Later Stages Of Maturation Research Articles

Milk from cows fed clover-rich silage compared with cows fed grass silage is higher in n-3 fatty acids

The present study investigated the effect of a high proportion of different forage species in the diet, parity, milking time, and days in milk (DIM) on milk fatty acid (FA) profile, and transfer efficiency of C18:2n-6, C18:3n-3, n-6, and n-3 in dairy cows. Swards with perennial ryegrass [early maturity stage (EPR) and late maturity stage (LPR)], festulolium, tall fescue (TF), red clover (RC), and white clover (WC) were cut in the primary growth, wilted, and ensiled without additives. Thirty-six Danish Holstein cows in an incomplete Latin square design were fed ad libitum with total mixed rations containing a high forage proportion (70% on dry matter basis). The total mixed rations differed only in forage source, which was either 1 of the 6 pure silages or a mixture of LPR silage with either RC or WC silage (50:50 on dry matter basis). Proportion of C18:2n-6 in milk FA was affected by diet, and RC and WC diets resulted in the highest proportion of C18:2n-6 in milk FA (21.6 and 21.8 g/kg of FA, respectively). The highest and lowest milk C18:3n-3 proportion was observed in WC and LPR, respectively. In addition, WC diet resulted in highest transfer efficiency of C18:3n-3 from feed to milk (12.2%) followed by RC diet (10.7%), whereas EPR diet resulted in the lowest transfer efficiency of C18:3n-3 (3.45%). The highest milk proportion of cis-9,trans-11 conjugated linoleic acid (CLA) was observed in cows fed TF (3.20 g/kg of FA), which was 23 to 64% higher than the proportion observed in the cows fed the other diets. The highest α-tocopherol concentration (µg/mL) in milk was observed in EPR (1.15), LPR (1.10), and festulolium (1.06). Primiparous cows showed higher proportion of cis-9,trans-11 CLA (2.63 g/kg of FA) than multiparous cows (2.21 g/kg of FA). Cows early in lactation had a higher proportion of long-chain FA in milk than cows later in lactation, as long-chain FA decreased with 0.184 g/kg of FA per DIM, whereas medium-chain FA increased with 0.181 g/kg of FA per DIM. Proportion of C18:2n-6 in milk from evening milking was higher than in milk from morning milking (16.7 vs. 15.8 g/kg of FA). In conclusion, the results showed that milk FA profile of cows was affected by forage source in the diet, and RC and WC increased the health-promoting FA components, particularly n-3, whereas the TF diet increased proportion of CLA isomers in milk. Proportion of CLA isomers in milk FA from primiparous cows was higher than in milk from multiparous cows. In addition, evening milk contained more FA originating from diets compared with morning milk.

Diurnal and seasonal CO2 exchange and yield of maize cropland under different irrigation treatments in semiarid Inner Mongolia

Different irrigation treatments in plants significantly influence CO2 emissions. This study investigated the net ecosystem CO2 exchange (NEE) and yield of maize cropland under different irrigation treatments to determine the optimal irrigation treatment at different maize growth stages. Different degrees of water deficit irrigation were carried out in four fields: full irrigation, 100% evapotranspiration (ET); moderate deficit irrigation, 65% ET; and severe deficit irrigation, 40% ET. Automated static and dynamic chambers were used to measure CO2 fluxes. We found that NEE showed a “V” -shaped trend during the day and slowly declined at night. The light response parameters were calculated using the rectangular hyperbola model, and the leaf area index accounted for 85% of the light use efficiency. In the late vegetative stage, severe deficit irrigation led to a reduction in CO2 emissions but significantly reduced the crop growth rate (p < 0.05). In the reproductive stage, full irrigation not only reduced CO2 emissions but also significantly increased the crop growth rate. In the maturation stage, moderate deficit irrigation reduced CO2 emissions, and the crop growth rate decreased compared with full irrigation, but the decrease was not significant (p > 0.05). Compared with full irrigation throughout the growth period, full irrigation in the reproductive stage and moderate deficit irrigation in the late vegetative and reproductive stages both increased the maize carbon sink capacity and light use efficiency at higher leaf area index, and the harvest index did not decrease significantly. However, inadequate irrigation in the reproductive stage or severe deficit irrigation in the late vegetative and maturation stages significantly reduced the maize harvest index. Thus, full irrigation during the reproductive stage, and moderate deficit irrigation during the late vegetative and reproductive stages were the optimal irrigation strategies for saving water, reducing CO2 emissions and ensuring a satisfactory maize yield.

