Tobacco Mutant Research Articles

A tobacco mutant with a reduced cell number in embryo sacs

The semisterile tobacco plant BG-141.4 was produced using both irradiation with X-rays and anther culture. The embryological investigation of the progeny of this plant following selfing (R1) revealed disturbances in the structure of the mature embryo sacs (ESs). The major abnormality was a decrease in cell number in the ESs. Other abnormalities included disturbances in the distribution of nuclei and in cell differentiation. The overall frequency of abnormal ESs in individual plants investigated varied from 8% to 83%, while the frequency of ESs with a reduced number of cells varied from 5% to 72%. Since the examination of three subsequent generations - R2, R3 and R4 - showed similar results, it is concluded that a mutation has been induced. Its main phenotypic manifestation is most likely related to its lacking one or two nuclear division(s) during the coenocytic stage of megagametogenesis. The penetrance of the mutation varies widely.

A plant cation-chloride co-transporter promoting auxin-independent tobacco protoplast division.

Although auxin plays a central role in plant development, little is known about the signal transduction pathways triggered by auxin regulating cell elongation, division and differentiation. We describe the molecular analysis of the mutant tobacco line axi 4/1, which was regenerated from an auxin-independent callus created by activation T-DNA tagging. Transcriptional enhancer-mediated deregulated expression of the tagged plant gene axi 4 uncouples division of axi 4/1 protoplasts from external auxin stimuli, whereas in untransformed protoplasts expression of axi 4 and cell division are auxin dependent. axi 4 encodes a 109 kDa protein with significant homology to a family of electroneutral cation-chloride co-transporters (CCCs). We show that overexpression of AXI 4 or another member of the CCC family, the Na+/K+/2Cl--co-transporter from shark, triggers auxin-independent growth of tobacco protoplasts. We suggest that Na+/K+/2Cl--co-transporters may play a role in signalling cell division and that this function is highly conserved between AXI 4 and the shark Na+/K+/2Cl--co-transporter. We also demonstrate that a C-terminal fragment of AXI 4 is sufficient to promote auxin-independent cell division, showing that the C-termini of CCCs are functional subunits triggering cell division. This may allow a molecular dissection of this process not only in plants but also in animal cells.

Differential Gene Expression in Response to Auxin Treatment in the Wild Type and rac, an Adventitious Rooting-Incompetent Mutant of Tobacco.

Histological analyses of auxin-treated cuttings from the wild type and the rac mutant of tobacco (Nicotiana tabacum cv Xanthii) previously revealed that some rac phloem parenchyma or inner cortical parenchyma cells form callus in response to exogenous auxin treatment but these cells never undergo the organized divisions associated with adventitious root initiation in the wild type. Here we report the effect of the rac mutation on the temporal and spatial expression patterns of three genes previously shown to be associated with adventitious root meristems, HRGPnt3, iaa4/5, and gh3. Using histochemical staining analyses of HRGPnt3-GUS transformant cuttings, we determined that the rac mutation blocks auxin activation of the HRGPnt3 promoter. Thus, activation of the HRGPnt3 promoter occurs specifically during adventitious root initiation in tobacco cuttings. Histochemical staining analyses of iaa4/5-GUS and gh3-GUS transformant cuttings revealed that the rac mutation does not repress the auxin activation of the iaa4/5 and gh3 promoters. Based on our histochemical staining analyses, we conclude that differential gene expression occurs in response to auxin treatment during adventitious root initiation in the wild type compared with callus formation in rac cuttings. We also determined that HRGPnt3 mRNA accumulation occurs in response to components of our root-induction protocol other than auxin, indicating that HRGPnt3 expression is regulated both developmentally and environmentally.

Systemic acquired silencing: transgene-specific post-transcriptional silencing is transmitted by grafting from silenced stocks to non-silenced scions.

Using grafting procedures, we investigated the transmission of co-suppression of nitrate reductase and nitrite reductase host genes and transgenes and of post-transcriptional silencing of a uidA transgene encoding glucuronidase in tobacco. We demonstrate that silencing is transmitted with 100% efficiency from silenced stocks to non-silenced scions expressing the corresponding transgene. Transmission is unidirectional from stock to scion, transgene specific, locus independent and requires the presence of a transcriptionally active transgene in the target scion. The transmission of co-suppression occurs when silenced stocks and non-silenced target scions are physically separated by up to 30 cm of stem of a non-target wild-type plant. Taken together, these results suggest that a non-metabolic, transgene-specific, diffusable messenger mediates the propagation of de novo post-transcriptional silencing through the plant.

