Inheritance of nuclear DNA content in leaf epidermal cells of Zea mays L.

 The nuclear DNA content (ploidy level) of maize leaf-epidermal cells was investigated by Feulgen cytophotometry in two lines, Illinois High Protein (IHP) and Illinois Low Protein (ILP), their reciprocal hybrids, and their F₂s. Epidermal cells have a 2C, 4C or 8C nuclear DNA content. The mean DNA content per nucleus in IHP was significantly higher than in ILP; the mean DNA content per nucleus in hybrids was intermediate between the parental lines, and the same DNA content was measured in reciprocal crosses. In F₂s the same mean DNA content as in F₁s was observed but with larger variability than in the F₁, possibly indicating genetic segregation. It is inferred that the ploidy level in the leaf epidermis is inherited, and incomplete dominance occurs in hybrids. The same behaviour in the different genotypes was observed for epidermal cell-surface area, except that an increase of mean surface area occurred in the F₁, probably due to heterotic effects. The difference in the accumulation of 4C and 8C nuclei in leaf epidermis parallels that reported between two genotypes for the endosperm tissue: to the greater chromosome endoreduplication found in the endosperm there were correspondingly higher frequencies of 4C and 8C nuclei in the leaf epidermis, indicating a higher general tendency to chromosome endoreduplication in IHP than in ILP. It is suggested that the accumulation of 4C nuclei (G₂-block) in the leaf epidermis may be regarded as the initial step of chromosome endoreduplication, the two phenomena being related to the control of the sequence DNA synthesis-mitosis, possibly involving the same genes in both endosperm and leaf. However, the inheritance of DNA content per nucleus in epidermal tissue seems to be different from that observed in endosperm tissue of the same genotypes, suggesting that differences may occur in the regulation of the activity of these genes. Received: 19 November 1996 / Accepted: 29 November 1996     

Photoinhibition of isolated mesophyll cells from cold-hardened and non-hardened winter rye

Cold-hardened rye leaves have been shown to be more resistant to low temperature photoinhibition than non-hardened rye leaves. Isolated mesophyll cells from winter rye (Secale cereale L. cv. Musketeer) were exposed to photoinhibitory light conditions to estimate the importance of leaf morphology and leaf optical properties in the resistance of cold-hardened rye leaves to photoinhibition. Cold-hardened rye cells showed more resistance to photoinhibition than non-hardened rye cells when monitored with chlorophyll a variable to maximal fluorescence ratio (F_v/F_m). Thus, leaf morphology does not contribute to the resistance of cold-hardened rye leaves to low temperature photoinhibition. However, cold-hardened and non-hardened rye cells showed a similar extent of photoinhibition when photsynthetic CO₂ fixation rates were measured. They also showed the same capacity to recover from photoinhibition. During both photoinhibition and recovery, F_v/F_m and light limited CO₂ fixation rates showed different kinetics. We propose that inactivation and subsequent reactivation during recovery of some light activated Calvin cycle enzymes explain the greater extent of photoinhibition of light limited CO₂ fixation and its faster recovery compared to F_v/F_m kinetics during photoinhibition.     

Analysis of DNA size,content and cell cycle in leaves of Napier grass (Pennisetum purpureum Schum.)

Summary Mesophyll cell nuclei isolated from leaves of Pennisetum purpureum were analysed by flow cytometry to determine the nuclear DNA content and the percentage of cells in different phases of the cell cycle. Samples taken from base, middle and tip regions of leaves 2 to 8 (leaf 1, which was adjacent to the meristem, was too small to sample) showed no significant differences in the amount of DNA per G₁ nucleus due to either age or position. The average amount of DNA per G₁ nucleus was 5.78 pg. Although the majority of cells for each sample were in G₁, samples taken from older leaves had higher percentages of cells in G₂ and S phases. More specifically, base and middle regions of older leaves had a higher percentage of cells in G₂ than all three positions in younger leaves. Electrophoretic analysis of nuclear DNA from leaves 2 to 7 showed no evidence of degradation or difference in fragment size for any sample or position. This study was compared to previous work on the relationship between leaf age and embryogenic competence in Pennisetum purpureum. The results suggest that changes in the cell cycle, and/or a loss or fragmentation of the nuclear DNA, are not responsible for loss of embryogenic competence in mature leaf tissue.     

Nuclear DNA content in differentiated tissues of sunflower (Helianthus annuus L.)