Proteomic analysis of pikeperch seminal plasma provides novel insight into the testicular development of domesticated fish stocks

Control of the reproduction of domesticated stocks is considered a prerequisite for aquaculture development of pikeperch. However, knowledge about the physiology of the captive pikeperch male reproductive system and the biology of semen is very limited, especially regarding protein characteristics. The aims of our study were to characterize pikeperch sperm quantity and quality parameters and to analyze changes in the proteome of the same males spawned for the first and second times. Moreover, attempts were made to generate the first proteomic library of seminal plasma proteins. Semen collected during the first spawning season was characterized by lower sperm concentration and volume than for the second season. Using mass spectrometry-based label-free quantitative proteomics, we identified 850 proteins in the seminal plasma of pikeperch from both spawning seasons, and 65 seminal proteins were found to be differentially abundant between the first and second spawning seasons. The majority of differentially abundant proteins were involved in stress and immune responses, developmental processes, cofactor metabolic processes, proteolysis, cellular oxidant detoxification and organization of the extracellular matrix (ECM). In addition, several proteins unique to pikeperch seminal plasma were identified, including antifreeze proteins, hibernation-specific plasma proteins, lectins and vitellogenin. In summary, our results indicate that males that spawned for the first time were characterized by incompletely mature gonads and the expression of proteins associated with the early phase of spermatogenesis and ECM organization. On the other hand, males that spawned for the second time exhibited advanced gonadal maturation and expression of proteins related to the late stage of spermatogenesis and sperm maturation, including regulation of reactive oxygen species generation, bicarbonate production, sperm elongation and separation. The identification of a large number of seminal plasma proteins provides a valuable resource for understanding the functions of seminal plasma and the molecular mechanisms involved in testicular development and maturation in domesticated fish, which is a prerequisite for better control of reproduction in captivity.

Mitochondrial localization and moderated activity are key to murine erythroid enucleation

Mammalian red blood cells (RBCs), which primarily contain hemoglobin, exemplify an elaborate maturation process, with the terminal steps of RBC generation involving extensive cellular remodeling. This encompasses alterations of cellular content through distinct stages of erythroblast maturation that result in the expulsion of the nucleus (enucleation) followed by the loss of mitochondria and all other organelles and a transition to anaerobic glycolysis. Whether there is any link between erythroid removal of the nucleus and the function of any other organelle, including mitochondria, remains unknown. Here we demonstrate that mitochondria are key to nuclear clearance. Using live and confocal microscopy and high-throughput single-cell imaging, we show that before nuclear polarization, mitochondria progressively move toward one side of maturing erythroblasts and aggregate near the nucleus as it extrudes from the cell, a prerequisite for enucleation to proceed. Although we found active mitochondrial respiration is required for nuclear expulsion, levels of mitochondrial activity identify distinct functional subpopulations, because terminally maturing erythroblasts with low relative to high mitochondrial membrane potential are at a later stage of maturation, contain greatly condensed nuclei with reduced open chromatin–associated acetylation histone marks, and exhibit higher enucleation rates. Lastly, to our surprise, we found that late-stage erythroblasts sustain mitochondrial metabolism and subsequent enucleation, primarily through pyruvate but independent of in situ glycolysis. These findings demonstrate the critical but unanticipated functions of mitochondria during the erythroblast enucleation process. They are also relevant to the in vitro production of RBCs as well as to disorders of the erythroid lineage.

Nutritive value for ruminants of different fresh and ensiled sorghum (Sorghum bicolor (L.) Moench) varieties harvested at varying maturity stages