Cuttings of a tobacco mutant, rae, undergo cell divisions but do not initiate adventitious roots in response to exogenous auxin

Numerous studies have shown that auxin induces adventitious root initiation in stem explants from a variety of species, including tobacco. A dominant, monogenic mutation previously identified in tobacco (Nicotiana tabacum cv. Xanthii), rac, confers tenfold auxin resistance to mesophyll‐derived cell suspensions and an impaired primary root development phenotype to seedlings. Results presented here demonstrate that adventitious root formation does not occur when heterozygous and homozygous rac stem cuttings are treated in vitro with indole‐3‐butyric acid (IBA) concentrations ranging from 0.5 μM to 500 μM. Histological analysis showed that some phloem parenchyma or inner cortical parenchyma cells in wild‐type stem cuttings undergo adventitious root morphogenesis when they are treated with 5 μM IBA. The same cell types in heterozygous and homozygous rac stem cuttings undergo mitoses in response to auxin, but never form adventitious root meristems. The lack of adventitious root initiation in rac stem cuttings is phenotypically distinct from the aberrant primary root development in rac seedlings. The rac mutation appears to block an essential process for auxin induction of adventitious root initiation but not cell division in phloem parenchyma or inner cortical parenchyma cells. Comparisons of rac heterozygous and homozygous seedling primary root length and callus formation in response to auxin in stem cuttings indicate that rac copy number is correlated to the degree of expression of these two phenotypes.

A single amino acid change in acetolactate synthase confers resistance to valine in tobacco.

The metabolic control of branches chain amino acid (BCAA) biosynthesis involves allosteric regulation of acetolactate synthase (ALS) by the end-products of the pathway, valine, leucine and isoleucine. We describe here the molecular basis of valine resistance. We cloned and sequenced an ALS gene from the tobacco mutant Valr-1 and found a single basepair substitution relative to the wild-type allele. This mutation causes a serine to leucine change in the amino acid sequence of ALS at position 214. We then mutagenized the wild-type allele of the ALS gene of Arabidopsis and found that it confers valine resistance when introduced into tobacco plants. Taken together, these results suggest that the serine to leucine change at position 214 of ALS is responsible for valine resistance in tobacco.

Non-Destructive Determination of Chlorophyll Content of Leaves of a Green and an Aurea Mutant of Tobacco by Reflectance Measurements

Summary Reflectance spectra from 400 to 800 nm with a spectral resolution of 1 nm and the content of photosynthetic pigments were acquired for leaves of an aurea mutant and a green tobacco ( Nicotiana tabacum L.) covering a range of chlorophyll a content from 7 to 30.8 μ cm -2 and total chlorophyll a + b level from 8.4 to 41.4 μg cm -2 leaf area. At a chlorophyll a content of more than 7 μg cm -2 leaf area the reflectance near 670 to 680 nm was not sensitive to a variation in chlorophyll content due to saturation of the relationship ≪absorption vs. chlorophyll content≫. The wavelength position of the red reflectance minimum (Rmin) in the main red absorption bands of in vivo chlorophyll a proved to be independent of the chlorophyll content of leaves and thus is not suitable for chlorophyll determination. Maximum sensitivity of leaf reflectance to variation in chlorophyll content was, however, found in a wide spectral range from 530 to 630 nm and near 700 nm. The wavelength of the red edge position of the reflectance spectrum (inflection point IP) in the range of 694 to 706 nm correlated very closely with the leaves' chlorophyll content in a curvilinear manner. The Ip is also closely correlated in a linear manner with the reflectance signals at 550 and 700 nm. Reflectances near 700 nm and 550 nm were found to be sensitive indicators of the red edge position and the chlorophyll content of leaves as well. The ratios of reflectances in the near infra-red range of the spectrum (above 750 nm) to that at 700 nm and at 550 nm, R NiR /R700 (or R750/R700) and R nir /R550 (or R750/R550), were directly proportional (r 2 >0.93) to the leaves' chlorophyll content. These two novel indices for chlorophyll determination (R750/R700 and R750/R550) allowed the estimation of leaf chlorophyll content with an error of less than 2.1 μg cm -2 . The two new vegetation indices, R750/R700 and R750/R550, are linearily correlated to the chlorophyll content. They exhibit a more than 6 times wider dynamic range than the widely used Normalized Difference Vegetation Index NDVT. The new vegetation index ratios are very sensitive to changes in chlorophyll content at low, medium and high chlorophyll content which is not the case for the NDVI. This suggests the application of the new indices R750/R700 and R750/R550 in the near and remote sensing of the chlorophyll content of terrestrial vegetation.

