bluuebunny/crossref_metadata_2025 huggingface dataset:
The Root-Knot Nematode Calreticulin Mi-CRT Is a Key Effector in Plant Defense Suppression
Authors: Jaouannet M. , Magliano M. , Arguel M. J. , Gourgues M. , Evangelisti E. , Abad P. , Rosso M. N.
Published: August 2012

Root-knot nematodes (RKN) are obligate biotrophic parasites that settle close to the vascular tissues in roots, where they induce the differentiation of specialized feeding cells and maintain a compatible interaction for 3 to 8 weeks. Transcriptome analyses of the plant response to parasitic infection have shown that plant defenses are strictly controlled during the interaction. This suggests that, similar to other pathogens, RKN secrete effectors that suppress host defenses. We show here that Mi-CRT, a calreticulin (CRT) secreted by the nematode into the apoplasm of infected tissues, plays an important role in infection success, because Mi-CRT knockdown by RNA interference affected the ability of the nematodes to infect plants. Stably transformed Arabidopsis thaliana plants producing the secreted form of Mi-CRT were more susceptible to nematode infection than wild-type plants. They were also more susceptible to infection with another root pathogen, the oomycete Phytophthora parasitica. Mi-CRT overexpression in A. thaliana suppressed the induction of defense marker genes and callose deposition after treatment with the pathogen-associated molecular pattern elf18. Our results show that Mi-CRT secreted in the apoplasm by the nematode has a role in the suppression of plant basal defenses during the interaction.
Molecular Analysis of a Plum pox virus W Isolate in Plum Germplasm Hand Carried into the USA from the Ukraine Shows a Close Relationship to a Latvian Isolate
Authors: Mavrodieva Vessela , James Delano , Williams Karen , Negi Sarika , Varga Aniko , Mock Ray , Levy Laurene
Published: August 2012

Four of 19 Prunus germplasm accessions hand carried from the Ukraine into the United States without authorization were found to be infected with Plum pox virus (PPV). Of the three isolates characterized, isolates UKR 44189 and UKR 44191 were confirmed to be isolates of PPV strain W, and UKR 44188 was confirmed to be an isolate of PPV strain D. UKR 44189 and UKR 44191 are very closely related to the PPV strain W isolate LV-145bt (HQ670748) from Latvia. Nucleotide and amino acid sequence identities between these three isolates were greater than 99%. This indicates that the isolates are very closely related and likely originated from a common source. The high genetic diversity among PPV-W strain isolates allowed the identification of potential recombination events between PPV isolates. It appears also that GF 305 peach and Prunus tomentosa are not hosts for the PPV isolate UKR 44189.
First Report of Phomopsis Seed Decay of Soybean Caused by Phomopsis longicolla in South China
Authors: Shan Z. , Li S. , Liu Y. , Yang Z. , Yang C. , Sha A. , Chen H. , Chen S. , Zhou X-A.
Published: August 2012

Phomopsis seed decay of soybean (Glycine max (L.) Merr.) causes poor seed quality and suppresses yield in most soybean-growing areas in the world. The disease is caused primarily by Phomopsis longicolla Hobbs. During the spring of 2010, soybean seeds without symptoms were planted in the fields but emergence was poor and the emergence rates ranged from 30 to 70% in south China. Approximately 3,000 symptomless seeds were randomly collected from seven fields at three locations in Guangzhou, Nanchang, and Wuhan. Seeds were surface disinfected with 1% sodium hypochlorite for 12 min, rinsed in sterile distilled water three to four times, and placed on 2% agar. Plates were then incubated at 26°C under 16/8-h photoperiod for 3 to 4 days. About 10 to 20% of the seeds produced white hyphae that spread rapidly and covered the whole seed. The hyphae from fungal isolates were transferred to potato dextrose agar (PDA) and incubated at 26°C in the dark. After 3 to 4 weeks, conidia were elliptical with two oil drops at both ends and hyaline (6.2 to 7.2 × 2.6 to 3.2 μm). The cultural and morphological characteristics of the isolates corresponded with the description of P. longicolla (2). Colonies on PDA were floccose, dense, and white. Stromata were large, black, and spreading. To confirm the morphological identification, the ribosomal internal transcribed spacers (ITS1-5.8S-ITS2) from three isolates were sequenced (GenBank Accession Nos. JQ899030, JQ899031, and JQ899032). BLAST analysis indicated that the isolates had 99% nucleotide sequence identity with P. longicolla (GenBank Accession Nos. AY857868.1, EF026104, and HQ130441.1). Pathogenicity tests were conducted on 2-week-old soybean seedlings (3). A mycelial plug (3 mm in diameter) from the margin of 1-week-old PDA culture of the Wuhan isolate was individually placed mycelial side down directly on the top of cut stem 1 to 2 cm above cotyledon node of the soybean seedling. PDA plugs without the fungus was used as the negative control. All seedlings were kept in a growth chamber at 26°C with 92 to 94% relative humidity. After 2 weeks, all inoculated seedlings showed browning, stem wilt, and the lesions were 0.3 to 2.0 cm long. No symptoms were observed in the control plants. P. longicolla was reisolated from the infected seedlings. The pathogenicity test was repeated three times. Soybean stem blight caused by P. longicolla has been reported in northeast China (1). To our knowledge, this is the first report of P. longicolla causing Phomopsis seed decay of soybean in south China. This report will establish a foundation for developing a program for screening germplasm for resistance to this disease in south China.

