Tian-Zhen Zhang

Publications (2)5.54 Total impact

• Article: Agrobacterium-mediated co-transformation of multiple genes in upland cotton

  Fei-Fei Li, Shen-Jie Wu, Tian-Zi Chen, Jie Zhang, Hai-Hai Wang, Wang-Zhen Guo, Tian-Zhen Zhang
  [show abstract] [hide abstract]
  ABSTRACT: Two cotton genotypes, Simian 3 (SM 3) and WC, were co-transformed using a mixture of four Agrobacterium tumefaciens cultures of strain LBA4404, each carrying a plasmid harboring the following genes, Bt+sck (for Bacillus thuringenesis protein and modified Cowpea trypsin inhibitor), bar (for glufosinate), keratin, and fibroin. The frequency of callus induction, embryogenesis, and plant regeneration were notably different between the two genotypes. However, there were no differences between the two genotypes for number of plantlets carrying multiple gene copies of different gene combinations as well as transformation frequency for different gene combinations. PCR analysis indicated that more than 80% of plantlets carried the nptII gene for kanamycin resistance. Overall, the co-transformation frequency of two or more genes was about 35%. Southern blot analysis confirmed integration of target genes into the cotton genome, and the number of copies of the transgene(s) varied from one to four. Multiple transgene expression was confirmed by RT-PCR analysis in some transgenic lines. Further analysis of T1 plants demonstrated that multiple transgenes were inherited and expressed in progenies.
  Plant Cell Tissue and Organ Culture 04/2012; 97(3):225-235. · 3.09 Impact Factor
• Article: Enhanced Agrobacterium-mediated Transformation of Embryogenic Calli of Upland Cotton via Efficient Selection and Timely Subculture of Somatic Embryos

  Shen-Jie Wu, Hai-Hai Wang, Fei-Fei Li, Tian-Zi Chen, Jie Zhang, Yan-Jie Jiang, Yezhang Ding, Wang-Zhen Guo, Tian-Zhen Zhang
  [show abstract] [hide abstract]
  ABSTRACT: Agrobacterium-tumefaciens-mediated transformation of cotton embryogenic calli (EC) was enhanced by choosing appropriate EC and improving efficiency of coculture, selection cultivation, and plant regeneration. After 48-h cocultivation, the number of β-glucuronidase (GUS)-positive calli characterized by yellow, loose, and fine-grained EC was twofold greater than that of gray, brown, and coarse-granule EC. It indicated that efficiency of transient transformation was affected by EC morphology. And transient transformation efficiency was also improved by cocultivation on the medium adding 50mg l−1 acetosyringone at 19°C for 48h. Subculturing EC on the selection medium with low cell density was beneficial to production of more kanamycin-resistant (Km-R) calli lines. From an original 0.3-g EC, an average of 20 Km-R calli lines were obtained from a selection dish and the GUS-positive rate of Km-R clones was 81.97%. A large number of normal plants were rapidly regenerated on the differentiation medium with dehydration treatments and the GUS-positive rate of regeneration plants was about 72.60%. Polymerase chain reaction analysis of GUS-positive plantlets revealed a 100% positive detection rate for neomycin phosphotransferase II gene and uidA. Southern blot of transgenic plants regenerated from different Km-R calli lines demonstrated that the target gene, mostly with the low copy number, has been integrated into the cotton genome.
  Plant Molecular Biology Reporter 04/2012; 26(3):174-185. · 2.45 Impact Factor