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Establishment of Tissue Culture Regeneration System of Spiraea×bumalda ’Gold Mound’

Author: ChengLeiJie
Tutor: HongBo
School: Northeast Forestry University
Course: Ornamental Plants and Horticulture
Keywords: Spiraea×bumalda’Gold Mound’ Regeneration Somatic embryos Receptor
CLC: S682.19
Type: Master's thesis
Year: 2009
Downloads: 114
Quote: 1
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Abstract

Spiraea×bumalda’Gold Mound’in Rosaceace is a plant which has high ornamental value with orderly plant type, colcorful leaves and flourish flower.The mass propagation of Spiraea×bumalda’Gold Mound’was established by using nodal segment explants. The high regeneration of Spiraea×bumalda’Gold Mound’was achieved via organogenesis, callus indirect organogenesis and somatic embryogenesis by using cotyledons , hypocotyls and leaves. It will lay the basis for the genetic transformation of Spiraeaxbumalda’Gold Mound’, supply the technical director. The primary research results as follows:1. Nodal segment explants of Spiraea×bumalda’Gold Mound’were cultured on Murashige and Skoog(MS) medium supplemented with different plant growth regulators(6-BA、NAA) for mass propagation. The results showed that the MS medium containing 2.0 mg/L 6-BA and 0.2 mg/L NAA was suitable for the germination of axillary buds. The germination rate was 93.3% and the axillay buds grew healthily. The proliferation of axillary buds on MS medium containing 0.5 mg/L and 0.1 mg/L NAA was achieved 3.6.2. The plant regeneration of Spiraea×bumalda’Gold Mound’was achieved via organogenesis by cotyledons and hypocotyls. The results showed that the highest regeneration rate from the cotyledons of seedlings of Spiraea×bumalda’Gold Mound’was achieved 60% on MS medium containing 1.0 mg/L 6-BA, 1.0 mg/L IBA and 1.0 mg/L TDZ, and the adventitious shoots grew healthily. The highest regeneration rate from the hypocotyls of seedlings of Spiraea×bumalda’Gold Mound’was achieved 50% on MS medium supplemented with 1.0 mg/L 6-BA, 1.0 mg/L IBA and 0.5 mg/L TDZ, and the adventitious shoots grew healthily.3. The plant regeneration of Spiraea×bumalda’Gold Mound’was achieved via callus indirect organogenesis and somatic embryogenesis by using leaves. The results showed that the frequency of callus from the leaves was achieved 96.7% on the MS medium containing 0.4 mg/L 6-BA, 0.04 mg/L IBA and 2.0 mg/L 2,4-D. The highest embryogenic callus rate was achieved 50% on MS medium supplemented with 0.4 mg/L 6-BA and 0.04 mg/L NAA. The frequency of callus differentiation was 90% on MS medium containing 0.8 mg/L 6-BA and 0.04 mg/L NAA. MS medium supplemented with 0.8 mg/L 6-BA and 0.1 mg/L NAA was applied for the induction and germination of somatic embryos.4. 1/2MS supplemented with 0.5 mg/L NAA and 0.1% AC was applied for the root developed.5. The receptor regeneration system for the genetic transformation of Spiraeaxbumalda’Gold Mound’ was achieved via the experiment about antibiotic sensitivities. The results demonstrated that the optimum selectivity antibiotic of Spiraea×bumalda’Gold Mound’was Kanamycin, the optimum concentration was 75 mg/L. The 300 mg/L Carbenicillin and 300mg/L Cefotaxime were the optimum bacteriostasis and could controlled the growth ofagrobacterium tumefaciens, and the rate of regeneration was achieved respectively 55% and60%.

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