Molecular and biological characterization of <Emphasis Type="Italic">Chrysanthemum stem necrosis virus</Emphasis> isolates from distinct regions in Japan

Three isolates of Chrysanthemum stem necrosis virus (CSNV) were obtained from chrysanthemum plants in distinct regions of Japan in 2006 and 2007. All the original host plants showed severe necrotic symptoms on the leaves and stems. Amino acid sequence data of the nucleocapsid protein genes of the three isolates (CbCh07A, TcCh07A, and GnCh07S) showed high identities with those of two other CSNV isolates, HiCh06A L1 from Japan and Chry1 from Brazil. Furthermore, for the first time the complete nucleotide sequence of the S RNA was determined for CSNV (isolate HiCh06A). In phylogenetic analysis based on the non-structural protein genes from the genus Tospovirus, HiCh06A L1 was placed in the same genetic group as Tomato spotted wilt virus (TSWV) and Impatiens necrotic spot virus. Host range examination for isolates HiCh06A L1 and CbCh07A showed that green pepper (cv. ‘Kyoyutaka’, ‘Saitamawase’, ‘Tosakatsura’, ‘L3 sarara’ and ‘L3 miogi’) and tomato (cv. ‘Sekaiichitomato’) were systemically susceptible hosts, whereas TSWV-resistant Solanaceae species, Capsicum chinense, Lycopersicon peruvianum and a TSWV-resistant cultivar of green pepper (cv. TSR miogi), were resistant.     

Application of the rpoS gene for the detection of Vibrio anguillarum in flounder and prawn by polymerase chain reaction

Vibrio anguillarum , an opportunistic fish pathogen, is the main species responsible for vibriosis, a disease that affects feral and farmed fish and shellfish, and causes considerable economic losses in marine aquaculture. In this study, we used polymerase chain reaction (PCR) to detect V. anguillarum . PCR specificity was evaluated by amplifying the rpoS gene, a general stress regulator, in six strains of V. anguillarum and 36 other bacterial species. PCR amplified a species-specific fragment (689 bp) from V. anguillarum . Furthermore, the PCR assay was sensitive enough to detect rpoS expression from 3 pg of genomic DNA , or from six colony-forming units (CFU) mL⁻¹ of cultured V. anguillarum . However, the assay was less sensitive when genomic DNA from the infected flounder and prawn was used (limit of detection, 50 ng and 10 ng g⁻¹ tissue, respectively). These data demonstrate that PCR amplification of the rpoS gene is a sensitive and species-specific method to detect V. anguillarum in practical situations.     

鲁南典型区域土地利用变化研究——以台儿庄区为例

以台儿庄区为研究区域,通过搜集2005、2009、2014年3个年份的土地利用状况数据为基础,研究2005~2014年该区域各土地利用类型面积、分布及程度在时间和区域空间的变化,以期对区域土地利用方向和资源管理提供理论和技术支持。结果表明,台儿庄区近十年来农用地总量减少4.23 km~2,建设用地增加6.03 km~2;各乡镇街道中运河街道变化较大,结构变化指数为30.45,土地利用程度变化率高于全区整体水平。     

Glutathione functions on physiological characters to increase copper-induced corn production

Glutathione (GSH) abundantly presents in plant and affects plants growth and development. To justify if GSH affects copper (Cu)-induced corn production, photosynthesis (Pn) rate, and photosynthetically active radiation (PAR), chlorophyll (Chl) content and Chl fluorescence, relative water content (RWC) and yields were evaluated. Different Cu concentrations (0, 0.2, 1.5 and 3.0 ppm of Zn) with or without 100 µm of N-acetyl cysteine (NAC) were arranged as completely randomize design with 5 replications. Results showed that both NAC and Cu affected plant height and leaf numbers. Copper reduced Pn rate, and PAR and increased RWC but no effect was observed on Chl content and Chl fluorescence in leaves. On the other hand, NAC application increased Pn, PAR, Chl parameters regardless of Cu treatment. In the presence of 1.5 ppm of Cu, corn plants showed improve yield and cob length irrespective to NAC treatment. Taken together, this study suggests that NAC might improve some physiological functions in plants to enhance Cu-induced corn production.     

Powdery mildew resistance gene (<Emphasis Type="Italic">Pm</Emphasis>-<Emphasis Type="Italic">AN</Emphasis>) located in a segregation distortion region of melon LGV

Powdery mildew is one of the most important melon pathogens all over the world. So far, many genes conferring resistance to powdery mildew of melon have been described, but few of these have been finely mapped or cloned. Two F₂ populations derived from Ano2 × Hami413 and Ano2 × Queen were used to map the powdery mildew resistance gene by methods of Bulked Segregation Analysis (BSA), comparative genomics and Resistance Gene Analogues (RGA) mapping. It was found that the resistance to powdery mildew in Ano2 was conferred by a dominant gene, and the gene was named Pm-AN. The genetic analysis revealed that Pm-AN located between two codominant markers RPW and MRGH63B in linkage groupV. The genetic distances between Pm-AN and these two markers were 1.4–1.8 and 1.6–2 cM. No recombination was found between Pm-AN and markers ME/E1, SRAP23. Pm-AN was located in a RGA-rich region and cosegregated with the RGA marker MRGH5 and the resistance gene Vat. Synteny analysis showed that markers in this region were collinear between melon and cucumber. Segregation distortion was found in this region using both Ano2 × Hami413 and Ano2 × Queen F₂ populations, and the distortion was more distinct in Ano2 × Hami413 F₂ population. The center of segregation distortion was located in the RGA rich region harboring Pm-AN.     

