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Nitrogen (N) deficiency and heat stress (HS) are major abiotic stresses that affect the quantity and quality of wheat grains. This study was conducted to examine wheat varietal response to RSI and RNDVI at the anthesis stage and their relationship to yield and yield-related traits under variable N supply and terminal heat stress. Twelve wheat varieties were evaluated in 2016–2017 and 2017–2018 at the National Agricultural Research Centre (NARC), Islamabad, Pakistan. The experiment was divided into three sets, i.e., N120 (120 kg N/ha), N60 (60 kg N/ha) and N0 (0 kg N/ha), based on the nitrogen fertilizer application. The physiological and yield-related parameters were recorded. Mean grain yield for all twelve varieties, averaged from two years of data, ranged between 1655.0 and 3890.1 kg/ha. Maximum RSI (0.99), RNDVI (1.03) and GY (3890.9 kg/ha) were recorded for FSD-08, while AARI-11 showed minimum RSI (0.50), RNDVI (0.56) and GY (1396.40 kg/ha). In the present study, mean CTD was lower, at N0 (3.57 °C), followed by N60 (5.07 °C) and N120 (5.47 °C) on average for the two years of data. The strong positive correlation of RSI and RNDVI with grain yield at R2 = 0.73 and R2 = 0.49 suggest that these parameters can be used as efficient and precise selection criteria for identifying nitrogen-use-efficient wheat varieties under terminal heat-stress conditions. This work will help the researchers to identify and develop nitrogen-use-efficient and thermos-tolerant wheat cultivars by minimizing the negative impacts of heat stress at the anthesis stage.

The present study aimed to reveal the impact of the stay-green trait in bread wheat under terminal heat stress. Field experiments (early and late sowing; for two consecutive years) were conducted to investigate the influence of terminal heat stress on the morpho-physiological traits in different stay-green types i.e., non-stay-green, moderately non-stay-green, moderately stay-green, and stay-green. In addition, the greenhouse experiment was performed to dissect the stay-green trait in functional stay-green, non-functional stay-green, and non-stay-green genotypes. The results of the field experiments confirmed that genotypes exhibiting the stay-green trait have a significantly high chlorophyll content, normalized difference vegetative index, grain yield, biological yield, kernel weight, and low canopy temperature under control and heat stress conditions. In the greenhouse experiment, functional stay-green and non-functional stay-green genotypes showed a high chlorophyll content and photochemical efficiency, whereas biological yield and grain yield showed a significant relation with the functional stay-green genotype under control and terminal heat stress treatments. The sequencing and expression analysis of chlorophyllide a oxygenase (CaO), light-harvesting complex (Cab), stay-green (SGR), and red chlorophyll catabolite reductase (RCCR) in functional stay-green, non-functional stay-green, and non-stay-green genotypes revealed variations in the exons of CaO and RCCR; and significant difference in the regulation of CaO and Cab at 7 days after anthesis under terminal heat stress. This study confirms that genotypes displaying the stay-green trait can aid wheat breeders to cope with increasing temperature in the impending decades.