Xin‐Jian He

Nuclear factor Y (NF-Y) is a ubiquitous transcription factor composed of three distinct subunits (NF-YA, NF-YB, and NF-YC). We found that the Arabidopsis thaliana NFYA5 transcript is strongly induced by drought stress in an abscisic acid (ABA)-dependent manner. Promoter:beta-glucuronidase analyses showed that NFYA5 was highly expressed in vascular tissues and guard cells and that part of the induction by drought was transcriptional. NFYA5 contains a target site for miR169, which targets mRNAs for cleavage or translational repression. We found that miR169 was downregulated by drought stress through an ABA-dependent pathway. Analysis of the expression of miR169 precursors showed that miR169a and miR169c were substantially downregulated by drought stress. Coexpression of miR169 and NFYA5 suggested that miR169a was more efficient than miR169c at repressing the NFYA5 mRNA level. nfya5 knockout plants and plants overexpressing miR169a showed enhanced leaf water loss and were more sensitive to drought stress than wild-type plants. By contrast, transgenic Arabidopsis plants overexpressing NFYA5 displayed reduced leaf water loss and were more resistant to drought stress than the wild type. Microarray analysis indicated that NFYA5 is crucial for the expression of a number of drought stress-responsive genes. Thus, NFYA5 is important for drought resistance, and its induction by drought stress occurs at both the transcriptional and posttranscriptional levels.

An NAC-type transcription factor gene AtNAC2 was identified from Arabidopsis thaliana when expression patterns of the genes from a microarray analysis were examined. The AtNAC2 expression was induced by salt stress and this induction was reduced in magnitude in the transgenic Arabidopsis plants overexpressing tobacco ethylene receptor gene NTHK1. AtNAC2 is localized in the nucleus and has transcriptional activation activity. It can form a homodimer in yeast. AtNAC2 was highly expressed in roots and flowers, but less expressed in other organs examined. In addition to the salt induction, the AtNAC2 can also be induced by abscisic acid (ABA), ACC and NAA. The salt induction was enhanced in the ethylene overproducer mutant eto1-1, but suppressed in the ethylene-insensitive mutants etr1-1 and ein2-1, and in the auxin-insensitive mutant tir1-1when compared with that in wild-type plants. However, the salt induction of AtNAC2 was not significantly affected in the ABA-insensitive mutants abi2-1, abi3-1 and abi4-1. These results indicate that the salt response of AtNAC2 requires ethylene signaling and auxin signaling pathways but does not require ABI2, ABI3 and ABI4, intermediates of the ABA signaling pathway. Overexpression of AtNAC2 in transgenic Arabidopsis plants resulted in promotion of lateral root development. AtNAC2 also promoted or inhibited downstream gene expressions. These results indicate that AtNAC2 may be a transcription factor incorporating the environmental and endogenous stimuli into the process of plant lateral root development.

Abstract Ethylene signaling plays important roles in multiple aspects of plant growth and development. Its functions in abiotic stress responses remain largely unknown. Here, we report that alteration of ethylene signaling affected plant salt-stress responses. A type II ethylene receptor homolog gene NTHK1 (Nicotiana tabacum histidine kinase 1) from tobacco (N. tabacum) conferred salt sensitivity in NTHK1-transgenic Arabidopsis (Arabidopsis thaliana) plants as judged from the phenotypic change, the relative electrolyte leakage, and the relative root growth under salt stress. Ethylene precursor 1-aminocyclopropane-1-carboxylic acid suppressed the salt-sensitive phenotype. Analysis of Arabidopsis ethylene receptor gain-of-function mutants further suggests that receptor function may lead to salt-sensitive responses. Mutation of EIN2, a central component in ethylene signaling, also results in salt sensitivity, suggesting that EIN2-mediated signaling is beneficial for plant salt tolerance. Overexpression of the NTHK1 gene or the receptor gain-of-function activated expression of salt-responsive genes AtERF4 and Cor6.6. In addition, the transgene NTHK1 mRNA was accumulated under salt stress, suggesting a posttranscriptional regulatory mechanism. These findings imply that ethylene signaling may be required for plant salt tolerance.