Asymmetric wall ingrowth deposition in Arabidopsis phloem parenchyma transfer cells is tightly associated with sieve elements

In Arabidopsis, polarized deposition of wall ingrowths in phloem parenchyma (PP) transfer cells (TCs) occurs adjacent to cells of the sieve element/companion cell (SE/CC) complex. However, the spatial relationships between these different cell types in minor veins, where phloem loading occurs, are poorly understood. PP TC development and wall ingrowth localization were compared to other phloem cells in leaves of Col-0 and the transgenic lines AtSUC2::AtSTP9-GFP and AtSWEET11::AtSWEET11-GFP that identify CCs and PP respectively. The development of PP TCs in minor veins, indicated by deposition of wall ingrowths, proceeded basipetally in leaves. However, not all PP develop ingrowths and higher levels of wall ingrowth deposition occur in abaxial- compared to adaxial-positioned PP TCs. Furthermore, the deposition of wall ingrowths was exclusively initiated on and preferentially covered the PP TC/SE interface, rather than the PP TC/CC interface, and only occurred in PP cells that were adjacent to SEs. Collectively, these results demonstrate the dominant impact of SEs on wall ingrowth deposition in PP TCs and suggest the existence of two sub-types of PP cells in leaf minor veins. Compared to PP cells, PP TCs showed more abundant accumulation of AtSWEET11-GFP, indicating functional differences in phloem loading between PP and PP TCs.

Effective ways of keeping queen bees before and after instrumental insemination

The article presents theoretical and experimental data on the results of the instrumental insemination and alimentation of queen bees. Queen bees were bred and inseminated. It has been established that when using various methods of keeping queen bees, a dynamic pattern is observed when there is safekeeping of queen bees. So, in 2019, before insemination in all experimental groups, the death of 1 to 4 infertile queens was observed. The highest indicator was recorded in the first experimental group; this is due to the fact that there were not enough young bees in the transfer cells to fully feed the infertile queen. In 2020, when using various methods of keeping queen bees before and after insemination, a similar pattern is observed with respect to the safety of queen bees. In a comparative aspect, before and after insemination, a high death rate of queens was recorded in the I-experimental group - 4 pcs, respectively fetal - 16 pcs. Whereas in the II-experimental and IV-experimental groups, the safest keeping of 18-19 queens was recorded respectively. Whereas in 2021, the greatest safety of fetal queen bees was in the IV-experimental group - 90%, and in the II-experimental and III-experimental groups, 85% each in comparison with the I-experimental group - 75%. Consequently, when using the method of keeping infertile queens in the transfer cells with accompanying bees, 5 pieces each, there is a high death rate of queen bees. It was also confirmed that the method of keeping queens in a nursery frame without accompanying bees is less effective compared to the method of keeping queens in a nursery frame without accompanying bees. Also, when obtaining fetal queens by instrumental insemination, attention should be focused on phenotypic indicators when selecting infertile queens, since the size of the queen bee is directly correlated with the number of egg tubes and, accordingly, affects the egg production of the fetal queen bee.

The Placenta of Physcomitrium patens: Transfer Cell Wall Polymers Compared across the Three Bryophyte Groups

Following similar studies of cell wall constituents in the placenta of Phaeoceros and Marchantia, we conducted immunogold labeling TEM studies of Physcomitrium patens to determine the composition of cell wall polymers in transfer cells on both sides of the placenta. Sixteen monoclonal antibodies were used to localize cell wall epitopes in the basal walls and wall ingrowths in this moss. In general, placental transfer cell walls of P. patens contained fewer pectins and far fewer arabinogalactan proteins AGPs than those of the hornwort and liverwort. P. patens also lacked the differential labeling that is pronounced between generations in the other bryophytes. In contrast, transfer cell walls on either side of the placenta of P. patens were relatively similar in composition, with slight variation in homogalacturonan HG pectins. Compositional similarities between wall ingrowths and primary cell walls in P. patens suggest that wall ingrowths may simply be extensions of the primary cell wall. Considerable variability in occurrence, abundance, and types of polymers among the three bryophytes and between the two generations suggested that similarity in function and morphology of cell walls does not require a common cell wall composition. We propose that the specific developmental and life history traits of these plants may provide even more important clues in understanding the basis for these differences. This study significantly builds on our knowledge of cell wall composition in bryophytes in general and in transfer cells across plants.

