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.     

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.     

Improved genetic parameter estimations in zoysiagrass by implementing post hoc blocking

Randomized complete block (RCB) design is the most widely used experimental design in biological sciences. As number of treatments increases, the block size become larger and it looses the capacity to control the variance within block, which is its original purpose. A method known as post hoc blocking could be used in these cases to improve the genetic parameter estimation and thus obtain an unbiased assessment of the performance of a given treatment. In trufgrass breeding, as other breeding program, this is a common challenge. The goal of this study was to test the capacity of different post hoc blocking designs to improve the genetic parameter estimation of zoysiagrass (Zoysia spp.). We evaluated two post hoc blocking designs; row–column (R–C) and incomplete block (IB) designs on five genotype trials located in Florida. The results showed that post hoc R–C design had superior model fitting than both the original RCB and the post hoc IB designs when studied at the single measurement level and at the site level. The narrow-sense heritability (0.24–0.40) and the genotype-by-measurement correlation (0.57–0.99) did not change significantly when R–C was compared to the original RCB design. The ranking of the top performing genotypes changed considerably when comparing RCB to R–C design, but the degree depended on the location analyzed. We conclude that the change in the ranking of the top (potentially select individuals) is coming from the better control of intra-block environmental variation, and this could potentially have a significant impact on the breeding selection process.     

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.     

Association mapping reveals loci associated with multiple traits that affect grain yield and adaptation in soft winter wheat

Genome-wide association studies (GWAS) are useful to facilitate crop improvement via enhanced knowledge of marker-trait associations (MTA). A GWAS for grain yield (GY), yield components, and agronomic traits was conducted using a diverse panel of 239 soft red winter wheat (Triticum aestivum) genotypes evaluated across two growing seasons and eight site-years. Analysis of variance showed significant environment, genotype, and genotype-by-environment effects for GY and yield components. Narrow sense heritability of GY (h ²  = 0.48) was moderate compared to other traits including plant height (h ²  = 0.81) and kernel weight (h ²  = 0.77). There were 112 significant MTA (p < 0.0005) detected for eight measured traits using compressed mixed linear models and 5715 single nucleotide polymorphism markers. MTA for GY and agronomic traits coincided with previously reported QTL for winter and spring wheat. Highly significant MTA for GY showed an overall negative allelic effect for the minor allele, indicating selection against these alleles by breeders. Markers associated with multiple traits observed on chromosomes 1A, 2D, 3B, and 4B with positive minor effects serve as potential targets for marker assisted breeding to select for improvement of GY and related traits. Following marker validation, these multi-trait loci have the potential to be utilized for MAS to improve GY and adaptation of soft red winter wheat.     

Characterization of resistance to the wheat stem sawfly in spring wheat landrace accessions from targeted geographic regions of the world

Plant landraces have long been recognized as potential gene pools for biotic and abiotic stress-related genes. This research used spring wheat landrace accessions to identify new sources of resistance to the wheat stem sawfly (WSS) (Cephus cinctus Norton), an important insect pest of wheat in the northern Great Plains of North America. Screening efforts targeted 1409 accessions from six geographical areas of the world where other species of grain sawflies are endemic or where a high frequency of accessions possesses the resistance characteristic of solid stems. Resistance was observed in approximately 14% of accessions. Half of the lines displayed both antixenosis and antibiosis types of resistance. Among the resistant accessions, 41% had solid or semi-solid stems. Molecular genetic screening for haplotypes at the solid stem QTL, Qss.msub.3BL, showed that 15% of lines shared the haplotype derived from ‘S-615’, the original donor of the solid stem trait to North American germplasm. Other haplotypes associated with solid stems were also observed. Haplotype diversity was greater in the center of origin of wheat. Evaluation of a representative set of resistant landrace accessions in replicated field trials at four locations over a three year period identified accessions with potential genes for reduced WSS infestation, increased WSS mortality, and increased indirect defense via parasitoids. Exploitation of distinct types of plant defense will expand the genetic diversity for WSS resistance currently present in elite breeding lines.     

Heterosis and combining ability of seven maize germplasm populations

Understanding the combining ability and heterosis of available germplasm is a prerequisite for successful maize improvement and breeding. The objectives of this study were to analyze the combining ability and heterosis of seven representative maize germplasm populations, and further, to evaluate their potential utility in germplasm improvement. A total of 21 crosses were made among these seven populations in a complete diallel without reciprocals. The parental populations and 21 crosses were evaluated for days to silking (DS), ear height (EH), and grain yield (GY) in the Northeast and Yellow and Huai River maize growing areas in China in 2012. Csyn5, Csyn7, Cpop.11, and Cpop.12 had desirable general combining ability (GCA) effects for DS and EH in both the Northeast China mega-environment (NCM) and the Yellow and Huai River Regions of China mega-environment (YHCM). Cpop.11 possessed a favorable GCA effect for GY in the NCM, as did Csyn5, Cpop.17, and Cpop.18 in the YHCM. Csyn6 and Csyn7 exhibited tremendous yield-enhancing potential in both mega-environments. Additionally, six combinations including Csyn7 × Csyn6, Csyn5 × Csyn6, Cpop.11 × Cpop.18, Cpop.12 × Cpop.17, Csyn7 × Cpop.17, and Csyn5 × Csyn7 exhibited better specific combining ability effects for GY, yield performance, and mid-parent heterosis in the appropriate mega-environment. These results indicated that the seven populations would be very useful for the improvement of related agronomic traits, and the six candidate combinations possessed great potential for further improvement and utilization.     

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.     

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.