Suppression of mRNAs for lipoxygenase (LOX), allene oxide synthase (AOS), allene oxide cyclase (AOC) and 12-oxo-phytodienoic acid reductase (OPR) in pea reduces sensitivity to the phytotoxin coronatine and disease development by Mycosphaerella pinodes

Using a recently developed model pathosystem involving Medicago truncatula and Mycosphaerella pinodes, causal agent of Mycosphaerella blight on pea to understand host molecular response to a fungal suppressor, we applied the suppressor to leaves of M. truncatula and identified 151 nonredundant cDNA fragments as newly expressed genes. These included genes encoding lipoxygenase (LOX) and enoyl-CoA hydratase, which are presumably involved in jasmonic acid (JA) synthesis. Potential genes encoding plastidic enzymes, including allene oxide synthase (AOS) and allene oxide cyclase (AOC), and other peroxisomal enzymes involved in β-oxidation were predicted from the Medicago Gene Index EST database and tested for altered expression by semiquantitative RT-PCR. The coordinated expression of genes encoding both plastidic and peroxisomal enzymes showed that the suppressor likely conditions certain cellular process(es) through the JA synthesis in M. truncatula. To explore the role of JA or JA-regulated cellular process(es) in conditioning susceptibility, we used an Apple latent spherical virus (ALSV)-based virus-induced gene silencing (VIGS) technology to silence pea genes including LOX, AOS, AOC and 12-oxo-phytodienoic acid reductase (OPR). In LOX-, AOS-, AOC- or OPR-silenced pea plants, disease development induced by M. pinodes was remarkably reduced. Similarly, silencing of mRNA for LOX, AOS, AOC or OPR reduced the sensitivity to a phytotoxin, coronatine, which is believed to act through a JA-dependent process. On the basis of these results, it is conceivable that M. pinodes has evolved a strategy to condition susceptibility by manipulating the physiology of host cells, in particular JA-regulated cellular process(es), to promote disease development in pea.     

Occurrence and characterization of tomato spotted wilt orthotospovirus isolated from cucumber

European Journal of Plant Pathology - Diseases associated with tomato spotted wilt orthotospovirus (TSWV) are a serious threat to vegetable production worldwide. In 2019, leaf necrosis disease was...     

A secondary symbiosis in progress?

Algae have acquired plastids by developing an endosymbiotic relationship with either a cyanobacterium (primary endosymbiosis) or other eukaryotic algae (secondary endosymbiosis). We report a protist, which we tentatively refer to as Hatena, that hosts an endosymbiotic green algal partner but inherits it unevenly. The endosymbiosis causes drastic morphological changes to both the symbiont and the host cell architecture. This type of life cycle, in which endosymbiont integration has only partially converted the host from predator to autotroph, may represent an early stage of plastid acquisition through secondary symbiosis.     

Human CHN1 mutations hyperactivate alpha2-chimaerin and cause Duane's retraction syndrome

Duane's retraction syndrome (DRS) is a complex congenital eye movement disorder caused by aberrant innervation of the extraocular muscles by axons of brainstem motor neurons. Studying families with a variant form of the disorder (DURS2-DRS), we have identified causative heterozygous missense mutations in CHN1, a gene on chromosome 2q31 that encodes alpha2-chimaerin, a Rac guanosine triphosphatase-activating protein (RacGAP) signaling protein previously implicated in the pathfinding of corticospinal axons in mice. We found that these are gain-of-function mutations that increase alpha2-chimaerin RacGAP activity in vitro. Several of the mutations appeared to enhance alpha2-chimaerin translocation to the cell membrane or enhance its ability to self-associate. Expression of mutant alpha2-chimaerin constructs in chick embryos resulted in failure of oculomotor axons to innervate their target extraocular muscles. We conclude that alpha2-chimaerin has a critical developmental function in ocular motor axon pathfinding.     

Chondrulelike objects in short-period comet 81P/Wild 2

The Stardust spacecraft returned cometary samples that contain crystalline material, but the origin of the material is not yet well understood. We found four crystalline particles from comet 81P/Wild 2 that were apparently formed by flash-melting at a high temperature and are texturally, mineralogically, and compositionally similar to chondrules. Chondrules are submillimeter particles that dominate chondrites and are believed to have formed in the inner solar nebula. The comet particles show oxygen isotope compositions similar to chondrules in carbonaceous chondrites that compose the middle-to-outer asteroid belt. The presence of the chondrulelike objects in the comet suggests that chondrules have been transported out to the cold outer solar nebula and spread widely over the early solar system.     

Eastern blotting and immunoaffinity concentration using monoclonal antibody for ginseng saponins in the field of traditional chinese medicines

Ginsenosides separated by silica gel TLC blotted to a PVDF membrane that was treated with a NaIO4 solution followed by bovine serum albumin (BSA) resulted in a ginsenoside-BSA conjugate on a PVDF membrane. The blotted spots were stained by anti-ginsenoside Rb1 (G-Rb1) and -Rg1 (G-Rg1) monoclonal antibodies (MAbs). The newly established immunostaining method, Eastern blotting, was applied for the determination of ginsenosides possessing protopanaxadiol and/or protopanaxatriol in the traditional Chinese medicine (TCM). This method developed a new way to separate the ginsenoside molecule into two functional parts using a simple and well-known chemical reaction. The sugar parts were oxidized by NaIO4 to give dialdehydes, which reacted with amino groups of the protein and covalently bound to the adsorbent PVDF membrane. The MAb bound to the aglycon part of the ginsenoside molecule for immunostaining. Double staining of Eastern blotting for ginsenosides using anti-G-Rb1 and -Rg1 MAbs promoted complete identification of ginsenosides in Panax species. The immunoaffinity concentration of G-Rb1 was deteremined by immunoaffinity column conjugated with anti-G-Rb1 MAb leading to the knock-out extract, which will be useful for the pharmacological investigation. To concentrate and determine G-Rb1 in P. japonicus, the crude extract of P. japonicus was fractionated by immunoaffinity column conjugated with anti-G-Rb1 MAb. Two ginsenosides, chikusetsusaponins III and IV having protopanaxadiol as an aglycon, were identified by Eastern blotting, although it was expected that G-Rb1 might be a component of P. japonicus by enzyme-linked immunosorbent assay (ELISA) analysis.