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    A conserved HSF:miR169:NF-YA loop involved in tomato and Arabidopsis heat stress tolerance
    (John Wiley & Sons, 2022) Rao, Sombir; Gupta, Apoorva; Bansal, Chandni; Sorin, Celine; Crespi, Martin; Mathur, Saloni
    Heat stress transcription factors (HSFs) and microRNAs (miRNAs) regulate different stress and developmental networks in plants. Regulatory feedbacks are at the basis of these networks. Here, we report that plants improve their heat stress tolerance through HSF-mediated transcriptional regulation of MIR169 and post-transcriptional regulation of Nuclear Factor- YA (NF-YA) transcription factors. We show that HSFs recognize tomato and Arabidopsis MIR169 promoters using yeast-one-hybrid/ChIP-qPCR. Silencing tomato HSFs using virus induced gene silencing (VIGS) reduced Sly-MIR169 levels and enhanced Sly-NF- YA9/A10 target expression. Further, Sly-NF-YA9/A10-VIGS knock-down tomato plants and Arabidopsis plants overexpressing At-MIR169d or At-nf-ya2 mutants showed a link with increased heat tolerance. In contrast, Arabidopsis plants overexpressing At-NF-YA2, or those expressing a non-cleavable At-NF-YA2 form (miR169d-resistant At-NF-YA2) as well as plants inhibited for At-miRNA169d regulation (miR169d mimic plants) were more sensitive to heat stress, highlighting NF-YA as negative regulator of heat tolerance. Furthermore, post-transcriptional cleavage of NF-YA by elevated miR169 levels resulted in alleviating the repression of heat stress effectors HSFA7 in tomato and Arabidopsis revealing a retroactive control of HSFs by the miR169:NF-YA node. Hence, a regulatory feedback loop involving HSFs, miR169s and NF-YAs plays a critical role in the regulation of heat stress response in tomato and Arabidopsis plants.
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    Exploring the master regulator heat stress transcription factor HSFA1a-mediated transcriptional cascade of HSFs in the heat stress response of tomato
    (Springer Nature Publishing AG, 2021) Rao, Sombir; Das, Jaishri Rubina; Mathur, Saloni
    The tomato heat stress transcription factor A1a (Sly-HSFA1a) acts as the master regulator of the heat stress (HS) by directly regulating the transcription of Sly-HSFA2. However, it is unclear whether the activation of Sly-HSFA2 alone is sufficient to trigger the entire transcriptional cascade downstream of Sly-HSFA1a. Therefore, the present study aims to delineate the Sly-HSFA1a governed downstream HSFs cascade regulating the tomato heat stress response. The study identified several HSFs with common and specific roles in different HS regimes as well as in HS memory. Furthermore, the study established Sly-HSFA7, Sly-HSFA6b, Sly-HSFA4c, Sly-HSFB1 and Sly-HSFB2b as new downstream targets of SlyHSFA1a during heat stress by using virus-induced-gene-silencing (VIGS) of Sly-HSFA1a. Moreover, the silencing of downstream target Sly-HSFA7 and Sly-HSFB1 revealed the orchestration of downstream transcriptional cascade of HSFs regulated individually or in a synergistic manner by Sly-HSFA1a and Sly-HSFA7 along with co-activator Sly-HSFB1. This complex transcriptional cascade of HSFs sheds light on regulatory mechanisms that enable tomato plants to respond to various heat stress conditions to maintain cellular homeostasis.