ABSTRACT The occurrence of the Western corn rootworm and increasing frequency of summer droughts have reduced forage maize yields and quality in Central Europe in recent years. Therefore, sorghum (Sorghum bicolor (L.) Moench) has been increasingly used as alternative forage in ruminant feeding, although information on the nutritive value of whole crop sorghum silage (WCSS) under Central European climate conditions is scarce. This research project aimed to study differences between sorghum varieties and harvest dates regarding dry matter (DM) yield, chemical composition of fresh panicle, fresh stover, fresh whole plant and WCSS as well as whole tract digestibility and energy content of WCSS. For this purpose, six sorghum varieties, classified by their predominant use (biomass, silage, grain), were cultivated and harvested at three maturity stages (late milk stage, dough stage, full maturity). Furthermore, WCSS quality was compared with feed value of fresh and ensiled forage maize. Dry matter yield was higher (p < 0.05) in biomass sorghum (24.9 t/ha) compared to other sorghum varieties (12.4 to 16.7 t/ha). However, panicle proportion was lowest in biomass sorghum (10.5%), highest in grain sorghum (59.1%) and intermediate in silage sorghum. Fibre content declined and starch content increased with rising panicle proportion in fresh panicle, fresh whole plant and WCSS. Therefore, whole tract organic matter digestibility (66.8%) and metabolisable energy (ME) content (9.49 MJ/kg DM) were highest in grain sorghum silage. Especially low ME content was found in the biomass sorghum silage (7.43 MJ/kg DM). Dry matter yield of sorghum increased between late milk and dough stage of maturity, while it remained constant afterwards. Furthermore, ME content of WCSS rose tendentially (0.05 < p < 0.10) between late milk and dough stage of maturity and dropped again between dough stage and full maturity. Highest nutritive value of WCSS is achieved if grain sorghum varieties are used and both nutritive value and DM yield are optimised if harvest is carried out at dough stage of maturity. However, compared to forage maize, grain sorghum varieties have disadvantages both in DM yield and in nutritive value under common Central European climate conditions.

Acquisition of the physiological quality of peanut (Arachis hypogaea L.) seeds during maturation under the influence of the maternal environment.

The scarcity of information on the maturation physiology of the peanut seed (Arachis hypogaea L.; Virgínia group) makes harvesting high quality seeds a challenge for the seed industry. During two consecutive crop seasons, we studied the acquisition of physiological quality of peanut seeds during maturation in tropical conditions. We bring new insights about the period of late maturation of seeds and the influence of the maternal environment on physiological quality. We monitored water content, dry weight, ability of germination, desiccation tolerance, vigor and longevity. In addition, we monitored temperature and precipitation throughout plant growth. We demonstrate that the physiological quality of peanut seeds is acquired during development, with a maximum between 57 and 76 days after flowering in the late stage of maturation. This final period represents about 25% of the development, considered the best time to harvest peanut seeds with the highest quality. Our findings also support the idea that the adequate proportion of rainfall and thermal sum in the maternal environment are factors that favor the acquisition of peanut seed longevity.

LRP10 interacts with SORL1 in the intracellular vesicle trafficking pathway in non-neuronal brain cells and localises to Lewy bodies in Parkinson\u2019s disease and dementia with Lewy bodies

Loss-of-function variants in the low-density lipoprotein receptor-related protein 10 (LRP10) gene have been associated with autosomal-dominant Parkinson’s disease (PD), PD dementia, and dementia with Lewy bodies (DLB). Moreover, LRP10 variants have been found in individuals diagnosed with progressive supranuclear palsy and amyotrophic lateral sclerosis. Despite this genetic evidence, little is known about the expression and function of LRP10 protein in the human brain under physiological or pathological conditions. To better understand how LRP10 variants lead to neurodegeneration, we first performed an in-depth characterisation of LRP10 expression in post-mortem brains and human-induced pluripotent stem cell (iPSC)-derived astrocytes and neurons from control subjects. In adult human brain, LRP10 is mainly expressed in astrocytes and neurovasculature but undetectable in neurons. Similarly, LRP10 is highly expressed in iPSC-derived astrocytes but cannot be observed in iPSC-derived neurons. In astrocytes, LRP10 is present at trans-Golgi network, plasma membrane, retromer, and early endosomes. Interestingly, LRP10 also partially co-localises and interacts with sortilin-related receptor 1 (SORL1). Furthermore, although LRP10 expression and localisation in the substantia nigra of most idiopathic PD and DLB patients and LRP10 variant carriers diagnosed with PD or DLB appeared unchanged compared to control subjects, significantly enlarged LRP10-positive vesicles were detected in a patient carrying the LRP10 p.Arg235Cys variant. Last, LRP10 was detected in Lewy bodies (LB) at late maturation stages in brains from idiopathic PD and DLB patients and in LRP10 variant carriers. In conclusion, high LRP10 expression in non-neuronal cells and undetectable levels in neurons of control subjects indicate that LRP10-mediated pathogenicity is initiated via cell non-autonomous mechanisms, potentially involving the interaction of LRP10 with SORL1 in vesicle trafficking pathways. Together with the specific pattern of LRP10 incorporation into mature LBs, these data support an important mechanistic role for disturbed vesicle trafficking and loss of LRP10 function in neurodegenerative diseases.

