Lina Wang, Bing Zhang, Jinrui Li, Xiaoyu Yang, Zhonghai Ren^*

Show more
College of Horticulture Science and Engineering, Shandong Agricultural University, Tai’an, China.
School of Life Sciences, Faculty of Science, The Chinese University of Hong Kong, Hong Kong, China.
Show less

Abstract: Ethyl methanesulfonate (EMS) is a stable and effective chemical mutagen. In this study, cucumber (Cucumis sativus L. cv. “Shannong No. 5”) seeds were treated by 1% EMS for 12 h, 24 h and 48 h to optimize EMS mutagenesis and determined median lethal dose of EMS (1% EMS and 24 h) for “Shannong No. 5”. After treated by 1% EMS for 24 h, 541 M1 plants were grown in greenhouse for phenotype investigation. The fertility of M1 cucumbers was very low, and only 79 lines produced seeds after self crossing. 60 independent M2 families comprising 600 M2 plants were investigated for phenotypic alteration, and 11 individual mutant lines were isolated into six groups: short-fruit mutants, long-fruit mutants, small-flower mutants, big-flower mutants, opposite-tendril mutants and clustered-leaf mutants. The mutation frequency was 18.3%. Two selected representatives, short-fruit mutants and clustered-leaf mutants, showed 1:3 of segregation ratio in M2 populations. This ratio is consistent with classic Mendelian model, indicating that the two kinds of mutants may be controlled by a single recessive gene, respectively. Long-fruit phenotype was stably inherited and no segregation was observed in M3 generation, indicating that this mutant line may be homozygous.

Keywords: Cucumber, Ethyl Methanesulfonate, Mutagenesis

Cite this paper: Wang, L. , Zhang, B. , Li, J. , Yang, X. and Ren, Z. (2014) Ethyl Methanesulfonate (EMS)-Mediated Mutagenesis of Cucumber (Cucumis sativus L.). Agricultural Sciences, 5, 716-721. doi: 10.4236/as.2014.58075.

References

[1]   Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, G., Heller, C., Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C. and Ecker, J.R. (2003) Genome-Wide Insertional Mutagenesis of Arabidopsis Thaliana. Science, 301, 653-657.
http://dx.doi.org/10.1126/science.1086391

[2]   Rosso, M.G., Li, Y., Strizhov, N., Reiss, B., Dekker, K. and Weisshaar, B. (2003) An Arabidopsis Thaliana T-DNA Mutagenized Population (GABI-Kat) for Flanking Sequence Tag-Based Reverse Genetics. Plant Molecular Biology, 53, 247-259.

[3]   An, G., Jeong, D.H., Jung, K.H. and Lee, S. (2005) Reverse Genetic Approaches for Functional Genomics of Rice. Plant Molecular Biology, 59, 111-123.
http://dx.doi.org/10.1007/s11103-004-4037-y

[4]   Uchida, N., Sakamoto, T., Kurata, T. and Tasaka, M. (2011) Identification of Ems-Induced Causal Mutations in a Non-Reference Arabidopsis Thaliana Accession by Whole Genome Sequencing. Plant and Cell Physiology, 52, 716-722.
http://dx.doi.org/10.1093/pcp/pcr029

[5]   Girija, M., Dhanavel, D. and Gnanamurthy, S. (2013) Gamma Rays and EMS Induced Flower Color and Seed Mutants in Cowpea (Vigna unguiculata L. Walp). Advances in Applied Science Research, 4, 134-139.
http://connection.ebscohost.com/c/articles/90260513/gamma-rays-ems-induced-flower-color-seed-mutants-cowpea-vigna-unguiculata-l-walp

[6]   Martín, B., Ramiro, M., Martínez-Zapater, J.M. and Alonso-Blanco, C. (2009) A High-Density Collection of EMS-Induced Mutations for TILLING in Landsberg Erecta Genetic Background of Arabidopsis. BMC Plant Biology, 9, 147.
http://dx.doi.org/10.1186/1471-2229-9-147

[7]   Østergaard, L. and Yanofsky, M.F. (2004) Establishing Gene Function by Mutagenesis in Arabidopsis Thaliana. Plant Journal, 39, 682-696.
http://dx.doi.org/10.1111/j.1365-313X.2004.02149.x

[8]   Henikoff, S., Till, B.J. and Comai, L. (2004) TILLING. Traditional Mutagenesis Meets Functional Genomics. Plant Physiology, 135, 630-636.
http://dx.doi.org/10.1104/pp.104.041061

[9]   Vora, J.D., Rane, L. and Kumar, S.A. (2014) Biochemical, Anti-Microbial and Organoleptic Studies of Cucumber (Cucumis sativus). International Journal of Science and Research, 3, 662-664.
http://www.ijsr.net/archive/v3i3/MDIwMTMxMjc0.pdf

[10]   Garcia, D., Saingery, V., Chambrier, P., Mayer, U., Jürgens, G. and Berger, F. (2003) Arabidopsis Haiku Mutants Reveal New Controls of Seed Size by Endosperm. Plant Physiology, 131, 1661-1670.
http://dx.doi.org/10.1104/pp.102.018762

[11]   Qu, L.J. and Qin, G. (2014) Generation and Identification of Arabidopsis EMS Mutants. Methods in Molecular Biology, 1062, 225-239.
http://dx.doi.org/10.1007/978-1-62703-580-4_12

[12]   Serrat, X., Esteban, R., Guibourt, N., Moysset, L., Nogués, S. and Lalanne, E. (2014) EMS Mutagenesis in Mature Seed-Derived Rice Calli as a New Method for Rapidly Obtaining TILLING Mutant Populations. Plant Methods, 10, 5.
http://dx.doi.org/10.1186/1746-4811-10-5

[13]   Uchida, N., Sakamoto, T., Tasaka, M. and Kurata, T. (2014) Identification of EMS-induced Causal Mutations in Arabidopsis Thaliana by Next-Generation Sequencing. Methods in Molecular Biology, 1062, 259-270.
http://dx.doi.org/10.1007/978-1-62703-580-4_14

[14]   Kim, Y., Schumaker, K.S. and Zhu, J.K. (2006) EMS Mutagenesis of Arabidopsis. Methods in Molecular Biology, 323, 101-103.
http://dx.doi.org/10.1385/1-59745-003-0:101

[15]   Kharkwal, M.C. (1999) Induced Mutations in Chickpea (Cicer arietinum L.) III. Frequency and Spectrum of Viable Mutations. Indian Journal of Genetics and Plant Breeding, 59, 451-464.
http://www.indianjournals.com/ijor.aspx?target=ijor:ijgpb&volume=59&issue=4&article=010

[16]   Waghmare, V.N. and Mehra, R.B. (2001) Induced Chlorophyll Mutations, Mutagenic Effectiveness and Efficiency in Lathyrus sativus L. Indian Journal of Genetics and Plant Breeding, 61, 53-56.
http://www.indianjournals.com/ijor.aspx?target=ijor:ijgpb&volume=61&issue=1&article=014