Pollen exine is essential for protection from the environment of the male gametes of seed-producing plants, but its assembly and composition remain poorly understood. We previously characterized Arabidopsis (Arabidopsis thaliana) mutants with abnormal pollen exine structure and morphology that we named kaonashi (kns). Here we describe the identification of the causal gene of kns4 that was found to be a member of the CAZy glycosyltransferase 31 gene family, identical to UNEVEN PATTERN OF EXINE1, and the biochemical characterization of the encoded protein. The characteristic exine phenotype in the kns4 mutant is related to an abnormality of the primexine matrix laid on the surface of developing microspores. Using light microscopy with a combination of type II arabinogalactan (AG) antibodies and staining with the arabinogalactan-protein (AGP)-specific b-Glc Yariv reagent, we show that the levels of AGPs in the kns4 microspore primexine are considerably diminished, and their location differs from that of wild type, as does the distribution of pectin labeling. Furthermore, kns4 mutants exhibit reduced fertility as indicated by shorter fruit lengths and lower seed set compared to the wild type, confirming that KNS4 is critical for pollen viability and development. KNS4 was heterologously expressed in Nicotiana benthamiana, and was shown to possess b-(1,3)-galactosyltransferase activity responsible for the synthesis of AG glycans that are present on both AGPs and/or the pectic polysaccharide rhamnogalacturonan I. These data demonstrate that defects in AGP/pectic glycans, caused by disruption of KNS4 function, impact pollen development and viability in Arabidopsis.Pollen, the male gametophyte of all seed plants, is crucial for reproductive success. Due to the harsh environmental conditions that pollen must survive in, it has developed an elaborate and specialized cell wall. However, despite its importance our knowledge of the fine structure and assembly of the pollen grain wall is poorly understood (Newbigin et al., 2009;Ariizumi and Toriyama, 2011;Quilichini et al., 2015;Shi et al., 2015). Arabidopsis (Arabidopsis thaliana) provides an ideal genetic and cell biological model to dissect the assembly of the components of the pollen grain wall. Arabidopsis pollen grains have a typical surface structure that consists of an inner intine layer, which is usually made up of cellulose and pectin, and the outer exine, which is largely composed of sporopollenin (Li et al., 1995). The exine is further subdivided into inner nexine and outer sexine. The sexine has a three-dimensional (3D) structure composed of many columns called baculae and a roof called tectum. These two structures give the Arabidopsis pollen surface a reticulate appearance with a uniform mesh size. The nexine is composed of outer nexine I (foot layer), which contains sporopollenin, and inner nexine II (endexine; Ariizumi and Toriyama, 2011;Jiang et al., 2013). A recent study suggests that arabinogalactan proteins (AGPs) are key constituents of nexine with express...

Integral subcellular and cellular functions ranging from gene expression, protein targeting and nutrient supply to cell differentiation and cell death require proteases. Plants have unique organelles such as chloroplasts composed of unique proteins that carry out the unique process of photosynthesis. Hence, along with proteases common across kingdoms, plants contain unique proteases. Improved knowledge on proteases can lead to a better understanding of plant development, differentiation and death. Because of their importance in multiple processes, plant proteases are actively studied. However, root proteases specifically are not as well studied. The associated rhizosphere, organic matter and/or inorganic matter make roots a difficult system. Yet recent research conclusively demonstrated the occurrence of endocytosis of proteins, peptides and even microbes by root cells, which, hitherto known for specialized pathogenesis or symbiosis, was unsuspected for nutrient uptake. These results reinforced the importance of root proteases in endocytosis or root exudate-mediated nutrient uptake. Rhizoplane, rhizosphere or in planta protease action on proteins, peptides and microbes generates sources of nitrogen, especially during abiotic stresses such as drought. This article highlights the recent research on root proteases for nitrogen uptake and the connection of the two to drought-tolerance mechanisms. Drought-induced proteases in rice roots, as known from rice expression databases, are discussed for future research on certain M50, Deg, FtsH, AMSH and deubiquitination proteases. The recent emphasis on linking drought and plant hydraulics to nutrient metabolism is illustrated and connected to the value of a systematic study of root proteases in crop improvement.

