QUANTITATIVE AND QUALITATIVE ANALYSES OF PROTEIN IN STIGMA-STYLES OF SELF-INCOMPATIBLE LILIUM LONGIFLORUM THUNB

Quantitative and qualitative analyses of total soluble protein in stigma-styles of self-incompatible Lilium longiflorum Thunb. 'Ace' and 'Nellie White' were conducted to examine the effect of aging, pollination, heat treatment, protein inhibitor, or nucleic acid inhibitor on both total protein and changing protein patterns. Slab electrophoresis and colorimetric procedures were used in the analyses to determine whether changes in incompatibility are related to changes in specific proteins.;Manuscript I. Protein was extracted with 30% DMSO from maturing, senescing stigma-styles of Lilium longiflorum Thunb. 'Ace' and 'Nellie White' collected from 2 days pre-anthesis to 10 days post-anthesis some styles were left unpollinated, pollinated just before extraction, cross pollinated, or self pollinated and incubated for 0, 12, 24, or 48 hr prior to extraction. Protein extraction was separated by slab electrophoresis on polyacrylamide gel. In 'Ace' 15 to 16 bands were obtained in 'NW' 14 to 15 bands. No specific bands were observed to be correlated with aging or type of pollination.;Protein was also extracted with 2N NaOH from freeze-dried powder obtained from the above pollination experiment and analyzed by colorimetric test. Pollination and subsequent pollen tube growth caused an increase in total protein after 12 hr of incubation, just before differentiation into compatible or incompatible pollen tubes.;It was concluded that alteration in protein patterns during flower maturation, senescence, and pollen tube growth is primarily quantitative rather than qualitative.;Results are presented in manuscript form for publication in the journal, Theoretical and Applied Genetics.;Manuscript II. Stigma-styles of Lilium longiflorum Thunb. 'Ace' and 'Nellie White' were collected one, two, or three days after anthesis, heat treated in 50(DEGREES)C water bath for 0. 2.5, 5, or 7.5 min and left unpollinated or heat treated for 5 min followed by self or cross pollination and incubated for 0, 12, 24, or 48 hr at 24(DEGREES)C. Protein was then extracted with 30% DMSO and separated by slab electrophoresis on 7.5% polyacrylamide gel resulting in 15 bands in 'Ace' and 14 bands in 'NW'. No specific band was observed to be correlated with any degree of heat treatment.;Protein was also extracted with 2N NaOH from freeze-dried powder obtained from the above experiments and analyzed by colorimetric test. Heat treatment of unpollinated styles for 5 min caused a slight increase in protein after 12 and 24 hr of incubation, followed by a slow degradation in the following 24 hr; but heat treatment for 7.5 min caused no protein changes in the first 24 hr. Heat treatment following pollination gave a considerable increase at 12 hr and a slow degradation after that.;Alteration of protein due to heat or heat following pollination is primarily quantitative rather than qualitative.;Manuscript III. Stigma-styles of Lilium longiflorum Thunb. 'Ace' and 'Nellie White' from pre-anthesis to 2 days post anthesis were injected with protein synthesis inhibitor (puromycin) or glass redistilled water and then incubated at 24(DEGREES)C for 0, 12, 24, or 48 hr. Separately, stigma-styles collected one day post-anthesis were injected with nucleic acid synthesis inhibitors (6-methylpurine or Actynomycin-D) or glass redistilled water; and then each treatment was left unpollinated, compatibly cross pollinated, or self pollinated, then incubated at 24(DEGREES)C for 0, 12, 24, or 48 hr.;After incubation protein was extracted with 30% DMSO, then separated by slab-electrophoresis on 7.5% polyacrylamide gel, resulting in 15 bands in 'Ace' and 14 bands in 'NW'. No specific bands were observed to be correlated with any type of inhibitor, pollination, or time of incubation.