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Bulletin of Botanical Research ›› 2003, Vol. 23 ›› Issue (2): 198-203.

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THE STUDY ON AGROBACTERIUM RHIZOGENES MEDIATED CHITINASE AND β-1, 3-GLUCANASE TRANSFORMATION FOR TOBACCO

ZHANG Yan-Ni1, QU Min2, XU Xiang-Lang3, LI Xin-Ling3   

  1. 1. College of Landscape, Northeast Forest University. Harbin 150040;
    2. Tourism School of Heilongjiang Province, Harbin 150086;
    3. Department of Biology, Harbin Normai University, Harbin 150080
  • Received:2002-12-30 Online:2003-06-15 Published:2016-06-14

Abstract: The plasmid pBLGC which constained chitinase gene and β-1, 3-glucanase gene and NTP-II gene (resistansing kanamycin) was transformed into Agrobacterium rhizogenes with root-in-ducing plasmid by using freeze-thaw method and triparental crossing method. Chitinase gene and β-1, 3-glucanase gene were placed under the control of the Cauliflower MosaicVirus 35S promoter and introduced into three varieties of tobacco by Agrobacterium rhizogenes mediated leaf-plated trans-formation method. 126 regenerated plants were selected by Kanamycin. Among them, 119 plants have been tested by 35S, chitinase gene and β-1, 3-glucanase gene primer separatly through PCR elec-trophoresis assay, of which 72 plants showed chitinase gene positive, 47β-1, 3-glucanase gene postive and 36 postive plants containing both chitinase gene and β-1, 3-glucanase gene. The trans-ferring frequency of foreign gene has relation with plant variety and the length of foreign gene. Chitinase gene seemed to inherit easier than β-1, 3-glucanase gene. 24 plants among PCR positive plants have been carried out by PCR-Southern. The result was that they showed positive. The study indicated chitinase gene and β-1, 3-glucanase gene respectively or together introducing and cointe-grating into plant genenome.

Key words: Agrobacterium rhizogenes, tobacco, leaf-mediated method, chitinase gene, β-1, 3-glucanase gene