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In our previous research, we showed that the cyclin-dependent kinase regulatory subunit (CKS2) in maize (Zea mays L.) was induced by water deficit and cold stress. To elucidate its expression patterns under adversity, we isolated and characterized its promoter (PZmCKS2). A series of PZmCKS2-deletion derivatives, P0–P3, from the translation start code (-1,455, -999, -367, and -3 bp) was fused to the β-glucuronidase (GUS) reporter gene, and each deletion construct was analyzed by Agrobacterium-mediated steady transformation into Arabidopsis. Leaves were then subjected to dehydration, cold, abscisic acid (ABA), salicylic acid (SA), and methyl jasmonic acid (MeJA). Sequence analysis showed that several stressrelated cis-acting elements (MBS, CE3, TGA element, and ABRE) were located within the promoter. Deletion analysis of the promoter, PZmCKS2, suggested that the -999 bp promoter region was required for the highest basal expression of GUS, and the -367 bp sequence was the minimal promoter for ZmCKS2 activation by low temperature, ABA, and MeJA. The cis-acting element ABRE was necessary for promoter activation by exogenous ABA.
Small heat shock proteins (sHSPs) are the critical components of responses to various environmental stresses. However, few have been functionally characterised in Primula. In this study, we cloned a sHSP gene, PfHSP17.1, which is highly up-regulated in the leaves of Primula forrestii exposed to thermal stress (42 °C for 2 h). Sequence alignment and phylogenetic analysis indicated that PfHSP17.1 is a member of the plant cytosolic class I sHSPs. This gene was basally and ubiquitously expressed in different plant organs. The expression of PfHSP17.1 was also triggered remarkably by salt, drought and oxidative stress conditions but was only slightly induced by abscisic acid. Transgenic Arabidopsis thaliana constitutively expressing PfHSP17.1 displayed increased thermotolerance and higher resistance to salt and drought compared with wild-type plants. These results highlight the important role that PfHSP17.1 plays in diverse physiological and biochemical processes related to adverse conditions.
Nuclear-encoded chloroplast small heat shock proteins (Cp-sHSPs) play important roles in plant stress tolerance due to their abundance and diversity. Their functions in Primula under heat treatment are poorly characterized. Here, expression analysis showed that the Primula Cp-sHSP gene, PfHSP21.4, was highly induced by heat stress in all vegetative and generative tissues in addition to constitutive expression in certain development stages. PfHSP21.4 was introduced into Arabidopsis, and its function was analysed in transgenic plants. Under heat stress, the PfHSP21.4 transgenic plants showed increased heat tolerance as shown by preservation of hypocotyl elongation, membrane integrity, chlorophyll content and photosystem II activity (Fv/Fm), increased seedling survival and increase in proline content. Alleviation of oxidative damage was associated with increased activity of superoxide dismutase and peroxidase. In addition, the induced expression of HSP101, HSP70, ascorbate peroxidase and Δ1-pyrroline-5-carboxylate synthase under heat stress was more pronounced in transgenic plants than in wild-type plants. These results support the positive role of PfHSP21.4 in response to heat stress in plants.
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