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In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.)

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Abstract

A protocol for in vitro induction of tetraploids via colchicine-treated somatic embryos from immature zygotic embryos of diploid grapevine (Vitis vinifera L.) is reported. Embryogenic callus was initiated from immature zygotic embryos cultured on Nitsch and Nitsch (NN) medium supplemented with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D). The callus was transferred to NN medium containing 1.0 mg/l α-naphthalene acetic acid (NAA) and 0.5 mg/l benzyladenine (BA) to establish somatic embryogenesis. The vigorously growing globular embryos were selected and treated by 0, 10 or 20 mg/l colchicine for 1, 2 or 3 days, and then immediately transferred to NN medium supplemented with 0.03 mg/l NAA and 0.5 mg/l BA, for somatic embryo conversion and plant regeneration. The number of surviving embryos and regenerated plantlets following colchicine treatment decreased with increasing colchicine concentration and treatment time. Among 29 randomly investigated plantlets regenerated from colchicine-treated somatic embryos, five solid tetraploids (2n = 4× = 76) were identified by chromosome counting analysis; all others were diploid (2n = 2× = 38). Ploidy level of plant regenerated was also determined from leaves using flow cytometry. No chimeras with both 2C and 4C nuclei was produced from colchicine-treated somatic embryos. Significant differences in leaf stomata parameters were observed between diploid and induced tetraploid plantlets.

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Acknowledgments

This research was financially supported by the National Natural Science Foundation of China (Nos. 90302010, 30570271) and the project of database platform construction for basis science and technology resource of the Ministry of Education of China (No. 505016). The authors would like to express their appreciation to Y.C. Xiong and S.J Xu for their reviewing the manuscript. Authors are grateful to J.L. Zhang for chromosome analysis, J. Yi for determination of the ploidy level by flow cytometry.

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Authors and Affiliations

  1. School of Life Science, Lanzhou University, Lanzhou, 730030, P.R. China

    X. M. Yang, L. Z. An & X. W. Fang

  2. Academy of Gansu Agriculture Science, Lanzhou, 730070, P.R. China

    X. M. Yang

  3. School of Agronomy, Gansu Agricultural University, Lanzhou, 730070, P.R. China

    Z. Y. Cao & Y. M. Wang

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  1. X. M. Yang
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  2. Z. Y. Cao
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  3. L. Z. An
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  4. Y. M. Wang
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  5. X. W. Fang
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Correspondence to L. Z. An.

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Yang, X.M., Cao, Z.Y., An, L.Z. et al. In vitro tetraploid induction via colchicine treatment from diploid somatic embryos in grapevine (Vitis vinifera L.) . Euphytica 152, 217–224 (2006). https://doi.org/10.1007/s10681-006-9203-7

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