MicroRNA-29 is an essential regulator of brain maturation through regulation of CH methylation

SUMMARYAlthough embryonic brain development and neurodegeneration have received considerable attention, the events that govern postnatal brain maturation are less understood. Here, we identify the miR-29 family to be strikingly induced during the late stages of brain maturation. Brain maturation is associated with a transient, postnatal period of de novo non-CG (CH) DNA methylation mediated by DNMT3A. We examine whether an important function of miR-29 during brain maturation is to restrict the period of CH methylation via its targeting of Dnmt3a. Deletion of miR-29 in the brain, or knockin mutations preventing miR-29 to specifically target Dnmt3a, result in increased DNMT3A expression, higher CH methylation, and repression of genes associated with neuronal activity and neuropsychiatric disorders. These mouse models also develop neurological deficits and premature lethality. Our results identify an essential role for miR-29 in restricting CH methylation in the brain and illustrate the importance of CH methylation regulation for normal brain maturation.

Effects of forest succession on the richness and composition of Zingiberales in a forest fragment in the southwestern Amazon

Tropical forests have been increasingly modified by human activities and the forest recovery through succession has caused an increase in the amount of secondary forests in the landscape. Little is known about changes in the richness and composition of non-woody species throughout the succession in tropical forests; therefore, the objective of this work was to evaluate the effects of forest succession on a community of herbs in the order Zingiberales. Zingiberales and soil were sampled and data on vegetation structure, including bamboo abundance, canopy opening, and basal area were collected from nine 80 × 90 m plots installed on the Catuaba Experimental Farm, Acre, representing three successional stages: early secondary forest, late secondary forest, and mature forest. Early secondary forest areas were experimentally deforested, burned, and sprayed with herbicide in the dry season of 2001 and the late secondary forest areas suffered deforestation followed by burning for agricultural cultivation between 1985 and 1986. A total of 3,831 individuals and 26 species distributed in nine genera and five families was registered. Both soil characteristics and vegetation structure variables showed significant differences among the successional stages. The richness of Zingiberales was lower in the early secondary forest areas and did not differ among the other successional stages. Canopy opening was the variable that best explained the variation in the richness of Zingiberales among the areas. The composition of Zingiberales differed among the successional stages, with the late secondary forest and mature forest stages being most similar. The soil characteristics, canopy opening, bamboo abundance and basal area were the variables that best explained a variation in the composition of Zingiberales. In the areas of initial secondary forest the group of dominant species was composed of Renealmia breviscapa, Calathea altissima and Heliconia rostrata, in the areas of late secondary forest by Chamaecostus lanceolatus Calathea polytricha, Calathea sp.1 and in the areas of mature forest by Heliconia acuminata, Calathea mansonis and Phenakospermum guyannense. These results show that the disturbance caused by cutting, burning, and herbicide application alters the community of Zingiberales. These data also highlight the importance of late secondary and mature forests in maintaining and recovering biodiversity.

Oil properties variation among the China Erlian Rift Basin petroleum systems: Correlation with organofacies and maturity

Petroleum bulk properties such as API gravity are important parameters in petroleum system analysis. Three sags (the Bayindulan, Saihantala and Wuliyasitai Sags) in the Erlian rift basin of northern China provide interesting insights into the correlations between organofacies, maturity and oil fluid properties. The API gravity of oils in the three sags shows obvious differences, ranging from 17.0 to 30.9° for Bayindulan oils, 25.7–40.5° for Saihantala oils, and 32.8–41.9° for Wuliyasitai oils. Oil geochemical parameters indicate that the differences in oil fluid properties among the three sags can be attributed to the variations in organofacies and thermal maturity, consistent with the analysis of geochemistry and maceral composition of the source rocks in the three sags. Bayindulan oils were expelled from organofacies C with an extremely high initial hydrogen index (HIo) of 600–900 mg/g TOC in the early mature stage (<0.8% Ro). Wuliyasitai oils were expelled from mixed algal and terrigenous organofacies D/E with HIo of 200–400 mg/g TOC in the maturation stage with 0.8–1.1% Ro. Saihantala oils were expelled from organofacies C (HIo ranging from 400 to 550 mg/g TOC) and D/E in the maturation stage with 0.7–0.9% Ro. The HIo controls the temperature of oil expulsion from lacustrine source rocks. Organofacies C source rocks with high HIo values expelled oil in the early mature stage, while organofacies D/E source rocks expelled oil in the middle and late mature stages, which leads to obviously different oil bulk properties in the three sags.

