Extracting Venom from the Parasitoid Wasp <em>Trichogramma dendrolimi</em> Using an Artificial Host

Parasitoid wasps are a diverse group of hymenopteran insects that serve as invaluable resources for pest biocontrol. To ensure successful parasitism, ...

Nanjing Agricultural University

Ministry of Agriculture

Three wheat spindle streak mosaic viruses (WSSMV) resistant cultivars ('Yining Xiaomai', 'Xu87-633', and 'Xifeng') and one susceptible cultivar ('Zhen9523') were used as parents of 3 crosses in this experiment. WSSMV resistance of the parents, F1, and F2 was evaluated under field condition. Based on the segregation ratios of resistant and susceptible plants in F, and F2 populations, it was deduced that the resistance to WSSMV was dominant and the inheritable factors controlling WSSMV resistance were encoded by the nuclear genome. WSSMV resistances in 'Yining Xiaomai' and 'Xifeng' were controlled by two pairs of alleles, which showed complementary effects. However the resistance in 'Xu-87633' was controlled by a single dominant gene. 266 pairs of SSR primers located on 21 wheat chromosomes were used for polymorphic analysis of the two resistant and the susceptible parents 'Yining Xiaomai' and 'Zhen9523', and 108 of them amplified polymorphic DNA products. By Bulk Segregant Analysis of resistant and susceptible pools, one pair of primer located on chromosome arm 2DS, Xgwm261, were found being linked to WSSMV resistance. The 224 F2 individuals were then amplified with marker Xgwm261. The statistic genetic distance between Xgwm261 and the resistance locus was calculated to be 22.9 cM using the software Mapmaker 3.0.

[Inheritance analysis and molecular marker selection of genes for wheat spindle streak mosaic disease resistance].

In order to determine the fragment size of Triticum timopheevii chromosome segments introduced into wheat background and physically map the Pm6 gene, a total of 72 primers (including SSR and STS primers) were used to analyze the eight introduced introgression lines containing Pm6 gene. Referring to the available mapping information of the analyzed markers on chromosome 2B, Pm6 was physically located in distal part of the long arm of chromosome 2B at the region of Bin 2BL-6. The introgressed fragment sizes of the chromosome 2G in different introgression lines was determined to be as follow (from short to long): IGV1-465<IGV1-464<IGV1-463<IGV1-468<(IGV1-448,IGV1-458)<IGV1-474< IGV1-466. The results were coincide with the RFLP results and further confirmed that line IGV1-465 contained the minimum introgression segment. Using the 10 polymorphism PCR markers identified in the present research, it is feasible to differentiate seven introgression lines. This information will be helpful for marker assistant identification of these lines in wheat breeding program and for further isolation of the Pm6 gene.

[Discrimination of the Triticum aestivum-T. timopheevii introgression lines using PCR-based molecular markers].

A total of 135 EST, 27 STS, and 253 SSR primer pairs located in 7 homoeologous groups of wheat and barly were used for amplification of 24 possible Wheat-R. ciliaris disomic addition lines. Fifty-five primer pairs could amplified polymorphic bands between common wheat variety "Inayama Komugi" and the Inayama Komugi- R. ciliaris amphiploid, and 31 of the 55 could amplify the specific bands of R. ciliaris in the addition lines. According to the PCR (polymerase Chain Reaction) results, the added R. ciliaris chromosomes in lines 07K02, 07K06, 07K39, 07K201, 07K202, 07K255, and 07K256 belong to homoeologous group 1 of wheat; the added R. ciliaris chromosomes in lines 07K07, 07K08, 07K09, 07K11, 07K14, and 07K17 belong to homoeologous group 2 of wheat; the added R. ciliaris chromosomes in lines 07K15, 07K16, 07K21, and 07K47 belong to homoeologous group 6 of wheat.

[Molecular marker analysis of wheat-Roegneria ciliaris additions lines.].

