Optimization of system for Agrobacterium tumefaciens -mediated AtMYB118 genetic transformation into friable embryogenic calli in Hevea brasiliensis

Bi Zheng-Hong, Li Zhe, Wang Zhi-Qiu, Dai Xue-Mei, Fang Jia-Lin, Huang Huasun, Lin Wei-Fu, Lardet Ludovic, Montoro Pascal, Carron Marc-Philippe, Leclercq Julie, Zuo Jian-Ru, Mu Jin-Ye, Yang Xiao-Hui, Li Xiang. 2014. Optimization of system for Agrobacterium tumefaciens -mediated AtMYB118 genetic transformation into friable embryogenic calli in Hevea brasiliensis. Journal of Southern Agriculture, 45 (7) : 1137-1146.

Résumé : [Objective]: An orthogonal design L25 (56) was used to optimize Agrobacterium -mediated AtMYB118 gene genetic transformation system of friable embryogenic calli in Hevea brasiliensis in order to provide references for genetic improvement of its clones. [Method]: Kanamycin at a concentration of 75.0 mg/L was used to select trans - formed calli. The effects of six factors viz . , preculture time, Agrobacterium growth phase (OD600) , acetosyringone (AS) concentration, infection time, co-culture temperature and co-culture time on genetic transformation were evaluated by GUS transient expression detection. [Result]: Six factors were found to significantly affect transformation frequency of long-term cultural friable embryogenic calli. The highest transformation efficiency was achieved by 0-day preculture, long-term cultural friable embryogenic calli as recipients infected by Agrobacterium cultures corresponding to OD600=0.7 for 7 min, followed by co-culture for 5 days in a co-culture medium containing 200 ?mol/L AS at 25 ?. After 4-6 months, 17 GUS positive callus lines were achieved. Polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR) analysis results of transgenic tissues further confirmed that uidA, nptII and AtMYB118 genes had been inserted into genome of calli and expressed. At last , 164 transgenic embryoids were obtained from 1539 cultured embryoids. A transformation efficiency of 10.6% was achieved for longterm cultural friable embryogenic calli using this protocol. Four AtMYB118 transgenic plantlets were obtained. [Conclusion]: The optimized genetic transformation system using friable embryogenic calli of rubber tree as recipients was effective and available,which would provide technological supports on genetic improvement of clones.

Mots-clés Agrovoc : Hevea brasiliensis

Mots-clés géographiques Agrovoc : Chine

Classification Agris : F30 - Génétique et amélioration des plantes
F60 - Physiologie et biochimie végétale

Champ stratégique Cirad : Axe 1 (2014-2018) - Agriculture écologiquement intensive

Auteurs et affiliations

• Bi Zheng-Hong, Hainan University (CHN)
• Li Zhe, Rubber Research Institute (CHN)
• Wang Zhi-Qiu, Hainan University (CHN)
• Dai Xue-Mei, Rubber Research Institute (CHN)
• Fang Jia-Lin, Rubber Research Institute (CHN)
• Huang Huasun, Rubber Research Institute (CHN)
• Lin Wei-Fu, Rubber Research Institute (CHN)
• Lardet Ludovic, CIRAD-BIOS-UMR AGAP (FRA)
• Montoro Pascal, CIRAD-BIOS-UMR AGAP (FRA)
• Carron Marc-Philippe, CIRAD-PERSYST-UPR Systèmes de pérennes (FRA)
• Leclercq Julie, CIRAD-BIOS-UMR AGAP (FRA) ORCID: 0000-0001-8110-3762
• Zuo Jian-Ru, Institute of Genetics and Developmental Biology (CHN)
• Mu Jin-Ye, Institute of Genetics and Developmental Biology (CHN)
• Yang Xiao-Hui, Institute of Genetics and Developmental Biology (CHN)
• Li Xiang, Hainan University (CHN)

Source : Cirad - Agritrop (https://agritrop.cirad.fr/575434/)

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