Effects of hormonal treatment before water warming on synchronisation of spawning time, oocyte size, and egg quality in pikeperch (Sander lucioperca)

Inducing reproduction during periods of the year when spawning typically does not occur is an important goal for the feasibility of commercial fish farming. Pre-seasonal propagation of pikeperch generally occurs about 3 months before the natural spawning season. The objective of this study was to assess effects of imposing a thermal schedule for control of water temperature and differing salmon gonadotropin releasing hormone analogue (sGnRHa) dosages on final stages of oocyte growth, and egg quality by optimizing protocol duration and synchronizing spawning time. In Experiment 1, there was analysis of thermal schedule effects for water temperature control when hormonal administrations occurred before or after water warming (WARMING and STABLE, respectively). In Experiment 2, there was assessment of the sGnRHa dosage effects during the warming schedule. In both experiments there was analysis of oocyte diameter from time of sGnRHa administration until the late stages of maturation. There was greater synchrony in time of spawning in specimens of the WARMING group with lesser variability in time from sGnRHa administration to spawning. In Experiment 2, values for reproductive variables were variable among the different groups, without any differences between treatments. Oocyte diameter at the time of sGnRHa administration was correlated with embryo survival. For effective pre-seasonal pikeperch propagation, the selection of breeders based on oocyte diameter, and administration of 5 μg/kg sGnRHa immediately upon transport to hatchery, followed by a 1 °C/d temperature increase to 10 °C, are effective methods for induction of spawning during periods when spawning does not naturally occur.

The influence of 17β-oestradiol on lymphopoiesis and immune system ontogenesis in juvenile sea bass, Dicentrarchus labrax

The female sex steroid 17β-oestradiol (E2) is involved in the regulation of numerous physiological functions, including the immune system development and performance. The role of oestrogens during ontogenesis is, however, not well studied. In rodents and fish, thymus maturation appears to be oestrogen-dependent. Nevertheless, little is known about the function of oestrogen in immune system development. To further the understanding of the role of oestrogens in fish immune system ontogenesis, fingerlings of European sea bass (Dicentrarchus labrax) were exposed for 30 days to 20 ng E2·L−1, at two ages tightly related to thymic maturation, i.e., 60 or 90 days post hatch (dph). The expression of nuclear and membrane oestrogen receptors was measured in the thymus and spleen, and the expression of several T cell-related gene markers was studied in both immune organs, as well as in the liver. Waterborne E2-exposure at 20.2 ± 2.1 (S.E.) ng·L−1 was confirmed by radioimmunoassay, leading to significantly higher E2-contents in the liver of exposed fish. The majority of gene markers presented age-dependent dynamics in at least one of the organs, confirming thymus maturation, but also suggesting a critical ontogenetic window for the implementation of liver resident γδ and αβ T cells. The oestrogen receptors, however, remained unchanged over the age and treatment comparisons with the exception of esr2b, which was modulated by E2 in the younger cohort and increased its expression with age in the thymus of the older cohort, as did the membrane oestrogen receptor gpera. These results confirm that oestrogen-signalling is involved in thymus maturation in European sea bass, as it is in mammals. This suggests that esr2b and gpera play key roles during thymus ontogenesis, particularly during medulla maturation. In contrast, the spleen expressed low or non-detectable levels of oestrogen receptors. The E2-exposure decreased the expression of tcrγ in the liver in the cohort exposed from 93 to 122 dph, but not the expression of any other immune-related gene analysed. These results indicate that the proliferation/migration of these innate-like T cell populations is oestrogen-sensitive. In regard to the apparent prominent role of oestrogen-signalling in the late thymus maturation stage, the thymic differentiation of the corresponding subpopulations of T cells might be regulated by oestrogen. To the best of our knowledge, this is the first study investigating the dynamics of both nuclear and membrane oestrogen receptors in specific immune organs in a teleost fish at very early stages of immune system development as well as to examine thymic function in sea bass after an exposure to E2 during ontogenesis.

