Post-thaw survival and proliferation of friable and embryogenie callus in Hevea brasiliensis by liquid nitrogen vitrification tryopreservation method [Preprint]

Li Zhe, Ni Yanmei, Zhou Quannan, Sun Aihua, Dai X., Huang Tiandai, Huang Huasun, Weifu Lin, Lardet Ludovic, Carron Marc-Philippe, Montoro Pascal. 2010. Post-thaw survival and proliferation of friable and embryogenie callus in Hevea brasiliensis by liquid nitrogen vitrification tryopreservation method [Preprint]. In : Climate change, low carbon economy and sustainable natural rubber industry : IRRDB International Rubber Conference, Sanya, China, 18-19 October 2010. s.l. : s.n., 435-436. IRRDB International Rubber Conference, Sanya, Chine, 18 Octobre 2010/19 Octobre 2010.

Résumé : Friable and embryogenic calli could be induced from the inner integument of the rubber tree clone Reyan 88-13. And they had been proliferated and maintained for more than 2 years. Somatic embryos and further regeneration plantlets had been obtained from these calli. In order to preserve these calli for a long time and maintain their genetic characters, a liquid nitrogen vitrification cryopreservation method had been studied with these calli as the material. These calli were inoculated on the pre-culture medium, and had been cultured for 2-3 days in the dark, the temperature was 25-28 oC. Then the calli were immersed for 10-30 min in the 60% PVS2 solution at room temperature. After dehydration treatment, these calli were immersed for 20-40 min in PVS2 solution (28%-32% glycerol + 12%-18% hexanediol + 12%-18% dimethyl sulphoxide + 136.9g1L sucrose) at O°C. Then the former PVS2 solution was removed, and new one was added. Afterward, these calli were thrown into liquid nitrogen (-196 OC) rapidly. The recovery of the calli was to take out of them from the liquid nitrogen quickly, immersed them in water for 2-3 min in which the temperature was 40 oC. After thaw and rinse, these calli were transferred on the subculture medium, and was cultured for 2-3 days in the dark, the temperature was 25-28 oC. Then they Were transferred on the fresh subculture medium and were cultured for 30 days. Most ofthe cryopreserved calli tumed brown and died. But sorne cell survived. So 30 days later, sorne tiny calli emerged, and grew. And they could be proliferated in the further subculture cycles. The post-thaw and survival calli could be used to induce somatic embryogenesis and further plant regeneration. The present results of the studies had the advantages that the experimental procedure was not complex, expensive apparatus could be avoided, repeatability and effect was good. So this method would be useful for long-term preservation of valuable germplasm materials, and would be beneficial for tissue culture and genetictransformation studies in Hevea brasiliensis.

Mots-clés Agrovoc : Hevea brasiliensis

Mots-clés géographiques Agrovoc : Chine

Classification Agris : F62 - Physiologie végétale - Croissance et développement
F02 - Multiplication végétative des plantes
F30 - Génétique et amélioration des plantes

Auteurs et affiliations

• Li Zhe, SCATC (CHN)
• Ni Yanmei
• Zhou Quannan, SCATC (CHN)
• Sun Aihua, SCATC (CHN)
• Dai X.
• Huang Tiandai
• Huang Huasun
• Weifu Lin
• Lardet Ludovic, CIRAD-BIOS-UMR DAP (FRA)
• Carron Marc-Philippe, CIRAD-BIOS-UMR DAP (FRA)
• Montoro Pascal

Source : Cirad - Agritrop (https://agritrop.cirad.fr/558768/)

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