R. Harold Brown

Novel characteristics of cassava, Manihot esculenta Crantz, a reputed C3-C4 intermediate photosynthesis species

The cassava plant, Manihot esculenta, grows exceptionally well in low fertility and drought prone environments, but the mechanisms that allow this growth are unknown. Earlier, and sometimes contradictory, work speculated about the presence of a C4-type photosynthesis in cassava leaves. In the present work we found no evidence for a C4 metabolism in mature attached cassava leaves as indicated i) by the low, 2 to 8%, incorporation of 14CO2 into C4 organic acids in short time periods, 10 s, and the lack of 14C transfer from C4 acids to other compounds in 12CO2, ii) by the lack of C4 enzyme activity changes during leaf development and the inability to detect C4 acid decarboxylases, and iii) by leaf CO2 compensation values between 49 and 65 μl of CO2 1−1 and by other infrared gas exchange photosynthetic measurements. It is concluded that the leaf biochemistry of cassava follows the C3 pathway of photosynthesis with no indication of a C3-C4 mechanism.However, cassava leaves exhibit several novel characteristics. Attached leaves have the ability to effectively partition carbon into sucrose with nearly 45% of the label in sucrose in about one min of 14CO2 photosynthesis, contrasting with 34% in soybean (C3) and 25% in pigweed (C4). Cassava leaves displayed a strong preference for the synthesis of sucrose versus starch. Field grown cassava leaves exhibited high rates of photosynthesis and curvilinear responses to increasing sunlight irradiances with a tendency to saturate only at high irradiances, above 1500 μmol m−2 s−1. Morphologically, the cassava leaf has papillose epidermal cells on its lower mesophyll surface that form ‘fence-like’ arrangements encircling guard cells. It is proposed that the active synthesis of sugars has osmotic functions in the cassava plant and that the papillose epidermal cells function to maintain a healthy leaf water status in various environments.

Immunocytochemical localization of phosphoenolpyruvate carboxylase and photosynthetic gas-exchange characteristics in ears of Triticum durum Desf.

The presence and distribution of phosphoenolpyruvate carboxylase (PEPCase) in the glumes and immature grains of durum wheat (Triticum durum Desf.) were studied by electron-microscopical immunolabeling of PEPCase with polyclonal antibodies followed by protein A-gold. Plants were grown under mediterranean field conditions and samples were obtained two weeks after anthesis. In the kernels, high gold label was associated with the unstained areas of the protein bodies of aleurone cells, whereas labeling in the cytoplasm and chloroplasts of the pericarp was slight, although significantly above the background. In the glumes, high gold label was only located in cytoplasmic granules (vesicles) of the mesophyll cells, although labeling in the cytoplasm and chloroplasts was also significantly above the background. These observations in immature kernels and glumes are in accordance with the anaplerotic role of this enzyme, as evidenced in C3 plants. Measurements of apparent photosynthesis and its O2 dependence and CO2 compensation concentration were made on ears and flag leaves of durum wheat. In addition, an analog of phosphoenolpyruvate, 3,3-dichloro-2-dihydroxy-phosphinoylmethyl-2-propenoate, was used to inhibit PEPCase and, thereby, to assess the contribution of the PEPCase to photosynthesis in detached ears. There was no effect of the inhibitor on the apparent photosynthesis of ears. Whereas inhibition of apparent photosynthesis by 210 mL · L−1 O2 in flag leaves was typical of C3 species, inhibition in ears was even greater. The CO2 compensation concentrations in different ear parts were similar to or higher than in flag leaves and the O2 dependence was also comparable (about 70%). Therefore, gas-exchange data give further support to the assumption that a C4 cycle is absent or limited to very low rates in ears of durum wheat.

