PHOTOOXIDATION REACTIONS BY CHLOROPLASTS SOLUBILIZED WITH SURFACE-ACTIVE AGENTS

1. Solubilization of chioroplasts with a mixture of 1 per centDuponol C and 1 per cent Span 80 (3: 1) caused a destructionof activity in the HILL reaction, but the treatment broughtabout an increase by about 60 per cent in the rate of ascorbatephotooxidation in the presence of DPIP. Heating the broken chloroplastscaused a marked decrease in the photooxidation activity. Byadding surface- active agents to the boiled preparation, theactivity was restored up to almost 80 per cent of the originallevel.
2. With colloidal suspensions of isolated chiorophylls,ascorbatewas only slightly photooxidized in the presence ofDPIP. Byaddi tion of the surface-active agents, the activitywas greatlyenhanced.
3. Dependency of the photooxidation bywhole and solubilized chloroplastsand isolated chlorophylla on the presence of DPIP was examined.DPIP can serve as anintermediate electron carrier in solubilizedchloroplasts aswell as in whole chloroplasts.
4. Effect of o-phenanthrolineon ascorbate photooxidation by thesethree preparations wastested. With solubilized chloroplastsand isolated chlorophylls,the addition of the inhibitor hadno influence on their ascorbatephotooxidation either in thepresence or absence of DPIP.
5. Treatmentof whole chloroplasts with the surface-active agentsinducedan activity of photooxidation of cytochrome c. The electron-flowpattern for the photooxidation of ascorbate by whole and solubilizedchloroplasts was briefly discussed.

¹ Contribution No. 130 from the Department of Biology, Facultyof Science, Kyushu University. Aided in part by Grant-in-Aidfor Fundamental Scientific Research from the Ministry of Education. (Received August 23, 1962; )     

CHLOROPHYLL-SENSITIZED REDUCTION OF PLASTOQUINONE BY ASCORBIC ACID AND QUENCHING OF CHLOROPHYLL-FLUORESCENCE BY PLASTOQUINONE

1. Photochemical reduction of plastoquinone by ascorbic acid inethanol was sensitized with some derivatives of chlorophyll.The order of effectiveness was as follows: allomerized chlorophylla > chlorophyllin a > chlorophyll a > chlorophyll b> pheophytin a.
2. Quenching of the fluorescence of chlorophylla by plastoquinonewas observed. The quenching constant calculatedwas 71 litreper mole.

¹Contribution No. 159 from the Department of Biology, Facultyof Science, Kyushu University. Supported in part by a grant-in-aidfor Fundamental Scientific Research from the Ministry of Education. ²Present address: Biological Laboratory, General Education Department,Kyushu University, Ropponmatsu, Fukuoka. ³Present address: Biological Institute, Daiichi College of PharmaceuticalSciences, Tamagawa-machi, Takamiya, Fukuoka.     

EFFECT OF LIPASE-DIGESTION ON PHOTOCHEMICAL ACTIVITIES OF ISOLATED CHLOROPLASTS

In order to elucidate the role of lipids in photosynthesis,chloroplasts were digested with lipase, and the effect of lipase-digestionon some photochemical activities was studied. The HILL reactionwas sensitive to the digestion, but chloroplasts having intactmembrane were somewhat resistant to the action of lipase. Theinactivation by lipase digestion seems to be due to the destructionof a component necessary for the Hill reaction to proceed. Thechloroplasts treated with lipase showed the following activities. (1) Active photooxidation of reduced cytochrome c and menadione. (2) Photooxidation of ascorbate, which was enhanced in the presenceof DPIP, and retarded in the absence of the dye. (3) NADP-photoreduction in the presence of the DPIP-ascorbatecouple, as the electron donor. These facts suggested that the site attacked with lipase wasresponsible for the photochemical oxygen evolution. The decrease in the fluorescence intensity of chlorophyll awas also observed during the digestion. ¹Present address : Biological Laboratory, General EducationDeparment, Kyushu University, Otsubo-machi, Fukuoka.     