Alteration of Gene Expression by Jasmonate and ABA in Tobacco and Tomato

Summary The synthesis of jasmonate-induced proteins in leaves of tobacco ( Nicotiana plumbaginifolia ) and tomato ( Lycopersicon esculentum ) was studied in order to find a possible functional link in the actions of abscisic acid (ABA) and jasmonates. ABA-deficient mutants of tobacco (CKR1) and of tomato ( sitiens, flacca ), and their corresponding wild-types, were compared with respect to endogenous contents of jasmonates and ABA, and polypeptide and transcript patterns in water- or jasmonate-floated leaves, leaves stressed by floating on sorbitol, or by weak desiccation. Our results indicate that in tobacco the synthesis of proteins induced by jasmonate differed from those induced by ABA, whereas in tomato some jasmonate-induced proteins were also induced by ABA. The results provide further evidence that different signalling pathways exist for jasmonate/ABA-responsive gene expression in various plant species.

Consequences of High CO2-Concentrations in Air on Growth and Gas-Exchange Rates in Tobacco Mutants

Abstract Wild type tobacco N. tabacum var. John William’s Broadleaf and the tobacco aurea mutant Su/su were permanently grown under 700 ppm CO2 in air. In comparison to plants grown under 350 ppm CO2 in air but under otherwise identical conditions growth was substantially enhanced. Gas exchange measurements carried out by mass spectrometry show that the rate of photosynthesis in the wild type and in the mutant is increased by more than 100%. The photorespiratory rate in the wild type measured as 18O2-uptake in the light in the “700 ppm CO2-plants” is not reduced to the extent expected or deduced from experiments in which the 350 ppm system responds under in vitro conditions to 700 ppm CO2. An analysis of the induction kinetics of room temperature fluorescence kinetics of the adapted (700 ppm CO2) system and the control system (350 ppm CO2) under various CO2-partial pressures shows that permanent growth under the elevated CO2-partial pressure leads to a structural modification of the photosynthetic apparatus.

Isolation and Partial Characterization of a Manganese Requiring ʟ-Arginine Metabolizing Enzyme Being Present in Photosystem II Complexes of Spinach and Tobacco

Abstract A low ʟ-arginine metabolizing enzyme (L-AME) activity leading to ornithine, urea and additional products not identified so far could be detected in photosystem II (PS II) membranes of spinach and of the chlorophyll deficient tobacco mutant Su/su. The detectable L-AME activity was very low in untreated PS II membranes, but increased significantly (about 10 fold) when the extrinsic peptides (psbO, P and Q gene products) were removed - suggesting that the L-AME is exposed at the lumen side of PS II. It was possible to isolate the detergent-solubilized protein from CaCl2-washed PS II membranes of spinach by a combination of anion and cation exchange columns. On the basis of SDS PAGE the protein was homogenous and had an apparent molecular mass of 7 kDa. N-terminal sequencing of the polypeptide gave a contiguous sequence of 20 amino acids showing no homologies to PS II polypeptides as yet sequenced. After chromatography of the L-AME on an anion exchange column at pH 9.5 (last purification step) a completely inactive enzyme was obtained. Maximal reactivation was achieved by dialyzing the protein against Hepes-NaOH buffer in the pH range of 6.5 to 7.5 containing 100 mᴍ chloride or sulfate (being the most effective anions). The L-AME activity was totally dependent on manganese added to the reaction mixture. Moreover, there were indications of a second cation binding site being more sequestered and requiring bound Ca2+ or Mn2+ for activity (Sr2+ was less effective and Mg2+ was ineffective). There are indications that the protein contains a redox active group - possibly an aminoacid- derived quinonoid (based on a redox cycling assay with glycine and nitroblue tetrazolium). The capability of this PS II associated protein to bind the cofactors of water oxidation and having a redox active group (preliminary results) suggests that this protein might be functional in photosynthetic water oxidation. This is further supported by the fact that the isolated L-AME has a low catalase activity

Sterol acyl transferase and steryl ester hydrolase activities in a tobacco mutant which overproduces sterols

