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Regeneration and shoot multiplication of common bean (Phaseolus vulgaris L. 'ICA Pijao') from half-cotyledonary nodes was achieved on modified Murashige and Skoog (1962) basal medium amended with 5 $ mu$M 6-benzylaminopurine. Histological studies confirmed the adventitious origin of the regenerated buds. Shoots were rooted ex vitro and developed into morphologically normal plants compared with seed-grown controls. The relative susceptibility of bean tissues to infection by a collection of wild-type Agrobacterium strains was tested. Positive transformation events were evaluated based on morphological and biochemical changes observed following Agrobacterium infection. The A. tumefaciens strain C58 was particularly virulent on greenhouse-grown plants, in vitro-derived stem sections, half-cotyledonary nodes and seedlings. A sensitive and rapid method was developed to detect opines using thin layer chromatography. Transient $ beta$-glucuronidase (GUS) gene expression was detected in 'ICA Pijao' bean buds regenerated from half-cotyledonary nodes following Agrobacterium-mediated gene transfer with the binary vector pGV1040 or p35SGUSINT. Four out of eight putative transformants contained the chimeric GUSINT gene following polymerase chain reaction (PCR) analysis. This was confirmed by Southern analysis of blotted PCR gels. However, there was no stable integration of the GUSINT gene as none of the R1 progeny showed an amplified GUSINT fragment with PCR.

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Red kidney bean seeds (Phaseolus vulgaris) contain a variety of bioactive proteins including lectins, enzyme inhibitors, hydrolytic enzymes and antifungal proteins. The aim of this research was to investigate activities of selected low pH and heat-stable bioactive proteins extracted from different parts of red kidney bean seed, seedling and pod as well as seed and root exudates. Crude red kidney bean seed extracts inhibited growth of Alternaria alternata as well as its protease activity, but not its amylase activity. A protein with inhibitory activity against growth of A. alternata was purified from extracts of the red kidney bean cotyledons and embryonic axis. This purified bean protein was devoid of chitinase and β-1, 3- glucanase activities. Also, it did not inhibit porcine pancreatic α-amylase, bovine trypsin, amylase and protease of A. alternata suggesting that the antifungal activity of the protein is not related to these activities. Proteinaceous extracts of red kidney bean cotyledons induced melanin and conidia formation in mycelium of A. alternata. A protein responsible for this conidiation inducing effect was shown for the first time to be a mannose-binding lectin which is also known as PvFRIL (Phaseolus vulgaris fetal liver tyrosine kinase 3-receptor interacting lectin). An unexpected finding was that extracts of the embryonic axis stimulated rather than inhibited porcine α-amylase activity. Phytohemagglutinin (PHA-L in particular), co-extracted with α-amylase inhibitor from red kidney bean seeds, was implicated as an α-amylase stimulator with the potential of greatly assisting digestion of starch. In cotyledonary extracts, amylase stimulatory activity was masked by amylase inhibitory activity that was inactivated when the extracts were boiled for 10 min. An in-gel non-denaturing electrophoretic method was used to show presence of porcine α-amylase isoinhibitors in extracts of the cotyledons and embryonic axis. All other seedling parts as well as seed and root exudates had amylase stimulatory activity. Another improved non-denaturing electrophoretic method with immobilized azoalbumin was developed for in-gel detection of isoinhibitors of bovine trypsin in seed parts. It eliminates the need for both time-consuming and labourious staining, destaining or renaturation steps used in other methods. Accumulation of most of the selected bioactive proteins during seed development in different seed parts appeared to start at 20 days after flower abscission. The activities of these proteins decreased to lower levels after 11 days of germination. Besides these observed developmental changes, under abiotic (UV-C irradiation) and biotic (seedlings co-cultured with A. alternata) stress, increased activity of some of the selected bioactive proteins were detected. In conclusion, this study has contributed to a better understanding of antifungal activity and the selected bioactive proteins in extracts of red kidney bean.

After a wild-type isolate of Epicoccum purpurascens was exposed to shortwave ultraviolet light, several new strains were recovered which were improved in sporulation, fungicide tolerance, and performance in suppression of white mold caused by Sclerotinia sclerotiorum. The efficacy of E. purpurascens in controlling white mold of snap bean (Phaseolus vulgaris) was assessed in greenhouse and field trials. White mold was significantly reduced in both greenhouse and field trials when 2-4 sprays of E. purpurascens conidial suspensions (in 1% malt extract) were sprayed onto the plant surface during the flowering period. Germination of E. purpurascens conidia on senescent petals was greater than on younger flowers. Addition of malt extract to conidial suspensions improved germination on flowers and increased colonization of emerging flowers. Application of E. purpurascens did not accelerate senescence of bean leaves or affect pod yield of bean in greenhouse trials. Mycoparasitism of S. sclerotiorum by E. purpurascens was found only rarely in in vitro tests and was not observed on flower disks. Production of inhibitory compounds by E. purpurascens was the most important mechanism in suppression of white mold but competition for nutrients also appeared to play a role in biocontrol. The influence of nutrients on conidial germination, growth, sporulation and production of antifungal compounds by E. purpurascens were also investigated.

Typescript. / Thesis (Ph. D.)--University of Hawaii, 1965. / Bibliography: leaves [104]-107. / viii, 107 leaves mounted illus., tables

Thesis (M.S.)--University of Wisconsin--Madison, 1970. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (Ph. D.)--University of Wisconsin--Madison, 1969. / Typescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliography.

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