Image facilitated assessment of intra-spike variation in grain size in wheat under high temperature and drought stress

In wheat (Triticum aestivum L.), the grain size varies according to position within the spike. Exposure to drought and high temperature stress during grain development in wheat reduces grain size, and this reduction also varies across the length of the spike. We developed the phenomics approach involving image-based tools to assess the intra-spike variation in grain size. The grains were arranged corresponding to the spikelet position and the camera of smart phone was used to acquire 333 images. The open-source software ImageJ was used to analyze features of each grain and the image-derived parameters were used to calculate intra-spike variation as standard deviation (ISVAD). The effect of genotype and environment were highly significant on the ISVAD of grain area. Sunstar and Raj 4079 contrasted in the ISVAD of grain area under late sown environment, and RNA sequencing of the spike was done at 25 days after anthesis. The genes for carbohydrate transport and stress response were upregulated in Sunstar as compared to Raj 4079, suggesting that these play a role in intra-spike assimilate distribution. The phenomics method developed may be useful for grain phenotyping and identifying germplasm with low intra-spike variation in grain size for their further validation as parental material in breeding.