The tobacco mutant LAB 1–4 overproduces sterols, most of which are esterified and stored as lipid droplets in the cytoplasm (P. Maillot-Vernier, L. Gondet, H. Schaller, P. Benveniste and G. Belliard, Mol. Gen. Genet. 231 (1991) 33–40). We studied the enzymes involved in the esterification of sterols and hydrolysis of steryl esters in tobacco calli and leaves. These activities were shown to be bound to microsomal membranes. The microsomes were delipidated by acetone treatment so as to discard endogenous sterols and steryl esters; optimal conditions were set up which allowed the selective determination of either the sterol acyl transferase (SATase) activity or the steryl ester hydrolase (SEHase) activity. The SATase activity of the LAB 1–4 mutant calli was shown to be twice that of the wild type calli whereas the activities were the same in the leaves of both genotypes. The SEHase activity of LAB 1–4 calli and leaves was only slightly higher than that of the wild type tissues. The results strongly suggest that the activity of the SATase is increased when the ratio sterols/enzyme exceeds a certain threshold. Both enzymes might be involved in the control of the free sterol content of the sterol-overproducing mutant and, more generally, in the homeostasis of free sterols in the plant cells.

In Vivo Photomodification of Ribulose-1,5-Bisphosphate Carboxylase/Oxygenase Holoenzyme by Ultraviolet-B Radiation (Formation of a 66-Kilodalton Variant of the Large Subunit).

Increased levels of solar ultraviolet (290-320 nm) (UV-B) radiation could have profound effects on plant proteins because the aromatic amino acids in proteins absorb strongly in this spectral region. We have investigated the effects of UV-B radiation on plant proteins and have observed a novel 66-kD protein. This product was formed in vivo when Brassica napus L. plants grown for 21 d in 65 [mu]mol m-2 s-1 photosynthetically active radiation were subsequently exposed to 65 [mu]mol m-2 s-1 photosynthetically active radiation plus UV-B radiation (1.5 [mu]mol m-2 s-1). The protein appeared after 4 h of UV-B irradiation and accumulated during the next 16 h in UV-B. The 66-kD protein cross-reacted with an antiserum against the ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) holoenzyme. Analysis of soluble leaf proteins revealed that the 66-kD product had a number of isoforms corresponding closely to those of the large subunit of Rubisco (LSU). Partial proteolytic digests of the LSU and the 66-kD protein resulted in an equivalent pattern of protein fragments, leading to the conclusion that the 66-kD protein was a photomodified form of the LSU. A similar high molecular mass variant of Rubisco was observed in soluble protein extracts from leaves of tomato (Lycopersicon esculentum), tobacco (Nicotiana tabacum), and pea (Pisum sativum L.) plants treated in vivo with UV-B, suggesting that it might be a common product, at least among C3 plants. It is interesting that the 66-kD product appears to be generated after incorporation of the LSU into holoenzyme complexes. This conclusion was drawn from two lines of evidence. First, the LSU variant co-purified with holoenzyme complexes isolated by nondenaturing polyacrylamide gel electrophoresis. Second, a UV-B-specific 66-kD protein did not accumulate in a tobacco mutant that synthesizes the Rubisco subunits but does not assemble them into normal holoenzyme complexes.

Partial Purification and Characterization of a Calcium-Dependent Protein Kinase and an Inhibitor Protein Required for Inactivation of Spinach Leaf Nitrate Reductase

Evidence is accumulating that the activity of spinach (Spinacia oleracea L.) leaf NADH:nitrate reductase (NR) is modulated both in vitro and in vivo by protein phosphorylation. From the present study we report the partial purification of the two protein factors needed for NR inactivation. We identified NR-protein kinase (NR-PK) as a calcium-dependent and metabolite-regulated protein kinase and have provided additional evidence that phosphorylation of NR is necessary but not sufficient to inactivate the enzyme. The inhibitor protein required for inactivation of phospho-NR was purified 625-fold by polyethylene glycol fractionation and sequential column chromatography. Using partially purified inhibitor protein and NR-PK, we characterized NR inactivation (increased sensitivity to Mg2+ inhibition) in a reconstituted in vitro system. NR-PK activity was inhibited by a variety of metabolic phosphate esters including di-hydroxyacetone phosphate, glucose-6-phosphate, and fructose-1,6-bisphosphate. Light-to-dark transition experiments with a starchless tobacco (Nicotiana sylvestris) mutant, which accumulates phosphate esters during the photoperiod, indicated that NR inactivation in vivo might, indeed, be down-regulated by metabolites. Additionally, we postulate that cytosolic free calcium could play an important role in the regulation of NR activity in vivo.

