Segregation distortion, identified as a strong evolutionary force, is common in genetic mapping research. In plants, the percentage, degree, origin and genetic effects of segregation distortion vary significantly with species, population types, crosses and marker types. The exhibition, common features, causes, methods of mapping segregation distortion loci, effects of segregation distortion on map construction and corresponding mapping strategies in plants, mainly in crops, are reviewed in this paper.

Random amplified polymorphic DNA (RAPD) analyses were performed with random primer H16 for the B biotype and two non-B populations of Bemisia tabaci collected from Zhejiang (China). The specific sequence fragments containing 446, 390 and 1317 nucleotides were amplified for the B biotype, ZHJ-1, ZHJ-2 populations, respectively. The three specific fragments were cloned and sequenced, and three pairs of SCAR primers were designed according to the sequences determined. With improvement of the conditions of the polymerase chain reaction (PCR), the specific fragments of B biotype, ZHJ-1 and ZHJ-2 populations, namely 439, 366 and 1238 nucleotides, respectively, were amplified with the sequence characterized amplified region (SCAR) primer of the corresponding population, while specific fragments of the other populations of B. tabaci or Trialeurodes vaporariorum could not be amplified.

Genetic diversity in 122 accessions of pummelo (Citrus grandis Osbeck) and its related varieties was assessed using simple sequence repeat (SSR) markers. Thirty-one pairs of SSR informative primers generated a total of 335 alleles. The average number of alleles per locus was 9.85. The value of allelic polymorphism information content (PIC) ranged from 0.1939 to 0.9073, with an average of 0.7085 per primer. The 122 accessions of pummelo and its related varieties could be clustered into seven groups by the unweighted pair-group method arithmetic average (UPGMA), in which the 110 pummelo accessions could be divided into 18 subgroups at similarity coefficient of 0.712. These subgroups were mainly composed of the Shatian pummelo variety group, the Wendan variety group and many of the hybrid pummelo groups. The classification method can be used in targeting many varieties in order to widen the genetic background of pummelo.

In plants, acid invertases are known to be the key enzymes cleaving sucrose into reducing sugars (RS) (glucose and fructose). To improve the quality of potato (Solanum tuberosum L.) chips, which is largely influenced by RS accumulation in tubers stored at low temperature, a part of acid invertase cDNA with hairpin RNA (hpRNA) structure was transformed into potato cv. N2. Detection of polymerase chain reaction (PCR) amplification and Northern blotting suggested that the RNA interference (RNAi) vector was successfully transformed into cv. N2. The analysis of acid invertase activity in the plantlets and microtubers of RNAi transgenic lines indicated that the expression of the acid invertase was significantly repressed by the activity of RNAi of plantlets by an average 69.8% (with the exception of line Ni-1) with a maximal decrease of 78% (line Ni-4), and the highest decrease of activity in microtubers of 68%. Compared with that of well-inhibited antisense inv transgenic plants, the comparative downregulation of RNAi suggests a distinct alteration of endogenous acid invertase activity and a potential strategy for post-transcriptional gene silencing (PTGS) in modulation of cold-sweetening in potato.

An efficient AHL (N-acyl-homoserine lactone) bioassay strain, JZA1, of Agrobacterium tumefaciens was used to detect the AHL production from Acidovorax avenae subsp. citrulli [the pathogen causing bacterial fruit blotch (BFB) of melons], and the results showed that A. avenae subsp. citrulli produced a 3-O-C8-homoserine (HSL) type signal molecule. Gene aiiA, which could degrade AHL molecules, was transformed into A. avenae subsp. citrulli strain NJF10, creating strain NJF10-aiiA. The AHL production from NJF10-aiiA was significantly reduced compared with wild-type NJF10. Inoculation tests showed that NJF10-aiiA had an obvious reduction of virulence on watermelon fruits. Our finds showed that AHL production by A. avenae subsp. citrulli was related to its pathogenicity. This work might provide a novel way to control BFB by QS (quorum sensing) interference.

