Programmed cell death (PCD) in animals depends on caspase protease activity. Plants also exhibit PCD, for example as a response to pathogens, although plant remains elusive. Here we show that vacuolar processing enzyme (VPE) is essential virus-induced hypersensitive involves PCD. VPE deficiency prevented tobacco plants. structurally unrelated caspases, has caspase-1 Thus, plants have evolved regulated cellular suicide strategy that, unlike PCD of animals, mediated by and the vacuole.

The creation of transgenic plants has contributed extensively to the advancement plant science. Establishing homozygous lines is time-consuming and laborious, using antibiotics or herbicides select transformed may adversely affect growth some plants. Here we describe a novel technology, which have named FAST (fluorescence-accumulating seed technology), that overcomes these difficulties. Although this technology was designed for use in Arabidopsis thaliana, it be adapted other based on...

Plants exhibit an ultimate case of the intracellular motility involving rapid organelle trafficking and continuous streaming endoplasmic reticulum (ER). Although it was long assumed that ER dynamics is actomyosin-driven, responsible myosins were not identified, characterized quantitatively. Here we developed software to generate a detailed velocity-distribution map for GFP-labeled ER. This revealed in most peripheral plane relatively static, whereas inner rapidly with velocities up ∼3.5...

Photosynthetic rate is determined by CO2 fixation and entry into the plant through pores in leaf epidermis called stomata. However, effect of increased stomatal density on photosynthetic remains unclear. This work investigated alteration capacity Arabidopsis thaliana. Stomatal was modulated overexpressing or silencing STOMAGEN, a positive regulator development. Leaf growth were examined transgenic plants. Increased STOMAGEN-overexpressing plants enhanced 30% compared to wild-type Transgenic...

Abstract The nuclear pore complex (NPC) facilitates nucleocytoplasmic transport, a crucial process for various cellular activities. NPC comprises ~30 nucleoporins and is well characterized in vertebrates yeast. However, only eight plant have been identified, little information available about the complete molecular structure of NPCs. In this study, an interactive proteomic approach was used to identify Arabidopsis thaliana nucleoporins. A series five cycles analysis performed using green...

Plants have developed their own defense strategies because they no immune cells. A common plant strategy involves programmed cell death (PCD) at the infection site, but how PCD-associated cell-autonomous immunity is executed in plants not fully understood. Here we provide a novel mechanism underlying immunity, which fusion of membranes large central vacuole with plasma membrane, resulting discharge vacuolar antibacterial proteins to outside cells, where bacteria proliferate. The...

Targeted genome modification technologies are key tools for functional genomics. The clustered regularly interspaced short palindromic repeats (CRISPR)-associated endonuclease Cas9 system (CRISPR/Cas9) is an emerging technology targeted modification. CRISPR/Cas9 consists of a guide RNA (gRNA), which specifies the target sequence, and protein, has activity. been applied to model animals flowering plants, including rice, sorghum, wheat, tobacco Arabidopsis. Here, we report application...

Marchantia polymorpha is one of the model species basal land plants. Although CRISPR/Cas9-based genome editing has already been demonstrated for this plant, efficiency was too low to apply functional analysis. In study, we show establishment CRISPR/Cas9 vectors with high both construction and editing. Codon optimization Cas9 Arabidopsis achieved over 70% at two loci tested. Systematic assessment revealed that guide sequences 17 nt or shorter dramatically decreased efficiency. We also a...

Novel vesicles that accumulate large amounts of proprotein precursors storage proteins were purified from maturing pumpkin seeds. These designated precursor-accumulating (PAC) and had diameters 200 to 400 nm. They contained an electron-dense core surrounded by electron-translucent layer, some also small vesicle-like structures. Immunocytochemical analysis revealed numerous aggregates within the endoplasmic reticulum. It is likely these develop into cores PAC then leave showed complex glycans...

The seeds of higher plants accumulate large quantities storage protein. During seed maturation, protein precursors synthesized on rough endoplasmic reticulum are sorted to vacuoles, where they converted into the mature forms and accumulated. Previous attempts determine sorting machinery for proteins have not been successful. Here we show that a type I membrane protein, AtVSR1/AtELP, Arabidopsis functions as receptor proteins. atvsr1 mutant missorts by secreting them from cells, resulting in...