Rubisco, rubisco activase and ribulose-5-phosphate kinase gene expression and polypeptide accumulation in a tobacco mutant defective in chloroplast protein synthesis

Expression of the genes for ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; rbcS and rbcL), Rubisco activase (rca) and ribulose-5-phosphate (Ru5-P) kinase (prk) and accumulation of the polypeptides was examined in chlorophyllous and chlorotic sectors of the DP1 mutant of Nicotiana tabacum. Plastids from chlorotic sectors of this variegated plastome mutant contained 30S and 50S ribosomal subunits, but had abnormally low levels of plastid polysomes. Consequently, mutant plastids were translationally repressed, unable to synthesize plastid-encoded polypeptides including the large subunit of Rubisco despite the presence of the corresponding mRNAs. Transcripts of rbcS accumulated to near wild type levels in chlorotic sectors, but there was little accumulation of the Rubisco small subunit (SS) polypeptide or holoenzyme. Messenger-RNA isolated from chlorotic sectors effectively directed the synthesis of Rubisco SS in vitro suggesting that posttranslational factors were responsible for the decrease in Rubisco SS abundance. Transcripts of rca and prk also accumulated to near wild type levels in chlorotic sectors and a diurnal rhythm in the abundance of rca mRNA was detected in green and chlorotic sectors. Despite the low abundance of Rubisco holoenzyme in chlorotic sectors, Rubisco activase and Ru5-P kinase polypeptides accumulated to significant levels. Activities of Rubisco and Ru5-P kinase paralleled protein levels, indicating that active forms of these enzymes were present in chlorotic sectors. The data indicate that the developmental events governing the accumulation of Rubisco activase and Ru5-P kinase polypeptides and the diurnal regulation of rca expression were not dependent on the attainment of photosynthetically competent plastids or the accumulation of Rubisco.

Photosynthetic Carbon Metabolism and Translocation in Wild-Type and Starch-Deficient Mutant Nicotiana sylvestris L

A high rate of daytime export of assimilated carbon from leaves of a starch-deficient mutant tobacco (Nicotiana sylvestris L.) was found to be a key factor that enabled shoots to grow at rates comparable to those in wild-type plants under a 14-h light period. Much of the newly fixed carbon that would be used for starch synthesis in leaves of wild-type plants was used instead for sucrose synthesis in the mutant. As a result, export doubled and accumulation of sucrose and hexoses increased markedly during the day in leaves of the mutant plants. The increased rate of export to sink leaves appeared to be responsible for the increase in the proportion of their growth that occurred during the day compared to wild-type plants. Daytime growth of source leaves also increased, presumably as a result of the increased accumulation of recently assimilated soluble carbon in the leaves. Even though starch accumulation did not occur in the leaves of mutant plants, nearly all the sugar that accumulated during the day was exported in the period of decreasing irradiance at the end of the diurnal light period. Changes in carbon allocation that occurred in leaves of wild-type and mutant plants near the end of the light period appeared to result from endogenous diurnal regulation associated with the day-night transition.

Regulation of Photosynthesis during Leaf Development in RbcS Antisense DNA Mutants of Tobacco.

We have previously characterized RbcS antisense DNA mutants of tobacco that have drastic reductions in their content of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco; S.R. Rodermel, M.S. Abbott, L. Bogorad [1988] Cell 55: 673-681). In this report we examine the impact of Rubisco loss on photosynthesis during tobacco (Nicotiana tabacum) leaf development. Photosynthetic capacities are depressed in the antisense leaves, but the patterns of change in photosynthetic rates during the development of these leaves are similar to those in wild-type plants: after attaining a maximum in young leaves, photosynthetic capacities undergo a prolonged senescence decline in older leaves. The alterations in photosynthetic capacities in both the wild type and mutant are closely correlated with changes in Rubisco activity and content. During wild-type leaf development, Rubisco accumulation is regulated by coordinate changes in RbcS and rbcL transcript accumulation, whereas in the antisense leaves, Rubisco content is a function of RbcS, but not rbcL, transcript abundance. This indicates that large subunit protein production is controlled posttranscriptionally in the mutants. The antisense leaves accumulate near-normal levels of chlorophyll and representative photosynthetic proteins throughout development, suggesting that photosynthetic gene expression is not feedback regulated by Rubisco abundance. Considered together, the data in this paper indicate that leaf developmental programs are generally insensitive to sharp reductions in Rubisco content and emphasize the metabolic plasticity of plant cells in achieving optimal photosynthetic rates.

