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We have examined the complexity of the phosphoenolpyruvate carboxylase kinase (PPCK) gene family in the C(4) monocots maize and sorghum. Maize contains at least four PPCK genes. The encoded proteins are similar to other phosphoenolpyruvate carboxylase (PEPC) kinases, in that they comprise a protein kinase domain with minimal extensions, except that two of the proteins contain unusual acidic insertions. The spatial and temporal expression patterns of the genes provide information about their presumed functions. Expression of ZmPPCK1 in leaves is mesophyll cell-specific and light-induced, indicating that it encodes the PEPC kinase that is responsible for the phosphorylation of leaf PEPC during C(4) photosynthesis. Surprisingly, ZmPPCK2 is expressed in leaf bundle sheath cells, preferentially in the dark. This suggests that a main function of the ZmPPCK2 gene product is to allow PEPC to function anaplerotically in bundle sheath cells in the dark without interfering with the C(4) cycle. ZmPPCK2, ZmPPCK3 and ZmPPCK4 are all induced by exposure of tissue to cycloheximide, whereas ZmPPCK1 is not. This suggests that the ZmPPCK2, ZmPPCK3 and ZmPPCK4 genes share the property that their expression is controlled by a rapidly turning over repressor. Sequence and expression data show that sorghum contains orthologues of ZmPPCK1 and ZmPPCK2.

Previous analysis of transcriptional changes after elicitation of Cf-9 transgenic tobacco (Nicotiana tabacum) by Avr9 peptide revealed a rapidly upregulated gene, ACRE276. We show that ACRE276 is transiently induced in wounded leaves within 15 min, but upon Avr9 elicitor treatment, this upregulation is enhanced and maintained until cell death onset in Cf-9 tobacco. ACRE276 RNA interference (RNAi) silencing in tobacco results in loss of hypersensitive response (HR) specified by Cf resistance genes. ACRE276 RNAi plants are also compromised for HR mediated by the tobacco mosaic virus defense elicitor p50. Silencing tomato (Lycopersicon esculentum) ACRE276 leads to breakdown of Cf-9-specified resistance against Cladosporium fulvum leaf mold. We confirmed that tobacco ACRE276 is an E3 ubiquitin ligase requiring an intact U-box domain. Bioinformatic analyses revealed Arabidopsis thaliana PLANT U-BOX17 (PUB17) and Brassica napus ARC1 as the closest homologs of tobacco ACRE276. Transiently expressing PUB17 in Cf-9 tobacco silenced for ACRE276 restores HR, while mutant PUB17 lacking E3 ligase activity fails to do so, demonstrating that PUB17 ligase activity is crucial for defense signaling. Arabidopsis PUB17 knockout plants are compromised in RPM1- and RPS4-mediated resistance against Pseudomonas syringae pv tomato containing avirulence genes AvrB and AvrRPS4, respectively. We identify a conserved class of U-box ARMADILLO repeat E3 ligases that are positive regulators of cell death and defense across the Solanaceae and Brassicaceae.

The Oryza sativa (rice) resistance gene Pia confers resistance to the blast fungus Magnaporthe oryzae carrying the AVR-Pia avirulence gene. To clone Pia, we employed a multifaceted genomics approach. First, we selected 12 R-gene analog (RGA) genes encoding nucleotide binding site-leucine rich repeats (NBS-LRRs) proteins from a region on chromosome 11 that shows linkage to Pia. By using seven rice accessions, we examined the association between Pia phenotypes and DNA polymorphisms in the 10 genes, which revealed three genes ( Os11gRGA3-Os11gRGA5) exhibiting a perfect association with the Pia phenotypes. We also screened ethyl methane sulfonate (EMS)-treated mutant lines of the rice cultivar 'Sasanishiki' harboring Pia, and isolated two mutants that lost the Pia phenotype. DNA sequencing of Os11gRGA3-Os11gRGA5 from the two mutant lines identified independent mutations of major effects in Os11gRGA4. The wild-type 'Sasanishiki' allele of Os11gRGA4 ( SasRGA4) complemented Pia function in both mutants, suggesting that SasRGA4 is necessary for Pia function. However, when the rice cultivar 'Himenomochi' lacking Pia was transfected with SasRGA4, the Pia phenotype was not recovered. An additional complementation study revealed that the two NBS-LRR-type R genes, SasRGA4 and SasRGA5, that are located next to each other and oriented in the opposite direction are necessary for Pia function. A population genetics analysis of SasRGA4 and SasRGA5 suggests that the two genes are under long-term balancing selection. [ABSTRACT FROM AUTHOR]