Peptide Transporter OsNPF8.1 Contributes to Sustainable Growth under Salt and Drought Stresses,and Grain Yield under Nitrogen Deficiency in Rice

Peptide transport is important for plant tissues where rapid proteolysis occurs,especially during germination and senescence,to enhance redistribution of organic nitrogen(N).However,the biological role of peptide transporters is poorly investigated in rice.We characterized the function of the peptide transporter OsNPF8.1 of rice nitrate transporter 1/peptide transporter family(NPF).Ectopic expression of OsNPF8.1 in yeast revealed that OsNPF8.1 encoded a high-affinity di-/tri-peptide transporter,and the osnpf8.1 mutants had a lower uptake rate of the fluorescent-labelled dipeptide c in leaves of rice seedlings.Histochemical assays showed that OsNPF8.1 was highly expressed in mesophyll cells and vascular parenchyma cells,but not detected in root hairs and epidermises.Expression of OsNPF8.1 was induced by N deficiency,drought,Na Cl and abscisic acid,and kept at a high level in senescing leaves.Under N deficiency conditions,compared with the wild type Zhonghua 11,the osnpf8.1 mutants grew slower at the seedling stage,and had lower grain yield and lower N content in the grains.In contrast,OsNPF8.1-over-expressing rice(OsNPF8.1-OE)grew faster at the seedling stage and had a higher grain yield.The osnpf8.1 seedlings were less tolerant to salt and drought stresses.These results suggested that stress-induced organic N transportation mediated by OsNPF8.1 might contribute to balance plant growth and tolerate to salt/drought stress and N-deficiency.

Real-Time Observation of Protein Transport across Membranes by Femtosecond Sum Frequency Generation Vibrational Spectroscopy

Characterization of conformation kinetics of proteins at the interfaces is crucial for understanding the biornolecular functions and the mechanisms of interfacial biological action. But it requires to capture the dynamic structures of proteins at the interfaces with suffi- cient structural and temporal resolutions. Here, we demonstrate that a ferntosecond sum frequency generation vibrational spectroscopy （SFG-VS） system developed by our group provides a powerful tool for monitoring the real-tirne peptide transport across the membranes with time resolution of less than one second. By probing the real-time SFG signals in the arnide I and arnide A bands as WALP23 interacts with DMPG lipid bilayer, it is found that WALP23 is initially absorbed at the gel-phase DMPG bilayer with a random coil structure. The absorption of WALP23 on the surface leads to the surface charge reversal and thus changes the orientation of rnembrane-bound water. As the DMPG bilayer changes from gel phase into fluid phase, WALP23 inserts into the fluid-phase bilayer with its N-terminal end moving across the membrane, which causes the membrane dehydration and the transition of WALP23 conformation from random coil to mixed helix/loop structure and then to pure α-helical structure. The established system is ready to be employed in characterizing other interracial fast processes, which will be certainly helpful for providing a clear physical picture of the interracial phenomena.

An emerging role of vitamin D_(3) in amino acid absorption in different intestinal segments of on-growing grass carp(Ctenopharyngodon idella)

Vitamin D_(3)(VD_(3)),an essential nutrient for animals,has been demonstrated to stimulate the uptake of certain amino acids.However,the role of VD_(3) in the intestine,the primary site for digestion and absorption of nutrients,remains poorly characterized.Here,the grass carp(Ctenopharyngodon idella)was studied to assess the influence of different doses of VD_(3)(15.2,364.3,782.5,1,167.9,1,573.8,and 1,980.1 IU/kg)on growth performance,intestinal morphology,digestive absorption,amino acid transport,and potential signaling molecule levels in a feeding experiment.As a result,dietary VD_(3) improved growth performance,intestinal structure,and digestive and brush border enzyme activities.Additionally,most intestinal free amino acids and their transporters were upregulated after VD_(3) intake,except for Ala,Lys,Asp,Leu,solute carrier(SLC)7A7,SLC1A5,and SLC1A3 mRNA in different segments,Leu and SLC6A14 mRNA in the proximal intestine,and SLC7A5 mRNA in the mid and distal intestine.In the crucial target of rapamycin(TOR)signal pathway of amino acid transport,the gene and protein expression of TOR,S6 kinase 1,and activating transcription factor 4 were elevated,whereas 4E-binding protein 1 was decreased,further suggesting an advanced amino acid absorption capacity in the fish due to VD_(3) supplementation.Based on percentage weight gain,feed efficiency,and trypsin activity,the VD_(3) requirements of on-growing grass carp were estimated to be 968.33,1,005.00,and 1,166.67 IU/kg,respectively.Our findings provide novel recommendations for VD_(3) supplementation to promote digestion and absorption capacities of fish,contributing to the overall productivity of aquaculture.