Effects of Organic Solvents on the Coupling Between ATP Hydrolysis and Sliding of Microtubules in Axonemes from Chlamydomonas Flagella

ABSTRACT. The effects of organic solvents on the ATPase activity and the sliding disintegration of axonemes from Chlamydomonas were investigated. The axonemal ATPase was markedly activated by methanol accompanying with marked inhibition of the sliding disintegration of axonemes. On the contrary, glycerol inhibited the ATPase activity without serious inhibition of the sliding disintegration. As far as the axonemes are not irreversibly denatured by extremely high concentration of solvents, the effects of solvents both on the ATPase and the ability of sliding are reversible. Therefore, the inhibition of sliding accompanied by the activation of ATPase is probably due to an inability to couple the hydrolysis of ATP to sliding between dynein and microtubule in the presence of methanol. The axonemal ATPase was less sensitive to vanadate inhibition after exposure to methanol. This indicates that methanol makes the dyneinADP.Pi complex unstable and increases product release. On the other hand, glycerol and ethylene glycol seem to stabilize the force generation responsible for the sliding through stabilizing the dynein.ADP.Pi complex.     

The translocation of photosynthetic products from mesophyll into midrib in tobacco plant III. The transformation of translocated 14C-sugars in the veins

¹⁴C-U-sugars were introduced into tobacco plants through themesophyll, the veins of the first order of branching, and themidrib, and ¹⁴C-compounds in the veins and the midrib whichtranslocated towards the base of the midrib were traced duringthe period of 120 min after the ¹⁴C-sugar introductions. 1) When ¹⁴C-U-sucrose was introduced into the leaf, no matterwhat the means of feeding was, most of the ¹⁴C which translocatedbasipetally in the veins and the midrib was found in the formof sucrose. 2) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was administered tothe leaf dirough the cut vein of the first order of branching,most of the ¹⁴C which translocated basipetally in the veinsand the midrib was found in the form of sucrose. 3) ¹⁴C-U-glucose or ¹⁴C-U-fructose injected into the vascularbundles of the midrib was translocated basipetally, as such,10 and 30 min after injection; and at 30 min, the amount ofthe ¹⁴C-sucrose in the midrib attained 9–22% of the 80%ethanol-soluble ¹⁴C in the midrib. 4) When ¹⁴C-U-glucose or ¹⁴C-U-fructose was supplied to themesophyll, the radioactivities of these hexoses were predominantin the first and second veins soon after application, then decreasingwith a concomitant increase in the radioactivity of the ¹⁴C-sucrose. From these results, it was inferred that in the veins of thefirst and second order of branching, glucose and fructose whichmoved from the mesophyll did not translocate as such, but wereutilized for the synthesis of sucrose available for translocationvia the midrib to the stem. ¹A part of this paper was presented at the Crop Science Societyof Japan, in April, 1969 (Received December 8, 1969; )     

Mesophyll conductance decreases in the wild type but not in an ABA‐deficient mutant (aba1) of Nicotiana plumbaginifolia under drought conditions

Under drought conditions, leaf photosynthesis is limited by the supply of CO₂. Drought induces production of abscisic acid (ABA), and ABA decreases stomatal conductance (g_s). Previous papers reported that the drought stress also causes the decrease in mesophyll conductance (g_m). However, the relationships between ABA content and g_m are unclear. We investigated the responses of g_m to the leaf ABA content [(ABA)_L] using an ABA‐deficient mutant, aba1, and the wild type (WT) of Nicotiana plumbaginifolia. We also measured leaf water potential (Ψ_L) because leaf hydraulics may be related to g_m. Under drought conditions, g_m decreased with the increase in (ABA)_L in WT, whereas both (ABA)_L and g_m were unchanged by the drought treatment in aba1. Exogenously applied ABA decreased g_m in both WT and aba1 in a dose‐dependent manner. Ψ_L in WT was decreased by the drought treatment to ?0.7 MPa, whereas Ψ_L in aba1 was around ?0.8 MPa even under the well‐watered conditions and unchanged by the drought treatment. From these results, we conclude that the increase in (ABA)_L is crucial for the decrease in g_m under drought conditions. We discuss possible relationships between the decrease in g_m and changes in the leaf hydraulics.     