Summary Scanning cytophotometry following Feulgen-staining was used to determine nuclear DNA content in many differentiated tissues of nine cultivars, hybrids or selfed lines ofHelianthus annuus. Apart from such ephemeral tissues as endosperm and anther tapetum, it was found that tissue differentiation in sunflower occurs in the diploid condition, cells being arrested in the DNA presynthetic phase (G₁). In certain cases, however, the nuclear DNA content of differentiated G₁ cells does not exactly match the 2C DNA content found in meristematic cells, but may be either higher or lower. In endosperm and anther tapetum cells, nuclear DNA content may be as high as 24 C and 32 C, respectively. Cytological and autoradiographic analyses after³H-thymidine incorporation reveal that polyploidy in the tapetal cells is due to chromosome endoreduplication. No detectable difference between male-fertile and male-sterile plants exists as far as occurrence and level of cell polyploidy are concerned. The results are discussed in the context of previous investigations on the nuclear condition of differentiatedHelianthus annuus tissue.     

Changes in nuclear and nucleolar protein content during the growth and differentiation of root parenchyma cells in plant species with different DNA-endoreplication dynamics

Summary Using cytophotometric procedures, we measured the nuclear and nucleolar protein content of successive zones of growth and differentiation in consecutive (1–7 mm) root segments obtained from eight species of the Angiospermae after staining the preparations with Feulgen-Naphthol Yellow S (F-NYS). In meristematic cells the nuclear and nucleolar protein content was found to double during the cell cycle. In species in which differentiation occurs at the same time as nuclear DNA endoreplication, i.e. Vicia faba subsp. minor, V. faba subsp. major, Pisum sativum, Hordeum vulgare and Amaryllis belladonna, the pool of nuclear proteins observed during the G₂ phase of the cell cycle was seen in the differentiated zone in nuclei containing 8C DNA. Species in which differentiation is not accompanied by the process of nuclear DNA endoreplication, i.e. Levisticum officinale, Tulipa kaufmanniana and Haemanthus katharinae, exhibited the highest nuclear proteins content during the G₂ phase of the cell cycle; comparably high values were not found in the differentiated zone. A decrease in nucleolar protein content was observed during the process of differentiation, this tendency being more evident in the studied species that do not exhibit endoreplication.This work was supported by the Polish Academy of Sciences as a part of project no 09.7.1.4.5     

Calcium influence on the dry mass content and surface area of nuclei and cytoplasm during differentiation of cortex cells in pea (Pisum sativum L.) roots treated with heavy metals

The effect of Ca addition to Cd, Cr and Pb solutions on the nuclear and cytoplasmic dry mass content and its concentration as well as on these organelles dimensions were studied in cortex cells of pea roots. Ca alone, at the concentration 10(-8)M brought about a decrease (in comparison to water) in the dry mass content of nuclei and its concentration, but the increment was almost twice in the dry mass content of cytoplasm; however, it has no significant effect on its concentration. Ca ions addition does not change the surface area of nuclei except the 1st and 5th mm segments but causes a doubling of the area occupied by cytoplasm. In response to Ca addition to Cd or Cr solutions a further diminution of nuclear dry mass content takes place. Only in the case of Pb nuclear dry mass increases in the 7th mm or is similar in remaining root segments. The diminution of nuclear dry mass content due to Ca presence in metal solutions is accompanied by a lowering in its concentration, although in the presence of Cd and Pb the diminution is not significant. Ca ions addition results in an increase in cytoplasmic dry mass content. No such regularities were observed in the 1st (Cd, Pb) and 3rd (Cr) root segments. In response to Ca ions the concentration of cytoplasmic dry mass content increased insignificantly in differentiation zone and underwent reducation in the meristematic zone--in the 1st mm (Cr) and 3rd (Cd). After Ca addition to studied metal solutions the decrease in nuclear dimensions was visible only in Cd or Pb treated cells in the 3rd and 7th or in the 1st mm, respectively. An increase in nuclear size occurred only in Cr treated cells in the 7th mm. Enrichment of heavy metals with Ca caused the marked enlargement in cytoplasmic area in differentiation zone but the increment in it in meristematic zone was observed only in Cd (1st, 3rd) and Cr (1st) treated cells.     

