Rapid identification of mutations caused by fast neutron bombardment in Medicago truncatula.
Rapid identification of mutations caused by fast neutron bombardment in Medicago truncatula.
复制标题
快速鉴定蒺藜苜蓿快中子轰击引起的突变
DOI：
10.1186/s13007-021-00765-y
复制
发表时间：
2021-06-16
期刊：
影响因子：
5.1
通讯作者：
Ge L
中科院分区：
文献类型：
--
作者：
Du H;Jiao Z;Liu J;Huang W;Ge L
背景
Background
Fast neutron bombardment (FNB) is a very effective approach for mutagenesis and has been widely used in generating mutant libraries in many plant species. The main type of mutations of FNB mutants are deletions of DNA fragments ranging from few base pairs to several hundred kilobases, thus usually leading to the null mutation of genes. Despite its efficiency in mutagenesis, identification of the mutation sites is still challenging in many species. The traditional strategy of positional cloning is very effective in identifying the mutation but time-consuming. With the availability of genome sequences, the array-based comparative genomic hybridization (CGH) method has been developed to detect the mutation sites by comparing the signal intensities of probes between wild-type and mutant plants. Though CGH method is effective in detecting copy number variations (CNVs), the resolution and coverage of CGH probes are not adequate to identify mutations other than CNVs.
Results
We report a new strategy and pipeline to sensitively identify the mutation sites of FNB mutants by combining deep-coverage whole-genome sequencing (WGS), polymorphism calling, and customized filtering in Medicago truncatula. Initially, we performed a bulked sequencing for a FNB white nodule (wn) mutant and its wild-type like plants derived from a backcross population. Following polymorphism calling and filtering, validation by manual check and Sanger sequencing, we identified that SymCRK is the causative gene of white nodule mutant. We also sequenced an individual FNB mutant yellow leaves 1 (yl1) and wild-type plant. We identified that ETHYLENE-DEPENDENT GRAVITROPISM-DEFICIENT AND YELLOW-GREEN 1 (EGY1) is the candidate gene for M. truncatula yl1 mutant.
Conclusion
Our results demonstrated that the method reported here is rather robust in identifying the mutation sites for FNB mutants.
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批量隔离分析与整个基因组测序（BSA-SEQ）耦合，以在玉米中快速基因克隆。
DOI：
10.1534/g3.118.200499
发表时间：
2018-11-06
期刊：
G3 (Bethesda, Md.)
影响因子：
--
作者：
Klein H;Xiao Y;Conklin PA;Govindarajulu R;Kelly JA;Scanlon MJ;Whipple CJ;Bartlett M
通讯作者：
Bartlett M
影响因子：
7.4
作者：
Rogers, Christian;Wen, Jiangqi;Oldroyd, Giles
通讯作者：
Oldroyd, Giles
影响因子：
56.9
作者：
Alonso, JM;Stepanova, AN;Ecker, JR
通讯作者：
Ecker, JR
使用下一代 DNA 测序数据进行变异发现和基因分型的框架。
影响因子：
30.8
作者：
通讯作者：
--
DOI：
10.1073/pnas.1003954107
发表时间：
2010-06-08
影响因子：
11.1
作者：
Chen, Jianghua;Yu, Jianbin;Chen, Rujin
通讯作者：
Chen, Rujin