References: (1) Y. L. Cui et al. Plant Pathol. 58:799, 2009. (2) T. W. Hobbs et al. Mycologia 77:535,1985. (3) S. Li et al. Plant Dis. 85:1031, 2001.
First Report of Stem Blight of Blueberry Caused by Botryosphaeria dothidea in China
Authors: Yu L. , Rarisara I. , Xu S. G. , Wu X. , Zhao J. R.
Published: August 2012

Blueberry (Vaccinium spp.) production in southwest and northeast China has grown to over 100,000 ha in the last 20 years thanks to the fruit's high nutritional and economic value. As blueberry acreage increases, the diversity of diseases and challenges for control are gaining more attention. In August 2010, stem and branch blight occurred on Highbush Blueberries (Vaccinium corymbosum L.) at commercial farms in Lijiang and Zongdian, Yunnan Province (southwestern China), with crop damage ranging from 10 to 15%. Typical symptoms of the disease were blight and dieback on the stems with lesions extending along entire branches. Diseased samples (phloem and xylem sectors in the wood) were washed with running tap water, disinfected with 2% sodium hypochlorite, then 70% alcohol, rinsed in sterile distilled water, plated on potato dextrose agar (PDA), and incubated at 28°C. Fungal isolates developed copious, white aerial mycelium that became dark gray after 4 to 5 days and formed black pycnidia after 18 days. Conidia were hyaline, aseptate, thin walled, fusiform, and measured 21 to 27 × 4 to 6 μm. Identity was confirmed by analysis of the ribosomal DNA internal transcribed spacer region ITSI-5.8S -ITS2 with primers ITS1 and ITS4. BLAST searches showed 99% identity with Botryosphaeria dothidea isolates from GenBank (Accession Nos. AB693904 and JF800139). Representative sequences of B. dothidea from Highbush Blueberries from China were deposited into GenBank (Accession No. JX096631). On the basis of morphological and molecular results, the fungus isolated from diseased Highbush Blueberries stem was confirmed to be B. dothidea. Pathogenicity tests were conducted on 2-year-old blueberry seedlings (Highbush Blueberries). Mycelial plugs (2 to 3 mm in diameter) of B. dothidea from actively growing colonies (PDA) were applied to same-size bark wounds in the center of the stems. Inoculation wounds were wrapped with Parafilm. Control seedlings received sterile PDA plugs. Inoculated and control seedlings (five each) were kept in a greenhouse and watered as needed. After 12 days, all of the inoculated but none of the control blueberry seedlings showed dark vascular stem tissue. B. dothidea was reisolated from symptomatic tissues, thus fulfilling Koch's postulates. No symptoms were visible in the control seedlings. B. dothidea has been reported as a pathogen of sycamore (3), olives (1), and peach (2). However, no research has been conducted on stem blight of blueberry caused by B. dothidea in southwest or mainland China. To our knowledge, this is the first report of B. dothidea on blueberry in southwest China.

References: (1) M. Chattaoui, et al. Plant Dis. 95:770, 2011. (2) Y. Ko et al. Plant Pathol. Bull. 1:70, 1992. (3) E. Turco, et al. Plant Dis. 90:1106, 2006.
First Report of Potato Gangrene Caused by Phoma foveata in China
Authors: Yang C.-D. , Chen X.-R. , Jiang H.-X. , Pu C.-J.
Published: August 2012

A potato tuber rot disease of unknown cause, affecting 5 to 15% of the potato tuber, was observed at Gansu Province of China in March 2010. Sunken, round, oval, or irregular lesions formed at the umbilicus or buds of potato tubers after 30 days of storage at 4°C. These lesions gradually expanded to form khaki, lavender sunken lesions ranging from 1 to 3 cm. Small black bodies were observed in the center of the lesions after 45 days. Twenty-six diseased tubers were collected and surface sterilized with 75% alcohol. Diseased tissue was then directly transferred to potato dextrose agar (PDA) medium for isolation of pathogenic fungi. Eight fungal isolates from disease tubers were obtained and pathogenicity was evaluated. Conidial suspensions (106 CFU/ml) of per isolate were sprayed on 20 potato tubers, respectively. These potato tubers were stabbed about 20 times with five wounds in a row along the tuber and maximum distance between each row. Wounds were made 2 mm deep and 0.5 mm in diameter with a no. 4 insect needle. Control tubers received water without conidia. The inoculated tubers were put in an incubator at 15°C after 72 h with relative humidity 100%. Assays were repeated three times. Typical symptoms of the disease were observed 14 days after inoculation. Pycnidia sharing the characteristics of the inoculated isolates were retrieved from new lesions after 6 weeks, whereas symptoms did not occur on control tubers. Eight isolates were cultured on PDA medium for 7 days at 20°C and then at 5°C for approximately 30 days to determine cultural and morphological characteristics. Pycnidia were black brown, spherical or oblate, scattered or clustered, and ranged from 82 to 210 × 64 to 175 μm. Conidia were unicellular and colorless, and 2.1 to 4.4 × 5.8 to 11.5 μm. Chlamydospores were spherical and 27 to 81 × 18 to 63 μm. The fungi shared morphological characteristics of P. foveata described in the literature. On oat medium (OA), yellow-green, needle-like crystals were formed. The growth rate of the pathogen on MA and OA was 1.0 cm/day. The pathogens were identified as P, foveata based on the symptoms, morphology, and growth rate (1, 2, 3). Genomic DNA was extracted with UNIQ-10 column fungal genomic DNA extraction kit and ribosomal DNA was amplified with ITS1(TCCGTAGGTGAACCTGCGG) and ITS4 (TCCTCCGCTTATTGATATGC) primers. The nucleotide sequence of the 539-bp amplicon (GenBank Accession No. JQ804843) was 99% identical to the ITS sequence from P. foveata available from GenBank (GU237742). Management strategies for potato disease control must be adjusted for the presence and control of gangrene disease in Gansu Province.

