Method for N-methyl-N-nitrosourea Mutagenesis on Hermaphroditic Germ Cells of Rice
Received date: 2018-11-13
Accepted date: 2019-06-18
Online published: 2019-06-18
N-methyl-N-nitrosourea (MNU) has been used in mutagenesis of fertilized eggs of rice (Oryza sativa). In this study, the method for MNU mutagenesis on hermaphroditic germ cells of rice was established by in vivo synchronous MNU treatment of mature germ-organs in the rice cultivar Liaoyan 6 and the screening of the progeny populations. As compared with MNU treatment of fertilized eggs in Liaoyan 6, MNU treatment of hermaphroditic germ cells under each group of conditions significantly decreased the growth and development indexes of M1 populations and increased the incidence of mutant characters in M1-M2 populations. When the hermaphroditic germ cells were treated for 60 min in MNU buffer (pH 4.8) containing 1.5 mmol∙L -1 MNU and 10 mmol∙L -1 phosphate, the incidence of mutant characters was about 3 times higher than that based on the MNU treatment of fertilized eggs. A series of homozygous mutants including novel plant and grain mutants were obtained by further screening M3 populations. The method for MNU mutagenesis on mature hermaphroditic germ-cells of rice significantly improved the efficiency of broad-spectrum mutagenesis. Its application can provide efficient technical support for large-scale development of various mutants for identifying unknown functional genes and breeding rice.
Key words： rice; hermaphroditic germ cells; N-methyl-N-nitrosourea; mutagenesis
Huaidong Tian,Jing Li,Baohua Tian,Pengfei Niu,Zhen Li,Zhongxiao Yue,Yajuan Qu,Jianfang Jiang,Guangyuan Wang,Huihui Cen,Nan Li,Feng Yan . Method for N-methyl-N-nitrosourea Mutagenesis on Hermaphroditic Germ Cells of Rice[J]. Chinese Bulletin of Botany, 2019 , 54(5) : 625 -633 . DOI: 10.11983/CBB18243
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