Variation Analysis of Root System Development in Wheat Seedlings Using Root Phenotyping System

Root system architecture is a vital part of the plant that has been shown to vary between species and within species based on response to genotypic and/or environmental influences. The root traits of wheat seedlings are critical for their establishment in soil and evidently linked to plant height and seed yield. However, plant breeders have not efficiently developed the role of RSA in wheat selection due to the difficulty of studying root traits. We set up a root phenotyping platform to characterize RSA in 34 wheat accessions. The phenotyping pipeline consists of the germination paper-based moisture replacement system, image capture units, and root-image processing software. The 34 accessions from two different wheat ploidy levels (hexaploids and tetraploids), were characterized in ten replicates. A total of 19 root traits were quantified from the root architecture generated. This pipeline allowed for rapid screening of 340 wheat seedlings within 10 days. At least one line from each ploidy (6× and 4×) showed significant differences (p \u3c 0.05) in measured traits, except for mean seminal count. Our result also showed a strong correlation (0.8) between total root length, maximum depth and convex hull area. This phenotyping pipeline has the advantage and capacity to increase screening potential at early stages of plant development, leading to the characterization of wheat seedling traits that can be further examined using QTL analysis in populations generated from the examined accessions

Assessing Root System Architecture of Wheat Seedlings Using A High-Throughput Root Phenotyping System

Background and aims Root system architecture is a vital part of the plant that has been shown to vary between species and within species based on response to genotypic and/or environmental influences. The root traits of wheat seedlings is critical for the establishment and evidently linked to plant height and seed yield. However, plant breeders have not efficiently developed the role of RSA in wheat selection due to the difficulty of studying root traits. Methods We set up a root phenotyping platform to characterize RSA in 34 wheat accessions. The phenotyping pipeline consists of the germination paper-based moisture replacement system, image capture units, and root-image processing software. The 34 accessions from two different wheat ploidy levels (hexaploids and tetraploids), were characterized in ten replicates. A total of 19 root traits were quantified from the root architecture generated. Results This pipeline allowed for rapid screening of 340 wheat seedlings within 10days. Also, at least one line from each ploidy (6x and 4x) showed significant differences (P \u3c 0.05) in measured traits except in mean seminal count. Our result also showed strong correlation (0.8) between total root length, maximum depth and convex hull area. Conclusions This phenotyping pipeline has the advantage and capacity to increase screening potential at early stages of plant development leading to characterization of wheat seedling traits that can be further examined using QTL analysis in populations generated from the examined accessions

Detection of novel allelic variations in soybean mutant population using Tilling by Sequencing

One 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

Development and Characterization of Soybean with Improved Fatty Acid Profile Through Mutation Breeding and Genome Editing

Soybean (Glycine max (L.) Merrill) is considered a key crop of modern agriculture due to its seed’s high protein and oil content. However, the high percentage of polyunsaturated fatty acids in soybean oil lim¬its its stability and shelf life. Modification in the fatty acid composition can improve its stability and nutritional quality. Mutagenesis is a useful tool to induce genetic and phenotypic variation for trait improvement and discovery of novel genes. A mutant population of soybean cultivar JTN-5203 (MG V) was generated using ethyl methane sulfonate (EMS) mutagenesis and was for screening novel fatty acid alleles by using forward and reverse genetics approaches. Optimum concentration of EMS was used to treat 15,000 bulk JTN-5203 seeds producing a total of 1,820 M2 population. Fatty acid profiles such as oleic acid, linoleic, and linolenic acid were measured in more than 5900 M2 plants using near-infrared spectroscopy. DNA was extracted, normalized and pooled from these individuals, and specific primers were designed for genes involved in fatty acid biosynthesis pathway for further analysis using Sanger and next-generation sequencing technologies. Forward genetics screening revealed mutants that contained either an elevated oleic acid or a reduction in linoleic or linolenic acid content, and mutants that carried all the three important phenotypes. Single nucleotide polymorphisms were observed in the genes for fatty acid biosynthesis pathway including, FAD2-1B, FAD2-2B, and FAD2-2C. Mutants with a range of genetic variations were also identified. In addition, the high throughput mutation discovery through TILLING-by-Sequencing approach has been successfully applied. Several mutations were detected that have consequences on FAD2-1A and FAD2-1B gene functions. This includes GC to AT transition mutations that consist of 15.89% in FAD2-1A and 69% in FAD2-1B. 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 an improved and heart-healthy-soybeans. Moreover, a modified version of Stupar’s whole plant transformation from half-seed explant in soybean has been used for genome editing, and putative transformants were successfully recovered

Rapid Profiling of Soybean Aromatic Compounds Using Electronic Nose

Soybean (Glycine max (L.)) is the world’s most important seed legume, which contributes to 25% of global edible oil, and about two-thirds of the world’s protein concentrate for livestock feeding. One of the factors that limit soybean’s utilization as a major source of protein for humans is its characteristic soy flavor. This off-flavor can be attributed to the presence of various chemicals such as phenols, aldehydes, ketones, furans, alcohols, and amines. In addition, these flavor compounds interact with protein and cause the formation of new off-flavors. Hence, studying the chemical profile of soybean seeds is an important step in understanding how different chemical classes interact and contribute to the overall flavor profile of the crop. In our study, we utilized the HERCALES Fast Gas Chromatography (GC) electronic nose for identification and characterization of different volatile compounds in five high-yielding soybean varieties, and studied their association with off-flavors. With aroma profiling and chemical characterization, we aim to determine the quantity and quality of volatile compounds in these soybean varieties and understand their effect on the flavor profiles. The study could help to understand soybean flavor characteristics, which in turn could increase soybean use and enhance profitability