Identification of QTLs for phenolic compounds in oilseed rape (<Emphasis Type="Italic">Brassica napus L.</Emphasis>) by association mapping using SSR markers

Association mapping identifies quantitative trait loci (QTLs) by examining the marker-trait associations that can be attributed to the strength of linkage disequilibrium between markers and functional polymorphisms across a set of diverse germplasm. In this study, association mapping was performed to detect QTL-linked and genome wide SSR markers linked to phenolic compounds of extraction meal in a population of 49 genetically diverse oilseed rape cultivars of dark-seeded, winter-type oilseed rape accessions. Correction for population structure was performed using 559 genome wide SSR markers. Results showed that seed colour is an important contributor to seed meal quality. Totally, 52 SSR markers linked to phenolic compounds were detected, five of them being QTL linked markers. Some of these markers were already mapped on Brassica napus chromosomes that contain known QTL controlling oilseed rape meal quality traits. Our results demonstrate that association mapping is a useful approach to complement and enhance previous QTL information for marker-assisted selection.     

Brenneria spp. and Rahnella victoriana associated with acute oak decline symptoms on oak and hornbeam in Iran

Decline diseases of forest trees are complex syndromes not attributable to single causal factors. In Iran, symptoms of decline disease have been observed in a number of native forest species including Quercus castaneifolia (chestnut‐leaved oak), Q. brantii (Persian oak) and Carpinus betulus (hornbeam). The symptoms are prevalent in the northern forests and the Zagros mountain forests. There are parallels between the disease in Iran and acute oak decline (AOD) reported in the UK, specifically the presence of weeping cankers, which have been attributed to a polybacterial complex wherein Brenneria goodwinii is considered a key necrogen. Based on the AOD symptomatology, and as a first step towards discovering potential causal agents of the stem weeping symptoms of affected trees in Iran, necrotic tissues were tested primarily for the presence of B. goodwinii. Symptomatic Q. castaneifolia and C. betulus from the Mazandaran Province and symptomatic Q. brantii from Kohgiluyeh and Boyerahmad Province were sampled. Isolation and culture on a selective medium yielded uniform bacterial colonies. Isolates were characterized using phenotypic and genotypic (DNA sequencing) tests. The isolates were phenotypically identical to members of Pectobacteriaceae and Yersiniaceae, specifically Brenneria and Rahnella spp. The nucleotide sequences of the 16S rRNA and housekeeping genes gyrB, infB and atpD (MLSA) amplified via PCR demonstrated that the isolates from the trees in Iran were indeed B. goodwinii, B. roseae subsp. roseae, Rahnella victoriana and an unknown species of Brenneria. Most bacteria isolated from non‐symptomatic trees were Gram‐positive, and Pseudomonas spp. were dominant, but bacterial species isolated from the diseased trees were not detected in healthy trees. Hypersensitivity response tests were positive, but inoculation on saplings was more variable with internal necrosis developing only once in the test period. Therefore, further testing is required. This is the first report of the incidence of B. goodwinii, B. roseae subsp. roseae, R. victoriana and Brenneria sp. associated with acute oak decline‐like symptoms on Q. castaneifolia, Q. brantii and C. betulus across the western forests of Iran and in the world.     

Temperature-dependent functional response of two aphid parasitoids, Aphidius colemani and Aphidius matricariae (Hymenoptera: Aphidiidae), on the cotton aphid

The functional response of two aphid parasitoids, Aphidius colemani Viereck and Aphidius matricariae (Haliday) (Hymenoptera: Aphidiidae), on the cotton aphid, Aphis gossypii Glover (Homoptera: Aphididae), was examined. Five constant temperatures (10, 15, 20, 25, and 30°C) and six host densities (2, 4, 8, 16, 32, and 64) were used during a 24-h period. At each temperature, 2–64 third-instar nymphs of A. gossypii were exposed to individual wasp mating pairs. A type II functional response model for both parasitoid wasps was fit separately for each temperature. The results showed that instantaneous attack rate (a) in A. colemani increased linearly as the temperature increases from 10 to 30°C, but in A. matricariae, (a) increased almost linearly with temperature to reach a maximum at 25°C, and then decreased at 30°C, displaying an asymmetrical dome-shaped pattern. The highest instantaneous attack rate for A. colemani and A. matricariae were calculated 0.940 ± 0.144 day⁻¹ at 30°C and 0.687 ± 0.157 day⁻¹ at 25°C, respectively. Handling time (T _h) for A. colemani was inversely proportional to temperature and ranged from 0.093 ± 0.026 day at 10°C to 0.032 ± 0.004 day at 30°C, but in A. matricariae, (T _h) was between 0.078 ± 0.015 day at 10°C and 0.036 ± 0.014 day at 20°C. A. colemani achieved higher parasitism rate than A. matricariae at higher temperatures. This observation suggests that A. colemani maybe more effective for biological control of the cotton aphid during warmer periods.     

