B. Todd Campbell

Identification of the family of aquaporin genes and their expression in upland cotton (Gossypium hirsutum L.)

BackgroundCotton (Gossypium spp.) is produced in over 30 countries and represents the most important natural fiber in the world. One of the primary factors affecting both the quantity and quality of cotton production is water. A major facilitator of water movement through cell membranes of cotton and other plants are the aquaporin proteins. Aquaporin proteins are present as diverse forms in plants, where they function as transport systems for water and other small molecules. The plant aquaporins belong to the large major intrinsic protein (MIP) family. In higher plants, they consist of five subfamilies including plasma membrane intrinsic proteins (PIP), tonoplast intrinsic proteins (TIP), NOD26-like intrinsic proteins (NIP), small basic intrinsic proteins (SIP), and the recently discovered X intrinsic proteins (XIP). Although a great deal is known about aquaporins in plants, very little is known in cotton.ResultsFrom a molecular cloning effort, together with a bioinformatic homology search, 71 upland cotton (G. hirsutum) aquaporin genes were identified. The cotton aquaporins consist of 28 PIP and 23 TIP members with high sequence similarity. We also identified 12 NIP and 7 SIP members that showed more divergence. In addition, one XIP member was identified that formed a distinct 5th subfamily. To explore the physiological roles of these aquaporin genes in cotton, expression analyses were performed for a select set of aquaporin genes from each subfamily using semi-quantitative reverse transcription (RT)-PCR. Our results suggest that many cotton aquaporin genes have high sequence similarity and diverse roles as evidenced by analysis of sequences and their expression.ConclusionThis study presents a comprehensive identification of 71 cotton aquaporin genes. Phylogenetic analysis of amino acid sequences divided the large and highly similar multi-gene family into the known 5 aquaporin subfamilies. Together with expression and bioinformatic analyses, our results support the idea that the genes identified in this study represent an important genetic resource providing potential targets to modify the water use properties of cotton.

Genome-wide identification of differentially expressed genes under water deficit stress in upland cotton (Gossypium hirsutum L.).

BackgroundCotton is the world’s primary fiber crop and is a major agricultural commodity in over 30 countries. Like many other global commodities, sustainable cotton production is challenged by restricted natural resources. In response to the anticipated increase of agricultural water demand, a major research direction involves developing crops that use less water or that use water more efficiently. In this study, our objective was to identify differentially expressed genes in response to water deficit stress in cotton. A global expression analysis using cDNA-Amplified Fragment Length Polymorphism was conducted to compare root and leaf gene expression profiles from a putative drought resistant cotton cultivar grown under water deficit stressed and well watered field conditions.ResultsWe identified a total of 519 differentially expressed transcript derived fragments. Of these, 147 transcript derived fragment sequences were functionally annotated according to their gene ontology. Nearly 70 percent of transcript derived fragments belonged to four major categories: 1) unclassified, 2) stress/defense, 3) metabolism, and 4) gene regulation. We found heat shock protein-related and reactive oxygen species-related transcript derived fragments to be among the major parts of functional pathways induced by water deficit stress. Also, twelve novel transcripts were identified as both water deficit responsive and cotton specific. A subset of differentially expressed transcript derived fragments was verified using reverse transcription-polymerase chain reaction. Differential expression analysis also identified five pairs of duplicated transcript derived fragments in which four pairs responded differentially between each of their two homologues under water deficit stress.ConclusionsIn this study, we detected differentially expressed transcript derived fragments from water deficit stressed root and leaf tissues in tetraploid cotton and provided their gene ontology, functional/biological distribution, and possible roles of gene duplication. This discovery demonstrates complex mechanisms involved with polyploid cotton’s transcriptome response to naturally occurring field water deficit stress. The genes identified in this study will provide candidate targets to manipulate the water use characteristics of cotton at the molecular level.

RNA-Seq transcriptome profiling of upland cotton (Gossypium hirsutum L.) root tissue under water-deficit stress.

