The Puccinia striiformis effector Hasp98 facilitates pathogenicity by blocking the kinase activity of wheat TaMAPK4

The obligate biotrophic fungus Puccinia striiformis f. sp. tritici (Pst) employs virulence effectors to disturb host immunity and causes devastating stripe rust disease. However, our understanding of how Pst effectors regulate host defense responses remains limited. In this study, we determined that the Pst effector Hasp98, which is highly expressed in Pst haustoria, inhibits plant immune responses triggered by flg22 or nonpathogenic bacteria. Overexpression of Hasp98 in wheat (Triticum aestivum) suppressed avirulent Pst-triggered immunity, leading to decreased H₂O₂ accumulation and promoting P. striiformis infection, whereas stable silencing of Hasp98 impaired P. striiformis pathogenicity. Hasp98 interacts with the wheat mitogen-activated protein kinase TaMAPK4, a positive regulator of plant resistance to stripe rust. The conserved TEY motif of TaMAPK4 is important for its kinase activity, which is required for the resistance function. We demonstrate that Hasp98 inhibits the kinase activity of TaMAPK4 and that the stable silencing of TaMAPK4 compromises wheat resistance against P. striiformis. These results suggest that Hasp98 acts as a virulence effector to interfere with the MAPK signaling pathway in wheat, thereby promoting P. striiformis infection.     

Production of l-Threonine by Auxotrophic Mutants of Escherichia coli

Two auxotrophic mutants of Escherichia coli have been shown to accumulate significant amounts of l-threonine in the culture medium. One mutant, 13071, is deficient in α,ε-diaminopimelic acid (DAP), and the other, 13070, is deficient in both DAP and methionine.     

N-Glycan structures of squid rhodopsin.

To determine the glycoforms of squid rhodopsin, N-glycans were released by glycoamidase A digestion, reductively aminated with 2-aminopyridine, and then subjected to 2D HPLC analysis [Takahashi, N., Nakagawa, H., Fujikawa, K., Kawamura, Y. & Tomiya, N. (1995) Anal. Biochem.226, 139-146]. The major glycans of squid rhodopsin were shown to possess the alpha1-3 and alpha1-6 difucosylated innermost GlcNAc residue found in glycoproteins produced by insects and helminths. By combined use of 2D HPLC, electrospray ionization-mass spectrometry and permethylation and gas chromatography-electron ionization mass spectrometry analyses, it was revealed that most (85%) of the N-glycans exhibit the novel structure Manalpha1-6(Manalpha1-3)Manbeta1-4GlcNAcbeta1-4(Galbeta1-4Fucalpha1-6)(Fucalpha1-3)GlcNAc.     

Low root zone temperature favours shoot B partitioning into young leaves of oilseed rape (Brassica napus)

In previous studies with tropical plant species, low root zone temperature (RZT) induced boron (B) deficiency, but it is not known if the same response to RZT will be expressed in temperate species, like oilseed rape, that are more tolerant of low temperature. The present experiments investigated the effect of RZT (10 and 20°C) on oilseed rape ( Brassica napus L. cv. Hyola 42) response to B in solution culture, in summer and winter. Regardless of canopy growth conditions, low RZT (10°C) promoted the partitioning of shoot B to the actively growing leaves, especially when B supply was low. However, low RZT did not significantly alter net B uptake rates or plant biomass. Low RZT decreased the shoot-to-root ratio, countering the effects of low B which increased it, leading to a decreased demand for B in the shoot at low RZT. At low B supply, B-deficiency symptoms appeared later at 10 than at 20°C, corresponding with higher B concentrations in the youngest fully opened leaves (YOLs) at 10°C RZT. Thus 10°C RZT increased the tolerance to low B supply. As a result, it is concluded that the effect of decreasing RZT on the responses of the temperate species, oilseed rape, to low B supply depends on whether the low RZT is above or below the optimal root temperature for growth.     

Increased cyclin B1/CDC 2 kinase activity and phosphorylation of Bcl-2 associated with paclitaxel-induced apoptosis in human nasopharyngeal carcinoma cells

Paclitaxel is a potential cancer chemotherapeutic agent for ovary, breast, and head and neck cancers; its effects on nasopharyngeal carcinoma (NPC) have not been reported previously. This study investigated the cytotoxic mechanism of paclitaxel in two NPC cell lines, NPC-TW01 and NPC-TW04. NPC cells treated with pacli-taxel showed convoluted nuclei, condensed chromatin and decreased cellular and nuclear volume, and also exhibited genomic DNA degradation into multiple oligonucleosomal fragments, suggesting that pacli-taxel induced apoptosis in these cells. The effects of paclitaxel on apoptosis-related proteins including Bcl-2, Bax and CDC 2 were also detected. Although the levels of Bcl-2 and Bax were not changed in NPC cells following treatment with 5 nM-1 μM of paclitaxel, phosphorylation of Bcl-2 was significantly observed in the cells treated with 1 μM of paclitaxel for 12 hours. In addition, cyclin B1-associated CDC 2 kinase was highly activated in the NPC cells exposed to paclitaxel even at low (5 nM) concentration, and this result is associated with the finding that low concentration of paclitaxel is able to induce apoptosis in NPC cells.     

