Using unsupervised learning techniques to assess interactions among complex traits in soybeans

Soybean yield components and agronomic traits are connected through physiological pathways that impose tradeoffs through genetic and environmental constraints. Our primary aim is to assess the interdependence of soybean traits by using unsupervised machine learning techniques to divide phenotypic associations into environmental and genetic associations. This study was performed on large scale, jointly analyzing 14 quantitative traits in a large multi-parental population designed for genetic studies. We collected phenotypes from 2012 to 2015 from a soybean nested association panel with 40 families of approximately 140 individuals each. Pearson and Spearman correlations measured phenotypic associations. A multivariate mixed linear model provided genotypic and environmental correlations. To evaluate relationships among traits, the study used principal component and undirected graphical models from phenotypic, genotypic, and environmental correlation matrices. Results indicate that high phenotypic correlation occurs when traits display both genetic and environmental correlations. In genetic terms, length of reproductive period, node number, and canopy coverage play important roles in determining yield potential. Optimal grain yield production occurs when the growing environment favors faster canopy closure and extended reproductive length. Environmental associations found among yield components give insight into the nature of yield component compensation. The use of unsupervised learning methods provides a good framework for investigating interactions among various quantitative traits and defining target traits for breeding.     

Identification and validation of root length QTLs for water stress resistance in hexaploid wheat (<Emphasis Type="Italic">Titicum aestivum</Emphasis> L.)

Thorough understanding of the genetic mechanisms governing drought adaptive traits can facilitate drought resistance improvement. This study was conducted to identify chromosome regions harbouring QTLs contributing for water stress resistance in wheat. A RIL mapping population derived from a cross between W7984 (Synthetic) and Opata 85 was phenotyped for root length and root dry weight under water stress and non-stress growing conditions. ANOVA showed highly significant (p ≤ 0.01) variation among the RILs for both traits. Root length and root dry weight showed positive and significant (p ≤ 0.01) phenotypic correlation. Broad sense heritability was 86% for root length under stress and 65% for root dry weight under non-stress conditions. A total of eight root length and five root dry weight QTLs were identified under both water conditions. Root length QTLs Qrln.uwa.1BL, Qrln.uwa.2DS, Qrln.uwa.5AL and Qrln.uwa.6AL combined explained 43% of phenotypic variation under non-stress condition. Opata was the source of favourable alleles for root length QTLs under non-stress condition except for Qrln.uwa.6AL. Four stress specific root length QTLs, Qrls.uwa.1AS, Qrls.uwa.3AL, Qrls.uwa.7BL.1 and Qrls.uwa.7BL.2 jointly explained 47% of phenotypic variation. Synthetic wheat contributed favourable alleles for Qrls.uwa.1AS and Qrls.uwa.3AL. Two stable root dry weight QTLs on chromosomes 4AL and 5AL were consistently found in both water conditions. Three validation populations were developed by crossing cultivars Lang, Yitpi, and Chara with Synthetic W7984 to transfer two of the QTLs identified under stress condition. The F_2.3 and F_3.4 validation lines were phenotyped under the same level of water stress as RILs to examine the effect of these QTLs. There were 13.5 and 14.5% increases in average root length due to the inheritance of Qrls.uwa.1AS and Qrls.uwa.3AL, respectively. The result indicated that closely linked SSR markers Xbarc148 (Qrls.uwa.1AS) and Xgwm391 (Qrls.uwa.3AL) can be incorporated into MAS for water stress improvement in wheat.     

Development and application of a molecular marker for TMV resistance based on <Emphasis Type="Italic">N</Emphasis> gene in tobacco (<Emphasis Type="Italic">Nicotiana tabacum</Emphasis>)

Tobacco mosaic virus (TMV) caused serious loss in yield and quality of tobacco every year. It is a long-term goal to improve the tobacco resistance against TMV by tobacco breeding. N gene was the firstly reported TMV-resistant gene, which showed resistance against all Tobamoviruses except the Ob stain and belonged to the toll-interleukin-1 receptor/nucleotide-binding site/leucine-rich repeat class of plant resistance (R) genes. At present, N gene had already been widely used in tobacco conventional breeding, but there is rare available molecular maker used in marker-assisted selection of TMV resistance. In this study, we designed a pair of primers that specific amplify N gene fragment based on the sequence of N gene intron III, named N-marker. Then, we identified TMV resistance by two selecting methods, PCR with N-marker and inoculated with the TMV-C strain. Results from the two method showed that (1) 13 varieties among 67 tobacco varieties displayed hypersensitive reaction when inoculated with the TMV-C strain, also contained N gene fragments screened by PCR with N-marker; (2) 105 strains of 200 BC1 strains showed resistance against TMV when inoculated with TMV-C strain, meanwhile, 103 of the 105 strains contained N gene fragment verified by PCR with N-marker. Therefore, the N-marker is reliable for high throughput screening of germplasm resources and tobacco breeding materials in selection of N-mediated TMV resistance. Our study not only developed a molecular marker for tobacco breeding, but also identified new germplasm resources that are resistant to TMV.     

