Efficient somatic embryogenesis from meristematic explants in grapevine (Vitis vinifera L.) cv. Chardonnay: an improved protocol

Published Date
August 2016, Volume 30, Issue 4, pp 1377–1387

Title 

Efficient somatic embryogenesis from meristematic explants in grapevine (Vitis viniferaL.) cv. Chardonnay: an improved protocol

• Author 

• Pascale Maillot, 
• Laurence Deglène-Benbrahim, 
• Bernard Walter

• 
  

• Abstract 

Somatic embryogenesis (SE) has great importance for propagation and transformation of woody perennial plants. In Vitis vinifera L., the most cultivated grape species, somatic embryos are generally obtained from floral organs (stamens, ovaries, whole flowers) not easily available. Moreover, the efficiency of SE induction from reproductive organs greatly depends on the physiological state of explants which is hardly controlled. We report here on a culture system for the efficient and reproducible induction of SE and the production of good quality embryos, based on the use of nodal explants excised from in vitro grown plantlets easily obtained all year round. This method involves a first culture step on a callus inducing medium (CIM) containing unusually elevated concentrations of auxin and cytokinin. SE efficiencies from 10.3 to 20.2 % were reproducibly attained with a combination of 18 μM of 2,4-dichlorophenoxyacetic acid (2,4-D) and 9 μM of 6-benzylaminopurine (BAP) in CIM. Callus formation and embryogenesis induction were preferentially observed in the bud region. Moreover, we showed that internodal stem segments, petioles and leaf fragments were not amenable to SE under the same conditions, strongly suggesting that axillary meristems are involved in the embryogenic response. In addition, we verified that high concentrations of 2,4-D and BAP in CIM did not affect the viability of embryos further generated, nor hamper their regeneration into whole plants.

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