Transportation of the Foreign Bt-Cry1Ac Protein and Insect Resistance Detection in Transgenic Poplar (Populus spp.) with Different Anvil Grafting
CHEN Xing-Hao1, ZHANG Jun1, *, LI Zheng1, YU Xiao-Yue1, ZHANG De-Jian2, YANG Min-Sheng1
1 Hebei Key Laboratory for Tree Genetic Resources and Forest Protection / College of Forestry Science, Hebei Agricultural University, Baoding 071000, China; 2 College of Life Science, Inner Mongolia University, Hohhot 010021, China
Abstract：With the extensive planting of poplar (Populus spp.) in China, the pest problem of poplar comes along.Nowadays, the application of chemical insecticides can not only fail to effectively reduce the harm of pests to poplar trees, but also seriously damage the ecological environment.Therefore, it is feasible to improve the insect resistance of poplar through grafting and propagation of transgenic poplar and non-transgenic poplar.In this study, in order to understand the ecological safety of grafted transgenic poplar, hybrid Populus tomentosa and Hb1 transgenic Populus × euramericana cv.'Neva' with Bt-Cry1Ac gene were used as scions and rootstocks to study whether the mRNA and protein of Cry1Ac gene were transported between scions and rootstocks, and whether the leaves of grafted seedlings were resistant to the insect pest.The results of reverse transcription-PCR (RT-PCR) and third-generation digital PCR showed that no Cry1Ac gene mRNA was detected in the shoots and leaves of hybrid P.tomentosa as scions or rootstocks, and the copy number of Cry1Ac gene was much lower than that of positive control, indicating that the exogenous Cry1Ac gene mRNA could not be transported between rootstocks and scions.Cry1Ac protein was detected in leaves, phloem and xylem of scions and rootstocks in 2 grafting combination by enzyme-linked immuno sorbent assay (ELISA), which proved that Cry1Ac protein could be transported between rootstocks and scions by grafting.The non-transgenic poplar grafted on the transgenic poplar could inhibit the growth and development of the larvae of the Hyphantria cunea, and slow down the development of the larvae, showing a certain degree of insect resistance.This study clarified the expression, transmission and insect resistance of Cry1Ac gene expression products between transgenic and non-transgenic tissues, and could provide theoretical basis and scientific guidance for the rational use of grafted poplar in insect resistance practice.
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