NtHSP70-8b positively regulates heat tolerance and seed size in Nicotiana tabacum

Heat stress considerably restricts the geographical distribution of crops and affects their growth, development, and productivity. HSP70 plays a critical regulatory role in plant growth response to heat stress. However, the mechanisms of this regulatory remain poorly understood. Here, an HSP70 gene, NtHSP70-8b, which is involved in the heat stress response of tobacco, was cloned and identified. The expression of NtHSP70-8b was induced by exogenous abscisic acid (ABA) treatment and abiotic stress, including heat, drought, and salt. Notably, high NtHSP70-8b expression occurred under heat stress conditions, which was consistent with the β-glucuronidase histochemical analysis. Moreover, NtHSP70-8b overexpression markedly enhanced heat stress tolerance by changing the stomatal conductance and antioxidant capacity in tobacco leaves. qRT-PCR showed that the expression levels of ABA synthesis and response genes (NtNCED3 and NtAREB), stress defence genes (NtERD10C and NtLEA5), and other HSP genes (NtHSP90 and NtHSP26a) in NtHSP70-8b-overexpressing tobacco were high under heat stress. The interaction of NtHSP70-8b with NtHSP26a was further confirmed by a luciferase complementation imaging assay. In contrast, NtHSP70-8b knockout mutants showed significantly reduced antioxidant capacity compared to the wild type (WT) under heat stress conditions, suggesting that NtHSP70-8b acts as a positive regulator of heat stress in tobacco. Moreover, NtHSP70-8b overexpression increased the 1000-seed weight. Taken together, NtHSP70-8b is involved in the heat stress response, and NtHSP70-8b overexpression contributed to enhanced tolerance to heat stress, which is thus an essential gene with potential application value for developing heat stress-tolerant crops.