Relationships between specific leaf weight and mineral concentration among genotypes

Abstract Physiological functions are usually expressed on a leaf area basis, whereas leaf mineral concentrations are often expressed on a dry matter basis. If specific leaf weight (SLW; g DM m−2 leaf) differs among genotypes then variability in mineral concentration may depend on the basis of expression. Data from experiments with peanut (Arachis hypogaea L.) and pearl millet [Pennisetum glaucum (L.) R. Br.] lines and from the literature were used to examine relationships between leaf mineral concentration and SLW. Peanut and pearl millet were grown in pots in the greenhouse in soil and solution cultures. Specific leaf weight and ash and mineral concentrations were determined at the end of the experiments. Leaf ash concentration on a dry matter basis was negatively correlated with SLW and the correlation coefficients were significant in six of nine comparisons for the two species; r = −0.65 to −0.93. In the one peanut experiment in which mineral elements were determined, the correlations with ash were due mainly to correlations with Ca and Mg, while in pearl millet, correlations were due mainly to K. The slope of a plot of leaf constituents per unit of leaf area against SLW for a range of lines is a measure of the contribution of that leaf constituent to increased SLW. From data in the literature it appears that increased SLW is due mostly to the increase of cell wall components and nonstructural carbohydrates, and sometimes protein. Leaf mineral per unit of leaf area appears to be unrelated or only slightly increased with increased SLW and thus declines on a unit weight basis because of dilution by increased cell wall content or soluble carbohydrate.

Leaf anatomical characteristics in Flaveria trinervia (C4), Flaveria brownii (C4-like) and their F1 hybrid

Several leaf anatomical and ultrastructural characteristics usually related with photosynthetic capacity were examined in two Flaveria species with strong differences in anatomy and their F1 hybrid. Flaveria trinervia (Spreng.) Mohr (C4) was the female parent and F. brownii A.M. Powell (C4-like) was the male parent. Quantitative anatomical analysis was made on transverse sections of leaves at both the light and electron microscope level. Four kinds of photosynthetic tissues were considered: bundle sheath (BS), mesophyll adjacent to the BS, mesophyll not adjacent to the BS, and larger spongy mesophyll cells. Flaveria trinvervia partitioned a larger proportion of its photosynthetic cells to BS and the mesophyll layer adjacent to BS and also possessed larger chloroplasts, especially in BS, than did F. brownii. These results suggest that although F. brownii is very C4-like, its anatomy is not as completely C4 as is the case for F. trinervia. In the F1 hybrid the relative contribution of the different tissues to the total photosynthetic tissue volume and area per unit leaf area was quite similar to that of F. trinervia. On the other hand, the chloroplast density and size of the F1 hybrid were fairly similar to those of F. brownii, especially in BS. Thus, there was no evidence of maternal inheritance in the chloroplast characteristics studied. A negative correlation (P<0.05) between chloroplast size and density was observed among species and relicates within each kind of tissue. This correlation was highest (r=−0.94, P<0.001) for the BS and when values were plotted on a logarithmic scale. Thus, higher chloroplast numbers for F. brownii and the F1 hybrid were offset by larger chloroplasts in F. trinervia. Less complete C4 photosynthesis in F. brownii may be partially due to incomplete development of Kranz anatomy usually associated with C4 photosynthesis.

Leaf cavity CO2 concentrations and CO2 exchange in onion, Allium cepa L.

Onion (Allium cepa L.) plants were examined to determine the photosynthetic role of CO2 that accumulates within their leaf cavities. Leaf cavity CO2 concentrations ranged from 2250 μL L−1 near the leaf base to below atmospheric (<350 μL L−1) near the leaf tip at midday. There was a daily fluctuation in the leaf cavity CO2 concentrations with minimum values near midday and maximum values at night. Conductance to CO2 from the leaf cavity ranged from 24 to 202 μmol m−2 s−1 and was even lower for membranes of bulb scales. The capacity for onion leaves to recycle leaf cavity CO2 was poor, only 0.2 to 2.2% of leaf photosynthesis based either on measured CO2 concentrations and conductance values or as measured directly by 14CO2 labeling experiments. The photosynthetic responses to CO2 and O2 were measured to determine whether onion leaves exhibited a typical C3-type response. A linear increase in CO2 uptake was observed in intact leaves up to 315 μL L−1 of external CO2 and, at this external CO2 concentration, uptake was inhibited 35.4±0.9% by 210 mL L−1 O2 compared to 20 mL L−1 O2. Scanning electron micrographs of the leaf cavity wall revealed degenerated tissue covered by a membrane. Onion leaf cavity membranes apparently are highly impermeable to CO2 and greatly restrict the refixation of leaf cavity CO2 by photosynthetic tissue.