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Combined
overexpression of
chitinase and defensin genesin transgenic tomato
enhances resistance to Botrytis cinerea
Chen, S. C.1
*, Liu, A. R.1, Wang, F. H.1 and
Ahammed, G. J.2,3
1College
of Forestry, Henan University of Science and Technology,
Luoyang, Henan 471003,
China.
2College
of Horticulture, Bangladesh Agricultural University,
Mymensingh 2200, Bangladesh.
3Department
of Horticulture, Zhejiang University, Hangzhou, 310029, P.R.
China.
*Corresponding author. E-mail:
chen_shuangchen@126.com.
Tel:
+86-379-64361682
Accepted
3 September, 2009 |
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The rice chitinase gene (CHI), the alfalfa defensin
gene (alfAFP) and their bivalent gene (CHI-AFP)
were introduced into tomato line Micro-Tom via
Agrobacterium-mediated gene transfer method.
Transformants were obtained and confirmed by GFP, PCR and
Southern blot hybridization. One to four copies of transgene
were integrated into the tomato nuclear genome.
Transcription levels of chitinase, alfAFP and their
bivalent gene CHI-AFP in various transgenic lines
were determined using Northern blot and Western blot
analysis. Performance test of resistance analyses to
Botrytis cinerea with T1 generation
transgenic tomato lines showed the transgenic lines
exhibited higher resistance to the pathogens infected than
that of the non-transgenic plants and the resistance levels
were related to expression levels of the transgene, showing
dosage-effect. The transgenic tomato harboring CHI-AFP
cassette showed the highest disease resistance; it suggested
that co-transformation with alfAFP and chitinase gene
was more effective than individual transformations on the
resistance to B. cinerea. Some independent lines with
high disease resistance, low variability and stable
expression of transgenes could be selected for the further
studies and molecular breeding.
Key
words:
Transgenic tomato, rice chitinase gene, alfalfa defensin
gene, Botrytis cinerea. |
