Metacaspases are cysteine proteases from plants, fungi, and protozoans that have structural similarity to metazoan caspases. They play a critical role in programmed cell death (PCD) induced by developmental cues and environmental signals. In this study, a type I metacaspase (PhMC1) was identified and characterized from Petunia ×hybrida ‘Mitchell Diploid’ (MD) (petunia). The recombinant PhMC1 had activity against the metacaspase substrate Boc-GRR-AMC (GRR). Activity was highest at pH 7–9 and was dependent on the active site C237. Quantitative polymerase chain reaction (qPCR) showed that PhMC1 transcripts increased at a later stage of petal development, when corollas were visibly senescent in both pollinated and unpollinated flowers. Gene expression patterns were similar to that of the senescence-related gene PhCP10, a homolog of Arabidopsis thaliana (arabidopsis) AtSAG12. PhMC1 transcripts were upregulated in the petals by ethylene treatment. This ethylene regulation did not require protein synthesis, indicating that PhMC1 is a primary ethylene response gene. Metacaspase-like activity against Boc-GRR-AMC increased in protein extracts from senescing petals. RNAi was used to knock down the expression of PhMC1. Transgenic PhMC1 petunias had no abnormal, vegetative growth phenotypes under normal greenhouse conditions, but flower senescence was accelerated by an average of 2 days.
Laura J. Chapin, Youyoun Moon and Michelle L. Jones
Juan O. Quijia Pillajo, Laura J. Chapin and Michelle L. Jones
Autophagy allows for the degradation and recycling of macromolecules and organelles. It plays a significant role in cellular homeostasis, nutrient remobilization during leaf senescence, and abiotic stress responses. Autophagosomes are the hallmark feature of autophagy, and their formation is regulated by the AuTophaGy-related (ATG) genes. The expression profiles of ATG genes have been reported in several agronomic and model plants. To gain insight into the role of autophagy in senescence and abiotic stress responses in floriculture crops, we investigated the regulation of petunia (Petunia ×hybrida) ATG genes (PhATG4, PhATG5, PhATG6, PhATG7, PhATG8a, and PhATG13) during flower senescence and in response to low fertility, nutrient deficiency (-N, -P, and -K), and chronic (weeks) or acute (hours) salt stress using quantitative polymerase chain reaction (PCR). Age-induced corolla wilting coincided with the increased expression of all ATG genes. Petunia ATG genes were upregulated by low fertility and N and P deficiency. Acute salt stress rapidly increased the expression of the petunia ATG genes, but chronic salt stress treatments did not. This project provides insight into the role of autophagy in flower senescence and abiotic stress responses in floriculture crops.