Variation in nitrogen partitioning and reproductive stage nitrogen remobilization determines nitrogen grain production efficiency (NUEg) in diverse rice genotypes under varying nitrogen supply.

Nitrogen (N) is an important macronutrient needed for grain yield, grain N and grain protein content in rice. Grain yield and quality are significantly determined by N availability. In this study, to understand the mechanisms associated with reproductive stage N remobilization and N partitioning to grain 2 years of field experiments were conducted with 30 diverse rice genotypes during 2019-Kharif and 2020-Kharif seasons. The experiments were conducted with two different N treatments; N deficient (N0-no external N application, available soil N; 2019-234.15 kgha-1, 2020-225.79 kgha-1) and N sufficient (N120-120 kgha-1 external N application, available soil N; 2019-363.77 kgha-1, 2020-367.95 kgha-1). N application increased the NDVI value, biomass accumulation, grain yield, harvest index and grain N accumulation. Post-anthesis N uptake and N remobilization from vegetative tissues to grain are critical for grain yield and N harvest index. Rice genotypes, Kalinga-1, BAM-4234, IR-8384-B-B102-3, Sahbhagi Dhan, BVD-109 and Nerica-L-42 showed a higher rate of N remobilization under N sufficient conditions. But, under N deficiency, rice genotypes-83929-B-B-291-3-1-1, BVD-109, IR-8384-B-B102-3 and BAM-4234 performed well showing higher N remobilization efficiency. The total amount of N remobilization was recorded to be high in the N120 treatment. The harvest index was higher in N120 during both the cropping seasons. RANBIR BASMATI, BAM-832, APO, BAM-247, IR-64, Vandana, and Nerica-L-44 were more efficient in N grain production efficiency under N deficient conditions. From this study, it is evident that higher grain N accumulation is not always associated with higher yield. IR-83929-B-B-291-3-1-1, Kalinga-1, APO, Pusa Basmati-1, and Nerica-L-44 performed well for different N use efficiency component traits under both N deficient (N0) and N sufficient (N120) conditions. Identifying genotypes/donors for N use efficiency-component traits is crucial in improving the fertilizer N recovery rate and site specific N management.

CO2 Elevation Accelerates Phenology and Alters Carbon/Nitrogen Metabolism vis-à-vis ROS Abundance in Bread Wheat.

Wheat is an important staple food crop of the world and it accounts for 18-20% of human dietary protein. Recent reports suggest that CO2 elevation (CE) reduces grain protein and micronutrient content. In our earlier study, it was found that the enhanced production of nitric oxide (NO) and the concomitant decrease in transcript abundance as well as activity of nitrate reductase (NR) and high affinity nitrate transporters (HATS) resulted in CE-mediated decrease in N metabolites in wheat seedlings. In the current study, two bread wheat genotypes Gluyas Early and B.T. Schomburgk differing in nitrate uptake and assimilation properties were evaluated for their response to CE. To understand the impact of low (LN), optimal (ON) and high (HN) nitrogen supply on plant growth, phenology, N and C metabolism, ROS and RNS signaling and yield, plants were evaluated under short term (hydroponics experiment) and long term (pot experiment) CE. CE improved growth, altered N assimilation, C/N ratio, N use efficiency (NUE) in B.T. Schomburgk. In general, CE decreased shoot N concentration and grain protein concentration in wheat irrespective of N supply. CE accelerated phenology and resulted in early flowering of both the wheat genotypes. Plants grown under CE showed higher levels of nitrosothiol and ROS, mainly under optimal and high nitrogen supply. Photorespiratory ammonia assimilating genes were down regulated by CE, whereas, expression of nitrate transporter/NPF genes were differentially regulated between genotypes by CE under different N availability. The response to CE was dependent on N supply as well as genotype. Hence, N fertilizer recommendation needs to be revised based on these variables for improving plant responses to N fertilization under a future CE scenario.

Visualization of Nitric Oxide, Measurement of Nitrosothiols Content, Activity of NOS and NR in Wheat Seedlings.

Nitric oxide (NO), is a redox-active, endogenous signalling molecule involved in the regulation of numerous processes. It plays a crucial role in adaptation and tolerance to various abiotic and biotic stresses. In higher plants, NO is produced either by enzymatic or non-enzymatic reduction of nitrite and an oxidative pathway requiring a putative nitric oxide synthase (NOS)-like enzyme. There are several methods to measure NO production: mass spectrometry, tissue localization by DAF-FM dye. Electron paramagnetic resonance (EPR) also known as electron spin resonance (ESR) and spectrophotometric assays. The activity of NOS can be measured by L-citrulline based assay and spectroscopic method (NADPH utilization method). A major route for the transfer of NO bioactivity is S-nitrosylation, the addition of a NO moiety to a protein cysteine thiol forming an S-nitrosothiol (SNO). This experimental method describes visualization of NO using DAF-FM dye by fluorescence microscopy (Zeiss AXIOSKOP 2). The whole procedure is simplified, so it is easy to perform but has a high sensitivity for NO detection. In addition, spectrophotometry based protocols for assay of NOS, Nitrate Reductase (NR) and the content of S-nitrosothiols are also described. These spectrophotometric protocols are easy to perform, less expensive and sufficiently sensitive assays which provide adequate information on NO based regulation of physiological processes depending on the treatments of interest.