Sterol overproduction is the biochemical basis of resistance to a triazole in calli from a tobacco mutant

Sterol overproduction is shown to be the biochemical basis of resistance to the triazole LAB 170250F (2-(4-chlorophenyl)-3-phenyl-1-(1H-1,2,4-triazol-1-yl)-2,3-oxidopropane), an inhibitor of cytochrome P450-dependent-obtusifoliol-14α-demethylase, in the tobacco (Nicotiana tabacum L. cv. Xanthi) mutant LAB 1-4. Genetic analysis at the callus level indicates that the resistance and the biochemical phenotypes co-segregate during meiotic recombination and therefore result most probably from the same mutation. Analysis of the intracellular distribution of sterols shows that in LAB 1-4 calli containing tenfold the sterol amount of the wild type, the overproduced metabolites, mainly obtusifoliol, are esterified by fatty acids and stored in hyaloplasmic lipid droplets. Thus, the mutant calli maintain a concentration of free sterols in cell membranes, mainly end-products of the sterol-biosynthesis pathway, which corresponds to physiological requirements, whereas the level of free sterols in the wild-type calli treated with the triazole is too low to ensure viability. We also show that sterol overproduction confers resistance to other phytotoxic sterolbiosynthesis inhibitors.

Regulation of Sterol Content in Membranes by Subcellular Compartmentation of Steryl-Esters Accumulating in a Sterol-Overproducing Tobacco Mutant.

The study of sterol overproduction in tissues of LAB 1-4 mutant tobacco (Nicotiana tabacum L. cv Xanthi) (P. Maillot-Vernier, H. Schaller, P. Benveniste, G. Belliard [1989] Biochem Biophys Res Commun 165: 125-130) over several generations showed that the overproduction phenotype is stable in calli, with a 10-fold stimulation of sterol content when compared with wild-type calli. However, leaves of LAB 1-4 plants obtained after two steps of self-fertilization were characterized by a mere 3-fold stimulation, whereas calli obtained from these plants retained a typical sterol-overproducing mutant phenotype (i.e. a 10-fold increase of sterol content). These results suggest that the expression of the LAB 1-4 phenotype is dependent on the differentiation state of cells. Most of the sterols accumulating in the mutant tissues were present as steryl-esters, which were minor species in wild-type tissues. Subcellular fractionation showed that in both mutant and wild-type tissues, free sterols were associated mainly with microsomal membranes. In contrast, the bulk of steryl-esters present in mutant tissues was found in the soluble fraction of cells. Numerous lipid droplets were detected in the hyaloplasm of LAB 1-4 cells by cytochemical and cytological techniques. After isolation, these lipid granules were shown to contain steryl-esters. These results show that the overproduced sterols of mutant tissues accumulate as steryl-esters in hyaloplasmic bodies. The esterification process thus allows regulation of the amount of free sterols in membranes by subcellular compartmentation.

Antisense RNA inhibition of Rubisco activase expression

SummaryRibulose bisphosphate carboxylase (Rubisco) activase catalyzes the activation of Rubisco in vivo. Activase antisense DNA mutants of tobacco have been generated to explore the control that activase exerts on the photosynthetic process. These mutants have up to 90% reductions in activase protein levels as a consequence of an inhibition of activase mRNA accumulation. It is shown that photosynthesis, measured as the rate of CO2 exchange (CER), is modestly decreased in plants exposed to high irradiances. The decreases in CER in the transgenic plants are accompanied by corresponding decreases in Rubisco activation, indicating that activase has a direct effect on photosynthetic rates in the antisense plants by influencing the activation state of Rubisco. It is concluded that in high light conditions, control of photosynthesis is largely shared between Rubisco and activase. Plant growth is also impaired in mutant plants that have severe reductions in activase. The inhibition of activase in the antisense plants does not have an impact on the accumulation of Rubisco large subunit or small subunit mRNAs or proteins. This indicates that the concerted expression of the genes for activase (Rca) and Rubisco (rbcL and rbcS) in response to light, developmental factors and circadian controls is not due to feedback regulation of rbcL or rbcS by the amount of activase protein.

Quantum Yields of Photosystem II and Photosynthesis in an Aurea Mutant of Tobacco (C<sub>3</sub>) and an Oil Yellow Mutant of Maize (C<sub>4</sub>) Which Have High Capacities for Photosynthesis Despite Low Chlorophyll Contents

Quantum Yields of Photosystem II and Photosynthesis in an Aurea Mutant of Tobacco (C<sub>3</sub>) and an Oil Yellow Mutant of Maize (C<sub>4</sub>) Which Have High Capacities for Photosynthesis Despite Low Chlorophyll Contents