Proprotein precursors of vacuolar components are transported from the endoplasmic reticulum into vacuoles, where they proteolytically processed their mature forms. However, processing mechanism in plant vacuoles is very obscure. Characterization a purified enzyme required to determine whether single responsible for many proteins with large variability molecular structure. If this true, how can it recognize numerous varieties sites? We have now ( M t =37 000) castor bean seeds. Our results...

Summary Green fluorescent protein (GFP) makes it possible for organelles and transport pathways to be visualized in living cells. However, GFP fluorescence has not yet been observed the vacuoles of any organs higher plants. We found that a vacuole‐targeted was stably transgenic Arabidopsis plants under dark conditions, rapidly disappeared light conditions. The vacuolar degraded within 1 h light, especially blue light. An inhibitor type H + ‐ATPase, concanamycin A, an papain‐type cysteine...

Vacuolar processing enzyme (VPE) has been shown to be responsible for maturation of various seed proteins in protein-storage vacuoles. Arabidopsis three VPE homologues; betaVPE is specific seeds and alphaVPE gammaVPE are vegetative organs. To investigate the activity VPE, we expressed a pep4 strain yeast Saccharomyces cerevisiae found that ability cleave peptide bond at carbonyl side asparagine residues. An immunocytochemical analysis revealed localization lytic vacuoles leaves had treated...

Some compatible pathogens secrete toxins to induce host cell death and promote their growth. The toxin-induced is a pathogen strategy for infection. To clarify the executioner of death, we examined fungal toxin (fumonisin B1 (FB1))-induced Arabidopsis plants. FB1-induced was accompanied with disruption vacuolar membrane followed by lesion formation. features were completely abolished in processing enzyme (VPE)-null mutant, which lacks all four VPE genes genome. Interestingly, an inhibitor...

The surfaces of land plants are covered with a cuticle that is essential for retention water. Epidermal Arabidopsis thaliana embryos and juvenile were homozygous abnormal leaf shape1 (ale1) mutations defective, resulting in excessive water loss organ fusion young plants. In ale1 embryos, the was rudimentary remnants endosperm remained attached to developing embryos. Juvenile had similar cuticle. ALE1 gene isolated using transposon-tagged allele ale1-1. predicted amino acid sequence...

Plants degrade cellular materials during senescence and under various stresses. We report that the precursors of two stress-inducible cysteine proteinases, RD21 a vacuolar processing enzyme (VPE), are specifically accumulated in ~0.5 µm diameter × ~5 long bodies Arabidopsis thaliana. Such have previously been observed but their function was not known. They surrounded with ribosomes thus assumed to be directly derived from endoplasmic reticulum (ER). Therefore, we propose call them ER bodies....

Proteases can catalyze both peptide bond cleavage and formation, yet the hydrolysis reaction dominates in nature. This presents an interesting challenge for biosynthesis of backbone cyclized (circular) proteins, which are encoded as part precursor proteins require post-translational formation to reach their mature form. The largest family circular plant-produced cyclotides; extremely stable with applications bioengineering scaffolds. Little is known about mechanism by they vivo but a highly...

Vacuolar processing enzyme (VPE) is a Cys proteinase responsible for the maturation of vacuolar proteins. Arabidopsis thaliana deltaVPE, which was recently found in database, specifically and transiently expressed two cell layers seed coat (ii2 ii3) at an early stage development. At this stage, death accompanying shrinkage occurs ii2 layer followed by ii3 layer. In deltaVPE-deficient mutant, delayed. Immunocytochemical analysis localized deltaVPE to electron-dense structures inside outside...

Asparaginyl endopeptidase (AEP)/legumain, an asparagine-specific cysteine proteinase in animals, is ortholog of plant vacuolar processing enzyme (VPE), which processes the exposed asparagine residues various proteins. In search for its physiological role mammals, here we generated and characterized AEP-deficient mice. Although their body weights were significantly reduced, they normally born fertile. wild-type kidney where expression AEP was exceedingly high among organs, localization mainly...