Watermelon lycopene β-cyclase: promoter characterization leads to the development of a PCR marker for allelic selection

In the carotenoid biosynthetic pathway, lycopene β-cyclase (LCYB) catalyzes the cyclization that converts lycopene into β-carotene. Only a single copy of LCYB was identified and was suggested to encode a chromoplast-specific LCYB (CYCB type) in watermelon [Citrullus lanatus (Thunb.), Matsum & Nakai]. Splicing variants in the 5′-untranslated region were identified, but alternative splicing did not provide an explanation of the regulation of carotenoid accumulation in watermelon flesh. A quantitative assay using real time-PCR showed that differential expression was not detected between red- and canary yellow-fleshed watermelon cultivars. LCYB promoter regions were isolated and characterized, and a sequence difference was identified in the promoter region between red and canary yellow LCYB alleles. This polymorphism did not change the expression of LCYB, but does provide a reliable marker for discriminating LCYB alleles for red and canary yellow flesh. To develop a PCR-based marker to distinguish between the two LCYB alleles, we designed primers flanking the polymorphic region. The newly developed marker, designated Clcyb.600, co-segregated perfectly with flesh color phenotypes and single nucleotide polymorphism (SNP) markers developed in our previous study. Moreover, the Clcyb.600 marker offers easier discrimination of LCYB alleles than SNP or cleaved amplified polymorphic sequence markers, as it does not require restriction enzyme digestion for genotyping. Genotyping of LCYB promoter alleles in various commercial cultivars and plant introductions indicated that watermelon cultivars can be classified into two groups, those carrying a red LCYB allele or a canary yellow LCYB allele.     

A molecular technique for selection of self-compatible varieties of Japanese pear (Pyrus pyrifolia Nakai)

The pear cultivar ‘Osa-Nijisseiki’ (S₂S^sm ₄; sm = stylar-part mutant) has been used as a parent to breed self-compatible cultivars that produce excellent fruits. However, determination of the self-compatibility of ‘Osa-Nijisseiki’ offspring requires a lot of time, 6 years or more, by conventional cross breeding. We have designed a rapid reliable method for the identification of self-compatible varieties of ‘Osa-Nijisseiki’ offspring based on the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) with S-allele specific restriction endonucleases. By using this method, 8 self-compatible varieties were selected among 16 selections resulting from a cross between the self-compatible cultivar ‘Osa-Nijisseiki’ (S₂S^sm ₄) and the self-incompatible cultivars ‘Niitaka’ (S₃S₉), ‘Whasan’ (S₃S₅), ‘Chuwhangbae’ (S₄S₆). The S-genotypes of 16 ‘Osa-Nijisseiki’ offsprings were also determined. This revised version was published online in August 2006 with corrections to the Cover Date.     

Determination of Moisture Deficit and Heat Stress Tolerance in Corn Using Physiological Measurements and a Low‐Cost Microcontroller‐Based Monitoring System

In the southern United States, corn production encounters moisture deficit coupled with high‐temperature stress, particularly during the reproductive stage of the plant. In evaluating plants for environmental stress tolerance, it is important to monitor changes in their physical environment under natural conditions, especially when there are multiple stress factors, and integrate this information with their physiological responses. A low‐cost microcontroller‐based monitoring system was developed to automate measurement of canopy, soil and air temperatures, and soil moisture status in field plots. The purpose of this study was to examine how this system, in combination with physiological measurements, could assist in detecting differences among corn genotypes in response to moisture deficit and heat stress. Three commercial hybrids and two inbred germplasm lines were grown in the field under irrigated and non‐irrigated conditions. Leaf water potential, photosynthetic pigments, cell membrane thermostability (CMT) and maximum quantum efficiency of photosystem II (Fv/Fm) were determined on these genotypes under field and greenhouse conditions. Variations observed in air and soil temperatures, and soil moisture in plots of the individual corn genotypes helped explain their differences in canopy temperature (CT), and these variations were reflected in the physiological responses. One of the commercial hybrids, having the lowest CT and the highest CMT, was the most tolerant among the genotypes under moisture deficit and heat stress conditions. These results demonstrated that the low‐cost microcontroller‐based monitoring system, in combination with physiological measurements, was effective in evaluating corn genotypes for drought and heat stress tolerance.     

Interactions and self-organization in the soil-microbe complex

Soil is the most complicated biomaterial on the planet. As with any material, the physical habitat is of prime importance in determining and regulating biological activity. However, until recently the opaque nature of soil has meant that any interrogation of its interior architecture has been relatively rudimentary, restricted to simple qualitative expressions of the physical heterogeneity that fail to relate to any specific function. However, new techniques and insights into the biophysical and biochemical processes of this inner space are leading to the developments of theoretical frameworks and experimental approaches that will allow us to sustainably manage Earth's most important resource. We introduce the concept that the soil-microbe system is self-organized and suggest new priorities for research based on an integrative approach that combines biochemistry and biophysics.