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Rice is rich in starch but low in protein. In countries where rice is the staple food, people are at high risk of protein deficiency. Legumes (such as pea) and milk are an important part of the diet in many developing and developed countries, respectively. For this reason, pea protein isolate (PPI) and whey protein concentrate (WPC) are incorporated into basmati (%20-25% amylose) and glutinous (%0-3% amylose) starches to test the effect of protein incorporation on the pasting properties of rice starch, and on starch and protein digestibility. Increasing protein incorporation reduced the peak, breakdown, and final viscosities. The effect is more pronounced for basmati mixtures compared to glutinous mixtures. For starch digestibility, basmati starch mixtures exhibited an increasing trend in the amount of glucose released over 120 min, whereas glutinous starch mixtures show a decrease in the amount of glucose released after 60 min. In addition, samples with the same % protein incorporation (PPI and WPC) shows similar trend in terms of amount of glucose released at each time point. The amylose content appears to have an effect on starch digestibility over time. For pea and whey protein digestibility, basmati starch samples exhibited lower digestion after 120 min compared with glutinous starch samples. The starch amylose content appears to have an impact on protein digestibility. This provides a way of fortifying rice starch with proteins, in which the proteins are effectively digested without significantly increasing blood glucose levels.

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Black ear mushroom (Auricularia auricula) is an important genus of cultivated mushroom, which contains health benefits. Incorporating black ear (BE) mushroom into brown rice by extrusion changed the physicochemical, and more importantly, the nutritional characteristics of the extrudates. With increased incorporation of BE mushroom in the extrudates in vitro starch digestion of the different extrudates revealed significantly reduced starch digestion, suggesting a lower glycaemic index. In addition, incorporation of BE in brown rice extrudates increased the total phenolic concentration of the samples, which led to higher % scavenging effect against free-radicals in DPPH assay. In the ORAC assay for anti-oxidant activity, BE powder exhibited the highest anti-oxidant activity, followed by 10% BE and 15% BE, and 5% BE extruded products. The extruded brown rice control exhibited the lowest antioxidant activity. Inclusion of black ear mushroom was shown to improve the nutritional qualities of the food product illustrating the connection between plant bioactive ingredients and human health.

Consumption of underutilised ancient crops has huge benefits for our society. It improves food security by diversifying our staple foods and makes our agriculture more adaptable to climate change. The Philippines has a rich biodiversity and many plant species used as staple foods are native to the Philippines. An example of ancient Philippine crops is the kabog millet, an ecotype of Panicum miliaceum. There is a dearth of information about its uses and properties; hence, in this study, the nutritional quality of kabog millet was evaluated. The total starch, % amylose, ash, dietary fibre, proteins, essential amino acid profile, phenolic acids, carotenoids, tocopherols, and the antioxidant properties of its total phenolic acid extracts were compared to four types of rice (white, brown, red, and black) and a reference millet, purchased from local Swiss supermarkets. Our analyses showed that kabog millet has higher total dietary fibre, total protein, total phenolic acids, tocopherols, and carotenoids content than white rice. It also performed well in antioxidant assays. Our results indicate that kabog millet is a good alternative to rice. It is hoped that the results of this study will encourage consumers and farmers to diversify their food palette and address food insecurity.

Arabinogalactan-proteins (AGPs) are members of the hydroxyproline-rich glycoprotein (HRGP) superfamily, a group of highly diverse proteoglycans that are present in the cell wall, plasma membrane as well as secretions of almost all plants, with important roles in many developmental processes. The role of GALT8 (At1g22015), a Glycosyltransferase-31 (GT31) family member of the Carbohydrate-Active Enzyme database (CAZy), was examined by biochemical characterization and phenotypic analysis of a galt8 mutant line. To characterize its catalytic function, GALT8 was heterologously expressed in tobacco leaves and its enzymatic activity tested. GALT8 was shown to be a β-(1,3)-galactosyltransferase (GalT) that catalyzes the synthesis of a β-(1,3)-galactan, similar to the in vitro activity of KNS4/UPEX1 (At1g33430), a homologous GT31 member previously shown to have this activity. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed the products were of 2-6 degree of polymerisation (DP). Previous reporter studies showed that GALT8 is expressed in the central and synergid cells, from whence the micropylar endosperm originates after the fertilization of the central cell of the ovule. Homozygous mutants have multiple seedling phenotypes including significantly shorter hypocotyls and smaller leaf area compared to wild type (WT) that are attributable to defects in female gametophyte and/or endosperm development. KNS4/UPEX1 was shown to partially complement the galt8 mutant phenotypes in genetic complementation assays suggesting a similar but not identical role compared to GALT8 in β-(1,3)-galactan biosynthesis. Taken together, these data add further evidence of the important roles GT31 β-(1,3)-GalTs play in elaborating type II AGs that decorate AGPs and pectins, thereby imparting functional consequences on plant growth and development.