Three pairs of primers were designed for amplification of the adiponectin gene in geese (Anser demeatica) based on the published sequences of chicken, duck and other adiponectin homologues. The complete 1062 bp sequence of the adiponectin gene in geese, obtained by polymerase chain reaction (PCR) amplification and splicing, contained two exons and one intron. The coding sequence was 738 bp long and included codes for 245 amino acids (GenBank Accession No. EU370686). The coding sequence and the corresponding protein of goose adiponectin shared the highest homology with ducks, 94.85% and 95.51%, respectively; homology with chicken adiponectin was next highest, while the lowest homology was to be found with mammals. The predicted molecular weight (MW) and isoelectric point (pI) were 26,603.3 Da and pH 5.19, respectively, which were highly similar to those of species considered above. Reverse transcription-PCR revealed that goose adiponectin mRNA was highly expressed in skeletal muscle, fatty tissue, heart and stomach muscles. Medium levels of expression were detected in the small intestine, stomach glandular tissue, kidney and lung, while it was only weakly expressed in the liver, spleen, ovaries and diencephalon.

Amplified fragment length polymorphism (AFLP) markers linked to the stem nematode resistance gene were developed in sweet potato (Ipomoea batatas (L.) Lam.). Using bulked segregant analysis (BSA), 800 AFLP primer combinations were screened in the resistant and susceptible bulked DNA from the 186 progeny of an F1 single-cross population of Xu781 (resistant parent)×Xushu18 (susceptible parent), and 245 of these AFLP primers showed polymorphic bands between resistant and susceptible DNA. Primer combinations detecting polymorphism between the two bulks were used to screen the parents and eight individuals from each of the bulks. The results showed that E2M23 and E33M20 produced a specific band of about 500 bp and 200 bp in length, respectively, in the resistant plants but not in the susceptible plants, suggesting that the markers named E2M23500 and E33M20200 linked to a gene for stem nematode resistance. Amplified analysis of the 186 F1 individuals indicated that the genetic distance between these two markers and the stem nematode resistance gene was 6.9 cM and 11.1 cM, respectively, measured with Mapmaker 3.0. These two AFLP markers were used to identify ten sweet potato varieties planted widely in China and the results were consistent with those of conventional resistance identification, indicating that the two markers can be used in molecular marker-assisted breeding for stem nematode resistance in the sweet potato.

N6 medium, Murashige and Skoog (MS) medium and BNBN medium were adopted in tissue cultures of Nipponbare (japonica) scutellum for performance comparison. Results indicated that, among the three kinds of culture media, the NB medium worked best in callus induction and gene transformation and was most suitable for tissue culture of Nipponbare scutellum. Based on this, the elicitor-encoding gene pemG1 from Magnaporthe grisea was introduced into the genome of Nipponbare via the Agrobacterium-mediated method. Transgenic rice plants were obtained. The integration, transcription and expression of pemG1 in rice were confirmed by polymerase chain reaction (PCR), Northern blot and Western blot, respectively. Genetic analysis showed that the transgenes were segregated normally in the progenies.

Based on conserved homologous amino-acid sequences of the Gq protein α subunit in arthropods, a pair of degenerate primers were designed to amplify the gene from the English grain aphid (Sitobion avenae), using reverse transcriptase polymerase chain reaction (RT-PCR) and (3′/5′)-rapid amplification of cDNA ends (3′/5′ RACE) techniques. A Gqα protein was obtained from the alate adult aphids. The open reading-frame was 1062 bp, encoding 352 amino-acid residues with a calculated molecular weight of 40.8 kDa. The cDNA sequence was deposited in GenBank with accession no. EF638906. The deduced amino-acid sequence of Gqα shared a high identity (≥82.17%) with reported Gqα from other insects and even vertebrates, and had the typical characteristics of Gqα protein. In order to explore the function of the Gqα gene, a eukaryotic expressional system (baculovirus expression vector system, BEVS) was constructed by TOPO and Gateway techniques. After the recombinant reaction occurring between pUC-Gqα and the Gateway-adapted baculovirus DNA from Autographa californica nuclear polyhedrosis virus (AcMNPV), the construct recombinant viruses containing V5-His6Gqα were transfected singly into the insect cell line of Tn-5B1-4. After collecting the infected cell, detection was conducted by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting. The result showed that the system comprising recombinant baculovirus and Tn could express Gqα protein successfully.

