Aldehyde decarbonylase is a class of enzymes participate in plant alkane biosynthesis. In order to breed new varieties for fruits drying process, ECERIFERUM1 (CER1), an aldehyde decarbonylase coding gene was subjected into an RNA interference (RNAi) assay in xerophytic plant Lycium barbarum ssp. Bianguo. The segment between the 1,469 bp to 1,840 bp of LbCER1 coding aldehyde decarbonylase region in Lycium barbarum ssp. Bianguo was chosen as the optimal target of RNAi. An intermediate vector, pKANNIBAL-LbCER1(+)-PDK intron-LbCER1(-), was used to achieve the proper hairpin structure required for RNAi assay. The final hairpin sequence containing reverse complementary fragments of the chosen LbCER1 sequence flanking the PDK intron was inserted into the vector pART27. In addition, the final vector, pART27-LbCER1(+)-PDK intron-LbCER1(-), was chemically transformed into Agrobacterium tumefaciens GV3101. Restriction enzyme digestion and PCR methods were employed to screen positive intermediate and final vectors. The result showed that A. tumefaciens containing pART27-LbCER1(+)-PDK intron-LbCER1(-) was achieved. Hence, using this method to generate plasmids was correct, which laid the foundation for obtaining new varieties for dry processing through bioengineering of LbCER1- RNAi plants.
YUAN Huijun
,
GAO Ze
,
WANG Juanjuan
,
BAO Jingting
,
FENG Zaiping
. Construction of RNAi expression vector of ECERIFERUM1 gene relating to epidermal wax synthesis in Lycium barbarum ssp. Bianguo[J]. Food and Fermentation Industries, 2020
, 46(10)
: 14
-18
.
DOI: 10.13995/j.cnki.11-1802/ts.023403
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