References: (1) G. H. Boerema et al. Page 220 in: Phoma Identification Manual. CABI Publishing, Wallingford, UK, 2004. (2) EPPO. Quarantine pests for Europe University Press, Cambridge. 865, 1997. (3) W. R. Stevenson et al. Page 25 in: Compendium of Potato Diseases, 2nd Edition. APS Press, St. Paul, MN, 2004.
Resistance to Cyprodinil and Lack of Fludioxonil Resistance in Botrytis cinerea Isolates from Strawberry in North and South Carolina
Authors: Fernández-Ortuño Dolores , Chen Fengping , Schnabel Guido
Published: August 2012

Chemical control of gray mold of strawberry caused by Botrytis cinerea is essential to prevent pre- and postharvest fruit decay. For more than 10 years, the anilinopyrimidine (AP) cyprodinil and the phenylpyrrole fludioxonil (Switch 62.5WG) have been available to commercial strawberry producers in the United States for gray mold control. Both active ingredients are site-specific inhibitors and, thus, prone to resistance development. In this study, 217 single-spore isolates of B. cinerea from 11 commercial strawberry fields in North and South Carolina were examined for sensitivity to both fungicides. Isolates that were sensitive (53%), moderately resistant (30%), or resistant (17%) to cyprodinil were identified based on germ tube inhibition at discriminatory doses of cyprodinil at 1 and 25 mg/liter at 10 of the 11 locations. None of the isolates was fludioxonil resistant. Phenotypes that were moderately resistant or resistant to cyprodinil were not associated with fitness penalties for mycelial growth rate, spore production, or osmotic sensitivity. Detached fruit assays demonstrated cross resistance between the two AP fungicides cyprodinil and pyrimethanil, and that isolates that were characterized in vitro as moderately resistant or resistant were equivalent in pathogenicity on fruit sprayed with pyrimethanil (currently the only AP registered in strawberry as a solo formulation). This suggests that the in vitro distinction of moderately resistant and resistant isolates is of little if any field relevance. The absence of cross-resistance with fludioxonil, iprodione, cycloheximide, and tolnaftate indicated that multidrug resistance in the form of multidrug resistance phenotypes was unlikely to be involved in conferring resistance to APs in our isolates. Implications for resistance management and disease control are discussed.
Sources of Verticillium dahliae Affecting Lettuce
Authors: Atallah Zahi K. , Maruthachalam Karunakaran , Subbarao Krishna V.
Published: August 2012

Since 1995, lettuce in coastal California, where more than half of the crop in North America is grown, has consistently suffered from severe outbreaks of Verticillium wilt. The disease is confined to this region, although the pathogen (Verticillium dahliae) and the host are present in other crop production regions in California. Migration of the pathogen with infested spinach seed was previously documented, but the geographic sources of the pathogen, as well as the impact of lettuce seed sparsely infested with V. dahliae produced outside coastal California on the pathogen population in coastal California remain unclear. Population analyses of V. dahliae were completed using 16 microsatellite markers on isolates from lettuce plants in coastal California, infested lettuce seed produced in the neighboring Santa Clara Valley of California, and spinach seed produced in four major spinach seed production regions: Chile, Denmark, the Netherlands, and the United States (Washington State). California produces 80% of spinach in the United States and all seed planted with the majority infested by V. dahliae comes from the above four sources. Three globally distributed genetic populations were identified, indicating sustained migration among these distinct geographic regions with multiple spinach crops produced each year and repeated every year in coastal California. The population structure of V. dahliae from coastal California lettuce plants was heavily influenced by migration from spinach seed imported from Denmark and Washington. Conversely, the sparsely infested lettuce seed had limited or no contribution to the Verticillium wilt epidemic in coastal California. The global trade in plant and seed material is likely contributing to sustained shifts in the population structure of V. dahliae, affecting the equilibrium of native populations, and likely affecting disease epidemiology.
TheSaccharomyces cerevisiaeW303-K6001 cross-platform genome sequence: insights into ancestry and physiology of a laboratory mutt
Authors: Ralser Markus , Kuhl Heiner , Ralser Meryem , Werber Martin , Lehrach Hans , Breitenbach Michael , Timmermann Bernd
Published: August 2012

Saccharomyces cerevisiaestrain W303 is a widely used model organism. However, little is known about its genetic origins, as it was created in the 1970s from crossing yeast strains of uncertain genealogy. To obtain insights into its ancestry and physiology, we sequenced the genome of its variant W303-K6001, a yeast model of ageing research. The combination of two next-generation sequencing (NGS) technologies (Illumina and Roche/454 sequencing) yielded an 11.8 Mb genome assembly at an N50 contig length of 262 kb. Although sequencing was substantially more precise and sensitive than whole-genome tiling arrays, both NGS platforms produced a number of false positives. At a 378× average coverage, only 74 per cent of called differences to the S288c reference genome were confirmed by both techniques. The consensus W303-K6001 genome differs in 8133 positions from S288c, predicting altered amino acid sequence in 799 proteins, including factors of ageing and stress resistance. The W303-K6001 (85.4%) genome is virtually identical (less than equal to 0.5 variations per kb) to S288c, and thus originates in the same ancestor. Non-S288c regions distribute unequally over the genome, with chromosome XVI the most (99.6%) and chromosome XI the least (54.5%) S288c-like. Several of these clusters are shared withΣ1278B, another widely used S288c-related model, indicating that these strains share a second ancestor. Thus, the W303-K6001 genome pictures details of complex genetic relationships between the model strains that date back to the early days of experimental yeast genetics. Moreover, this study underlines the necessity of combining multiple NGS and genome-assembling techniques for achieving accurate variant calling in genomic studies.
Methanomassiliicoccus luminyensis gen. nov., sp. nov., a methanogenic archaeon isolated from human faeces
Authors: Dridi Bédis , Fardeau Marie-Laure , Ollivier Bernard , Raoult Didier , Drancourt Michel
Published: August 2012