LPS1, Encoding Iron-Sulfur Subunit SDH2-1 of Succinate Dehydrogenase, Affects Leaf Senescence and Grain Yield in Rice.

The iron-sulfur subunit (SDH2) of succinate dehydrogenase plays a key role in electron transport in plant mitochondria. However, it is yet unknown whether SDH2 genes are involved in leaf senescence and yield formation. In this study, we isolated a late premature senescence mutant, lps1, in rice (Oryza sativa). The mutant leaves exhibited brown spots at late tillering stage and wilted at the late grain-filling stage and mature stage. In its premature senescence leaves, photosynthetic pigment contents and net photosynthetic rate were reduced; chloroplasts and mitochondria were degraded. Meanwhile, lps1 displayed small panicles, low seed-setting rate and dramatically reduced grain yield. Gene cloning and complementation analysis suggested that the causal gene for the mutant phenotype was OsSDH2-1 (LOC_Os08g02640), in which single nucleotide mutation resulted in an amino acid substitution in the encoded protein. OsSDH2-1 gene was expressed in all organs tested, with higher expression in leaves, root tips, ovary and anthers. OsSDH2-1 protein was targeted to mitochondria. Furthermore, reactive oxygen species (ROS), mainly H2O2, was excessively accumulated in leaves and young panicles of lps1, which could cause premature leaf senescence and affect panicle development and pollen function. Taken together, OsSDH2-1 plays a crucial role in leaf senescence and yield formation in rice.

Abscisic Acid Inhibits Asymbiotic Germination of Immature Seeds of Paphiopedilum armeniacum.

Paphiopedilum armeniacum is a rare orchid native to China with high ornamental value. The germination of P. armeniacum seeds is difficult, especially for the mature seeds, which is the major limitation for their large-scale reproduction. This study explored the reasons for seed germination inhibition from the aspects of the important plant endogenous hormone—abscisic acid (ABA). The major endogenous hormone contents of seeds were determined at different developmental stages. The ABA content was 5.8 ng/g in 73 days after pollination (DAP) for the immature seeds, peaked at 14.6 ng/g in 129 DAP seeds, and dropped to 2.6 ng/g in the late mature stage of the 150 DAP seeds. The reduction of ABA content in the mature seed suggests a possible contribution to the increased expression of CYP707A, an ABA catabolism gene. The germination rate of the immature seeds was reduced to 9% from 69% when 5 μg/mL ABA was added to the Hyponex N026 germination medium. The result showed that ABA can inhibit the germination of P. armeniacum immature seeds. However, for the heavily lignified mature seeds, reduction in endogenous ABA level does not result in an increase in the germination rate. Lignin accumulation in the seed coat imposes the physical dormancy for P. armeniacum. In summary, the germination of P. armeniacum is regulated by both ABA and lignin accumulation.

Physiological Profiling of Embryos and Dormant Seeds in Two Arabidopsis Accessions Reveals a Metabolic Switch in Carbon Reserve Accumulation.

In flowering plants, sugars act as carbon sources providing energy for developing embryos and seeds. Although most studies focus on carbon metabolism in whole seeds, knowledge about how particular sugars contribute to the developmental transitions during embryogenesis is scarce. To develop a quantitative understanding of how carbon composition changes during embryo development, and to determine how sugar status contributes to final seed or embryo size, we performed metabolic profiling of hand-dissected embryos at late torpedo and mature stages, and dormant seeds, in two Arabidopsis thaliana accessions with medium [Columbia-0 (Col-0)] and large [Burren-0 (Bur-0)] seed sizes, respectively. Our results show that, in both accessions, metabolite profiles of embryos largely differ from those of dormant seeds. We found that developmental transitions from torpedo to mature embryos, and further to dormant seeds, are associated with major metabolic switches in carbon reserve accumulation. While glucose, sucrose, and starch predominantly accumulated during seed dormancy, fructose levels were strongly elevated in mature embryos. Interestingly, Bur-0 seeds contain larger mature embryos than Col-0 seeds. Fructose and starch were accumulated to significantly higher levels in mature Bur-0 than Col-0 embryos, suggesting that they contribute to the enlarged mature Bur-0 embryos. Furthermore, we found that Bur-0 embryos accumulated a higher level of sucrose compared to hexose sugars and that changes in sucrose metabolism are mediated by sucrose synthase (SUS), with SUS genes acting non-redundantly, and in a tissue-specific manner to utilize sucrose during late embryogenesis.

