ABSTRACT
KEY MESSAGE: The heat stress transcription factor HSFA2e regulates both temperature and drought response via hormonal and secondary metabolism alterations. High temperature and drought are the primary yield-limiting environmental constraints for staple food crops. Heat shock transcription factors (HSF) terminally regulate the plant abiotic stress responses to maintain growth and development under extreme environmental conditions. HSF genes of subclass A2 predominantly express under heat stress (HS) and activate the transcriptional cascade of defense-related genes. In this study, a highly heat-inducible HSF, HvHSFA2e was constitutively expressed in barley (Hordeum vulgare L.) to investigate its role in abiotic stress response and plant development. Transgenic barley plants displayed enhanced heat and drought tolerance in terms of increased chlorophyll content, improved membrane stability, reduced lipid peroxidation, and less accumulation of ROS in comparison to wild-type (WT) plants. Transcriptome analysis revealed that HvHSFA2e positively regulates the expression of abiotic stress-related genes encoding HSFs, HSPs, and enzymatic antioxidants, contributing to improved stress tolerance in transgenic plants. The major genes of ABA biosynthesis pathway, flavonoid, and terpene metabolism were also upregulated in transgenics. Our findings show that HvHSFA2e-mediated upregulation of heat-responsive genes, modulation in ABA and flavonoid biosynthesis pathways enhance drought and heat stress tolerance.
Subject(s)
Droughts , Gene Expression Regulation, Plant , Heat-Shock Response , Hordeum , Plant Growth Regulators , Plant Proteins , Plants, Genetically Modified , Hordeum/genetics , Hordeum/physiology , Plant Proteins/genetics , Plant Proteins/metabolism , Plants, Genetically Modified/genetics , Heat-Shock Response/genetics , Plant Growth Regulators/metabolism , Heat Shock Transcription Factors/genetics , Heat Shock Transcription Factors/metabolism , Chlorophyll/metabolism , Stress, Physiological/genetics , Secondary Metabolism/genetics , Metabolic Networks and Pathways/genetics , Drought ResistanceABSTRACT
Iron and zinc deficiencies are the most prevalent cause of global hidden hunger. Rice, being one of the most consumed crops worldwide, is suitable to target for Fe and Zn biofortification. In present study, we generated rice transgenic lines to meet the recommended dietary requirement of iron and zinc through endosperm specific expression of dicot (kidney bean) and monocot (pearl millet) Ferritins along with constitutive expression of rice nicotianamine synthase 2 (OsNAS2) gene. Visualization through perls' prussian staining and quantification by ICP-MS showed significant improvement in grain iron content in all the transgenic lines. The transgenic lines expressing any of the three selected gene combinations (PvFerrtin-OsNAS2, feedPgFerrtin-OsNAS2 and foodPgFerritin-OsNAS2), showed the potential to surpass the 30% of the estimated average requirement (13 µg/g Fe and 28 µg/g Zn) proposed for rice in HarvestPlus breeding program. Though the expression of PvFerritin along with OsNAS2 gene in IET10364 (indica) variety showed the best result, providing up to 4.2- and 3.5-fold increase in iron (30.56 µg/g) and zinc (60.1 µg/g) content, respectively; in polished grains compared to non-transgenic control. Thus, the lines developed in our study can be used for further breeding purpose to enhance the iron and zinc content in commercial rice varieties.