During attempts to obtain novel, human-associated species of the domainArchaea, a coccoid micro-organism, designated strain B10T, was isolated in pure culture from a sample of human faeces collected in Marseille, France. On the basis of its phenotypic characteristics and 16S rRNA andmcrA gene sequences, the novel strain was classified as a methanogenic archaeon. Cells of the strain were non-motile, Gram-staining-positive cocci that were approximately 850 nm in diameter and showed autofluorescence at 420 nm. Cells were lysed by 0.1 % (w/v) SDS. With hydrogen as the electron donor, strain B10Tproduced methane by reducing methanol. The novel strain was unable to produce methane when hydrogen or methanol was the sole energy source. In an atmosphere containing CO2, strain B10Tcould not produce methane from formate, acetate, trimethylamine, 2-butanol, 2-propanol, cyclopentanol, 2-pentanol, ethanol, 1-propanol or 2,3-butanediol. Strain B10Tgrew optimally with 0.5–1.0 % (w/v) NaCl, at pH 7.6 and at 37 °C. It required tungstate-selenite for growth. The complete genome of the novel strain was sequenced; the size of the genome was estimated to be 2.05 Mb and the genomic DNA G+C content was 59.93 mol%. In phylogenetic analyses based on 16S rRNA gene sequences, the highest sequence similarities (98.0–98.7 %) were seen between strain B10Tand several uncultured, methanogenicArchaeathat had been collected from the digestive tracts of a cockroach, a chicken and mammals. In the same analysis, the non-methanogenic ‘CandidatusAciduliprofundum boonei’ DSM 19572 was identified as the cultured micro-organism that was most closely related to strain B10T(83.0 % 16S rRNA gene sequence similarity). Each of the three treeing algorithms used in the analysis of 16S rRNA gene sequences indicated that strain B10Tbelongs to a novel order that is distinct from theThermoplasmatales. The novel strain also appeared to be distinct fromMethanosphaera stadtmanaeDSM 3091T(72.9 % 16S rRNA gene sequence similarity), another methanogenic archaeon that was isolated from human faeces and can use methanol in the presence of hydrogen. Based on the genetic and phenotypic evidence, strain B10Trepresents a novel species of a new genus for which the nameMethanomassiliicoccus luminyensisgen. nov., sp. nov. is proposed. The type strain of the type species is B10T( = DSM 24529T = CSUR P135T).
A baculovirus isolated from wild silkworm encompasses the host ranges of Bombyx mori nucleopolyhedrosis virus and Autographa californica multiple nucleopolyhedrovirus in cultured cells
Authors: Xu Yi-Peng , Gu Lin-Zhu , Lou Yi-Han , Cheng Ruo-Lin , Xu Hai-Jun , Wang Wen-Bing , Zhang Chuan-Xi
Published: August 2012

A baculovirus, named BomaNPV S2, was isolated from a diseased larva of the wild silkworm, Bombyx mandarina. Notably, BomaNPV S2 exhibited a distinguishing feature in that its host range covered that of both Bombyx mori nucleopolyhedrosis virus (BmNPV) and Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in cultured cells. It could replicate in cells of B. mori (Bm5 and BmN), Spodoptera frugiperda (Sf9) and Trichoplusia ni (Tn-5B1-4). However, occlusion-derived virions of BomaNPV S2 in B. mori cells contained only a single nucleocapsid, whereas they contained multiple nucleocapsids in Tn-5B1-4 cells. The complete genome sequence of BomaNPV S2, including predicted ORFs, was determined and compared with the genome sequence of its close relatives. The comparison results showed that most of the BomaNPV S2 genome sequence was shared with BmNPV (BmNPV T3) or BomaNPV S1, but several regions seemed more similar to regions of AcMNPV. This observation might explain why BomaNPV S2 covers the host ranges of BmNPV and AcMNPV. Further recombinant virus infection experiments demonstrated that GP64 plays an important role in BomaNPV S2 host-range determination.
DNA viruses and the cellular DNA-damage response
Authors: Turnell Andrew S. , Grand Roger J.
Published: August 2012

It is clear that a number of host-cell factors facilitate virus replication and, conversely, a number of other factors possess inherent antiviral activity. Research, particularly over the last decade or so, has revealed that there is a complex inter-relationship between viral infection and the host-cell DNA-damage response and repair pathways. There is now a realization that viruses can selectively activate and/or repress specific components of these host-cell pathways in a temporally coordinated manner, in order to promote virus replication. Thus, some viruses, such as simian virus 40, require active DNA-repair pathways for optimal virus replication, whereas others, such as adenovirus, go to considerable lengths to inactivate some pathways. Although there is ever-increasing molecular insight into how viruses interact with host-cell damage pathways, the precise molecular roles of these pathways in virus life cycles is not well understood. The object of this review is to consider how DNA viruses have evolved to manage the function of three principal DNA damage-response pathways controlled by the three phosphoinositide 3-kinase (PI3K)-related protein kinases ATM, ATR and DNA-PK and to explore further how virus interactions with these pathways promote virus replication.
Photodynamic inactivation of prions by disulfonated hydroxyaluminium phthalocyanine
Authors: Janouskova Olga , Rakusan Jan , Karaskova Marie , Holada Karel
Published: August 2012

