TILLING by Sequencing: A Successful Approach to Identify Rare Alleles in Soybean Populations
1
Department of Agronomy, Purdue University, 915 West State Street, West Lafayette, IN 47907, USA
2
USDA-ARS Crop Production and Pest Control Research Unit, 915 West State Street, West Lafayette, IN 47907, USA
*
Author to whom correspondence should be addressed.
†
Current address: BASF Corporation, 925 County Road 378, Beaver Crossing, NE 68313, USA.
Genes 2019, 10(12), 1003; https://doi.org/10.3390/genes10121003
Received: 29 October 2019 / Revised: 25 November 2019 / Accepted: 28 November 2019 / Published: 3 December 2019
(This article belongs to the Special Issue TILLING and CRISPR to design the varieties of tomorrow)
Soybean seeds produce valuable protein that is a major component of livestock feed. However, soybean seeds also contain the anti-nutritional raffinose family oligosaccharides (RFOs) raffinose and stachyose, which are not digestible by non-ruminant animals. This requires the proportion of soybean meal in the feed to be limited, or risk affecting animal growth rate or overall health. While reducing RFOs in soybean seed has been a goal of soybean breeding, efforts are constrained by low genetic variability for carbohydrate traits and the difficulty in identifying these within the soybean germplasm. We used reverse genetics Targeting Induced Local Lesions in Genomes (TILLING)-by-sequencing approach to identify a damaging polymorphism that results in a missense mutation in a conserved region of the RAFFINOSE SYNTHASE3 gene. We demonstrate that this mutation, when combined as a double mutant with a previously characterized mutation in the RAFFINOSE SYNTHASE2 gene, eliminates nearly 90% of the RFOs in soybean seed as a proportion of the total seeds carbohydrates, and results in increased levels of sucrose. This represents a proof of concept for TILLING by sequencing in soybean.
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited
MDPI and ACS Style
Thapa, R.; Carrero-Colón, M.; Rainey, K.M.; Hudson, K. TILLING by Sequencing: A Successful Approach to Identify Rare Alleles in Soybean Populations. Genes 2019, 10, 1003.
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