AMPKβ1 and AMPKβ2 define an isoform-specific gene signature in human pluripotent stem cells, differentially mediating cardiac lineage specification

AMP-activated protein kinase (AMPK) is a key regulator of energy metabolism that phosphorylates a wide range of proteins in order to maintain cellular homeostasis. AMPK consists of three subunits: α, β and γ. AMPKα and β are encoded by two genes, the γ subunit by three genes, all of which being expressed in a tissue-specific manner. If individual isoforms have different functions is not understood so far. Using RNA-Seq technology, we provide evidence that the loss of AMPKβ1 and AMPKβ2 lead to different gene expression profiles in human induced pluripotent stem cells (hiPSCs), indicating isoform-specific function. The knockout of AMPKβ2 was associated with a higher number of differentially regulated genes than the deletion of AMPKβ1, suggesting that AMPKβ2 has a more comprehensive impact on the transcriptome. Bioinformatics analysis identified cell differentiation as one biological function being specifically associated with AMPKβ2. Correspondingly, the two isoforms differentially affected lineage decision towards a cardiac cell fate. While the lack of PRKAB1 impacted differentiation into cardiomyocytes only at late stages of cardiac maturation, the availability of PRKAB2 was indispensable for mesoderm specification as shown by gene expression analysis and histochemical staining for cardiac lineage markers such as cTnT, GATA4, and NKX2.5. Ultimately, the lack of AMPKβ1 impairs, while deficiency of AMPKβ2 abrogates differentiation into cardiomyocytes. Finally, we demonstrate that AMPK affects cellular physiology by engaging in the regulation of hiPSC transcription in an isoform-specific manner, providing the basis for further investigations elucidating the role of dedicated AMPK subunits in the modulation of gene expression.

How do preharvest applications of nitrogen fertiliser affect branch growth, leaf morphology and fruit, inMangifera indica(mango) orchards?

The effect of preharvest applications of nitrogen (N) fertiliser to mango trees during the late fruit maturation stage was investigated over 3 years (2012-2014) to determine the effects on canopy growth in 8 year-old 'Kensington Pride' (KP) mango trees grown in Queensland. The experiment consisted of six treatments where 156 g N tree-1 was applied as a single treatment, as 340 g of urea or split between three different times. The N treatments were 1) 100% of N 2 weeks postharvest (control), 2) 50% of N 2 weeks preharvest, plus 50% 2 weeks postharvest, 3) 65% 2 weeks preharvest, plus 35% 2 weeks postharvest, 4) 35% 4 weeks preharvest plus 65% 2 weeks postharvest, and 5) 65% 4 weeks preharvest, plus 35% 2 weeks postharvest. Vegetative growth was investigated at the branch, sub-branch (shoots), growth unit (GU) and leaf scales. Branches from different aspects (north, south, east and west), in the upper and lower canopy were assessed, and for 2 branch types (apical or lateral) was considered. Fruit counts were also considered at the branch level. Results showed that when 50-65% of the N was applied 2 and 4 weeks before harvest with the balance 2 weeks postharvest, the number of branches, branch length and fruit count increased. When 35% N was applied 4 weeks preharvest with the balance 2 weeks postharvest, only stem biomass was increased. Generally, branches in the upper canopy had longer sub-branches, longer and thicker, first level growth units, shorter leaf petioles and shorter and thinner leaves compared to those in the lower canopy. The SPAD N index was lowest in the northern canopy aspect. Differences were also observed between lateral and apical branches. Lateral branches had longer leaves than apical branches while apical branches had a trend for higher leaf area, stem biomass and leaf counts. There were also negative relationships between fruit count and measurements for stem biomass, leaf count and leaf area for apical branches however for lateral branches these relationships were positive. In summary, when a higher proportion of N was applied 2 or 4 weeks prior to harvest the partitioning of growth to leaves and stem increased while low rates of N applied prior to harvest led to increased stem biomass only. Applications of N 2 or 4 weeks prior to harvest resulted in increased fruit count and more efficient nitrogen use efficiency (NUE), although the relationship between fruit count, and leaf and stem growth was dependent on branch type.