Sulfonated phthalocyanines (Pcs) are cyclic tetrapyrroles that constitute a group of photosensitizers. In the presence of visible light and diatomic oxygen, Pcs produce singlet oxygen and other reactive oxygen species that have known degradation effects on lipids, proteins and/or nucleic acids. Pcs have been used successfully in the treatment of bacterial, yeast and fungal infections, but their use in the photodynamic inactivation of prions has never been reported. Here, we evaluated the photodynamic activity of the disodium salt of disulfonated hydroxyaluminium phthalocyanine (PcDS) against mouse-adapted scrapie RML prions in vitro. PcDS treatment of RML brain homogenate resulted in a time- and dose-dependent inactivation of prions. The photodynamic potential of Pcs offers a new way to inactivate prions using biodegradable compounds at room temperature and normal pressure, which could be useful for treating thermolabile materials and liquids.
Patient-derived hepatitis C virus and JFH-1 clones differ in their ability to infect human hepatoma cells and lymphocytes
Authors: Sarhan Mohammed A. , Chen Annie Y. , Russell Rodney S. , Michalak Tomasz I.
Published: August 2012

Hepatitis C virus (HCV) is a hepatotropic virus that also infects cells of the immune system. HCV clones cultivated in human hepatoma Huh-7.5 cells have significantly advanced our understanding of HCV replication and candidate hepatocyte receptors. However, naturally occurring patient-derived HCV, in contrast to the HCV JFH-1 clone, is unable to infect Huh-7.5 cells, while it can replicate in human primary T-cells and selected T-cell lines. To better understand this incongruity, we examined the susceptibility of primary T-cells, PBMCs and T-cell lines to infection with patient-derived HCV, the classical HCV JFH-1 and a cell culture-adapted JFH1T known to be highly infectious to Huh-7.5 cells. We also tested whether Huh-7.5 cells are prone to virus readily infecting T-lymphocytes. The results revealed that while primary T-cells and Molt4 and Jurkat T-cell lines were susceptible to patient-derived HCV, they were resistant to infection with either JFH1T or JFH-1. However, the JFH1T clone interacted more firmly, although non-productively, with the cells than JFH-1. Further, Huh-7.5 cells robustly supported replication of JFH1T but not patient-derived, wild-type virus, despite using highly sensitive detection assays. In conclusion, JFH-1 and JFH1T clones were unable to establish productive infection in human primary T-cells, PBMCs and T-cell lines known to be prone to infection by patient-derived HCV, while Huh-7.5 cells were resistant to infection with naturally occurring virus infecting immune cells. The data showed that the ability to infect lymphocytes is a characteristic of native virus but not laboratory HCV clones.
Seed Yield of Mungbean (Vigna radiata(L.) Wilczek) in relation to Growth and Developmental Aspects
Authors: Mondal M. M. A. , Puteh A. B. , Malek M. A. , Ismail M. R. , Rafii M. Y. , Latif M. A.
Published: August 2012

Growth parameters such as leaf area (LA), total dry mass (TDM) production, crop growth rate (CGR), relative growth rate (RGR), and net assimilation rate (NAR) were compared in six varieties of mungbean under subtropical condition (24°8′ N 90°0′ E) to identify limiting growth characters for the efficient application of physiology breeding for higher yields. Results revealed that a relatively smaller portion of TDM was produced before flower initiation and the bulk of it after anthesis. The maximum CGR was observed during pod filling stage in all the varieties due to maximum leaf area (LA) development at this stage. Two plant characters such as LA and CGR contributed to the higher TDM production. Results indicated that high yielding mungbean varieties should possess larger LA, higher TDM production ability, superior CGR at all growth stages, and high relative growth rate and net assimilation rate at vegetative stage which would result in superior yield components.
Subjective Outcome Evaluation of the Project P.A.T.H.S. (Extension Phase) Based on the Perspective of Program Implementer
Authors: Shek Daniel T. L. , Yu Lu
Published: August 2012

A total of 231 schools participated in the Project P.A.T.H.S. in 2009/2010 school year. After completion of the Tier 1 Program, subjective outcome evaluation data were collected from 3,259 program implementers. Based on the consolidated data with schools as units, results showed that participants had positive perceptions of the program, implementers, and benefits of the program. More than four-fifth of the implementers regarded the program as helpful to the program participants. Multiple regression analyses revealed that perceived qualities of the program and the program implementers predicted perceived effectiveness of the program. Similar to previous studies, compared to implementers’ perception about their performance, the perceived program content appeared to be a stronger predictor of program success. The present study provides additional support for the effectiveness of the Tier 1 Program of the Project P.A.T.H.S. in Hong Kong.
mRNA Expression of EgCHI1, EgCHI2, and EgCHI3 in Oil Palm Leaves (Elaeis guineesisJacq.) after Treatment withGanoderma boninensePat. andTrichoderma harzianumRifai
Authors: Naher Laila , Tan Soon Guan , Ho Chai Ling , Yusuf Umi Kalsom , Ahmad Siti Hazar , Abdullah Faridah
Published: August 2012

