The Effects of Fusicoccin,Abscisic Acid,and Benzyladenine on the Transport and Partitioning of Substances as Related to Cytokinin Sink Activity

We tested the possible cytokinin effect on the functioning of the active transport system involved in the assimilate loading into the phloem as a cause for the cytokinin sink and retention effect. This effect is manifested in the deceleration of substance export from and the stimulation of substance import to the sites of local phytohormone application to the mature detached leaf from untreated leaf areas. To affect the membrane mechanisms of the substance transport, we used leaf treatment with the phytotoxin fusicoccin, an enhancer of plasmalemmal H⁺-ATPase and a potential stimulator of assimilates export, and with the phytohormone ABA affecting transport, metabolism, and plant growth. However, fusicoccin did not enhance ¹⁴C-sucrose export from the leaf blade and did not interfere with the cytokinin-induced export deceleration. ABA reduced substantially ¹⁴C export from the leaf but eliminated the cytokinin effect on this process. Similar results were obtained for broad bean (Vicia faba L.) leaves with apoplastic phloem loading, involving H⁺-ATPase activity, and pumpkin (Cucurbita pepo L.) leaves with symplastic phloem loading, that is, occurring without sucrose transmembrane translocation and without H⁺-ATPase involvement. The conclusion is that the cytokinin-induced development of sink zones in source leaves is not related to the membrane mechanisms of the substance transport in the mesophyll–phloem system. The data obtained support the idea that the cause for the cytokinin sink and retention effect is the enhancement of elongation growth and total activation of metabolism in the mesophyll cells of the detached leaf.     

Sink Effect of Cytokinin in Detached Leaves Is Not Caused by Structural Rearrangements of Phloem

The sink effect of cytokinin is manifested as a decrease in source capacity and the induction of sink activity in the phytohormone-treated region of a mature excised leaf. In order to find out whether this effect was due to the direct action of cytokinin on the phloem structure, two types of phloem terminals were examined. In pumpkin (Cucurbita pepo L.) leaves, the phloem terminals are open; i.e., they are linked to mesophyll by numerous symplastic connections, which are located in narrow areas called plasmodesmal pit fields. In broad bean (Vicia faba L.) leaves, the phloem terminals belong to the closed type and have no symplastic links with mesophyll. The electron microscopic study of terminal phloem did not reveal any structural changes in the companion cells, which could account for the suppression of assimilate export. The treatment of leaves with cytokinin neither disturbed the structure of plasmodesmal pit fields in pumpkin leaves nor eliminated the wall protuberances (the ingrowths promoting phloem loading) in bean leaves. No evidence was obtained that the cytokinin-induced import of assimilates in mature leaves is caused by the recovery of meristematic activity, i.e., by either formation of new phloem terminals having immature sieve elements capable of unloading or by the development of new sieve elements within the existing veins. Cytokinin did not induce de novo formation of phloem elements. Structural characteristics of the leaf phloem, such as the number of branching orders in the venation pattern, the number of vein endings per areole, the number of areoles per leaf, the area of one areole, and the number of sieve elements per bundle remained unaltered. It is concluded that the sink effect of cytokinin in excised leaves cannot be determined by alteration of the phloem structure.     

Cytokinin-Regulated Mesophyll Cell Division and Expansion during Development of Cucurbita pepo Leaves

The role of cytokinins in the development of mesophyll structure was studied in developing pumpkin Cucurbita pepo L. leaves. Leaves were treated with cytokinins at different stages of growth: when they reached 25 or 50% of their final size (S _max), immediately after leaf growth ceased, and during senescence. At the early stages of leaf development, treatment with exogenous benzyladenine accelerated division of mesophyll cells. At the later stages of development, BA treatment activated expansion of growing cells and those, which have just accomplished their growth. The exogenous cytokinin did not affect the senescent leaf cells. The content of endogenous cytokinins changed during mesophyll development. The juvenile leaves (25% of S _max) were characterized by low level of these phytohormones. In the expanding leaves (50% of S _max), the content of phytohormones increased and decreased when leaf growth ceased. In the senescent leaves, the cytokinin content decreased markedly. It was concluded that the response of mesophyll cells to cytokinin depended on the cell growth phase at the moment of hormone action. Furthermore, in the young leaves, lower cytokinin concentrations were required for division of mesophyll cells in vivo than for cell expansion at the final stage of leaf development.     

Structure of the photosynthetic apparatus of the bean Vicia faba L. when treated with 6-benzylaminopurine

The level of endogenous cytokinins changed with growth and development of faba bean (Vicia faba L.) leaves. Typical of juvenile leaves, amounting to 25% of the final leaf size (Smax) was a low content of these plant hormones. The level of cytokinins increased in growing leaves (50% of Smax) and decreased in the leaves that stopped growing. The content of cytokinins in senescent leaves dropped considerably. Exogenous treatment with 6-benzylaminopurine (BAP) had no effect on the structure of terminal phloem; however, it stimulated elongation of mesophyll cells; increases in the area and thickness of leaf blade, amount of photosynthetic pigments, and assimilation potential; and delayed senescence of leaves and defoliation, thereby increasing biomass of the aboveground plant part. It was inferred that BAP had a potential for induction of photosynthetic apparatus development and increase in the yield of faba bean green mass.     

The Effects of Arid Region Conditions on the Hormonal Status of Humans

The human hormonal status and its circadian and circannual rhythms that provide a background for human activities and diseases were studied under conditions of the extremely continental climate of the arid region with a long period of elevated ambient temperatures. Age- and sex-related features, biological rhythms (seasonal and circadian), and the dependence on the work specificity were studied for the activities and interrelations of the adrenal cortex, thyroid gland, and catecholamines. The peripheral blood plasma levels of the hormones and the urinary excretion of the hormone metabolites were determined by conventional methods. The findings show that the hormonal status depended on both the climatic and occupational conditions.     

Structural and Functional Rearrangements of Mesophyll as a Probable Basis for the Cytokinin-Dependent Assimilate Translocation in Detached Leaves

The effects of benzyladenine (BA) on the mesophyll functioning, such as osmotic potential (), the effect of the inhibitors of ⁺-ATPase on the influx of ¹⁴C-sucrose, the direction of carbon metabolism, and the rate of dark respiration, were followed in the detached leaves of pumpkin (Cucurbita pepo L.) and broad beans (Vicia faba L.). BA elevated and established a gradient of (_p) between the treated and untreated leaf regions. The inhibitors of H⁺-ATPase did not affect the BA-induced influx of ¹⁴C-sucrose. The changes were accompanied with the elevated synthesis of starch and other polymeric compounds and the diminished synthesis of the substances of relatively low molecular weight. The stimulation of dark respiration was short and inconsiderable. The author concludes that the BA-induced transport was a passive process related to a increase. Leaf expansion accompanied by the synthesis of high-molecular-weight substances essential for cell growth and by starch synthesis apparently increased the sink capacity of the BA-treated detached leaves. The diminished efflux from the leaf blade was probably related to a lowered level of the transportable carbon compounds restricting their entry into the phloem. The influx induction could result from the activation of growth and metabolic processes, the decline in the number of organic molecules per cell volume unit, and the development of _p between the source and sink leaf regions.