Study on inheritance of seed technological characteristics in sugar beet

Seed is an important input in production of field crops. The successful production of sugar from sugar beet depends on high yielding varieties, optimum plant establishment after sowing and good agricultural practices. Therefore, seed with the highest germination ability and vigour is necessary to ensure an early and high level of field emergence. Knowledge of the genetic control of seed characters may help breeders to efficiently select for improved germination. In this research, variance components and inheritance of sugar beet seed characteristics were studied. Five monogerm diploid O-Type lines were crossed with 15 monogerm, diploid cytoplasmic male sterile (CMS) lines on the basis of North Carolina Design II (Factorial Design), and 75 F1 hybrids (single crosses) were produced. The hybrids were then compared in three completely randomized designs of 25 entries each. Eight seed traits of F1 hybrids were evaluated in the laboratory in three replications. Based on genetic analysis, the additive variance for rootlet length, hypocotyle length, uniformity of germination and 1,000-germ weight was significant at 5% probability level. Also, the dominance variance was significant at 1% probability level for all the seed traits measured except for germination percentage. Narrow-sense heritabilities of rootlet and hypocotyle lengths were significant at 5% probability level. Correlation analysis showed that germination speed and hypocotyle length were positively and negatively correlated with germination uniformity, respectively, at 1% probability level. These results indicated that there was a remarkable genetic variation for the seed traits which could be exploited by selection and heterosis breeding.     

Ectopic expression of barley constitutively activated ROPs supports susceptibility to powdery mildew and bacterial wildfire in tobacco

ROPs (also called RACs) are RHO-like monomeric G-proteins of plants, well-known as molecular switches in plant signal transduction processes, which are involved in plant development and a variety of biotic and abiotic stress responses. The barley (Hordeum vulgare) ROPs RACB, RAC1 and RAC3 have been shown to be involved in cellular growth, polarity and in susceptibility to the biotrophic barley powdery mildew fungus Blumeria graminis f.sp. hordei. We produced transgenic tobacco (Nicotiana tabacum) plants expressing constitutively activated (CA) mutants of the barley ROPs RACB and RAC3 to monitor the impact of heterologous ROP expression on cell polarity and disease susceptibility of tobacco. CA HvROPs influenced leaf texture, disturbed root hair polarity and induced cell expansion in tobacco. Both barley ROPs induced super-susceptibility to the compatible powdery mildew fungus Golovinomyces cichoracearum but only CA HvRAC3 induced super-susceptibility to the bacterial leaf pathogen Pseudomonas syringae pv. tabaci. Data suggest involvements of ROPs in tobacco cell expansion, polar growth and in response to bacterial and fungal leaf pathogens.     

Efficient Process Development of Recombinant Human Granulocyte Colony-Stimulating Factor (rh-GCSF) Production in Escherichia coli

Background:

The protein hormone granulocyte colony-stimulating factor (GCSF) stimulates the production of white blood cells and plays an important role in medical treatment of cancer patients.

Methods:

An efficient process was developed for heterologous expression of the human GCSF in E. coli BL21 (DE3). The feeding rate was adjusted to achieve the maximum attainable specific growth rate under critical value. In this method, specific growth rate was maintained at the maximum value of 0.55 h^-1 at the beginning of feeding to 0.4 h^-1 at the induction time. Recombinant human GCSF (rh-GCSF) was produced as inclusion body. At first, inclusion bodies were released by cell disruption and then washed, solubilized and refolded. Finally, the rh-GCSF was purified by cation exchange chromatography.

Results:

Obviouly, higher specific growth rate decreases process time and consequently increases productivity. The final concentration of biomass and GCSF was achieved 126 g DCW.l^-1 and 32.1 g.l^-1. Also, the final specific yield (Y_P/X) and total productivity of rh-GCSF were obtained 254 mg.g^-1 DCW and 1.83 g.l^-1.h^-1, respectively. According to the available data, this is one of the highest Y_P/X and productivity that has been reported for any human protein which is expressed in E. coli. Recovery yield of purification process was %40 and purity of recombinant protein was over than 99%. The circular dichroism spectra of purified rh-GCSF, Neupogen® and PD-Grastim showed that all proteins have a similar secondary structure.

Conclusion:

Modified exponential feeding strategy for fed-batch cultivation of recombinant E. coli, results in minimum fed-batch duration and maximum productivity. Key Words: Escherichia coli, Granulocyte colony-stimulating factor (GCSF), Process development     

Temperature-dependent development of <Emphasis Type="Italic">Diglyphus isaea</Emphasis> (Hymenoptera: Eulophidae) on <Emphasis Type="Italic">Liriomyza sativae</Emphasis> (Diptera: Agromyzidae) on cucumber

The development of Diglyphus isaea (Walker), a parasitoid of leafminers, was studied under laboratory conditions at seven constant temperatures (10, 15, 20, 25, 30, 35, and 40°C) on Liriomyza sativae Blanchard reared on cucumber (Cucumis sativus L.). The total development period (oviposition to adult emergence) decreased with increasing temperature between 15 and 35°C. In 10 and 40°C no development rate was found in D. isaea, so that it may be claimed that these temperatures fall outside the temperature range for development. Linear regression was used to describe the relationship between development rate and temperature. For egg to adult development, males of D. isaea required 153.8 degree-days (DD) above the theoretical threshold of 9.2°C and females required 161.3 DD above 9.4°C. Data were fitted to four nonlinear temperature-dependent models. Evaluation of the models took place based on the following criteria: fit to data, number and biological value of the fitted coefficient, and accuracy on the estimation of the thresholds. It could be concluded that the Briere-1 and Briere-2 models are suitable for estimating the minimum, maximum and optimal temperature thresholds of D. isaea. Thermal requirements and temperature thresholds can be used to predict the occurrence, number of generations and population dynamics of D. isaea.