Pseudomonas fluorescens P5 was isolated in Yunnan Province and proved to be an effective biocontrol agent of various soil-borne plant diseases. The aim of the study was to enhance the biocontrol effect of this strain by introducing a chitinase gene. A 6.5 kb DNA fragment containing the chitinase gene chiB was inserted into pDSK519 vector to construct a new plasmid, pDSK51965. This plasmid was introduced into P. fluorescens P5 and resulted in a new strain named P5-1. Restriction enzyme digestion and chitin plate culture confirmed that P5-1 contained a functional chitinase gene. Compared with wild-type P5 in pot experiments, P5-1 had an increased effect against rice sheath blight and cotton damping-off caused by Rhizoctonia solani. It also increased the effect on suppression of wheat take-all caused by Gaeumannomyces graminis var. tritici.

A rapid and accurate real-time quantitative polymerase chain reaction (real-time PCR) method with SYBR Green I was established for detecting Escherichia coli O157:H7. A pair of primers were designed to amplify the eae gene. The dissociation curves showed that the amplification product was very specific. The optimal conditions and standard curve were established. The result indicated that real-time PCR was 1000 times more sensitive than ordinary PCR.

An immuno-capture polymerase chain reaction (IC-PCR) method for detection of Acidovorax avenae subsp. citrulli (AAC), the causal organism of bacterial fruit blotch (BFB) of watermelon, was developed by combining the immunosorbent enrichment (ISE) method with classical PCR and comparing with the direct PCR and growth check methods. The results showed that all A. avenae subsp. citrulli strains tested have produced 360 bp specific fragments using IC-PCR and direct PCR methods, while other strains from 10 different genera showed negative PCR results. The minimum detection concentration was about 50–100 cfu/ml and 104 cfu/ml, respectively. The IC-PCR sensitivity was 100 times higher than that of direct PCR. The examination of seven batches of different melon seeds from the markets by IC-PCR showed that one cantaloupe, two honeydew melon and two watermelon seed varieties carried the pathogen, indicating that the IC-PCR is an accurate, sensitive, rapid and low-cost technique.

Six DNA components of Banana bunchy top virus (BBTV) were amplified by polymerase chain reaction (PCR) using total DNA from banana tissues having the typical BBTV symptoms as template. The sequence analysis indicated that the six fragments (1104 nt, 1067 nt, 1059 nt, 1045 nt, 1014 nt and 1081 nt) were all full-length components of BBTV. The viral sequences from different countries were compared. Results revealed that the sequence of component 1 was more conservative than other components, DNA1 of the Hainan isolate shared 95.4% homology with that of the Vietnam isolate, while sequences of both components 2 and 4 showed less homology. DNA2 of the Hainan isolate only shared 83% homology with that of the Guangzhou isolate. According to the results of the DNA1 sequence comparison, the BBTV Hainan isolate should belong to the Asian sub-group.

A total of 791 microsatellites (SSRs) were isolated from 7055 Panax ginseng expressed sequence tags (ESTs). According to primer design criteria, 68 primer pairs for EST-SSR were designed. Under an appropriate polymerase chain reaction (PCR) system, all EST-SSR primer pairs were screened against genomic DNA of Ji'anchangbo and Fusong'ermaya from Panax ginseng, and 43 EST-SSR primer pairs out of the above 68 resulted in PCR products. Then, all 43 pairs were detected in nine P. ginseng, two Panax quinquefolius and two Acanthopanax senticosus cultivars for polymorphisms, and 26 pairs (60.47%) were found to be polymorphic, accounting for 38.23% of the total number of designed primer pairs. These results demonstrate the possibility of developing EST-SSR markers using P. ginseng ESTs.