Seventeen special loci were selected from 77 microsatellite markers to distinguish three varieties of tilapias, including the six differential loci UNH636, UNH117, UNH172, UNH738, UNH878 and UNH896 in Oreochromis aureus; five differential loci UNH913, UNH907, UNH222, UNH980 and UNH880 in O. niloticus; and six differential loci of UNH876, UNH899, UNH853, UNH932, UNH933 and UNH773 in O. mossambicus. Any one of the 17 loci could amplify particular bands to distinguish one tilapia from the other two. The genetic structure of O. aureus, O. niloticus and O. mossambicus stocks and their phylogenetic relationships were also analysed using these 17 loci. In total 142 alleles were detected, and the average number of alleles per locus was 8.35. Additionally, a clustering analysis was performed based on the result of the Popgen32 software package and phylogenetic trees were constructed by MEGA4 using the unweighted pair group method using arithmetic average (UPGMA). The results showed that the mean value of observed heterozygosity was 0.0941, 0.5490 and 0.2588, the mean value of expected heterozygosity was 0.1089, 0.7230 and 0.1965, and the polymorphism information content was 0.0869, 0.7149 and 0.1643, in O. aureus, O. niloticus and O. mossambicus, respectively. The UPGMA tree demonstrated that O. aureus was more closely related to O. mossambicus than to O. niloticus.

A pair of degenerate primers was designed based on a conserved domain of GA 20-oxidase reported in other plants. The full-length (1179 bp) carnation (Dianthus caryophyllus L. cv. Master) GA 20-oxidase cDNA (named Dc20ox) was cloned by reverse transcriptase-polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). BLAST analysis revealed that the deduced amino acid sequence had high homology (66–75%) with the GA 20-oxidase sequences from other plants. An RNAi vector (pART400) was constructed from a 400 bp fragment representing a highly conserved region of GA 20-oxidase.

Bacillus cereus can produce α-amylase, which has important industrial production value and can endure high temperatures. The enzymatic characteristics of two α-amylases from B. cereus (B905 and B904) were studied. The results showed that both preserved their activity at 90–100°C. Their thermal stability and enzymatic activity did not depend on Ca2+. However, their protein molecular weights were obviously different. The structural genes of amy905 and amy904 were cloned successfully using the polymerase chain reaction (PCR) and expressed in Escherichia coli. The amy904 and amy905 genes were 1362 bp and 1761 bp long and their protein molecular weights were about 55 kDa and 68 kDa, respectively.

With the rapid development of nanotechnology, the security of nanomaterials has been an increasing cause for concern. In this study, the impact of titanium dioxide nanoparticles (nano-TiO2) on the phyllosphere bacterial community were analysed by both a culturable-dependent method and a polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) method. The quantity of culturable phyllosphere bacteria was significantly reduced with an increased concentration of nano-TiO2. With increasing concentrations from 0.002 to 20 mg/ml of nano-TiO2, the quantity of culturable phyllosphere bacteria decreased from 1.8×106 to 3.1×105 cfu/g. The phyllosphere bacteria community was analysed by PCR-DGGE, and when the concentrations of nano-TiO2 were higher than 0.02 mg/ml, the DGGE bands were significantly lower than in the control. Sequencing results of the bands from the DGGE gel showed that there were at least seven genera in the phyllosphere bacteria. Only one uncultured bacterium was unaffected by the concentration of nano-TiO2.

The spatio-temporal expressed profiles of two kinds of chemical communication-related protein genes, the odorant-binding protein of Ac-ASP2 and chemosensory protein of Ac-ASP3, were identified by real-time polymerase chain reaction (PCR). Results obtained using the 2−ΔΔCt method showed that Ac-ASP2 was a gene coding antenna-specific protein that did not express in larvae and pupae, but had discontinuous high abundance periods at 1, 9, 15, 27 and 30 days. The expressing abundance at such periods was at least ten times higher than that at other periods. From the distribution of Ac-ASP3 mRNA observed in different tissues, the transcript levels seemed to be higher in the wings, abdomen and thorax (of order ~106), and lower in the legs, antennae and head (of order ~105). From highest to lowest, the original copy number was found in the various body parts in the following order: wings, abdomen, thorax, legs, antenna, and head. The results suggest that Ac-ASP3 has an intimate relation with the chemosensory behaviour of wings and abdomen in Apis cerana cerana.

