Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2008, Vol. 34 ›› Issue (07): 1285-1289.

• RESEARCH NOTES • Previous Articles     Next Articles

Transgenetic Research of Antigen VP4 Gene into Peanut (Arachis hypo-gaea L.) via Agrobacterium tumefaciens

LIU Feng123,WAN Shu-Bo1,BI Yu-Ping2,YAN Cai-Xia1,LI Chun-Juan1,Zhao Jin-Ping2,SHAN Shi-Hua1,*   

  1. 1 Shandong Peanut Research Institute, Qingdao 266100, Shandong; 2 High-Tech Research Center, Shandong Academy of Agricultural Sciences, Ji’nan 250100, Shandong; 3 Marine Biological Culture Collection, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, Shandong, China
  • Received:2007-09-29 Revised:1900-01-01 Online:2008-07-12 Published:2008-07-12
  • Contact: SHAN Shi-Hua

Abstract: Peanut (Arachis hypogaea L.) is an important commercial crop worldwide and provides an excellent source of protein and other nutrients. So it is one of the ideal vectors in oral vaccine of transgenic plant. The Agrobacterium strain LBA4404 harboring the binary vector pBI121 was used for transformation. The plasmid carries genes for VP4 (2 350 bp) of the human rotavirus (HRV) driven by CaMV 35S promoter and neomycin phosphotransferase (npt II). Two peanut cultivars were used as source of cotyledon explants in this experiment. Efficient transformation of cotyledons by A. tumefaciens strain LBA4404 carrying nptII and VP4 gene on binary vectors led to the production of a large percentage of transgenic plants. Transformed individuals were obtained through selection on medium containing 125 mg L-1 Kan. Integration of the transgenes was assessed by PCR amplification and Southern blot hybridizations. Taking pBG1VP4 or pBG1VP4 plasmid as positive control, non-transformed peanut as negative control, 22 plants among 26 plants grown up through selection on medium containing 125 mg L-1 Kan were assessed by PCR amplification of 620 bp fragment of npt II gene. Then all of 22 plants were assessed by PCR amplification of 2 350 bp fragment of VP4 gene. Taking VP4 gene with a-32P-dCTP maker as probe, five plants selected randomly from 22 positive plants were analysed by PCR-Southern blot hybridizations and showed DNA bloting bands. Then the genomic DNA of 4 plants chosen from PCR-Southern positive plants was further analyzed with Southern blot hybridizations and showed correspondent DNA blotting bands. The results showed that the foreign gene was integrated into genome of transformated peanuts. The total RNA from 11 plants of Luhua 14 was assessed by RT-PCR analysis and evidenced the expression of G1VP4 gene. In addition, expression of critical protein in 30 kD was assayed with Western-blot method. An edible vaccine based on the VP4 of human rotavirus (HRV) could provide a means to protect children from severe acute diarrhea, enabling intact antigen to reach the gut associated lymphoid tissue, as the rigid walls of the plant cell protect antigenic proteins from the acidic environment of stomach. Elevated expression of the rotavirus VP4 antigen in transgenic peanuts is a critical factor in the development of a safe and effective rotavirus vaccine.

Key words: Peanut, Cotyledon, Genetic transformation, VP4 gene

CLC Number: 