USE OF PURCHASE PREFERENCE OPTIONS TO INCREASE "NO PREFERENCE" FREQUENCIES IN PLACEBO PREFERENCE TESTS

Besides responses to the different stimuli being tested in a paired preference test, responses to identical "placebo" stimuli can be used as a screening tool to identify biased consumers. Those consumers who give preference responses to identical stimuli can be assumed to be biased. Accordingly, only the data from unbiased consumers need to be considered for the different stimuli. The problem with this procedure is that the sample size is reduced. The goal of the present research was to see whether using options associated with purchase intent, elicited a greater number of "No Preference" responses to identical "placebo" stimuli. It was found that they did. The increase was large when the preference options implied exclusivity. In conditions where the preference strength options were not so strong, the frequency of "No Preference" responses dropped accordingly.

PRACTICAL APPLICATIONS

A problem with paired preference testing is the tendency of consumers to give false preferences, which produces the seriously misleading overestimation of the proportion of consumers who have preferences for one or other of the products being assessed. The "placebo" condition is an important control for alleviating this problem. The statistical analysis can be improved by finding a protocol that maximizes the proportion of "No Preference" responses in the placebo condition. The key finding here is that using purchase intent questions rather than preference questions may possibly provide a way of achieving this aim.     

Testing the 'assortative mating' hypothesis on a variation maintenance mechanism for flowering time within a forest-floor population of Primula sieboldii

Flowering phenology and allozyme variation were studied to test the existence of positive assortative mating for flowering time in a natural population of Primula sieboldii E. Morren, a heterostylous perennial herb, consisting of approximately 180 genets in a deciduous forest. There was significant variation in flowering date among genets, but not between heterostylous morphs. The temporal order of the flowering time of genets was fairly constant for the two years of the study. The spatial heterogeneity of light availability at the study site was small during the flowering season of the species. In order to analyze the extent of genetic differentiation between early- and late-flowering genet groups, allozyme diversities were analyzed with 10 loci. The G_ST between the early- and late-flowering groups was not significantly different from zero. Evidence of positive assortative mating for flowering time was not detected. Prolongation of flowering duration due to pollen limitation may be one important factor preventing the genetic differentiation of early- and late-flowering groups by enhancing the overlap of flowering time among genets.     

FLUORESCENCE YIELD OF CHLOROPHYLL A AND PHOTOCHEMICAL ACTIVITIES OF ISOLATED CHLOROPLASTS

1. Effect of various treatments on the fluorescence yield of chlorophyllo in isolated chloroplasts was studied in relation to the activitiesof the HILL reaction and the ascorbate photooxidation.
2. CMUand o-phenanthroline showed no appreciable effect on thefluorescenceyield at the concentration which completely inhibited the HILLreaction.
3. 0.01% SDS was sufficient to inactivate the HILLreaction completelyand caused a 40% decrease in the fluorescenceyield, althoughno shift of the absorption and fluorescencebands could be observed.At higher concentrations of SDS, thefluorescence yield increasedby more than three times the originalvalue and the absorptionand fluorescence bands shifted by 5rap toward the shorter wavelengthside.
4. Ultraviolet irradiation,incubation at pH 9.2, heating at 40and digestion with trypsinresulted in loss of the HILL activityand decrease of the fluorescenceyield.
5. The DPIP-mediated photooxidation of ascorbate was enhancedbyheating broken chloroplasts at 55 for 5 min, and inactivatedat 70. The photooxidation of ascorbate in the absence of thedye was inhibited by 60% on heating at 55. The o-phenanthroline-sensitivephotooxidation of ascorbate was inactivated at 55. The o-phenanthroline-resistantphotooxidation of ascorbate was rather thermostable. The fluorescenceyield was reduced by 40–50% at 55.

¹ Present address: Biological Laboratory, General EducationDepartment, Kyushu University, Otsubo-machi, Fukuoka.