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In vitro mutagenesis and identification of mutants via ISSR in lily (Lilium longiflorum)

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Abstract

An efficient in vitro mutagenesis protocol for Lilium longiflorum Thunb. cv. White fox has been established. The effect of 6-BA and NAA on adventitious bud formation from the bulblet-scale thin cell layers was tested. Results showed that the optimal medium for adventitious bud induction is MS basal medium supplemented with 2.0 mg/l 6-BA and 0.1 mg/l NAA. The differentiation frequency and the average number of adventitious buds reached 95.55% and 3.00, respectively. Various doses (0.0, 0.5, 1.0, 1.5, 2.0, and 2.5 Gy) of gamma rays were applied to investigate the effect of radiation on adventitious bud formation from bulblet-scale thin cell layers. The forming capacity of the adventitious buds significantly decreased with the increase of radiation dose. The results suggested that the optimal irradiation dose is 1.0 Gy. Dose of 1.0 Gy treatment resulted in 55.33% survival of irradiated bulblet-scale thin cell layers and 39.27% mutagenesis rate. The genetic variations among the morphological mutants were evaluated by DNA fingerprinting using ISSR molecular marker. The genetic variation frequency reached 36.06% using seven ISSR primers. Out of the 50 mutant lines transferred to the greenhouse, 9 were observed to have significantly different morphological characters than those of the controls.

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Abbreviations

6-BA:

6-Benzylaminopurine

CTAB:

Hexadecyltrimethyl ammonium bromide

ISSR:

Inter simple sequence repeat

MS:

Murashige and Skoog (1962)

NAA:

a-Naphthaleneacetic acid

TCL:

Thin cell layers

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Acknowledgments

The authors wish to thank Professor Zhongen Xi (Chongqing University of Posts and Telecommunications, China) for his valuable assistance in preparation of the manuscript. Financial support from the National Natural Science Foundation of China (30972407), Jiangsu Provincial Key Basic Research foundation for Universities (07KJA21019), “948” project of the State Administration of Forestry of China (2007-4-06), and A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions.

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Authors and Affiliations

  1. Key Laboratory of Forest Genetics and Biotechnology of the Ministry of Education, Nanjing Forestry University, Nanjing, 210037, People’s Republic of China

    Mengli Xi, Lina Sun, Shuai Qiu, Juanjuan Liu, Jin Xu & Jisen Shi

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  1. Mengli Xi
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  2. Lina Sun
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  3. Shuai Qiu
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  4. Juanjuan Liu
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  5. Jin Xu
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Correspondence to Jisen Shi.

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Communicated by R. Schmidt.

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Xi, M., Sun, L., Qiu, S. et al. In vitro mutagenesis and identification of mutants via ISSR in lily (Lilium longiflorum). Plant Cell Rep 31, 1043–1051 (2012). https://doi.org/10.1007/s00299-011-1222-8

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  • DOI: https://doi.org/10.1007/s00299-011-1222-8

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