Copy numbers of chloroplast and nuclear genomes are proportional in mature mesophyll cells of Triticum and Aegilops species

The possibility of estimating the proportion of chloroplast DNA (ctDNA) and nuclear DNA (nDNA) in nucleic-acid extracts by selective digestion with the methylation-sensitive restriction enzyme PstI, was tested using leaf extracts from Spinacia oleracea and Triticum aestivum. Values of ctDNA as percentage nDNA were estimated to be 14.58%±0.56 (SE) in S. oleracea leaves and 4.97%±0.36 (SE) in T. aestivum leaves. These estimates agree well with those already reported for the same type of leaf material. Selective digestion and quantitative dot-blot hybridisation were used to determine ctDNA as percentage nDNA in expanded leaf tissue from species of Triticum and Aegilops representing three levels of nuclear ploidy and six types of cytoplasm. No significant differences in leaf ctDNA content were detected: in the diploids the leaf ctDNA percentage ranged between 3.8% and 5.1%, and in the polyploids between 3.5% and 4.9%. Consequently, nuclear ploidy and nDNA amount were proportional to ctDNA amount (r₍₁₉₎=0.935, P>0.01) and hence to ctDNA copy number in the mature mesophyll cells of these species. There was a slight increase in ctDNA copy numbers per chloroplast at higher ploidy levels. The balance between numbers of nuclear and chloroplast genomes is discussed in relation to polyploidisation and to the nuclear control of ctDNA replication.Abbreviations ctDNA chloroplast DNA - nDNA nuclear DNA - RuBPCase ribulose-1,5-bisphosphate carboxylase - DAPI 4,6-diamidine-2-phenylindole     

Nuclear DNA content and the control of chloroplast replication in wheat leaves

During development of the first leaf of breadwheat (Triticum aestivum L.) the number of chloroplasts per mesophyll cell increases between three- and four-fold. To establish if chloroplast replication is accompanied by endoreduplication, the nuclear DNA content of the cells was determined by chemical assay of isolated nuclei from mesophyll protoplasts and by microdensitometry of nuclei in mesophyll tissue. The DNA content of the nuclei was constant (27 to 32 pg) at each phase of chloroplast replication. Approximately 93% of the cells had a nuclear DNA content close to the 2C value of 32 pg. It is concluded that chloroplast replication is not dependent on nuclear endoreduplication in seedling leaves of wheat.     

Cessation of Nuclear DNA Synthesis in Differentiating Naegleria*

SYNOPSIS. DNA synthesis during growth and differentiation in Naegleria gruberi strain NEG populations has been studied. Autoradiography of cells labeled with [³H]thymidine revealed that grains are concentrated over the nuclei in logarithmically growing populations of cells, whereas in differentiating cells, grains are scattered over the cytoplasm; i.e. no significant nuclear labeling is detectable. It was established by MAK chromatographic analysis that [³H]thymidine is incorporated into double-stranded DNA in Naegleria and that the actual amount of incorporation in the logarithmically growing populations of cells is 20 times greater than that in differentiating cells. These results suggest that nuclear DNA synthesis is reduced markedly soon after the initiation of differentiation, while cytoplasmic DNA synthesis continues. It was established from cell cycle analysis that the approximate intervals of G₁, S, G₂, and M phases were 180, 183, 90, and 28 min, respectively. Hence, the reduction in the nuclear DNA synthesis in differentiating cells is not due to the inhibition of initiation of DNA replication, but rather to the termination of the DNA replicating process. Thus DNA synthesis is curtailed in the presence of RNA and protein synthesis which are required for differentiation.     

Comparative analysis of photosynthetic characteristics and productivity in F2 hybrids of winter rye

Diversity of photosynthetic characteristics determined by plant genotypes provides the grounds for the increase in potential crop productivity by means of producing plant forms whose photosynthetic apparatus (PSA) has optimal size and functional efficiency. Parental forms of winter rye (Secale cereale L.) and their interline and intravarietal reciprocal F₂ hybrids were compared in terms of photosynthetic pigment (PSP) content, characteristics of chlorophyll a (Chl) fluorescence, some morphological traits, and grain productivity. At the booting and anthesis stages, significant divergence in chlorophyllous pigment content and Chl a fluorescence parameters was observed for winter rye inbred lines, rye varieties, and hybrids. The hybrids were revealed whose elevated grain productivity correlated with high PSP content and the highest photosystem II (PSII) activity. Analysis of correlations in reciprocal F₂ hybrids at booting and anthesis stages of rye development showed that accumulation of PSII is related to stem-forming capacity and to the flag leaf surface area. In reciprocal hybrids, the correlations between morphological traits, grain productivity, plastid pigments, and photochemical activity were subject to variations. The relationships between PSA parameters and grain productivity in winter rye F₂ hybrids varied depending on the developmental stage as well as on crossing combination; this relation was largely determined by the choice of pairs for hybridization, by direction of crosses, and by genetic features of parental forms.     