An RNA-Seq experiment was performed using field grown well-watered and naturally rain fed cotton plants to identify differentially expressed transcripts under water-deficit stress. Our work constitutes the first application of the newly published diploid D5 Gossypium raimondii sequence in the study of tetraploid AD1 upland cotton RNA-seq transcriptome analysis. A total of 1,530 transcripts were differentially expressed between well-watered and water-deficit stressed root tissues, in patterns that confirm the accuracy of this technique for future studies in cotton genomics. Additionally, putative sequence based genome localization of differentially expressed transcripts detected A2 genome specific gene expression under water-deficit stress. These data will facilitate efforts to understand the complex responses governing transcriptomic regulatory mechanisms and to identify candidate genes that may benefit applied plant breeding programs.

Genetic dissection of yield and its component traits using high-density composite map of wheat chromosome 3A: bridging gaps between QTLs and underlying genes.

Earlier we identified wheat (Triticum aestivum L.) chromosome 3A as a major determinant of grain yield and its component traits. In the present study, a high-density genetic linkage map of 81 chromosome 3A-specific markers was developed to increase the precision of previously identified yield component QTLs, and to map QTLs for biomass-related traits. Many of the previously identified QTLs for yield and its component traits were confirmed and were localized to narrower intervals. Four novel QTLs one each for shoot biomass (Xcfa2262-Xbcd366), total biomass (wPt2740-Xcfa2076), kernels/spike (KPS) (Xwmc664-Xbarc67), and Pseudocercosporella induced lodging (PsIL) were also detected. The major QTLs identified for grain yield (GY), KPS, grain volume weight (GVWT) and spikes per square meter (SPSM) respectively explained 23.2%, 24.2%, 20.5% and 20.2% of the phenotypic variation. Comparison of the genetic map with the integrated physical map allowed estimation of recombination frequency in the regions of interest and suggested that QTLs for grain yield detected in the marker intervals Xcdo549-Xbarc310 and Xpsp3047-Xbarc356 reside in the high-recombination regions, thus should be amenable to map-based cloning. On the other hand, QTLs for KPS and SPSM flanked by markers Xwmc664 and Xwmc489 mapped in the low-recombination region thus are not suitable for map-based cloning. Comparisons with the rice (Oryza sativa L.) genomic DNA sequence identified 11 candidate genes (CGs) for yield and yield related QTLs of which chromosomal location of two (CKX2 and GID2-like) was confirmed using wheat aneuploids. This study provides necessary information to perform high-resolution mapping for map-based cloning and for CG-based cloning of yield QTLs.

Regression-Based Multi-Trait QTL Mapping Using a Structural Equation Model

Quantitative trait loci (QTL) mapping often results in data on a number of traits that have well-established causal relationships. Many multi-trait QTL mapping methods that account for the correlation among multiple traits have been developed to improve the statistical power and the precision of QTL parameter estimation. However, none of these methods are capable of incorporating the causal structure among the traits. Consequently, genetic functions of the QTL may not be fully understood. Structural equation modeling (SEM) allows researchers to explicitly characterize the causal structure among the variables and to decompose effects into direct, indirect, and total effects. In this paper, we developed a multi-trait SEM method of QTL mapping that takes into account the causal relationships among traits related to grain yield. Performance of the proposed method is evaluated by simulation study and applied to data from a wheat experiment. Compared with single trait analysis and the multi-trait least-squares analysis, our multi-trait SEM improves statistical power of QTL detection and provides important insight into how QTLs regulate traits by investigating the direct, indirect, and total QTL effects. The approach also helps build biological models that more realistically reflect the complex relationships among QTL and traits and is more precise and efficient in QTL mapping than single trait analysis.

Performance and combining ability in cotton (Gossypium hirsutum L.) populations with diverse parents

Improving fiber quality properties of cotton is important for increasing the efficiency of manufacturing textiles, including enhancing yarn quality and spinning performance. This study was conducted to determine if we could identify valuable cotton cultivars to use as parents in breeding programs with the goal of improving fiber properties. Seven parents were combined in a diallel design and selfed to obtain 21 F2 populations. Positive general and specific combining ability effects were observed for all traits. General combining ability tended to be larger than specific combining ability, indicating these traits are controlled primarily by additive genetic effects. Correlations among traits were generally positive except for lint yield correlations with fiber strength and length. For improving the fiber quality measures of strength and length, line 7235 shows excellent general combining ability effects. SG125 would provide elite germplasm to increase agronomic measures of lint yield and lint percent. The MD51 genotype has the highest potential among the genotypes tested here to provide germplasm combining both improved yield and fiber strength. These parents, or their selected progeny, should be useful in a breeding program to generate variability from which selection can be used to identify lines with improved fiber and/or agronomic properties.