In vitro bulb scales of Lilium longiflorum×L. formosanum were used as explants to develop a highly efficient regeneration system. A high regeneration rate (100%) was reached through organogenesis on basal Murashige and Skoog (MS) medium supplemented with 1.0 mg/l 6-benzylaminopurine (6-BA) and 1.0 mg/l naphthaleneacetic acid (NAA). A genetic transformation system for the lily was developed using an Agrobacterium tumefaciens-mediated method. An improved genetic transformation rate (12‰) was obtained when the explants were pre-cultured for 3 days, immersed in bacterial suspension (OD600≈0.8) for 5 min, and co-cultivated for 5 days. The binary vector pBI121 containing Zm401, a maize pollen-specific gene, was introduced into the Agrobacterium strain LBA4404 and transformed into the explants using the genetic transformation system. Gene integration into the lily genome was confirmed by polymerase chain reaction (PCR) and PCR–Southern analysis. These results could lead to the production of new pollenless lily plants.

The microsatellite, or short sequence repeat (SSR), is a powerful genetic marker, useful in many areas of fish genetics and breeding. Polymorphic microsatellite loci have been frequently applied to the analysis of genetic diversity, population genetic structure, and genomic mapping. These co-dominant markers have also been applied to the classification and systematics, parentage identification, germplasm conservation, and breeding programme of food fish.

Bacillus thuringiensis strain WZ-9, isolated from soil in Hebei province, China, was effective against Henosepilachna vigintioctomaculata larvae. The strain presented bipyramidal crystals with a protein band of 130 kDa in SDS–PAGE. The pH changes of the culture media showed important fluctuations during the 24 h growth cycle. The pH varied less in log and stationary phases than it did in the exponential phase. Bioassay results showed that the WZ-9 strain was only harmful to larvae of H. vigintioctomaculata and not to either adults of H. vigintioctomaculata or other several lepidopteran and coleopteran insects. LC50 to second-instar larvae of H. vigintioctomaculata was 2.95×107 cells/ml after 72 h. Genotypic investigations showed that this strain possessed the cry7 gene. Sequence analysis demonstrated that the encoding gene contained an open reading frame (ORF) of 3414 bp and encoded 1138 amino acid residues. The deduced amino acid sequence was 99.65% identical to that of the reported Cry7Ab2 sequences. This gene was designated by the Bt δ-endotoxin nomenclature committee as Cry7Ab3 with accession number BI 1015188 in the GenBank database.

The techniques used to detect genetically modified organisms (GMO), including qualitative polymerase chain reaction (PCR), quantitative PCR, enzyme-linked immunosorbent assay (ELISA) and many others, are systematically described and discussed. The application progress of GMO in species-specific detection, endogenous genes, standard substances and restraining factors influencing detection are reviewed. The ongoing problems and development prospects of detection techniques of GMO are also pointed out.

A total of six amplified fragment length polymorphism (AFLP) primer combinations were used to detect genetic variation of pooled DNA in a sample of 12 chicken breeds indigenous to China, and AFLP DNA fingerprinting of each chicken breed was constructed. Polymorphic bands, specific bands and genetic similarity coefficients of 12 chicken breeds were calculated from AFLP data. A total of 279 polymorphic bands were generated by the six primer combinations, giving, on average, 46.5 polymorphic markers detected per primer combination. Nine specific bands were produced in the pooled DNA of Jiuyuan black and Dongxiang black chickens. However, one specific band was produced in the pooled DNA of Wenchang and Xingyi bantam chickens. An unweighted-pair-group method using average linkages (UPGMA) cluster analysis revealed that the 12 chicken breeds could be divided into three groups. Genetic similarity coefficients and the UPGMA tree of the 12 chicken breeds were consistent with their breeding history as well as their geographical distribution. Based on AFLP DNA fingerprinting, genetic diversity, genetic relationship and identification of chicken breeds can be analysed.

