High temperature (HT) has an adverse effect on rice grain filling by inhibiting the accumulation of storage materials. However, the regulatory mechanism of this inhibition remains unknown. Here, we report that Opaque2 like transcription factor OsbZIP58 is a key factor regulating storage material accumulation under HT. The OsbZIP58 gene promotes expression of many seed storage protein genes and starch synthesis genes while inhibits expression of some starch hydrolyzing α‐amylase genes under HT. The loss of OsbZIP58 function leads to floury and shrunken endosperms and dramatically reduced storage materials in the seeds under HT. HT is found to affect alternative splicing of OsbZIP58, promoting the formation of the truncated OsbZIP58β protein form over the full‐length OsbZIP58α protein form. The OsbZIP58β form has a lower transcriptional activity than the OsbZIP58α form, especially under HT condition. Interestingly, rice varieties with less heat sensitivity have reduced alternative splicing of OsbZIP58. Therefore, OsbZIP58 is a crucial gene in regulating storage material accumulation under HT and lower alternative splicing of OsbZIP58 may contribute to heat tolerance during grain filling.
BTF3, which was originally recognized as a basal transcription factor, has been known to be involved in transcription initiation, translational regulation and protein localization in many eukaryotic organisms. However, its function remains largely unknown in plant species. In the present study, we analyzed a BTF3-related sequence in Oryza sativa L. subsp. japonica, which shares the conserved domain of a nascent polypeptide-associated complex with human BTF3, and was referred to as Osj10gBTF3. The expression of Osj10gBTF3 was primarily constitutive and generally modulated by salt, high temperature and exogenous phytohormone stress. The Osj10gBTF3::EGFP (enhanced green fluorescence protein) fusion protein was localized in both the nucleus and cytoplasmic membrane system. Inhibition of Osj10gBTF3 led to significant morphological changes in all detected tissues and organs, with a reduced size of between 25% and 52%. Furthermore, the pollen that developed was completely sterile, which was correlated with the altered expression of two Rf (fertility restorer)-like genes that encode pentatricopeptide repeat-containing proteins OsPPR676 and OsPPR920, translational initiation factors OseIF3e and OseIF3h, and the heat shock protein OsHSP82. These findings were verified through a yeast two-hybrid assay using a Nipponbare callus cDNA library as bait followed by the reverse transcription-PCR analysis of total leaf or anther RNAs. Our demonstration of the important role of Osj10gBTF3 in rice growth and development provides new insights showing that more complex regulatory functions are associated with BTF3 in plants.
In an earlier greenhouse screening, we identified a local indica cultivar HT54 tolerant to high temperature at both seedling and grain-filling stages. In this study, we develop an optimized procedure for fine assessment of this heat tolerance. The results indicated that HT54 seedlings could tolerate high temperature up to 48 °C for 79h. The genetic analysis of F(1) and F(2) offspring derived from the cross between HT54 and HT13, a heat-sensitive breeding line, reveals that the heat tolerance of HT54 was controlled by a dominant major locus, which has been designated as OsHTAS (Oryza sativa heat tolerance at seedling stage). This locus was mapped on rice chromosome 9 within an interval of 420kb between markers of InDel5 and RM7364. The determined candidate ZFP gene has been confirmed to be cosegregated with a single nucleotide polymorphism (SNP) developed PCR-restriction fragment length polymorphism (RFLP) marker RBsp1407 in its promoter region. Another heat tolerance-associated SNP was identified in the first intron of its 5'-untranslated region. The existence of these SNPs thereby indicated that the OsHTAS locus contains at least two alleles. We named the one from HT54 as OsHTAS ( a ) and the one from HT13 as OsHTAS ( b ). Further dynamic expression analysis demonstrated that OsHTAS ( a ) was actively responsive to 45 °C high temperature stress compared with the OsHTAS ( b ) allele.
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