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Light-induced absorbance changes and partial reactions of photosynthesis in mutants of Scenedesmus obliquus, strain D₃

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Title Light-induced absorbance changes and partial reactions of photosynthesis in mutants of Scenedesmus obliquus, strain D₃
Names Pratt, Lee Herbert (creator)
Bishop, Norman I. (advisor)
Date Issued 1967-08-10 (iso8601)
Note Graduation date: 1968
Abstract Partial reactions of photosynthesis were examined in the wild
type strain and six photosynthetic mutants (8, 11, 26, 40, 50, a') of
Scenedesmus obliquus, strain D₃. A reproducible procedure for the
isolation of active chloroplast fragments from Scenedesmus was
developed. Several chloroplast reactions were examined, as well
as two in vivo reactions (anaerobic glucose assimilation and non-photochemical nitrite reduction). In addition, an extensive examination
was made of the light-induced 520 nm absorbance change, both
in vivo and in chloroplast preparations.
Earlier results on mutant 8 which indicated that it was a system I mutant were confirmed. Chloroplast preparations of this mutant
would neither reduce NADP nor perform cyclic photophosphorylation,
although Hill reaction activity with low redox potential oxidants, such as DCIP and ferricyanide, was present.
Mutants 11, 40, and a' were found to be defective close to the
site of system II since they lacked Hill reaction activity but possessed
normal cyclic photophosphorylation, DCIP-ascorbate mediated reduction
of NADP, and in vivo photoreduction activities. (Photoreduction
is the light-dependent reduction of carbon dioxide by hydrogen gas
utilizing an adaptable hydrogenase.) Chloroplasts of mutants 26 and
50 performed the same chloroplast reactions as the system II mutants,
but are not typical system II mutants since they lacked in vivo
photoreduction activity, a process requiring only system I. This
lack of photoreduction was not the result of a defective hydrogenase
since both reduced nitrite non-photochemically. Although mutants
26 and 50 could produce ATP in vitro, they performed no in vivo
cyclic photophosphorylation as evidenced by their lack of anaerobic,
light-dependent glucose assimilation activity. Apparently, in vivo
and in vitro cyclic photophosphorylation require different cofactors
of the electron transport chain. Provisionally, the defects in mutants
26 and 50 were located in the electron transport chain between the
two light reactions.
Photosynthetic mutants of Scenedesmus were also used in an
attempt to clarify the nature of the 520 nm absorbance change. Difference
spectra of this change in the wild type and mutant strains of
Scenedesmus support the previously suggested hypothesis that two pigments are involved. One portion of the change (first phase) was
defined as a system I photooxidation with a difference peak near 520
nm. It is stimulated by anaerobic conditions (argon); quantum yield
determinations indicate that long wavelength absorbing pigments are
primarily responsible for its production. All six of the mutants
possess only this phase of the absorbance change in vivo.
Another portion of the change (second phase) was identified as
a system I photoreduction with a difference peak near 510 nm. It is
present in wild type Scenedesmus under oxygen and under an atmosphere
of 96% H₂ and 4% CO₂ after adaptation for photoreduction. It
is inhibited entirely by DCMU under oxygen. None of the mutants
exhibit this phase of the absorbance change in vivo. Quantum yield
measurements, as well as the absence of this phase of the change in
both system I and system II mutants, indicate the dependence of this
phase of the signal upon both photoreactions of photosynthesis under
normal aerobic conditions.
Evidence obtained with the photosynthetic mutants of Scenedesmus
indicates that the physical unit of photosystem I encompasses
more than just the photochemically active pigments. Mutants 26 and
50 have reduced system I activity while mutant 8 exhibits reduced
system II activity. To explain the reduced activities in these mutants
which are not specifically correlated with their known or hypothetical defects, it is suggested that system I encompasses the bulk of the
intermediates in the electron transport chain connecting the two
photoreaction centers.
Genre Thesis/Dissertation
Topic Scenedesmus obliquus
Identifier http://hdl.handle.net/1957/46983

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