  • 10.3724/SP.J.1006.2008.01285
[1] YANG Huan, ZHOU Ying, CHEN Ping, DU Qing, ZHENG Ben-Chuan, PU Tian, WEN Jing, YANG Wen-Yu, YONG Tai-Wen. Effects of nutrient uptake and utilization on yield of maize-legume strip intercropping system [J]. Acta Agronomica Sinica, 2022, 48(6): 1476-1487.
[2] LI Hai-Fen, WEI Hao, WEN Shi-Jie, LU Qing, LIU Hao, LI Shao-Xiong, HONG Yan-Bin, CHEN Xiao-Ping, LIANG Xuan-Qiang. Cloning and expression analysis of voltage dependent anion channel (AhVDAC) gene in the geotropism response of the peanut gynophores [J]. Acta Agronomica Sinica, 2022, 48(6): 1558-1565.
[3] FENG Ya, ZHU Xi, LUO Hong-Yu, LI Shi-Gui, ZHANG Ning, SI Huai-Jun. Functional analysis of StMAPK4 in response to low temperature stress in potato [J]. Acta Agronomica Sinica, 2022, 48(4): 896-907.
[4] DING Hong, XU Yang, ZHANG Guan-Chu, QIN Fei-Fei, DAI Liang-Xiang, ZHANG Zhi-Meng. Effects of drought at different growth stages and nitrogen application on nitrogen absorption and utilization in peanut [J]. Acta Agronomica Sinica, 2022, 48(3): 695-703.
[5] HUANG Li, CHEN Yu-Ning, LUO Huai-Yong, ZHOU Xiao-Jing, LIU Nian, CHEN Wei-Gang, LEI Yong, LIAO Bo-Shou, JIANG Hui-Fang. Advances of QTL mapping for seed size related traits in peanut [J]. Acta Agronomica Sinica, 2022, 48(2): 280-291.
[6] WANG Ying, GAO Fang, LIU Zhao-Xin, ZHAO Ji-Hao, LAI Hua-Jiang, PAN Xiao-Yi, BI Chen, LI Xiang-Dong, YANG Dong-Qing. Identification of gene co-expression modules of peanut main stem growth by WGCNA [J]. Acta Agronomica Sinica, 2021, 47(9): 1639-1653.
[7] WANG Jian-Guo, ZHANG Jia-Lei, GUO Feng, TANG Zhao-Hui, YANG Sha, PENG Zhen-Ying, MENG Jing-Jing, CUI Li, LI Xin-Guo, WAN Shu-Bo. Effects of interaction between calcium and nitrogen fertilizers on dry matter, nitrogen accumulation and distribution, and yield in peanut [J]. Acta Agronomica Sinica, 2021, 47(9): 1666-1679.
[8] SHI Lei, MIAO Li-Juan, HUANG Bing-Yan, GAO Wei, ZHANG Zong-Xin, QI Fei-Yan, LIU Juan, DONG Wen-Zhao, ZHANG Xin-You. Characterization of the promoter and 5'-UTR intron in AhFAD2-1 genes from peanut and their responses to cold stress [J]. Acta Agronomica Sinica, 2021, 47(9): 1703-1711.
[9] GAO Fang, LIU Zhao-Xin, ZHAO Ji-Hao, WANG Ying, PAN Xiao-Yi, LAI Hua-Jiang, LI Xiang-Dong, YANG Dong-Qing. Source-sink characteristics and classification of peanut major cultivars in North China [J]. Acta Agronomica Sinica, 2021, 47(9): 1712-1723.
[10] ZHANG He, JIANG Chun-Ji, YIN Dong-Mei, DONG Jia-Le, REN Jing-Yao, ZHAO Xin-Hua, ZHONG Chao, WANG Xiao-Guang, YU Hai-Qiu. Establishment of comprehensive evaluation system for cold tolerance and screening of cold-tolerance germplasm in peanut [J]. Acta Agronomica Sinica, 2021, 47(9): 1753-1767.
[11] XUE Xiao-Meng, WU JIE, WANG Xin, BAI Dong-Mei, HU Mei-Ling, YAN Li-Ying, CHEN Yu-Ning, KANG Yan-Ping, WANG Zhi-Hui, HUAI Dong-Xin, LEI Yong, LIAO Bo-Shou. Effects of cold stress on germination in peanut cultivars with normal and high content of oleic acid [J]. Acta Agronomica Sinica, 2021, 47(9): 1768-1778.
[12] HAO Xi, CUI Ya-Nan, ZHANG Jun, LIU Juan, ZANG Xiu-Wang, GAO Wei, LIU Bing, DONG Wen-Zhao, TANG Feng-Shou. Effects of hydrogen peroxide soaking on germination and physiological metabolism of seeds in peanut [J]. Acta Agronomica Sinica, 2021, 47(9): 1834-1840.
[13] ZHANG Wang, XIAN Jun-Lin, SUN Chao, WANG Chun-Ming, SHI Li, YU Wei-Chang. Preliminary study of genome editing of peanut FAD2 genes by CRISPR/Cas9 [J]. Acta Agronomica Sinica, 2021, 47(8): 1481-1490.
[14] DAI Liang-Xiang, XU Yang, ZHANG Guan-Chu, SHI Xiao-Long, QIN Fei-Fei, DING Hong, ZHANG Zhi-Meng. Response of rhizosphere bacterial community diversity to salt stress in peanut [J]. Acta Agronomica Sinica, 2021, 47(8): 1581-1592.
[15] HUANG Bing-Yan, SUN Zi-Qi, LIU Hua, FANG Yuan-Jin, SHI Lei, MIAO Li-Juan, ZHANG Mao-Ning, ZHANG Zhong-Xin, XU Jing, ZHANG Meng-Yuan, DONG Wen-Zhao, ZHANG Xin-You. Genetic analysis of fat content based on nested populations in peanut (Arachis hypogaea L.) [J]. Acta Agronomica Sinica, 2021, 47(6): 1100-1108.
Full text



No Suggested Reading articles found!