Two reported cytotypes of the emergent orchid model species <Emphasis Type="Italic">Erycina pusilla</Emphasis> are two different species

Each species is characterized by a specific set of chromosomes, which is described as the chromosome portrait or karyotype. In general, such a karyotype is the same for all individuals in the population. An exception to that rule has recently been found in the orchid Erycina pusilla, which has been reported to have two cytotypes with chromosome numbers of 2n = 10 and 2n = 12. Here, we examined the karyotypes of the two cytotypes and found differences in arm ratios and heterochromatin patterns as well as in the presence of satellite chromosomes and in the number and location of rDNA and telomeric repeat sites. These differences are extensive and would have required multiple chromosome rearrangements to generate the differences between the two karyotypes. We also found that F1 hybrids between the parents with the two different chromosome numbers resulted in sterile offspring, in accordance with our previous findings. The combination of hybrid sterility and extensively rearranged chromosomes supports the hypothesis that these two reported cytotypes are, in fact, two different species.     

Determining resistance to <Emphasis Type="Italic">Fusarium verticillioides</Emphasis> and fumonisin accumulation in African maize inbred lines resistant to <Emphasis Type="Italic">Aspergillus flavus</Emphasis> and aflatoxins

Fusarium verticillioides and Aspergillus flavus cause Fusarium ear rot (FER) and Aspergillus ear rot (AER) of maize, respectively. Both pathogens are of concern to producers as they reduce grain yield and affect quality. F. verticillioides and A. flavus also contaminate maize grain with the mycotoxins fumonisins and aflatoxins, respectively, which has been associated with mycotoxicosis in humans and animals. The occurrence of common resistance mechanisms to FER and AER has been reported. Hence, ten Kenyan inbred lines resistant to AER and aflatoxin accumulation were evaluated for resistance to FER, F. verticillioides colonisation and fumonisin accumulation; and compared to nine South African lines resistant to FER and fumonisin accumulation. Field trials were conducted at three localities in South Africa and two localities in Kenya. FER severity was determined by visual assessment, while F. verticillioides colonisation and fumonisin content were quantified by real-time PCR and liquid chromatography tandem mass spectrometry, respectively. Significant genotype x environment interactions was determined at each location (P ≤ 0.05). Kenyan inbred CML495 was most resistant to FER and F. verticillioides colonisation, and accumulated the lowest concentration of fumonisins across localities. It was, however, not significantly more resistant than Kenyan lines CML264 and CKL05015, and the South African line RO549 W, which also exhibited low FER severity (≤5%), fungal target DNA (≤0.025 ng μL^?1) and fumonisin levels (≤2.5 mg kg^?1). Inbred lines resistant to AER and aflatoxin accumulation appear to be promising sources of resistance to F. verticillioides and fumonisin contamination.     

Identifying and exploring significant genomic regions associated with soybean yield,seed fatty acids,protein and oil

Soybean [Glycine max (L.) Merrill] yield and seed fatty acids, protein, and oil content are important traits for which an improved understanding of significant genomic regions would be useful. To accomplish this, a soybean population consisting of 203 F5 derived recombinant inbred lines (RILs) was developed and genotyped with 11,633 polymorphic single nucleotide polymorphisms (SNPs). Each RIL was grown in a single plot at Knoxville, TN in 2010; followed by replicated, multi-location field trials in 2013 and 2014. The data from 2010, 2013, and 2014 were analyzed together in order to detect quantitative trait loci (QTL) for these traits, and 30 total QTLs were detected. Five QTLs are candidates for confirmed status and one QTL is a candidate for positional confirmation. Many of the genes with mutations in close proximity to the fatty acid QTLs are involved in biological processes for fatty acids and/or lipids and could be considered possible candidate genes. Similarly, genes with mutations in genomic regions near yield, protein, and oil QTLs were plentiful and may contribute to the variation observed in these traits. Except for yield and stearic acid, each trait displayed pleiotropic effects with other traits in this study. Notable are the pleiotropic effects for oleic and linolenic acid on chromosomes 9, 13, and 19. Overall, the findings from this research contribute new information to the genetic understanding of soybean yield and seed fatty acids, protein and oil content. This understanding will be useful in making trait improvements.     