The DagA gene and DagA(▽), which is a DagA gene encoding sequence without signal peptide, were cloned from genome DNA of Pseudoalteromonas atlantica 19262 by polymerase chain reaction (PCR). After ligation with pET21 vector, DagA and DagA(▽) were respectively expressed in Escherichia coli ER2566 using molecular chaperones DsbC and FkpA. A strain of ER2566-pET21a-DagA(▽)-DsbC was screened as a highly effective expressing system in the form of an inclusion body that had the target protein with up to 60% total bacterial protein. DagA protein was renatured and purified by dissolving it in 8 mol/l of urea, using Ni-NTA resin affinity chromatography and refolding using the urea gradient method. DagA with a molecular weight of ~30.8 kDa was identified by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and had the ability to digest agarose. In a pH range of 4.8–6.8, DagA maintained a bioactivity greater than 60%, with 5.8 being the optimum pH, and it exhibited activity at temperatures from 37°C to 60°C, with 55°C being the optimum temperature.

Simple sequence repeats (SSR) and random amplification of polymorphic DNA (RAPD) molecular markers were used to assess the genetic diversity of 80 isolates of Phytophthora infestans in potato (Solanum tuberosum) from Fujian, Heilongjiang, Hebei and Inner Mongolia Provinces in China. Polymorphism was identified by 13 SSR primers and 14 RAPD primers in the isolates of P. infestans in potato. A total of 76 bands were amplified by SSRs, with the percentage of polymorphic bands (PPB) being 78.9% and the similarity coefficient ranging between 0.00 and 0.42. A total of 189 bands were amplified by RAPDs, with the percentage of polymorphic bands being 95.2% and the similarity coefficient ranging between 0.04 and 0.66. Analysis of genetic diversity showed that there exists higher genetic variation in the Fujian population in comparison to the populations of Heilongjiang, Hebei and Inner Mongolia. Nei's genetic identity analysis indicates that the genetic similarity between populations of Heilongjiang and Inner Mongolia is the highest and that between Fujian and Hebei is the lowest. A cluster analysis revealed that isolates from Fujian, in the south of China, are distantly related to those from Heilongjiang, Hebei and Inner Mongolia in the north, and the Fujian population is distributed among more groups than the other three, exhibiting a higher genetic diversity.

BalB/C neonate mice and adult BalB/C mice were vaccinated using BCG (Bacille Calmette–Guerin). The pathogenic growth characteristics of BCG in in vitro culture on spleen cells (SPC) were observed and changes in induced expression of IFN (interferon)-γ and IL (interleukin)-4 in SPC were detected using the ELISPOT assay. The results showed that a low dose of BCG (2×103 cfu) exerted 100% immunoprotection on 7-day-old neonate mice and a high dose of BCG (4×104 cfu) exerted 75% immunoprotection. A low dose of BCG (2×103 cfu) exerted 67% immunoprotection on 35-day-old mice. It is also shown that Th1-type cell immunity dominated by IFN-γ was enhanced significantly in the neonate mice injected with a low dose of BCG (2×103 cfu), and Th2-type cell immunity dominated by IL-4 was depressed at the same time. IFN-γ and IL-4 induced by a high dosage of BCG (4×104 cfu) in neonate mice were both increased. IFN-γ and IL-4 induced by a low dose of BCG (2×103 cfu) in 35-day-old mice were also increased. The results indicate that there exists a marked correlation between immunoprotection by BCG in mice and both the immunizing dose and age of the immune animals, which might be relevant to the changes induced by BCG on Th1- and Th2-type cell immunity.

Six transgenic maize lines with the fully modified gene GFM Cry1A were obtained and polymerase chain reaction (PCR) analysis of their T5 generation plants indicated that foreign genes could be stably inherited. Three hybrids (Simi25Bt, Tongdan24Bt and Jidan209Bt) were created using the transgenic lines and combined with regular maize inbred lines. The results of resistance identification of the transgenic inbred lines and hybrids showed that the difference of resistance among transgenic lines was very significant; there also existed a difference among individuals of the same line and among the three hybrids. Compared with the control inbred line (CK), the average of four transgenic inbred lines in the number of tunnels per stalk, larval tunnel length per plant and length of surviving larvae were decreased by 70%, 80% and 70%, respectively. Enzyme-linked immunosorbent assay (ELISA) testing showed that the average of Bt Cry1A protein expression level for six transgenic inbred lines was 0.16% total protein and the highest expression level was 0.19%. At maturity, compared with the control variety (CK), the larval tunnel length per plant of the three hybrids (Simi25Bt, Tongdan24Bt and Jidan209Bt) was decreased by 39.97%, 36.20% and 53.83%, respectively, which was a decrease of 43.36% on average. The investigation of agronomic characters showed that there was no significant difference between the improved hybrids and the control in plant height, ear length, row number per ear, kernel number per row and 100-kernels weight. It is thought that the GFM Cry1A gene can be applied to improve resistance to corn-borer, and maize inbred lines with the Bt gene can be directly applied to conventional maize breeding.