Response of mitochondria to light intensity in the leaves of sun and shade species

The present authors have shown previously that both respiration rates and in vivo activities of the alternative oxidase (AOX) of leaves of Alocasia odora, a shade species, are lower than those in sun species, thereby optimizing energy production under limited light conditions (Noguchi et al., Australian Journal of Plant Physiology 28, 27–35, 2001). In the present study, mitochondria isolated from A. odora leaves were examined in order to investigate the biochemical basis for the differences in respiratory parameters. Alocasia odora and spinach plants were cultivated under both high and low light intensities, mitochondria were isolated from their leaves, and their respiratory properties compared. Mitochondrial content of leaf extracts from the two species was estimated using fumarase activities and antibody detection of porin (the voltage-dependent anion channel of the outer mitochondrial membrane). On a mitochondrial protein basis, spinach leaves showed higher capacities of the cytochrome pathway and cytochrome c oxidase (COX) than A. odora leaves. However, on a mitochondrial protein basis, A. odora showed higher capacities of AOX, which had a high affinity for ubiquinone when activated by pyruvate. Alocasia odora also had larger amounts of mitochondrial protein per leaf dry weight, even under severely shaded conditions, than spinach. Lower growth light intensity led to lower activities of most pathways and proteins tested in both species, especially glycine-dependent oxygen uptake. In the low light environment, most of the AOX protein in A. odora leaves was in its inactive, oxidized dimer form, but was converted to its reduced active form when plants were grown under high light. This shift may prevent over-reduction of the respiratory chain under photo-oxidative conditions.     

Phytochemical studies on tobacco alkaloids XIV. The occurrence and properties of putrescine N-methyltransferase in tobacco roots

Putrescine N-methyltransferase, a new enzyme catalyzing theformation of N-methylputrescine from putrescine and S-adenosyl-L-methioninewas found in roots of tobacco plants. The enzyme was purified30-fold from crude extracts of tobacco roots. NMethylputrescinewas identified as the reaction product by comparison with theauthentic compound. The enzyme had a pH optimum between pH 8and 9, and a molecular weight of about 60,000, as determinedby gel filtration. Km values for putrescine and 5-adenosyl-L-methioninewere 4.0 x 10^–4 M and 1.1 x 10^–4 M, respectively.Enzyme activity was inhibited by N-chloromercuribenzoate andAg⁺. No cofactors were required. Of the various substrates tested,only putrescine served as a methyl acceptor. The enzyme waslocalized exclusively in the roots and its activity was greadyenhanced by decapitation. The presence of putrescine N-methyltransferase in tobacco rootsstrongly suggests that N-methylputrescine participates as anintermediate in nicotine biosynthesis. (Received March 2, 1971; )     

Effect of respiratory homeostasis on plant growth in cultivars of wheat and rice

Some plants have the ability to maintain similar respiratory rates (measured at the growth temperature), even when grown at different temperatures, a phenomenon referred to as respiratory homeostasis. The underlying mechanisms and ecological importance of this respiratory homeostasis are not understood. In order to understand this, root respiration and plant growth were investigated in two wheat cultivars (Triticum aestivum L. cv. Stiletto and cv. Patterson) with a high degree of homeostasis, and in one wheat cultivar (T. aestivum L. cv. Brookton) and one rice cultivar (Oryza sativa L. cv. Amaroo) with a low degree of homeostasis. The degree of homeostasis (H) is defined as a quantitative value, which occurs between 0 (no acclimation) and 1 (full acclimation). These plants were grown hydroponically at constant 15 or 25 °C. A good correlation was observed between the rate of root respiration and the relative growth rates (RGR) of whole plant, shoot or root. The plants with high H showed a tendency to maintain their RGR, irrespective of growth temperature, whereas the plants with low H grown at 15 °C showed lower RGR than those grown at 25 °C. Among several parameters of growth analysis, variation in net assimilation rate per shoot mass (NAR_m) appeared to be responsible for the variation in RGR and rates of root respiration in the four cultivars. The plants with high H maintained their NAR_m at low growth temperature, but the plants with low H grown at 15 °C showed lower NAR_m than those grown at 25 °C. It is concluded that respiratory homeostasis in roots would help to maintain growth rate at low temperature due to a smaller decrease in net carbon gain at low temperature. Alternatively, growth rate per se may control the demand of respiratory ATP, root respiration rates and sink demands of photosynthesis. The contribution of nitrogen uptake to total respiratory costs was also estimated, and the effects of a nitrogen leak out of the roots and the efficiency of respiration on those costs are discussed.