| Copper is essential for plants to maintain normal life activities as one of the essential micronutrients,involved in many physiological processes in plants.Copper in leaf mainly exists in plastocyanin in the chloroplasts,participating in photosynthetic electron transport.Copper ion is a necessary cofactor for ethylene receptors involved in the process of ethylene signal transduction.Copper ions also play an important role in plant development,through affecting the pollen sterility in plants.The dynamic range between copper deficiency and toxicity is very narrow.In order to efficiently use copper ions,the plants very precisely control copper homeostasis.SPL7 is the central factor of copper homeostasis,which regulates the expression of many genes.At present,a large number of possible SPL7 target genes have been identified using different methods.Only a small fraction of these genes have explicit function associated with copper homeostasis.Here,I analyzed the function of a conserved copper protein,RAN1.Firstly,I observed the subcellular localization of RAN1 by tagging with the green fluorescent protein GFP and found that RAN1 localizes in cell membrane.Secondly,I detected gene expression in different tissues from Arabidopsis thaliana by real-time qPCR,I found that RAN1 expression level is highest in the mature pollen.Further use of GUS reporter gene revealed that RAN1 promoter is highly activated in the mature pollen grains.Then I observed the sterile phenotypes of ran1-2 which destroyed the binding site of copper ions,and the results showed that pollen development was defective and the fertility was significantly reduced.Furthermore,the distribution of copper ions in pollen cells was disordered in ran1-2 mutant Finally,we verified that RAN1 interacted with SPL7 which is known as a male sterility-related copper protein by using yeast two-hybrid and Split-LUC assay.These results indicate that RAN1 plays an important role in male development. |
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