<i>In vitro</i> conservation of native Chinese wild grape (<i>Vitis heyneana</i> Roem. & Schult) by slow growth culture
The aim of the present research work was to develop a protocol to preserve Chinese wild grape by slow growth conservation. Spectacular success was achieved in preserving shoot apices of Vitis heyneana under slow growth conditions. The optimized nutrient formulation to maintain slow growth of cultures was Murashige and Skoog (MS) media contained 5 g∙L-1 agar, 0.05 mg∙L-1 indole-3-butyric acid (IBA) and 0.1 mg∙L-1 indole acetic acid (IAA) and 0.5 mg∙L-1 abscisic acid (ABA). The best osmotic adjustment of nutrient medium was achieved by employing 10 g∙L-1 mannitol where 47.78 % cultures could be conserved up to 12 months without any subculture. Among different combination of air breathable film area (ABFA), light intensity and chlorocholine chloride (CCC) concentration, used for increasing the subculture period, 19.63 mm2 ABFA with 5.0 g∙L-1 CCC cultured under lower light intensity suited best for slow growth conservation with 48.00 % microplants were able to survive 10 months without subculture. Further tests showed that the CCC had a negative effect to grape conservation. Cultures responded better when incubated at 10 °C compared with the control (25 °C). Our study also found that the combination of factors were also more beneficial to grape conservation than that of a single factor. 100 % survived shoots by slow growth conservation could regenerate to normal plantlets and transplant successfully. Transplanting plantlets showed no obvious difference in morphology with the control and the maternal parent in the field.
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