Enabling Reverse Genetics in Medicago truncatula Using High-Throughput Sequencing for Tnt1 Flanking Sequence Recovery

Author(s):  
Xiaofei Cheng ◽  
Nick Krom ◽  
Shulan Zhang ◽  
Kirankumar S. Mysore ◽  
Michael Udvardi ◽  
...  
2016 ◽  
Vol 17 (6) ◽  
pp. 460-475 ◽  
Author(s):  
Anis Ben-Amar ◽  
Samia Daldoul ◽  
Götz M. Reustle ◽  
Gabriele Krczal ◽  
Ahmed Mliki

2019 ◽  
Author(s):  
Reneth Millas ◽  
Mary Espina ◽  
CM Sabbir Ahmed ◽  
Angelina Bernardini ◽  
Ekundayo Adeleke ◽  
...  

ABSTRACTOne of the most important tools in genetic improvement is mutagenesis, which is a useful tool to induce genetic and phenotypic variation for trait improvement and discovery of novel genes. JTN-5203 (MG V) mutant population was generated using an induced ethyl methane sulfonate (EMS) mutagenesis and was used for detection of induced mutations in FAD2-1A and FAD2-1B genes using reverse genetics approach. Optimum concentration of EMS was used to treat 15,000 bulk JTN-5203 seeds producing 1,820 M2 population. DNA was extracted, normalized, and pooled from these individuals. Specific primers were designed from FAD2-1A and FAD2-1B genes that are involved in the fatty acid biosynthesis pathway for further analysis using next-generation sequencing. High throughput mutation discovery through TILLING-by-Sequencing approach was used to detect novel allelic variations in this population. Several mutations and allelic variations with high impacts were detected for FAD2-1A and FAD2-1B. This includes GC to AT transition mutations in FAD2-1A (20%) and FAD2-1B (69%). Mutation density for this population is estimated to be about 1/136kb. Through mutagenesis and high-throughput sequencing technologies, novel alleles underlying the mutations observed in mutants with reduced polyunsaturated fatty acids will be identified, and these mutants can be further used in breeding soybean lines with improved fatty acid profile, thereby developing heart-healthy-soybeans.


2016 ◽  
Vol 44 (5) ◽  
pp. 1385-1393 ◽  
Author(s):  
Veronica Dezi ◽  
Chavdar Ivanov ◽  
Irmgard U. Haussmann ◽  
Matthias Soller

Modified nucleotides in messenger RNA (mRNA) have been discovered over 40 years ago, but until recently little was known about which transcripts contain them and what their function is. High-throughput sequencing approaches revealed a dynamic landscape of the ‘Epitranscriptome’ for many mRNA modifications in various organisms from yeast to humans. Meanwhile, also many genes encoding mRNA-modifying enzymes and auxiliary proteins have been identified yielding functional insights by reverse genetics into their role in development and disease.


BMC Genomics ◽  
2008 ◽  
Vol 9 (1) ◽  
pp. 593 ◽  
Author(s):  
Gyorgy Szittya ◽  
Simon Moxon ◽  
Dulce M Santos ◽  
Runchun Jing ◽  
Manuel PS Fevereiro ◽  
...  

BMC Genomics ◽  
2011 ◽  
Vol 12 (1) ◽  
Author(s):  
Tianzuo Wang ◽  
Lei Chen ◽  
Mingui Zhao ◽  
Qiuying Tian ◽  
Wen-Hao Zhang

Planta ◽  
2011 ◽  
Vol 235 (2) ◽  
pp. 375-386 ◽  
Author(s):  
Lei Chen ◽  
Tianzuo Wang ◽  
Mingui Zhao ◽  
Qiuying Tian ◽  
Wen-Hao Zhang

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