A new, harmless, high-throughput endosperm-based DNA extraction method for wheat
2019
In this study, a non-destructive, high-throughput, endosperm-based DNA extraction method was developed. To verify the nondestructive nature of this method, a germination test was performed on 288 seeds after sampling their endosperm, which gave a seedling
emergence rate that was higher (97.6%) than that of the control group (92%). To confirm the feasibility of the new method, DNA was extracted
from plants of a BC1F2 population by two different methods, namely, from endosperm using our rapid, high-throughput method (ER-DNAand from young leaves emerging from the same sampled seed using the CTAB method (LC-DNA). The ER-DNA was undetectable by agarose
gel electrophoresis, but was found to be an adequate replacement for LC-DNA for the amplification and detection of simple sequence repeats
(SSRs). Further analysis revealed that ER-DNA was generally suitable for the generation of specific 500–750-bp fragments, but not for the
amplification of 1,000–2,000-bp fragments. Our rapid, high-throughput method therefore has no deleterious effects on wheat seeds and yields
DNA for SSR genotyping that is a suitable alternative to traditionally obtained DNA.
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