Cultured human keratinocytes: discrimination of different cell cycle compartments based upon measurement of nuclear RNA or total cellular RNA content

Correlated measurements of total cellular RNA and DNA of cultured human keratinocytes by flow cytometry, followed by multivariate analysis, discriminate three distinct subpopulations of cells differing in RNA content. The first subpopulation is comprised of small cells resembling basal cells of epidermis, with low RNA content and long (100-300 h) generation times. The second subpopulation consists of keratinocytes resembling cells in the spinous layer of epidermis, characterized by increased RNA content and shorter (35-40 h) generation times. The third subpopulation consists of the largest, keratinohyalin-containing cells which remain in G1 and undergo terminal differentiation. In contrast to total cellular RNA, correlated measurements of DNA and nuclear RNA reveal that: (1) entrance of all cultured cells from G1 into S phase occurs only after accumulation of the same, threshold amount of nuclear RNA; hence there is only a single population of S + G2 + M-phase cells; (2) there are two distinct subpopulations in G1, one with minimal nuclear RNA content and another with increased RNA. Stathmokinetic experiments indicate that the G1-phase cells with low nuclear RNA have distinctly longer residence times in G1 compared to cells with high nuclear RNA content. Thus, measurements of the total cellular RNA versus nuclear RNA content reveal kinetically distinct cell subpopulations. Whereas total cellular RNA content correlates more with differentiation, nuclear RNA content reflects primarily the kinetic properties of the cell.     

Cultured human keratinocytes: discrimination of different cell cycle compartments based upon measurement of nuclear RNA or total cellular RNA content

Abstract Correlated measurements of total cellular RNA and DNA of cultured human keratinocytes by flow cytometry, followed by multivariate analysis, discriminate three distinct subpopulations of cells differing in RNA content. The first subpopulation is comprised of small cells resembling basal cells of epidermis, with low RNA content and long (100–300 h) generation times. The second subpopulation consists of keratinocytes resembling cells in the spinous layer of epidermis, characterized by increased RNA content and shorter (35–40 h) generation times. The third subpopulation consists of the largest, keratinohyalin-containing cells which remain in G₁ and undergo terminal differentiation. In contrast to total cellular RNA, correlated measurements of DNA and nuclear RNA reveal that: (1) entrance of all cultured cells from G₁ into S phase occurs only after accumulation of the same, threshold amount of nuclear RNA; hence there is only a single population of S + G₂+ M-phase cells; (2) there are two distinct subpopulations in G₁, one with minimal nuclear RNA content and another with increased RNA. Stathmokinetic experiments indicate that the G₁-phase cells with low nuclear RNA have distinctly longer residence times in G₁ compared to cells with high nuclear RNA content. Thus, measurements of the total cellular RNA versus nuclear RNA content reveal kinetically distinct cell subpopulations. Whereas total cellular RNA content correlates more with differentiation, nuclear RNA content reflects primarily the kinetic properties of the cell.     

Chromosome diminution in Ascaris suum. Two-fold increase of nucleosomal histone to DNA ratios during development

The swine intestinal nematode, Ascaris suum, eliminates chromatin material from its primordial somatic cells during early embryogenesis. A technique for isolation of nuclei from pre- and post-diminution stage embryos has been developed and these isolated nuclei were used in investigations of nuclear events during diminution. The amount of DNA per nucleus determined by diphenylamine assays and isotope dilutions was 0.66 pg and 0.29 pg in pre- and post-diminution nuclei, respectively. Thus, A. suum loses 56% of its nuclear DNA during diminution. The loss of nuclear DNA enabled in vivo examination of histone to DNA ratios as a function of changes in DNA quantities. Ascaris histones were identified by acid extractability and tryptic fingerprint comparison with rat liver histones. Measurement of histone quantities was accomplished using linearity of Coomassie blue binding to histones separated in dodecyl sulfate gels. Ascaris nucleosomal histones levels were relatively constant in pre- and post-diminution nuclei. However, nucleosomal histone to DNA ratios approximately doubled during diminution.     

Chromatin diminution in nematodes

The process of chromatin diminution in Parascaris and Ascaris is a developmentally controlled genome rearrangement, which results in quantitative and qualitative differences in DNA content between germ line and somatic cells. Chromatin diminution involves chromosomal breakage, new telomere formation and DNA degradation. The programmed elimination of chromatin in presomatic cells might serve as an alternative way of gene regulation. We put forward a new hypothesis of how an ancient partial genome duplication and chromatin diminution may have served to maintain the genetic balance in somatic cells and simultaneously endowed the germ line cells with a selective advantage.     

