Song Yion Yeu

Protein Disulfide Isomerase-Like Protein 1-1 Controls Endosperm Development through Regulation of the Amount and Composition of Seed Proteins in Rice

Protein disulfide isomerase (PDI) is a chaperone protein involved in oxidative protein folding by acting as a catalyst and assisting folding in the endoplasmic reticulum (ER). A genome database search showed that rice contains 19 PDI-like genes. However, their functions are not clearly identified. This paper shows possible functions of rice PDI-like protein 1-1 (PDIL1-1) during seed development. Seeds of the T-DNA insertion PDIL1-1 mutant, PDIL1-1Δ, identified by genomic DNA PCR and western blot analysis, display a chalky phenotype and a thick aleurone layer. Protein content per seed was significantly lower and free sugar content higher in PDIL1-1Δ mutant seeds than in the wild type. Proteomic analysis of PDIL1-1Δ mutant seeds showed that PDIL1-1 is post-translationally regulated, and its loss causes accumulation of many types of seed proteins including glucose/starch metabolism- and ROS (reactive oxygen species) scavenging-related proteins. In addition, PDIL1-1 strongly interacts with the cysteine protease OsCP1. Our data indicate that the opaque phenotype of PDIL1-1Δ mutant seeds results from production of irregular starch granules and protein body through loss of regulatory activity for various proteins involved in the synthesis of seed components.

Transcript Profile of Transgenic Arabidopsis Constitutively Producing Methyl Jasmonate

Using an Affymetrix GeneChip™ containing 8300 oligonucleotide probes, we measured transcript levels in transgenicArabidopsis overexpressing the jasmonate carboxyl methyltransferase gene (AtJMT). When compared with wild-type plants, 5-week-old transgenics exhibited significant alterations (more than a two-fold increase or decrease) in the expression levels of 168 genes. Among them, 80 were up-regulated, including those involved in defense, oxidative stress-tolerance, and senescence. In contrast, the expression of 88 genes, including those that function in photosynthesis and cold/drought-stress responses, was significantly down-regulated. Thus, endogenous generation of methyl jasmonate through AtjMT-overexpression modified the transcript levels of genes previously identified as being jasmonate-responsive. This result confirms that MeJA formation is a key control point for jasmonate-responsive gene expression in plants.

Identification of a Novel Jasmonate-Responsive Element in the AtJMT Promoter and Its Binding Protein for AtJMT Repression

Jasmonates (JAs) are important regulators of plant biotic and abiotic stress responses and development. AtJMT in Arabidopsis thaliana and BcNTR1 in Brassica campestris encode jasmonic acid carboxyl methyltransferases, which catalyze methyl jasmonate (MeJA) biosynthesis and are involved in JA signaling. Their expression is induced by MeJA application. To understand its regulatory mechanism, here we define a novel JA-responsive cis-element (JARE), G(C)TCCTGA, in the AtJMT and BcNTR1 promoters, by promoter deletion analysis and Yeast 1-Hybrid (Y1H) assays; the JARE is distinct from other JA-responsive cis-elements previously reported. We also used Y1H screening to identify a trans-acting factor, AtBBD1, which binds to the JARE and interacts with AtJAZ1 and AtJAZ4. Knockout and overexpression analyses showed that AtBBD1 and its close homologue AtBBD2 are functionally redundant and act as negative regulators of AtJMT expression. However, AtBBD1 positively regulated the JA-responsive expression of JR2. Chromatin immunoprecipitation from knockout and overexpression plants revealed that repression of AtJMT is associated with reduced histone acetylation in the promoter region containing the JARE. These results show that AtBBD1 interacts with JAZ proteins, binds to the JARE and represses AtJMT expression.

The serine proteinase inhibitor OsSerpin is a potent tillering regulator in rice

Tillering in rice (Oryza sativa L.) is an important agronomic trait that enhances grain production. A tiller is a specialized grain-bearing branch that is formed on a non-elongated basal internode that grows independently of the mother stem. Transgenic rice over-expressing the transcription factorOsTB1, a homologue of maizeTB1 (Teosinte Branched 1), exhibits markedly reduced lateral branching without the propagation of axillary buds being affected. However, the tillering mechanism remains unknown. Therefore, to further understand that mechanism, we applied proteomics methodology to isolate the proteins involved. Using two-dimensional gel electrophoresis and mass spectrometry, our analysis of the basal nodes from two rice cultivars that differ in their numbers of tillers showed that a rice serine proteinase inhibitor, OsSerpin, accumulates in great amounts in high-tillering ‘Hwachung’ rice. Northern blot analysis revealed that much moreOsSerpin transcript is found in ‘Hwachung’ than in relatively low-tillering ‘Hanmaeum’, likely because of high levels of transcription. Therefore, our data suggest that OsSerpin content determines the extent of lateral branching.