Cloning CPI from Camellia azalea and Overexpressing to Tobacco Enhances Insect Tolerance

doi:10.7668/hbnxb.2015.05.010

Abstract

Abstract: The majority of ornamental camellias are cultivated plants in greenhouse,but pests and diseases are likely to occur in hot-damp environment,which leads to detrimental effects on ornamental and economic value.On the basic of homologous sequences of Camellia japonica,a Cysteine proteinase inhibitor(CPI)gene was isolated from the tender leaf in C.azalea by the 3',5'-RACE technology named CaCPI to improve the resistance of plants to the insect pest.The full-length cDNA of CaCPI was 579 bp,containing a 306 bp ORF which encodes 101 amino acids.The relative molecular weight of CaCPI protein was 11.078 kDa,and its isoelectric point(pI) was 6.72.Expressions of CaCPI in root,stem and leaf of camellias were analyzed by fluorescent quantitative Real-time PCR.The results showed that the expression level of CaCPI in leaf was the highest,medium in stem and the lowest in root.It was also found that compared with stem and root,the expression level in leaf was about 1.53-fold and 1.61-fold,respectively.Furthermore,the expression levels of CaCPI in transgenic plants increased up to 55.84-174.83 times as high as wild type tobacco plants.In addition,overexpression of CaCPI enhanced insect tolerance in transgenic plants.Two genotypes plants were inoculated with aphids after 5 days indicated that overexpression of CaCPI gene significantly increased insect resistant to Homoptera Myzus persicae,cumulative mortality rate of aphids in transgenic tobacco plants was up to 90.75% and about 5.14 fold compared with wild type plants.

Key words: Camellia azalea, Gene clone, Insect tolerance, Expression analysis, Transgenic tobacco

CLC Number:

Q78
S685.14

WANG Jiang-ying, FAN Zheng-qi, YIN Heng-fu, WU Bin, LIU Wei-xin, LI Ji-yuan. Cloning CPI from Camellia azalea and Overexpressing to Tobacco Enhances Insect Tolerance[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(5): 57-64. doi: 10.7668/hbnxb.2015.05.010.

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