selected by the differentially expressed cDNAs andtheir DNA sequences determined. A gene encoding a pectinestera- likesequence, and the pod expressed cDNA used to select it , were found to betwo members of a small multigene family in pea. The second gene selectedproved to encode a protein containing two distinct domains; the N-terminal region being of a repetitive proline-rich nature and the C-terminal region being hydrophobic and cysteine rich . This gene waspresent as a single copy in the pea genome and its expression appearedto be linked to pigmentation.

Thesis (Ph.D.)--University of Western Australia, 2005.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1943. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 96-98).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1940. / Typescript. Includes abstract and vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1953. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 56-58).