Modification of target molecules by ubiquitin or ubiquitin-like (Ubl) proteins is generally reversible. Little is known, however, about the physiological function of the reverse reaction, deconjugation. Atg8 is a unique Ubl protein whose conjugation target is the lipid phosphatidylethanolamine (PE). Atg8 functions in the formation of double-membrane autophagosomes, a central step in the well-conserved intracellular degradation pathway of macroautophagy (hereafter autophagy). Here we show that the deconjugation of Atg8-PE by the cysteine protease Atg4 plays dual roles in the formation of autophagosomes. During the early stage of autophagosome formation, deconjugation releases Atg8 from non-autophagosomal membranes to maintain a proper supply of Atg8. At a later stage, the release of Atg8 from intermediate autophagosomal membranes facilitates the maturation of these structures into fusion-capable autophagosomes. These results provide new insights into the functions of Atg8-PE and its deconjugation.

Dual roles of Atg8-PE deconjugation by Atg4 in autophagy.

Secretory component (SC) is a component of secretory IgA that is designated sIgA to distinguish it from IgA. The monoclonal antibody (MAb) against SC has been shown to be an excellent tool for the detection of the level of sIgA and for the evaluation of the efficacy of mucosal immunity. To prepare a monoclonal antibody against porcine SC, a recombinant porcine SC was expressed and purified. To develop this recombinant SC, the gene encoding the porcine SC was ligated into the pCold I vector. The recombinant vector was then transformed into Escherichia coli BL 21 (DE3), and gene expression was successfully induced by isopropyl-β-D-thiogalactoside (IPTG). After affinity purification with Ni-NTA resin and gel recovery, the recombinant SC protein was used to immunize BALB/c mice. Finally, three hybridoma cell lines showing specific recognitions of both recombinant SC and native SC were used as stable secretors of MAbs against porcine SC and were confirmed to have no reaction to porcine IgA or IgG. The successful preparations of recombinant SC protein and MAbs provide valuable materials that can be used in the mucosal infection diagnosis for porcine disease and mucosal immune evaluation for porcine vaccine, respectively.

Preparation of the porcine secretory component and a monoclonal antibody against this protein.

Cytogenetics was established based on the "Chromosome theory of inheritance", proposed by Boveri and Sutton and evidenced by Morgan's lab in early stage of the 20  centrary. With rapid development of related research areas, especially molecular genetics, cytogenetics developed from traditional into a new era, molecular cytogenetics in late 1960s. Featured by an established technique named DNA in situ hybridization (ISH), molecular cytogenetics has been applied in various research areas. ISH provids vivid and straightforward figures showing the virtual presence of DNA, RNA or proteins. In combination with genomics and cell biology tools, ISH and derived techniques have been widely used in studies of the origin, evolution, domestication of human, animal and plant, as well as wide hybridization and chromosome engineering. The physical location and order of DNA sequences revealed by ISH enables the detection of chromosomal re-arrangments among related species and gaps of assembled genome sequences. In addition, ISH using RNA or protein probes can reveal the location and quantification of transcripted RNA or translated protein. Since the 1970s, scientists from universities or institutes belonging to the Jiangsu Society of Genetics have initiated cytogenetics researches using various plant species. In recent years, research platforms for molecular cytogenetics have also been well established in Nanjing Agricultural University, Yangzhou University, Nanjing Forestry University, Jiangsu Xuhuai Academy of Agricultural Sciences, and Jiangsu Normal University. The application of molecular cytogenetics in plant evolution, wide hybridization, chromosome engineering, chromosome biology, genomics has been successful. Significant progresses have been achieved, both in basic and applied researches. In this paper, we will review main research progresses of plant cytogenetics in Jiangsu province, and discuss the potential development of this research area.

Hai-yan Wang

Department of Entomology

Nanjing Agricultural University

Extracting Venom from the Parasitoid Wasp Trichogramma dendrolimi Using an Artificial Host

Hai-yan Wang

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Entomology

Nanjing Agricultural University

Extracting Venom from the Parasitoid Wasp Trichogramma dendrolimi Using an Artificial Host

Department of Entomology