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Bulletin of Botanical Research ›› 2017, Vol. 37 ›› Issue (4): 596-602.doi: 10.7525/j.issn.1673-5102.2017.04.016

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Specific Molecular Marker Analysis of the Somaclonal Mutant No.30 of Dendrocalamus farinosus through SLAF-seq

WANG Mao-Lin1,2, HU Shang-Lian1,2, CAO Ying1,2, LU Xue-Qin1,2, XU Gang1,2, HUANG Yan1,2, LONG Zhi-Jian1,2   

  1. 1. Lab of Plant Cell Engineering Southwest University of Science and Technology, Mianyang 621010;
    2. Engineering Research Center for Biomass Resource Utilization and Modification of Sichuan Province, Mianyang 621010
  • Received:2016-11-25 Online:2017-07-15 Published:2017-07-22
  • Supported by:
    National Natural Science Foundation of China(31400257,31400333);Breeding Program Fund Project by the 13th Five-Year Plan of Sichuan Province(16ZS212301);Fund of Engineering Research Center for Biomass Resource Utilizaiton and Modification of Sichuan Province(12zxsk07,13zxsk01,14tdgc05)

Abstract: Somaclonal mutant No.30 of Dendrocalamus farinosus has the specific characteristic of high cellulose and lignin contents, long fiber cells and high length-to-width ratio of fiber cells. Because vegetative propagation is the main way for D.farinosus multiplies, resulting in the failure of homozygous plants, molecular identification of the mutant has not been performed so far, which severely hinders the popularization and application of mutants. Specific molecular marker analysis of the somaclonal mutant No.30 of D.farinosus and identify the sources of character variations. A reduced-representation genome sequencing was performed on wild-type D.farinosus and the mutant No.30 using SLAF-seq technology. After sequencing, library construction, polymorphic sites development, specific screening of polymorphic sites, annotation analysis was carried out on the results of specific screening of polymorphic sites. GC content and homozygous InDel in the mutant No.30 were less than that in the wild type, while total InDel and heterozygous InDel in the mutant No.30 were more than that in the wild type. After specific screening of InDel and SNP sites obtained from SLAF-seq, specific stable InDel/SNP markers including 9381 C-T transition, 9472 A-G transition and 329 InDel were found in the mutant No.30. According to the annotation of these specific SLAF markers, there were 6 SNP markers and 1 InDel related to cellulose synthesis, 3 SNP markers related to lignin synthesis, as well as 4 SNP markers and 1 InDel related to fiber cell morphogenesis. The results revealed genome-level mutations in the mutant No.30 during somatic cell culture, and the sources of character variations were initially identified. However, the correlation between these specific SLAF markers and the characters of the mutant No.30, including cellulose and lignin contents, and fiber cell development, still requires further validation.

Key words: clonal plants, mutants identification, Dendrocalamus farinosus, somaclone, SLAF-seq

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