Exploring diversity among rice germplasms based on their physiological traits responses to salinity

Salinity is one of the major abiotic stresses that severely effects rice production throughout the world. Previously, there have not been many studies which focused on studying diversity among rice germplasm based on specific physiological traits for salt tolerance. Our diversity study was based on physiological traits such as Na+ concentration, K⁺ concentration, K⁺/Na⁺ ratio, osmotic potential, and biomass which are major components determining salt tolerance. This study has systematically analyzed phenotypic data of 191 germplasms in two different salt concentrations apart from the control. The current study identified salt tolerant germplasms based on their response to a single physiological trait as well as a combination of different physiological traits. Some of the germplasm identified outperformed known salt tolerant cultivar Pokkali. This study identifies correlation among various physiological traits. The salt tolerant germplasms can be taken forward into developing better varieties by conventional breeding and exploring genes for salt tolerance.     

Geospatial delineation of South Korea for adjusted barley cultivation under changing climate

Determining effective measures to alleviate the impact of climate change on crops under various regional environments is one of the most urgent issues facing agriculture. In this study, geographic regions of South Korea for future-adjusted barley cultivation were outlined and the impact of climate change on barley production in the next 100 years was evaluated under two greenhouse gas concentration trajectory scenarios: the representative concentration pathway (RCP) 4.5 and RCP 8.5. To achieve our intended study goals, a geospatial crop simulation modeling (GCSM) scheme was formulated using CERES-barley model of Decision Support System for Agricultural Technology (DSSAT) crop model package version 4.6 to simulate grid-based geospatial crop yields. Two experiments were carried out at an open field to obtain model coefficients for the nation and at temperature gradient field chambers to evaluate the performance of the CERES-barley model under elevated temperature conditions. Suitable cultivation regions for three different types of barley (naked, hooded, and malting) under changing climate were projected to expand to the northern regions under both RCP 8.5 and RCP 4.5. However, they were projected to expand more rapidly under RCP 8.5 than those under RCP 4.5. Projected yields of four barley varieties were increased with a slow phase as year progressed under RCP 4.5 scenario. However, they were rapidly increased under RCP 8.5 scenario. It appears that geospatial variation in barley yield under changing climate can be effectively outlined. Therefore, GCSM system might be useful for determining impacts of climate change on geospatial variations of crops, potentially providing means to impede food insecurity.     

Allelic composition and associated quality traits of the <Emphasis Type="Italic">Glu-1</Emphasis> and <Emphasis Type="Italic">Glu-3</Emphasis> loci in selected modern Ethiopian durum wheat varieties

Gluten protein determines the processing quality of both durum wheat and bread wheat. The glutenin subunits compositions and associated quality traits of 20 Ethiopian durum wheat varieties were systematically analyzed using SDS-PAGE and Payne numbers. A total of 16 glutenin patterns were identified. At the Glu-A1 locus, all varieties scored the null allele. The predominant glutenin alleles at the Glu-B1 locus were Glu-B1b (7+8) and Glu-B1e (20). In Glu-3, the most abundant glutenin subunits were Glu-A3a and Glu-B3c. Based on the Payne scores, the varieties Yerer, Ginchi, Candate, and Foka were identified to have allelic composition suitable for pasta making. The cluster analysis using agglomerative hierarchical clustering (AHC) method classified the varieties into four similarity classes. Based on the findings of this experiment, suggestions were made for allelic composition improvement through introgression of superior alleles from known Glu-1 and Glu-3 sources.     

Identification and validation of cold responsive microRNAs of <Emphasis Type="Italic">Hevea brasiliensis</Emphasis> using high throughput sequencing

Cold stress is one of the major abiotic factors that influence the productivity and geographical distribution of many agriculturally important crops like Hevea brasiliensis. Cultivation of H. brasiliensis in India is being extended to northeastern regions, where low temperature during winter adversely affects its survival, growth, and productivity. Developing cold-tolerant genotypes is a primary requisite to maximize the productivity under such challenging environmental conditions. However, lack of methods for early evaluation of cold tolerance in the newly developed clones and the extensive time required for assessing their tolerance in the field are major constraints for clonal selection. The present study was initiated with an objective to identify and characterize cold stress responsive miRNAs from H. brasiliensis that show stronger association with cold tolerance. Next generation sequencing using Illumina HiSeq method revealed the expression of 21 and 29 conserved miRNA (from clone RRIM 600) families in cold-stressed and control samples, respectively. Forty-two novel miRNAs were identified from this study. Upon differential expression analysis, eight conserved miRNAs were found commonly expressed in both the samples. When expression analyses were performed subsequently with six selected miRNAs in two Hevea clones (viz. RRII 105 and RRIM 600), miR169 showed a strong association with cold tolerance. miRNAs such as miR482 and miR159 also exhibited association with cold tolerance. This study suggests the possibility of employing these miRNAs as markers for cold tolerance after validation in more number of genotypes with varying levels of cold tolerance.