期刊
GM CROPS & FOOD-BIOTECHNOLOGY IN AGRICULTURE AND THE FOOD CHAIN
卷 12, 期 1, 页码 382-395出版社
TAYLOR & FRANCIS AS
DOI: 10.1080/21645698.2021.1944013
关键词
Cotton; transformation; eagle-2; agrobacterium; pectin methyl esterase enzyme; insecticidal cry proteins
资金
- National Key R&D Program of China [2017YFD0101603-11, 2016YFD0100500]
- Natural Science Foundation of China [31471538, 31371668]
- Agricultural Science and Technology Innovation Program for CAAS (CAAS-ASTIP-ICRCAAS)
- National High Technology Research and Development Program of China [2012AA101108, 2009AA101104]
- Central Level of the Scientific Research Institutes for Basic R&D Special Fund Business [1610162014008]
This study presents a strategy to improve insect pest control in cotton by enhancing methanol production from cell wall by pectin methyl esterase enzymes (PME) and expressing cry genes from Bacillus thuringiensis. Results showed that transgenic plants expressing cry and pme genes had increased methanol production and higher mortality rates for different insect pests compared to plants expressing only cry genes. This demonstrates that Bt. cry genes coupled with pme genes are an effective approach for controlling insect pests.
The idea of enhanced methanol production from cell wall by pectin methyl esterase enzymes (PME) combined with expression of cry genes from Bacillus thuringiensis as a strategy to improve insect pest control in cotton is presented. We constructed a cassette containing two cry genes (cry1Fa and Cry32Aa) and two pme genes, one from Arabidopsis thaliana (AtPME), and other from Aspergillus. niger (AnPME) in pCAMBIA1301 plant expression vector using CAMV-35S promoter. This construction was transformed in Eagle-2 cotton variety by using shoot apex-cut Agrobacterium-mediated transformation. Expression of cry genes and pme genes was confirmed by qPCR. Methanol production was measured in control and in the cry and pme transformed plants showing methanol production only in transformed plants, in contrast to the non-transgenic cotton plants. Finally, insect bioassays performed with transgenic plants expressing cry and pme genes showed 100% mortality for Helicoverpa armigera (cotton bollworm) larvae, 70% mortality for Pectinophora gossypiella (pink bollworm) larvae and 95% mortality of Earias fabia, (spotted bollworm) larvae, that was higher than the transgenic plants expressing only cry genes that showed 84%, 49% and 79% mortality, respectively. These results demonstrate that Bt. cry-genes coupled with pme genes are an effective strategy to improve the control of different insect pests.
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