Background. Basal stem rot (BSR) disease caused by the fungusGanoderma boninenseis the most serious disease affecting the oil palm; this is because the disease escapes the early disease detection. The biocontrol agentTrichoderma harzianumcan protect the disease only at the early stage of the disease. In the present study, the expression levels of three oil palm (Elaeis guineensisJacq.) chitinases encoding EgCHI1, EgCHI2, and EgCHI3 at 2, 5, and 8 weeks inoculation were measured in oil palm leaves from plants treated withG. boninenseorT. harzianumalone or both.Methods. The five-month-old oil palm seedlings were treated with Gano-wood blocks inoculum and trichomulch. Expression of EgCHI1, EgCHI2, and EgCHI3 in treated leaves tissue was determined by real-time PCR.Results. Oil palm chitinases were not strongly expressed in oil palm leaves of plants treated withG. boninensealone compared to other treatments. Throughout the 8-week experiment, expression of EgCHI1 increased more than 3-fold in leaves of plants treated withT. harzianumandG. boninensewhen compared to those of control and other treated plants.Conclusion. The data illustrated that chitinase cDNA expression varied depending on tissue and the type of treatment.
Potential CanonicalWntPathway Activation in High-Grade Astrocytomas
Authors: Schüle Rebecca , Dictus Christine , Campos Benito , Wan Feng , Felsberg Jörg , Ahmadi Rezvan , Centner Franz-Simon , Grabe Niels , Reifenberger Guido , Bermejo Justo L. , Unterberg Andreas , Herold-Mende Christel
Published: August 2012

Aberrantwntpathway activation through cytoplasmic stabilization ofβ-catenin is crucial for the development of various human malignancies. In gliomagenesis, the role of canonical (i.e.,β-catenin-dependent) signalling is largely unknown. Here, we studied canonicalwntpathway activation in 15 short-term cultures from high-grade gliomas and potential pathomechanisms leading to cytoplasmicβ-catenin accumulation. Furthermore, we assessed the prognostic relevance ofβ-catenin expression in a tissue microarray comprising 283 astrocytomas. Expression ofβ-catenin, its transcriptional cofactors TCF-1 and TCF-4 as well as GSK-3βand APC, constituents of theβ-catenin degradation complex was confirmed by RT-PCR in all cultures. A cytoplasmicβ-catenin pool was detectable in 13/15 cultures leading to some transcriptional activity assessed by luciferase reporter gene assay in 8/13. Unlike other malignancies, characteristic mutations ofβ-catenin and APC leading to cytoplasmic stabilization ofβ-catenin were excluded by direct sequencing or protein truncation test. In patient tissues,β-catenin expression was directly and its degradation product's (β-catenin-P654) expression was inversely correlated with WHO grade. Increasedβ-catenin expression and lowβ-catenin-P654 expression were associated with shorter survival. Altogether, we report on potential canonicalwntpathway activation in high-grade gliomas and demonstrate thatβ-catenin expression in astrocytomas is associated with increased malignancy and adverse outcome.
Characteristics ofStreptococcus pneumoniaeStrains Colonizing Upper Respiratory Tract of Healthy Preschool Children in Poland
Authors: Korona-Glowniak Izabela , Malm Anna
Published: August 2012

Antibiotic resistant and invasive pneumococci may spread temporally and locally in day care centers (DCCs). We examined 267 children attending four DCCs located in the same city and 70 children staying at home in three seasons (autumn, winter, and spring) to determine prevalence, serotype distribution, antibiotic resistance patterns, and transmission of pneumococcal strains colonizing upper respiratory tract of healthy children without antipneumococcal vaccination. By pheno- and genotyping, we determined clonality of pneumococci, including drug-resistant strains. The average carriage of pneumococci in three seasons was 38.2%. 73.4% and 80.4% of the isolates belonged to serotypes present in 10- and 13-valent conjugate vaccine, respectively. Among the pneumococcal strains, 33.3% were susceptible to all antimicrobial tested and 39.2% had decreased susceptibility to penicillin. Multidrug resistance was common (35.7%); 97.5% of drug-resistant isolates represented serotypes included to 10- and 13-valent conjugate vaccine. According to BOX-PCR, clonality definitely was observed only in case of serotype 14. Multivariate analysis determined DCC attendance as strongly related to pneumococcal colonization in all three seasons, but important seasonal differences were demonstrated. In children attending DCCs, we observed dynamic turnover of pneumococcal strains, especially penicillin nonsusceptible and multidrug resistant, which were mostly distributed among serotypes included to available pneumococcal conjugate vaccines.
In VitroChondrogenesis Transformation Study of Mouse Dental Pulp Stem Cells
Authors: Zainal Ariffin Shahrul Hisham , Kermani Shabnam , Megat Abdul Wahab Rohaya , Senafi Sahidan , Zainal Ariffin Zaidah , Abdul Razak Mohamad
Published: August 2012

A major challenge in the application of mesenchymal stem cells in cartilage reconstruction is that whether the cells are able to differentiate into fully mature chondrocytes before grafting. The aim of this study was to isolate mouse dental pulp stem cells (DPSC) and differentiate them into chondrocytes. For this investigation, morphological, molecular, and biochemical analyses for differentiated cells were used. To induce the chondrocyte differentiation, DPSC were cultured in chondrogenic medium (Zen-Bio, Inc.). Based on morphological analyses using toluidine blue staining, proteoglycan products appear in DPSC after 21 days of chondrocyte induction. Biochemical analyses in differentiated group showed that alkaline phosphatase activity was significantly increased at day 14 as compared to control (P<0.05). Cell viability analyses during the differentiation to chondrocytes also showed that these cells were viable during differentiation. However, after the 14th day of differentiation, there was a significant decrease (P<0.05) in the viability proportion among differentiated cells as compared to the control cells. In RT-PCR molecular analyses, mouse DPSC expressedCd146andCd166which indicated that these cells belong to mesenchymal stem cells.Coll IandColl IImarkers showed high expression after 14 and 21 days, respectively. In conclusion, this study showed that DPSC successfully differentiated into chondrocytes.
Reliability and Validity of the Greek QLQ-C30 and QLQ-MY20 for Measuring Quality of Life in Patients with Multiple Myeloma
Authors: Kontodimopoulos Nick , Samartzis Alexandros , Papadopoulos Angelos A. , Niakas Dimitris
Published: August 2012