Difference in light-induced increase in ploidy level and cell size between adaxial and abaxial epidermal pavement cells of Phaseolus vulgaris primary leaves

Changes in nuclear DNA content and cell size of adaxial andabaxial epidermal pavement cells were investigated using brightlight-induced leaf expansion of Phaseolus vulgaris plants. Inprimary leaves of bean plants grown under high (sunlight) ormoderate (ML; photon flux density, 163 µmol m^–2s^–1) light, most adaxial epidermal pavement cells hada nucleus with the 4C amount of DNA, whereas most abaxial pavementcells had a 2C nucleus. In contrast, plants grown under lowintensity white light (LL; 15 µmol m^–2 s^–1)for 13 d, when cell proliferation of epidermal pavement cellshad already finished, had a 2C nuclear DNA content in most adaxialpavement cells. When these LL-grown plants were transferredto ML, the increase in irradiance raised the frequency of 4Cnuclei in adaxial but not in abaxial pavement cells within 4d. On the other hand, the size of abaxial pavement cells increasedby 53% within 4 d of transfer to ML and remained unchanged thereafter,whereas adaxial pavement cells continuously enlarged for 12d. This suggests that the increase in adaxial cell size after4 d is supported by the nuclear DNA doubling. The differentresponses between adaxial and abaxial epidermal cells were notinduced by the different light intensity at both surfaces. Itwas shown that adaxial epidermal cells have a different propertythan abaxial ones. Key words: Cell enlargement, endopolyploidization, epidermal pavement cells, incident light intensity, leaf expansion, nuclear DNA content, Phaseolus vulgaris     

The influence of some environmental factors on cytological and biometric parameters and chlorophyll content of <Emphasis Type="Italic">Deschampsia antarctica</Emphasis> Desv. in the maritime Antarctic

Under the environmental conditions of the Point Thomas Oasis (King George Island, the South Shetland Islands), we studied the influence of month-long artificial treatment with fresh water, salt water, and guano solution on the biometric characteristics, chlorophyll content, as well as the nuclear area of leaf parenchymal cells and nuclear DNA content, in a maritime Antarctic aboriginal plant Deschampsia antarctica. The modeled factors induced an increase in the generative shoot height and the length of the largest leaf but did not influence the number of flowers. Treatment with guano caused an increase in the chlorophyll a and b contents, while fresh water treatment only led to some increase in chlorophyll a. Fluctuations of physiologically significant traits, such as the nuclear area and DNA content in the leaf parenchyma cells of D. antarctica, have been traced under the influence of the studied factors. Understanding of the hierarchy of influence of these factors as well as and sensitivity of plants of this species to external agents require further investigation.     

Photosynthesis, Leaf Anatomy, and Cellular Constituents in the Polyploid C(4) Grass Panicum virgatum

Photosynthetic gas exchange, activities of six key C₄ cycle enzymes, amounts of soluble protein, chlorophyll, and DNA, and various leaf anatomical and structural features were measured in naturally occurring tetraploid and octaploid plants of the NAD-malic enzyme type C₄ grass Panicum virgatum L. On a leaf area basis, the photosynthetic rate and concentrations of DNA, soluble protein, and chlorophyll were 40 to 50% higher, and enzyme activities 20 to 70% higher in the octaploid than in the tetraploid. Photosynthetic cells in the octaploid were only 17 to 19% larger in volume, yet contained 33 to 38% more chloroplasts than cells in the tetraploid. On a per cell basis the contents of DNA, soluble protein, and chlorophyll, activities of carboxylating photosynthetic enzymes, and carbon assimilation rate were all doubled in octaploid compared with tetraploid cells. Since cellular volume did not double with genome doubling, cellular constituents were more concentrated in the cells of the octaploid. The influences of polyploidy were balanced between mesophyll and bundle sheath cells since the changes in physical and biochemical parameters with ploidy level were similar in both cell types. We conclude that photosynthetic activity in these two polyploid genotypes of P. virgatum is determined by enzyme activities and concentrations of biochemical constituents, and that selection for smaller cell volume has led to higher photosynthetic rates per unit leaf area in the octaploid. The ratio of DNA content to cellular volume is a major factor determining the concentrations of gene products in cells. The number of chloroplasts, however, is controlled more by cellular volume than by the number of nuclear chromosomes.     