The development of near-isogenic lines (NILs) is an important fundamental step in the cloning of quantitative trait loci (QTL) and molecular marker assisted breeding. In the present study, NILs for two major QTLs of basal root thickness (BRT) and 1000-grain-weight (TGW) were obtained by molecular marker-assisted selection (MAS) through foreground selection for target QTL and background selection in three backcross generations (BC1F1, BC2F1 and BC3F1). After phenotypic evaluation in the BC3F2 generation, nine BRT QTL-NILs with BRT ranges of 1.07–1.16 mm (6.11–15.18% greater than the recurrent parent) and an average recovery ratio of genetic background (RRGB) of 97.22%, and 11 TGW QTL-NILs with ranges of 21.25–26.25 g (7.05–32.16% greater than the recurrent parent) and 95.97% of RRGB, were selected.

A bioassay method was developed to use the parasporal crystal protein of Bacillus thuringiensis against plant-parasitic nematodes. Using this method, the parasporal crystal proteins of ten Bt strains showed activity against plant-parasitic nematodes. The toxicity of YBT-021 against Meloidogyne hapla, Pratylenchus scribneri, Tylenchorhynchus sp., Ditylenchus destructor and Aphelenchoides sp. was also assayed. The resulting LC50 values were 35.62 μg/ml, 75.65 μg/ml, 94.31 μg/ml, 215.21 μg/ml and 128.76 μg/ml, respectively.

The population doubling number (70–80 times) of human fetal bone marrow mesenchymal stem cells (BMMSCs) is about two times higher than that (30–40 times) of adult BMMSCs, and their differentiation capacity is superior to that of their adult counterparts. In this study, BMMSCs were isolated from long bones of 2- to 3-month-old human abortuses by rinsing and selectively culturing whole marrow cells. Basic medium and serum concentration were optimized and growth curves plotted, both by MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-di-phenyltetrazolium bromide] reduction assay. Isolated cells were identified by flow cytometry and immunocytochemistry for their antigen markers. The biosafety of isolated cells was evaluated by karyotype analysis and a tumour-forming experiment. The results indicated that lengthwise scissoring of fetal long bones and rinsing of their marrow cells was practical and useful for recovery of BMMSCs from the investigated human abortuses. In this experiment, α-MEM (minimum essential medium, alpha medium)+20% FCS (fetal calf serum) was the best for in vitro culture of BMMSCs. The third-passage BMMSCs expressed Oct4, SSEA3 and SSEA4 besides the surface markers of their adult counterparts. The population doubling time of the BMMSCs of passage 6, 12 and 24 were 34, 36 and 40 h, respectively. Cells in all passages showed a diploid karyotype and formed no tumours in nude mice. The BMMSCs used in this study proved to be biologically safe and ideal seed cells for research on human tissue engineering and regeneration medicine.

Primary culture of goat mammary gland cells was achieved by outgrowth of migrating cells from fragments of tissue. The fibroblast and epithelial cells were purified according to their different sensitivity to trypsin and the characteristics of goat mammary epithelial cells were observed under a light microscope. The results showed that the purified goat mammary epithelial cells retained normal characteristics until the 15th passage. The purified mammary epithelial cells propagated and formed a dome-like structure that resembled a nipple and was called a ‘milk orb’. The mammary epithelial cells could produce and secrete milk. There were different cell types: the majority were short shuttle-like or polygons, resembling a beehive; while some were large, flat and round; and others were elongated. The enhanced green fluorescent protein (EGFP) gene was transferred successfully into the goat mammary epithelial cells using electrotransfection, and its expression was observed under a fluoroscope.

With the development of plant genetic engineering, many transformation methods can be used to transfer heterogeneous genes into plants to develop genetic crops. However, a lot of research results have shown that transgene expression remains largely unpredictable and there is great variation of expression level in different transgenic plant lines. Plant genetic engineering research is reviewed in the present paper. We analysed the reasons why low efficiency of heterogeneous gene expression has happened in transgenic plants in terms of DNA modification, localization of proteins and methods of transformation used. Some strategies for improving heterogeneous gene expression in transgenic plants are also discussed.