Objectives. The aim of this study was to assess the psychometric properties of the Greek EORTC QLQ-C30 and QLQ-MY20 instruments.Method. A sample of myeloma patients (N=89) from two tertiary hospitals were surveyed with the QLQ-C30, QLQ-MY20 and various demographic and disease related questions. The previously validated Greek SF-36 instrument was used as a “gold standard” for health-related quality of life (HRQoL) comparisons. Hypothesized scale structure, internal consistency reliability (Cronbach's alpha) and various forms of construct validity (convergent, discriminative, concurrent and known-groups) were assessed.Results. Multitrait scaling confirmed scale structure of the QLQ-C30 and QLQ-MY20, with good item convergence (96% and 72%) and discrimination (78% and 58%) rates. Cronbach'sαwas >0.70 for all but one scale (cognitive functioning). Spearman's correlations between similar QLQ-C30 and SF-36 scales ranged between 0.35–0.80 (P<0.001). Expected interscale correlations and known-groups comparisons supported construct validity. QLQ-MY20 scales showed comparatively lower correlations with QLQ-C30 functional scales, and higher correlations with conceptually related symptom scales.Conclusions. The observed psychometric properties of the two instruments imply suitability for assessing myeloma HRQoL in Greece. Future studies should focus on generalizability of the results, as well as on specific issues such as longitudinal validity and responsiveness.
Deriving Freshwater Quality Criteria for Iron, Lead, Nickel, and Zinc for Protection of Aquatic Life in Malaysia
Authors: Shuhaimi-Othman M. , Nadzifah Y. , Nur-Amalina R. , Umirah N. S.
Published: August 2012

Freshwater quality criteria for iron (Fe), lead (Pb), nickel (Ni), and zinc (Zn) were developed with particular reference to aquatic biota in Malaysia, and based on USEPA’s guidelines. Acute toxicity tests were performed on eight different freshwater domestic species in Malaysia which wereMacrobrachium lanchesteri(prawn), two fish:Poecilia reticulataandRasbora sumatrana, Melanoides tuberculata(snail),Stenocypris major(ostracod),Chironomus javanus(midge larvae),Nais elinguis(annelid), andDuttaphrynus melanostictus(tadpole) to determine 96 h LC50values for Fe, Pb, Ni, and Zn. The final acute value (FAV) for Fe, Pb, Ni, and Zn were 74.5, 17.0, 165, and 304.9 μg L−1, respectively. Using an estimated acute-to-chronic ratio (ACR) of 8.3, the value for final chronic value (FCV) was derived. Based on FAV and FCV, a criterion maximum concentration (CMC) and a criterion continuous concentration (CCC) for Fe, Pb, Ni, and Zn that are 37.2, 8.5, 82.5, and 152.4 μg L−1and 9.0, 2.0, 19.9, and 36.7 μg L−1, respectively, were derived. The results of this study provide useful data for deriving national or local water quality criteria for Fe, Pb, Ni, and Zn based on aquatic biota in Malaysia. Based on LC50values, this study indicated thatN. elinguis, M. lanchesteri, N. elinguis,andR. sumatranawere the most sensitive to Fe, Pb, Ni, and Zn, respectively.
An Emerging New Paradigm in Opioid Withdrawal: A Critical Role for Glia-Neuron Signaling in the Periaqueductal Gray
Authors: Ouyang Handong , Liu Shue , Zeng Weian , Levitt Roy C. , Candiotti Keith A. , Hao Shuanglin
Published: August 2012

The chronic use of opiates (i.e., narcotics such as the natural derivatives of opium including morphine or codeine) or opioids (i.e., semisynthetic derivatives of opium and other molecules that activate opioid receptors) induces dependence, which is associated with various specific behavioral and somatic signs after their withdrawal or after the administration of an opioid antagonist. Among the brain regions implicated in opiate dependence and withdrawal, the periaqueductal gray area (PAG) appears to be critical in regulating the complex signs and symptoms of opioid withdrawal. Numerous neurochemical mechanisms in the PAG have been identified that may contribute to the opioid withdrawal syndrome. Accumulating evidence suggests that glial activation leading to the release of proinflammatory molecules acting on neurons is important in the complex syndrome of opioid dependence and withdrawal. This paper focuses on the recent advances in our understanding of the vital role that glia-neuron interactions play in opioid dependence and withdrawal within the PAG. We summarize those neurochemical mechanisms associated with opioid withdrawal including the recently defined importance of TNFαrelease from activated glial cells that communicate with TNF receptors on PAG neurons.
Dietary Available Phosphorus Affected Growth Performance, Body Composition, and Hepatic Antioxidant Property of Juvenile Yellow CatfishPelteobagrus fulvidraco
Authors: Tang Qin , Wang Chunfang , Xie Congxin , Jin Jiali , Huang Yanqing
Published: August 2012

