Hu Zanmin

Multiple transgenes Populus xeuramericana ‘Guariento’ plants obtained by biolistic bombardment

A JERF36 regulation gene, a selection marker gene (NPT-II), and the foreign genes levansucrase (SacB), Vitreoscilla hemoglobin (vgb), and Binary coleopterus insect resistance (BtCry3A+OC-I) were co-transferred into Populus xeuramericana ‘Guariento’ using biolistic bombardment; 25 kanamycin resistant plants were obtained. The results of PCR and Southern hybridization showed that the foreign genes had been integrated into the genome of P. xeuramericana ‘Guariento’ and 5 genes were all transferred into 7 poplar plants. The results of a BtCry3A ELISA experiment indicated that the BtCry3A gene was expressed in the 7 transgenic popular plants, and these plants grew well on coastal saline land.

Chromosome microdissection by laser microbeam, chromosomal fragment isolation and amplificationin vitro in barley (Hordeum vulgare L.)

A method for microdissection, isolation and amplification of plant chromosomal fragments using laser microbeam and a glass microneedle was established. Firstly, 7H chromosome of barley (Hordeum vulgare L.) was dissected by Nd: YAG laserbeam with suitable parameters and the fragment comprising a satellite was isolated with a glass microneedle which was fixed on a micromanipulator. Then, the chromosomal fragment DNA was amplified by LA-PCR (linker adaptor PCR) for two rounds. The size of the DNA fragments of PCR products varied from 500–3 000 bp and the PCR products originated from the genome of barley were verified by Southern hybridization. Compared with previous reports, there are some advantages in this research. The performance is easier. the dissection is more precise and the cost is low. It also permits efficient amplification with only one single chromasome fragment. Laser microbeam-glass microneedle method may be useful in the microdissection of special chromosome regions, especially in plants with middle or small chromosomes.