梁山慈竹体细胞突变体No.30的SLAF-seq特异分子标记分析

doi:10.7525/j.issn.1673-5102.2017.04.016

植物研究 ›› 2017, Vol. 37 ›› Issue (4): 596-602.doi: 10.7525/j.issn.1673-5102.2017.04.016

梁山慈竹体细胞突变体No.30的SLAF-seq特异分子标记分析

王懋林^1,2, 胡尚连^1,2, 曹颍^1,2, 卢学琴^1,2, 徐刚^1,2, 黄艳^1,2, 龙志坚^1,2

1. 西南科技大学植物细胞工程实验室, 绵阳 621010;
2. 四川省生物质资源利用与改性工程技术研究中心, 绵阳 621010

收稿日期:2016-11-25 出版日期:2017-07-15 发布日期:2017-07-22
通讯作者: 胡尚连,E-mail:hushanglian@126.com E-mail:hushanglian@126.com
作者简介:王懋林(1989-),男,硕士研究生,主要从事植物遗传与品种改良研究。
基金资助:
国家自然科学基金（31400257，31400333）；四川省"十三五"重点公关资助项目（2016NYZ0038）；四川省生物质资源利用与改性工程技术研究中心基金资助（12zxsk07，13zxsk01，14tdgc05）

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

WANG Mao-Lin^1,2, HU Shang-Lian^1,2, CAO Ying^1,2, LU Xue-Qin^1,2, XU Gang^1,2, HUANG Yan^1,2, LONG Zhi-Jian^1,2

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)

摘要/Abstract

摘要： 梁山慈竹体细胞突变体No.30具有纤维素含量高、木质素含量高、纤维细胞长度长、纤维细胞长宽比大等特点，但由于梁山慈竹主要进行无性繁殖，不易获得纯合植株，突变体No.30的分子水平鉴定一直无法进行，严重阻碍了该突变体的推广利用。对梁山慈竹体细胞突变体No.30进行特异分子标记分析，并初步确定其性状变异来源。本研究利用SLAF-seq技术分别对突变体No.30以及同地区野生型梁山慈竹群体（CK）的基因组进行了简化基因组测序。经过了测序、建库、多态位点开发、特异多态性位点筛选等过程，最后对特异多态性位点筛选结果进行注释分析。结果发现突变体No.30的GC含量、纯合InDel数低于野生型梁山慈竹，而检测到的InDel总数以及杂合InDel数却高于野生群体。对SLAF-seq得到的InDel、SNP位点进行特异性筛选，获得了突变体No.30中稳定存在的特异InDel/SNP标签，包括特异C-T转换9381个、特异A-G转换9472个、特异InDel 329个。通过对这些特异SLAF标签进行注释，发现有6个特异SNP标签和1个特异InDel与纤维素合成相关，3个特异SNP标签与木质素合成相关，以及4个特异SNP标签和1个特异InDel与纤维细胞形态建成有关。这些结果表明突变体No.30在体细胞培养的过程中在基因组水平上发生了突变，并初步确定了其性状变异来源。但这些特异SLAF标签与突变体No.30在纤维素含量和木质素含量，以及纤维细胞发育等性状方面的相关性仍需进一步验证。

关键词: 克隆植物, 突变体鉴定, 梁山慈竹, 体细胞无性系, SLAF-seq

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

中图分类号:

S795.5

王懋林, 胡尚连, 曹颍, 卢学琴, 徐刚, 黄艳, 龙志坚. 梁山慈竹体细胞突变体No.30的SLAF-seq特异分子标记分析[J]. 植物研究, 2017, 37(4): 596-602.

WANG Mao-Lin, HU Shang-Lian, CAO Ying, LU Xue-Qin, XU Gang, HUANG Yan, LONG Zhi-Jian. Specific Molecular Marker Analysis of the Somaclonal Mutant No.30 of Dendrocalamus farinosus through SLAF-seq[J]. Bulletin of Botanical Research, 2017, 37(4): 596-602.

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梁山慈竹体细胞突变体No.30的SLAF-seq特异分子标记分析

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

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