An 8-week feeding trial was carried out with juvenile yellow catfish to study the effects of dietary available phosphorus (P) on growth performance, body composition, and hepatic antioxidant property. Six pellet diets were formulated to contain graded available P levels at 0.33, 0.56, 0.81, 1.15, 1.31, and 1.57% of dry matter, respectively. Triplicate tanks with each tank containing 60 juveniles (3.09 ± 0.03 g) were fed one of the six experimental diets for 8 weeks. Specific growth rate, feeding rate, and protein efficiency ratio were significantly higher at 0.81% dietary available P. Efficiency of P utilization distinctly decreased with increasing P level. Body lipid content significantly decreased while body ash and feces P content significantly increased with increasing P level. Quadratic regression analysis indicated that vertebrae P content was maximized at 1.21% dietary available P. Fish fed 1.57% dietary available P had highest activity of hepatic superoxide dismutase and catalase and malonaldehyde content. In conclusion, decreasing dietary available P increased P utilization efficiency and body lipid content while decreased vertebrae P content. Juvenile yellow catfish were subjected to oxidative damage under the condition of high dietary P content (1.57%), and the damage could not be eradicated by their own antioxidant defense system.
Spatial and functional relationships among Pol V-associated loci, Pol IV-dependent siRNAs, and cytosine methylation in the Arabidopsis epigenome
Authors: Wierzbicki Andrzej T. , Cocklin Ross , Mayampurath Anoop , Lister Ryan , Rowley M. Jordan , Gregory Brian D. , Ecker Joseph R. , Tang Haixu , Pikaard Craig S.
Published: August 2012

Multisubunit RNA polymerases IV and V (Pols IV and V) mediate RNA-directed DNA methylation and transcriptional silencing of retrotransposons and heterochromatic repeats in plants. We identified genomic sites of Pol V occupancy in parallel with siRNA deep sequencing and methylcytosine mapping, comparing wild-type plants with mutants defective for Pol IV, Pol V, or both Pols IV and V. Approximately 60% of Pol V-associated regions encompass regions of 24-nucleotide (nt) siRNA complementarity and cytosine methylation, consistent with cytosine methylation being guided by base-pairing of Pol IV-dependent siRNAs with Pol V transcripts. However, 27% of Pol V peaks do not overlap sites of 24-nt siRNA biogenesis or cytosine methylation, indicating that Pol V alone does not specify sites of cytosine methylation. Surprisingly, the number of methylated CHH motifs, a hallmark of RNA-directed de novo methylation, is similar in wild-type plants and Pol IV or Pol V mutants. In the mutants, methylation is lost at 50%–60% of the CHH sites that are methylated in the wild type but is gained at new CHH positions, primarily in pericentromeric regions. These results indicate that Pol IV and Pol V are not required for cytosine methyltransferase activity but shape the epigenome by guiding CHH methylation to specific genomic sites.
MdCOP1 Ubiquitin E3 Ligases Interact with MdMYB1 to Regulate Light-Induced Anthocyanin Biosynthesis and Red Fruit Coloration in Apple
Authors: Li Yuan-Yuan , Mao Ke , Zhao Cheng , Zhao Xian-Yan , Zhang Hua-Lei , Shu Huai-Rui , Hao Yu-Jin
Published: August 2012

MdMYB1 is a crucial regulator of light-induced anthocyanin biosynthesis and fruit coloration in apple (Malus domestica). In this study, it was found that MdMYB1 protein accumulated in the light but degraded via a ubiquitin-dependent pathway in the dark. Subsequently, the MdCOP1-1 and MdCOP1-2 genes were isolated from apple fruit peel and were functionally characterized in the Arabidopsis (Arabidopsis thaliana) cop1-4 mutant. Yeast (Saccharomyces cerevisiae) two-hybrid, bimolecular fluorescence complementation, and coimmunoprecipitation assays showed that MdMYB1 interacts with the MdCOP1 proteins. Furthermore, in vitro and in vivo experiments indicated that MdCOP1s are necessary for the ubiquitination and degradation of MdMYB1 protein in the dark and are therefore involved in the light-controlled stability of the MdMYB1 protein. Finally, a viral vector-based transformation approach demonstrated that MdCOP1s negatively regulate the peel coloration of apple fruits by modulating the degradation of the MdMYB1 protein. Our findings provide new insight into the mechanism by which light controls anthocyanin accumulation and red fruit coloration in apple and even other plant species.
Plasma Membrane Localization of Solanum tuberosum Remorin from Group 1, Homolog 3 Is Mediated by Conformational Changes in a Novel C-Terminal Anchor and Required for the Restriction of Potato Virus X Movement
Authors: Perraki Artemis , Cacas Jean-Luc , Crowet Jean-Marc , Lins Laurence , Castroviejo Michel , German-Retana Sylvie , Mongrand Sébastien , Raffaele Sylvain
Published: August 2012

The formation of plasma membrane (PM) microdomains plays a crucial role in the regulation of membrane signaling and trafficking. Remorins are a plant-specific family of proteins organized in six phylogenetic groups, and Remorins of group 1 are among the few plant proteins known to specifically associate with membrane rafts. As such, they are valuable to understand the molecular bases for PM lateral organization in plants. However, little is known about the structural determinants underlying the specific association of group 1 Remorins with membrane rafts. We used a structure-function approach to identify a short C-terminal anchor (RemCA) indispensable and sufficient for tight direct binding of potato (Solanum tuberosum) REMORIN 1.3 (StREM1.3) to the PM. RemCA switches from unordered to α-helical structure in a nonpolar environment. Protein structure modeling indicates that RemCA folds into a tight hairpin of amphipathic helices. Consistently, mutations reducing RemCA amphipathy abolished StREM1.3 PM localization. Furthermore, RemCA directly binds to biological membranes in vitro, shows higher affinity for Detergent-Insoluble Membranes lipids, and targets yellow fluorescent protein to Detergent-Insoluble Membranes in vivo. Mutations in RemCA resulting in cytoplasmic StREM1.3 localization abolish StREM1.3 function in restricting potato virus X movement. The mechanisms described here provide new insights on the control and function of lateral segregation of plant PM.