DNA-Feulgen cytophotometric determination of genome size for the freshwater-invading copepod Eurytemora affinis.

Variation in nuclear DNA content within some eukaryotic species is well documented, but causes and consequences of such variation remain unclear. Here we report genome size of an estuarine and salt-marsh calanoid copepod, Eurytemora affinis, which has recently invaded inland freshwater habitats independently and repeatedly in North America, Europe, and Asia. Adults and embryos of E. affinis from the St. Lawrence River drainage were examined for somatic cell DNA content and the presence or absence of embryonic chromatin diminution, using Feulgen-DNA cytophotometry to determine a diploid or 2C genome size of 0.6-0.7 pg DNA/cell. The majority of somatic cell nuclei, however, have twice this DNA content (1.3 pg/nucleus) in all of the adults examined and possibly represent a population of cells arrested at the G2 stage of the cell cycle or associated with some degree of endopolyploidy. Both suggestions contradict assumptions that DNA replication does not occur in adult tissues during the determinate growth characteristic of copepods. Absence of germ cell nuclei with markedly elevated DNA values, commonly found for species of cyclopoid copepods that show chromatin diminution, indicates that E. affinis lacks this trait. The small genome size and presumed absence of chromatin diminution increase the potential utility of E. affinis as a model for genomic studies on mechanisms of adaptation during freshwater invasions.     

The application of growth regulators for the physiological emasculation of rye(secale cereale l.) flowers

The present paper deals with the possibility of using growth regulators for the physiological emasculation of rye flowers and utilizes our knowledge of the higher sensibility of stamens to the auxin level. By means of bioassays it has been found that a relatively low content of auxins and a definite level of gibberellins and inhibitors are characteristic for normal rye spikes at the time of stamen differentiation. The higher level of auxins and expressive inhibitions occur in the later course of pistil differentiation. Two sprayings of plants with mixed solutions of MH and NAA and two further sprayings with a solution of NAA only before the stamen differentiation change slightly the natural character of endogenous regulators in spikes and cause the destruction of mother pollen cells in anthers and the sterility of anthers in the majority of rye florets. The anthers in the basis of spikes sometimes contain pollen grains after treatment, but their fertility is substantially lowered because the spray evidently disturbed the accumulation of reserve substances in pollen grains. The development of pistil was not affected by the exogenous application of growth regulators. Normal kernels arose in the majority of flowers after supplementary pollination. Some accidental effects of exogenous application of growth regulators, as for example reduced stem growth and disorder of lodicule function were lowered to a great extent by adding GA_S to the last two sprayings of NAA.     

Effects of Polyploidy on Photosynthetic Rates, Photosynthetic Enzymes, Contents of DNA, Chlorophyll, and Sizes and Numbers of Photosynthetic Cells in the C(4) Dicot Atriplex confertifolia

Photosynthetic rates, chlorophyll content, and activities of several photosynthetic enzymes were determined per cell, per unit DNA, and per unit leaf area in five ploidal levels of the C₄ dicot Atriplex confertifolia. Volumes of bundle sheath and mesophyll protoplasts were measured in enzymatic digestions of leaf tissue. Photosynthetic rates per cell, contents of DNA per cell, and activities of the bundle sheath enzymes ribulose 1,5-bisphosphate carboxylase (RuBPC) and NAD-malic enzyme per cell were correlated with ploidal level at 99% or 95% confidence levels, and the results suggested a near proportional relationship between gene dosage and gene products. There was also a high correlation between volume of mesophyll and bundle sheath cells and the ploidal level. Contents of DNA per cell, activity of RuBPC per cell, and volumes of cells were correlated with photosynthetic rate per cell at the 95% confidence level. The mesophyll cells did not respond to changes in ploidy like the bundle sheath cells. In the mesophyll cells the chlorophyll content per cell was constant at different ploidal levels, there was less increase in cell volume than in bundle sheath cells with an increase in ploidy, and there was not a significant correlation (at 95% level) of phosphoenolpyruvate carboxylase activity or content and pyruvate,Pi dikinase activity with increase in ploidy. The number of photosynthetic cells per unit leaf area progressively decreased with increasing ploidy from diploid to hexaploid, but thereafter remained constant in octaploid and decaploid plants. Numbers of cells per leaf area were not correlated with cell volumes. The mean photosynthetic rates per unit leaf area were lowest in the diploid, similar in 4×, 6×, and 8×, and highest in the decaploid. The photosynthetic rate per leaf area was highly correlated with the DNA content per leaf area.