Cutinsomes of Malus Mill. (Rosaceae) leaf and pericarp: genesis, localization, and transport.

New data were obtained on specific bionanostructures, cutinsomes, which are involved in the formation of cuticles on the surface of leaf blades and pericarp of Malus domestica Borkh (Malus Mill., Rosaceae)introduced to the mountains at the altitudes of 1200 and 1700 m above sea level. Cutinsomes, which are electron-dense structures of spherical shape, have been identified by transmission electron microscopy. It was demonstrated that plastids can be involved in the synthesis of their constituent nanocomponents. The greatest number of nanoparticles was observed in the granal thylakoid lumen of the chloroplasts in palisade mesophyll cells and pericarp hypodermal cells. The transmembrane transport of cutinsomes into the cell wall cuticle proper by exocytosis has been visualized for the first time. The plasma membrane is directly involved in the excretion of nanostructures from the cell. Nanoparticles of cutinsomes in the form of necklace-like formations line up in a chain near cell walls, merge into larger conglomerates and are loaded into plasmalemma invaginations, and then, in membrane packing, they move into the cuticle, which covers both outer and inner cell walls of external tissues. The original materials obtained by us supplement the ideas about the non-enzymatic synthesis of cuticle components available in the literature and expand the cell compartment geography involved in this process.

Zinc- and nickel-induced changes in fatty acid profiles in the zinc hyperaccumulator Arabidopsis halleri and non-accumulator Arabidopsis lyrata.

This pilot study aimed at comparing zinc (Zn) and nickel (Ni) effects on the fatty acid (FA) profiles, oxidative stress and desaturase activity in the Zn hyperaccumulator Arabidopsis halleri and the excluder Arabidopsis lyrata to allow a better picture of the physiological mechanisms which may contribute to metal tolerance or acclimation. The most significant changes in the FA composition were observed in the shoots of the hyperaccumulator and in the roots of the excluder, and were not only metal-dependent, but also species-specific, since the most significant changes in the shoots of A. halleri were observed under Ni treatment, though Ni, in contrast to Zn, was accumulated mainly in its roots. Several FAs appeared in the roots and shoots of A. lyrata only upon metal exposure, whereas they were already found in control A. halleri. In both species, there was an increase in oleic acid under Ni treatment in both organs, whereas in Zn-treated plants the increase was shown only for the shoots. A rare conjugated α-parinaric acid was identified only in the shoots of metal-treated A. halleri. In the shoots of the hyperaccumulator, there was an increase in the content of saturated FAs and a decrease in the content of unsaturated FAs, while in the roots of the excluder, the opposite pattern was observed. These metal-induced changes in FA composition in the shoots of A. halleri can lead to a decrease in the fluidity of membranes, which could diminish the penetration of ROS into the membrane and thus maintain its stability.

A leader peptide of the extracellular cyanobacterial carbonic anhydrase ensures the efficient secretion of recombinant proteins in Escherichia coli.

Several forms of EcaA protein, correspondent to the extracellular α-class carbonic anhydrase (CA) of cyanobacterium Crocosphaera subtropica ATCC 51142 were expressed in Escherichia coli. The recombinant proteins with no leader peptide (EcaA and its fusion with thioredoxin or glutathione S-transferase) were allocated inside cells in a full-length form; these cells did not display any extracellular CA activity. Soluble proteins (including that of periplasmic space) of E. coli cells that expressed both ЕсаА equipped with its native leader peptide (L-EcaA) as well as L-EcaA fused with thioredoxin or glutathione S-transferase at N-terminus, mainly contained the processed EcaA. The appearance of mature ЕсаА in outer layers of E. coli cells expressed leader peptide-containing forms of recombinant proteins, has been directly confirmed by immunofluorescent microscopy. Those cells also displayed high extracellular CA activity. In addition, the mature EcaA protein was detected in the culture medium. This suggests that cyanobacterial signal peptide is recognized by the secretory machinery and by the leader peptidase of E. coli even as a part of a fusion protein. The efficiency of EcaA leader peptide was comparable to that of PelB and TorA signal peptides, commonly used for biotechnological production of extracellular recombinant proteins in E. coli.

The features of the fatty acid composition of Pyrus L. total lipids are determined by mountain ecosystem conditions.

The composition of fatty acids (FAs) of total lipids of pericarp, seeds, and leaves of Pyrus caucasica Fed. and Pyrus communis L. growing in mountain ecosystems at different altitudes (300, 700 and 1200 m) was studied. It was found that the greatest differences in the relative content of FAs within a species, depending on the altitudes above sea level, were characteristic of the outer tissues of the pericarp (peel) and leaves, which were in direct contact with the external environment. Pericarp parenchyma to a lesser extent, and seeds practically did not differ in FA composition at different heights. At altitudes with increased UV radiation, conjugated octadecadienoates: rumenic acid (9,11-18:2) and 10,12-18:2 were registered in the pericarp and leaf of Purys L., the functions of which in plants were practically not studied. The wild P. caucasica at all growing altitudes was characterized by more very-long-chain FAs (VLCFAs) than the P. communis cultivar. At 700 m, most likely when exposed to fungal infections, the relative number of VLCFAs increased significantly, and new species of individual odd-chaine FAs appeared in their composition in both representatives. It was especially worth noting the appearance in peel and leaf melissic acid (30:0), which was rarely recorded in the plant. A characteristic feature of only P. communis at an altitude of 700 m was the large number of unsaturated individual VLCFAs. Based on the data obtained, a scheme of possible pathways for VLCFA biosynthesis in P. communis were proposed.

Polyunsaturated and Very-Long-Chain Fatty Acids Are Involved in the Adaptation of Maloideae (Rosaceae) to Combined Stress in the Mountains.

One of the mechanisms of plant adaptation to combined stress under conditions of altitudinal zonation is changing the lipid fatty acid (FA) composition. The main changes in the FA composition occurred in the outer cell layers of the pericarp, but not in the parenchyma. Adaptation was found to be species-specific. In Cydonia oblonga Mill. and Malus domestica Borkh., the ratio of polyunsaturated 18:2 and 18:3 lipid FAs changed with increasing height, while a constitutive level of the unsaturation index (UI) and low contents of very-long-chain fatty acids (VLCFAs) were maintained. Mespilus germanica L. was characterized by a higher level of VLCFAs due to the high content of 20:0. The sum of VLCFAs in medlar increased by up to 16 % with changing altitude, which was accompanied by the changes in the ultrastructure of chloroplasts and a noticeable decrease in the UI. We attribute the differences in the adaptive strategies in C. oblonga, M. domestica and M. germanica to specific structural features of the pericarp peel. Despite different adaptation mechanisms, the studied species can grow equally successfully at the altitudes from 300 to 1200 m.