Heat stress during the grain-filling stage seriously affects grain quality in rice. However, very limited information is available regarding the effects of short-term high nighttime temperature (HNT) on grain chalkiness formation in rice. In this paper, the effects of HNT at the early grain-filling stage (7 days after ear emergence) on rice chalkiness formation and the potential causes were investigated by using two rice varieties that differed in susceptibility to high temperature. Although the HNT treatment at night dramatically increased the grain chalkiness in Jiuxiangzhan (JXZ) and Huanghuazhan (HHZ), the increase was greater in JXZ compared to HHZ. The net photosynthetic rate and SPAD value were significantly reduced by HNT treatment in the flag leaves of JXZ, while no significant differences were observed in HHZ. Furthermore, HNT treatment reduced the antioxidant enzyme activity in the flag leaves of JXZ, while the opposite was observed in HHZ, exhibiting increased antioxidant enzyme activity. Moreover, HNT treatment altered the endogenous hormone levels, enhanced the enzymatic activities related to starch biosynthesis, and accelerated the filling rate in grains of JXZ when compared to HHZ. Scanning electron microscopy (SEM) observation exhibited that the starch granules in the endosperm of JXZ were loosely packed together and more starch granules with small pits were produced after HNT treatment. Based on these data, we inferred that HNT stress during the early stage of rice grain filling accelerated the grain-filling rate but shortened the grain-filling duration by changing the endogenous hormone levels and enhancing the enzymatic activities responsible for starch biosynthesis, resulting in significant changes in the morphological structure and arrangement of starch granules and eventually causing the occurrence of grain chalkiness.

The AP2/ERF transcription factor ABSCISIC ACID INSENSITIVE 4 (ABI4) plays diverse roles in plant development and responses to abiotic stress. However, its potential involvement in regulating anthocyanin biosynthesis is not fully understood. In this study, three different loss-of-function abi4 alleles (abi4-1, abi4-2, and abi4-101) were employed to investigate the role of ABI4 in the regulation of anthocyanin accumulation in Arabidopsis seedlings. These abi4 mutants exhibited significantly increased anthocyanin accumulation, which was associated with elevated expression of genes involved in anthocyanin biosynthesis. HY5 (LONG HYPOCOTYL 5), a central component of photomorphogenesis, acts as a key light-regulated molecular switch. Further analysis revealed that ABI4 requires HY5 to function as a negative regulator of anthocyanin biosynthesis. Additionally, loss of ABI4 resulted in heightened light sensitivity, leading to increased light-induced chlorophyll accumulation and chloroplast development, along with upregulation of photosynthesis-related genes. Interestingly, the light-hypersensitive phenotype of abi4 mutants was partially rescued by the loss of HY5 function. Taken together, these findings demonstrate that ABI4 negatively regulates anthocyanin accumulation in Arabidopsis seedlings through a HY5-dependent light signaling pathway.

Although the role of OsHY5L2 in promoting photomorphogenic development is well characterized, its function in regulating rice quality is poorly understood. In this study, we found that OsHY5L2 plays an important role in regulating starch metabolism and modulating its fine structure and physicochemical properties. Overexpression of OsHY5L2 significantly reduced the chalky grain rate and degree of chalkiness but dramatically increased the head rice rate. OsHY5L2 was found to negatively regulate the accumulation of starch in rice endosperm by inhibiting starch biosynthesis and promoting starch hydrolysis. Transcriptomic analysis revealed that OsHY5L2 mainly regulated the expression of genes encoding enzymes involved in starch and sucrose metabolism. Moreover, OsHY5L2 overexpression induced the formation of numerous pinhole structures on the surfaces of starch granules. Analysis of the amylopectin chain length distribution showed that overexpression of OsHY5L2 decreased the proportion of ultra-short chains (DP 6–7) and intermediate chains (DP 13–24) of amylopectin while increasing the proportion of short chains (DP 8–12) and long chains (DP 25–36). Further studies demonstrate that OsHY5L2 overexpression altered the pasting properties of rice starch by affecting its multi-level structure and function. The results of this study improve our understanding of the functions of OsHY5L2 in regulating rice quality.