Cell wall polymers in the Phaeoceros placenta reflect developmental and functional differences across generations

The placenta of hornworts is unique among bryophytes in the restriction of transfer cells that are characterized by elaborate wall labyrinths to the gametophyte generation. During development, cells around the periphery of the sporophyte foot elongate, forming smooth-walled haustorial cells that interdigitate with gametophyte cells. Using immunogold labeling with 22 antibodies to diverse cell wall polymers, we examined compositional differences in the developmentally and morphologically distinct cell walls of gametophyte transfer cells and sporophyte haustorial cells in the placenta of Phaeoceros. As detected by Calcofluor White fluorescence, cellulose forms the cell wall scaffolding in cells on both sides of the placenta. Homogalacturonan (HG) and rhamnogalacturonan I (RG-I) pectins are abundant in both cell types, and haustorial cells are further enriched in methyl-esterified HGs. The abundance of pectins in placental cell walls is consistent with the postulated roles of these polymers in cell wall porosity and in maintaining an acidic apoplastic pH favorable to solute transport. Xyloglucan hemicellulose, but not mannans or glucuronoxylans, are present in cell walls at the interface between the two generations with a lower density in gametophytic wall ingrowths. Arabinogalactan proteins (AGPs) are diverse along the plasmalemma of placental cells and are absent in surrounding cells in both generations. AGPs in placental cell walls may play a role in calcium binding and release associated with signal transduction as has been speculated for these glycoproteins in other plants. Callose is restricted to thin areas in cell walls of gametophyte transfer cells. In contrast to studies of transfer cells in other systems, no reaction to the JIM12 antibody against extensin was observed in Phaeoceros.

Localization, Gene Expression, and Functions of Glutamine Synthetase Isozymes in Wheat Grain (Triticum aestivum L.)

Glutamine synthetase (GS) plays a major role in plant nitrogen metabolism, but the roles of individual GS isoforms in grains are unknown. Here, the localization and expression of individual TaGS isozymes in wheat grain were probed with TaGS isoenzyme-specific antibodies, and the nitrogen metabolism of grain during the grain filling stage were investigated. Immunofluorescence revealed that TaGS1;1, TaGS1;3, and TaGS2 were expressed in different regions of the embryo. In grain transporting tissues, TaGS1;2 was localized in vascular bundle; TaGS1;2 and TaGS1;1 were in chalaza and placentochalaza; TaGS1;1 and TaGS1;3 were in endosperm transfer cells; and TaGS1;3 and TaGS2 were in aleurone layer. GS exhibited maximum activity and expression at 8 days after flowering (DAF) with peak glutamine content in grains; from then, NH4+ increased largely from NO3- reduction, glutamate dehydrogenase (GDH) aminating activity increased continuously, and the activities of GS and glutamate synthase (GOGAT) decreased, while only TaGS1;3 kept a stable expression in different TaGS isozymes. Hence, GS-GOGAT cycle and GDH play different roles in NH4+ assimilation of grain in different stages of grain development; TaGS1;3, located in aleurone layer and endosperm transfer cells, plays a key role in Gln into endosperm for gluten synthesis. At 30 DAF, grain amino acids are mainly transported from maternal phloem.

Structure and histochemistry of sorghum caryopsis in relation to grain-filling

Anatomical and histochemical studies of ovary and caryopsis of sorghum reveal the importance of the chalazal complex in transporting nutrients from maternal sources to the filial diploid embryo and triploid endosperm. The presence of starch, protein, lipid, Ca, K, Mg, and Fe in various tissues at different stages of development can be revealed by a variety of histochemical techniques. Vascular supply ends at the base of the ovary and transport occurs through vascular parenchyma, pigment strand and nucellar projection where symplastic continuity is broken. Nutrients unloaded into an apoplastic placental sac then enter the endosperm and embryo through the aleurone transfer cells. The later possess characteristic wall ingrowth. The single layer of aleurone surrounding the endosperm may also help in transport during later stages of grain-filling. Grain-filling in C4 sorghum is compared with other C4 and C3 grasses showing the variety of strategies evolved to transport nutrients into filial tissues. Standardization of terminologies to describe the tissues of the crease region will help in further research and communication.

Unraveling the puzzle of phloem parenchyma transfer cell wall ingrowth

This article comments on: Wei X, Nguyen ST, Collings DA, McCurdy DW. 2020. Sucrose regulates wall ingrowth deposition in phloem parenchyma transfer cells in Arabidopsis via affecting phloem loading activity. Journal of Experimental Botany 71, 4690–4702.