Comparative Proteomic Analysis of Walnut (Juglans regia L.) Pellicle Tissues Reveals the Regulation of Nut Quality Attributes.

Walnuts (Juglans regia L.) are a valuable dietary source of polyphenols and lipids, with increasing worldwide consumption. California is a major producer, with ’Chandler’ and ’Tulare’ among the cultivars more widely grown. ’Chandler’ produces kernels with extra light color at a higher frequency than other cultivars, gaining preference by growers and consumers. Here we performed a deep comparative proteome analysis of kernel pellicle tissue from these two valued genotypes at three harvest maturities, detecting a total of 4937 J. regia proteins. Late and early maturity stages were compared for each cultivar, revealing many developmental responses common or specific for each cultivar. Top protein biomarkers for each developmental stage were also selected based on larger fold-change differences and lower variance among replicates, including proteins for biosynthesis of lipids and phenols, defense-related proteins and desiccation stress-related proteins. Comparison between the genotypes also revealed the common and specific protein repertoires, totaling 321 pellicle proteins with differential abundance at harvest stage. The proteomics data provides clues on antioxidant, secondary, and hormonal metabolism that could be involved in the loss of quality in the pellicles during processing for commercialization.

Calcium Bioavailability in the Soluble and Insoluble Fibers Extracted from Opuntia ficus indica at Different Maturity Stages in Growing Rats.

Childhood and adolescence are crucial stages of life for bone health. Therefore, an adequate calcium intake and a healthy life style constitute the main strategies to prevent the risk of osteoporosis-related fractures during adulthood. It has been demonstrated that inclusion of indigestible carbohydrates in foods can help improve calcium absorption in growing stages. The objective of this study was to evaluate the effect of supplementation of soluble and insoluble fibers extracted from O. ficus indica cladodes on calcium bioavailability. Male Wistar rats 4-week old were fed diets added with soluble and insoluble fibers extracted from O. ficus indica cladodes at early and late maturity stages, as the only source of calcium. The mineral content, bone mineral density (BMD), physical, microstructural, and biomechanical properties of rat femurs were determined. The bones of rats fed with diets containing a soluble fiber extracted from O. ficus indica at early and late maturity stages exhibited better bone properties (resistance to fracture, microarchitecture, and calcium content) than control rats and rats fed with an insoluble fiber from O. ficus indica cladodes at both maturity stages. As expected, based on these results, the BMD values were higher in adolescent and pubertal rats fed with a diet containing the O. ficus indica soluble fiber. These results demonstrate that the soluble fiber from O. ficus indica cladodes is indeed a valuable source of bioavailable calcium, which contributes to improve physical, densitometric, biomechanical, and microstructural properties of bone in growing rats.

MYASTHENIA & RELATED DISORDERS: P.37 Autophagy affected in patients with hypokalemic periodic paralysis

Hypokalemic periodic paralysis (hypoPP) is an autosomal dominant rare disorder that is caused by mutations in the genes for voltage gated calcium channel CaV1.1 (CACNA1S) and NaV1.4 (SCN4A). Patients with hypoPP may suffer from periodic paralysis alone, periodic paralysis with weakness or permanent weakness. HypoPP has been known to be associated with vacuolar myopathy for decades but the cause of this has not been known. We have investigated a cohort of 14 hypoPP patients with the R528H mutation in the CACNA1S gene for vacuoles and their content. The R528H mutation has previously been demonstrated to cause a Ca2+ leak through CaV1.1 into the cytoplasm, possibly causing disturbance in the calcium homeostasis. Histology and transmission electron microscopy revealed that vacuoles had a very heterogenous content of glycogen, fibrils and autophagosomes/endosomes. Further investigation demonstrated autophagosomes and endosomes were arrested in the autophagic processing pathway between the early and late stages of maturation, as well as large non-fused lysosomes. Expression analysis of proteins associated with endosomes, autophagosomes and lysosomes showed a significant decrease in levels of proteins involved in autophagy such as Rab5, Rab7, p62/SQSTM1, LC3I, Atg5 and LAMP1 in patients with hypoPP suggesting a systemic effect on the protein expression at the gene level, which decreased mRNA expression levels confirmed, possibly through a master regulator of autophagy gene expression. The findings suggest that changes in calcium homeostasis leads to disruptions in autophagic flux and abnormally large endosomes, autophagosomes and lysosomes that contribute to the vacuoles seen in patients with hypoPP.