Crops ›› 2017, Vol. 33 ›› Issue (6): 50-54.doi: 10.16035/j.issn.1001-7283.2017.06.009

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EMS Mutant Screening and RAPD Analysis on Industrial Hemp (Cannabis sativa L.)

Jiang Ying,Sun Yufeng,Pan Dongmei,Li Qiuzhi   

  1. Daqing Branch of Heilongjiang Academy of Sciences,Daqing 163319,Heilongjiang,China
  • Received:2017-07-26 Revised:2017-09-13 Online:2017-12-15 Published:2018-08-26

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Abstract:

Mutant library is the basis of functional genomics research. In this study, seeds of hemp varieties Huoma No.1 were treated by chemical mutagen Ethyl sulfonate (EMS). Mutation plants were screened in M2 generation according to agronomic and biological traits. 37 mutants were acquired from seedling stage to adult stage. 27 mutants were found in seedling stage and 10 mutants were found in adult stage, correspondingly mutant percentages were 20.93% and 11.24%. Furthermore, mutants were screened by RAPD (Random Amplified Polymorphic DNA), 11 out of 26 random primers could detect DNA polymorphism. Results showed the mutants varied not only on phenotypic changes, but also on the DNA level, which provided a theoretical and practical basis for the research of functional genomics and breeding direction in EMS mutation population of industrial hemp.

Key words: Industrial hemp (Cannabis sativa L.), EMS, Mutants, RAPD

Table 1

The selected primers used in amplification and their sequences (5′-3′)"

引物Primer 序列Sequence 引物Primer 序列Sequence
OPU-05 TTGGCGGCCT OPS-11 AGTCGGGTGG
OPU-12 TCACCAGCCA OPS-17 TGGGGACCAC
OPU-15 ACGGGCCAGT OPV-06 ACGCCCAGGT
OPX-13 ACGGGAGCAA OPV-08 GGACGGCGTT
OPX-17 GACACGGACC OPW-09 GTGACCGAGT
OPY-05 GGCTGCGACA OPW-16 CAGCCTACCA
OPY-11 AGACGATGGG OPZ-06 GTGCCGTTCA
OPY-15 AGTCGCCCTT OPZ-13 GACTAAGCCC
OPT-07 GGCAGGCTGT OPH-18 GAATCGGCCA
OPT-13 AGGACTGCCA OPI-07 CAGCGACAAG
OPP-12 AAGGGCGAGT OPR-08 CCCGTTGCCT
OPP-16 CCAAGCTGCC OPR-13 GGACGACAAG
OPP-17 TGACCCGCCT OPO-12 CAGTGCTGTG

Fig.1

DNA extraction from mutations of hemp DNA marker：1kb plus DNA ladder"

Fig.2

Amplification of genomic DNAs using primer OPP-12 1, No mutagenic Huoma No.1 materials; 2-9, Mutagenic Huoma No.1 materials; the same below"

Fig.3

Amplification of genomic DNAs using primer OPV-08"

Fig.4

Amplification of genomic DNAs using primer OPY-11"

Fig.5

Amplification of genomic DNAs using primer OPU-05"

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