Variability in four diverse cotton ( Gossypium hirsutum L.) germplasm populations

A broad range of cotton (Gossypium hirsutum L.) germplasm resources exist with characteristics useful for improving modern cotton cultivars. However, much of this germplasm is not well utilized. The objective of this study was to evaluate agronomic and fiber traits of four germplasm populations to determine the effectiveness of pooling germplasm for generating variability to improve traits of interest. Four populations were developed with parents chosen based on (1) dwarfing genes, (2) a combination of fiber strength and length traits, (3) glandless genes, or (4) inclusion in the G. hirsutum center-of-origin, i.e. race, germplasm group. The dwarf germplasm population had smaller bolls, a smaller lint index, a smaller seed index, lower micronaire, and shorter fibers than the other three populations. There were no significant differences in lint yield, elongation, or strength among the germplasm populations. In contrast to the general lack of significant differences among populations for agronomic and fiber traits, within population variation was observed to be high. Therefore, selections could be made within the race population to raise lint yields and within the fiber population to increase fiber length. Likewise, selections within the glandless population could be made for boll size, lint index, seed index, micronaire, and strength. These results suggest that intercrossing multiple parents in complex populations generates a large amount of variability with potential uses in crop improvement.

Variation and relationship of quality and near infrared spectral characteristics of cotton fibers collected from multi-location field performance trials

Standardized instrument for testing of cotton (SITC) and advanced fiber information system (AFIS) measurements are increasingly being utilized as primary and routine means of acquiring fiber quality data by cotton breeders and fiber processors. A significant amount of information regarding fiber and yarn qualities is present, but little information exists about the compositional and chemical structure difference of cotton fibers harvested at different locations. Such information could prove useful in attempts to understand the variety selection of cotton cultivars. The purpose of this study was to characterize the fiber SITC and AFIS quality and also yarn skein strength of cottons harvested from various locations, and also to unravel the near infrared (NIR) spectral response to these differential environments. Moderate positive or negative relationships among fiber properties were observed. However, these relationships varied across experimental locations and years. Further, the analysis of variance tests indicated substantial variations among genotypes for most fiber properties, but less detectable variation among genotypes for yarn tenacity. Interestingly, principal component analysis of NIR spectra enhanced the similarity or dissimilarity of cotton fibers harvested at differing locations, implying the feasibility of the NIR technique for site selection in future cotton variety trials.

Genotypic comparisons of chromosomes 01, 04, and 18 from three tetraploid species of Gossypium in topcrosses with five elite cultivars of G. hirsutum L.

Upland cotton, Gossypium hirsutum L. is the most widely planted cultivated cotton in the United States and the world. The other cultivated tetraploid species G. barbadense L. is planted on considerable less area; however, it produces extra long, strong, and fine fibers which spins into superior yarn. The wild cotton tetraploid species G. tomentosum Nuttall ex Seemann, native to the Hawaiian Archipelago also exhibits traits, such as drought tolerance, that would also be desirable to transfer to Upland cotton. Long-term breeding efforts using whole genome crosses between Upland and these species have not been successful in transferring very many desirable alleles into Upland cotton. Our chromosome substitution lines (CSL) have one chromosome or chromosome arm from an alien species backcrossed into the Upland cotton line,TM-1, via aneuploid technology. Five Upland cultivars were crossed with CS-B01, CS-T01, CS-B04, CS-T04, CS-B18 and CS-T18 and TM-1 the recurrent parent of the CSLs. This provided an opportunity to determine the effects of chromosomes 01, 04, and 18 from the three species in crosses with the five cultivars. Predicted genotypic mean effects of the parents, F2, and F3 generations for eight agronomic and fiber traits of importance were compared. The predicted hybrid mean effects for the three chromosomes from each species were different for several of the traits across cultivars. There was no single chromosome or species that was superior for all traits in crosses. Parental and hybrid lines often differed in the effect of a particular chromosome among the three species. The predicted genotypic mean effects for F2 and F3, with a few exceptions, generally agree with our previous results for additive and dominance genetic effects of these CSL.