Production of chitinolytic enzymes from a novel species ofAeromonas

A bacterial strain secreting potent chitinolytic activity was isolated from shrimp-pond water by enrichment culture using colloidal crab-shell chitin as the major carbon source. The isolated bacterium, designated asAeromonas sp No. 16 exhibited a rod-like morphology with a polar flagellum. Under optimal culture conditions in 500-ml shaker flasks, it produced a chitinolytic activity of 1.4 U ml^–1. A slightly higher enzymatic activity of 1.5 U ml^–1 was obtained when cultivation was carried out in a 5-liter jar fermentor using a medium containing crystalline chitin as the carbon source. The secretion of the enzyme(s) was stimulated by several organic nitrogenous supplements. Most carbon sources tested (glucose, maltose, N-acetylglucosamine, etc) enhanced cell growth, but they slightly inhibited enzyme secretion. Glucosamine (0.5% w/v) severely inhibited cell growth (16% of the control), but it did not significantly affect enzyme secretion. The production of chitinolytic enzymes was pH sensitive and was enhanced by increasing the concentration of colloidal chitin to 1.5%. The observed chitinolytic activity could be attributed to the presence of -N-acetylglucosaminidase and chitinase. Chitinase was purified by ammonium sulfate fractionation and preparative gel electrophoresis to three major bands on SDS-PAGE. An in-gel enzymatic activity assay indicated that all three bands possessed chitinase activity. Analysis of the enzymatic products indicated that the purified enzyme(s) hydrolyzed colloidal chitin predominantly to N,N-diacetyl-chitobiose and, to a much lesser extent, the mono-, tri, and tetramer of N-acetylglucosamine, suggesting that they are mainly endochitinases.     

Cigarette advertising and onset of smoking in children: questionnaire survey.

OBJECTIVE: To investigate uptake of smoking in a cohort of 11 to 12 year olds related to awareness of advertised cigarette brands named. DESIGN: Self completed questionnaires administered to whole classes of schoolchildren in June 1993 and June 1994. SETTING: Primary, middle, and secondary schools in the north and south of England. SUBJECTS: 1450 pupils aged 11 and 12 years at the time of the first survey. MAIN OUTCOME MEASURES: Onset of smoking and brands smoked by the second survey related to cigarette brands named in the first one. Less advertised brands were used as the base for calculating odds ratios. RESULTS: Girls who named the most advertised brands-namely, Benson and Hedges alone (odds ratio = 2.50, 95% confidence interval = 1.18 to 5.30) or Benson and Hedges and Silk Cut (2.15, 1.04 to 4.42) in the first survey were at greatest risk of taking up smoking by the second one. The difference was similar but not significant for boys. Boys and girls who named the least advertised brands in the first survey were at no greater risk of taking up smoking by the second survey than those who named no brands (boys odds ratio = 0.49 (0.24 to 1.01); girls 0.79 (0.38 to 1.62)). New smokers were more likely to smoke any available brand (29.5%) or a less advertised brand such as Embassy (24.6%) than the most advertised ones, Benson and Hedges (19.7%) and Silk Cut (14.8%). Established smokers were more selective, only 15% smoking any available brand and 38.3% smoking Benson and Hedges. CONCLUSIONS: Cigarette advertising appears to increase children''s awareness of smoking at a generic level and encourages them to take up the behaviour, beginning with any cigarettes which are available and affordable.     

DISCOVERY OF VERTEBRATE FOSSILS AND PALEOLITHIC ARTIFACTS IN DANJIANG SUBMERGING AREA AND ITS IMPLICATIONS

IntheareatobefloodedinthesecondengineeringstagefortheDanjiangreservoir,wediscovered16vertebratefossillocalitiesand52Paleolithicsitesin1994,andcollected603artifactsandmanyfossils.Ofthel6new1yfoundvertebratefossillocalities,threearereptilesitesrepresentedbydinosaureggsandlimbbones,andl3producemamma1s,including4Pale0gene,2Ne0geneand7Quaternarysites'ThemammalianlocalitiesareofPale0cene,Eocene,EarlyMiocene,andPli0ceneorEarlyPleistocene,MiddleandIntePleistocene,respectively.Theyfillinsomestrat…