Jinping Hua

Analysis of Complete Nucleotide Sequences of 12 Gossypium Chloroplast Genomes: Origin and Evolution of Allotetraploids

Background Cotton (Gossypium spp.) is a model system for the analysis of polyploidization. Although ascertaining the donor species of allotetraploid cotton has been intensively studied, sequence comparison of Gossypium chloroplast genomes is still of interest to understand the mechanisms underlining the evolution of Gossypium allotetraploids, while it is generally accepted that the parents were A- and D-genome containing species. Here we performed a comparative analysis of 13 Gossypium chloroplast genomes, twelve of which are presented here for the first time. Methodology/Principal Findings The size of 12 chloroplast genomes under study varied from 159,959 bp to 160,433 bp. The chromosomes were highly similar having >98% sequence identity. They encoded the same set of 112 unique genes which occurred in a uniform order with only slightly different boundary junctions. Divergence due to indels as well as substitutions was examined separately for genome, coding and noncoding sequences. The genome divergence was estimated as 0.374% to 0.583% between allotetraploid species and A-genome, and 0.159% to 0.454% within allotetraploids. Forty protein-coding genes were completely identical at the protein level, and 20 intergenic sequences were completely conserved. The 9 allotetraploids shared 5 insertions and 9 deletions in whole genome, and 7-bp substitutions in protein-coding genes. The phylogenetic tree confirmed a close relationship between allotetraploids and the ancestor of A-genome, and the allotetraploids were divided into four separate groups. Progenitor allotetraploid cotton originated 0.43–0.68 million years ago (MYA). Conclusion Despite high degree of conservation between the Gossypium chloroplast genomes, sequence variations among species could still be detected. Gossypium chloroplast genomes preferred for 5-bp indels and 1–3-bp indels are mainly attributed to the SSR polymorphisms. This study supports that the common ancestor of diploid A-genome species in Gossypium is the maternal source of extant allotetraploid species and allotetraploids have a monophyletic origin. G. hirsutum AD1 lineages have experienced more sequence variations than other allotetraploids in intergenic regions. The available complete nucleotide sequences of 12 Gossypium chloroplast genomes should facilitate studies to uncover the molecular mechanisms of compartmental co-evolution and speciation of Gossypium allotetraploids.

The Complete Mitochondrial Genome of Gossypium hirsutum and Evolutionary Analysis of Higher Plant Mitochondrial Genomes

Background Mitochondria are the main manufacturers of cellular ATP in eukaryotes. The plant mitochondrial genome contains large number of foreign DNA and repeated sequences undergone frequently intramolecular recombination. Upland Cotton (Gossypium hirsutum L.) is one of the main natural fiber crops and also an important oil-producing plant in the world. Sequencing of the cotton mitochondrial (mt) genome could be helpful for the evolution research of plant mt genomes. Methodology/Principal Findings We utilized 454 technology for sequencing and combined with Fosmid library of the Gossypium hirsutum mt genome screening and positive clones sequencing and conducted a series of evolutionary analysis on Cycas taitungensis and 24 angiosperms mt genomes. After data assembling and contigs joining, the complete mitochondrial genome sequence of G. hirsutum was obtained. The completed G.hirsutum mt genome is 621,884 bp in length, and contained 68 genes, including 35 protein genes, four rRNA genes and 29 tRNA genes. Five gene clusters are found conserved in all plant mt genomes; one and four clusters are specifically conserved in monocots and dicots, respectively. Homologous sequences are distributed along the plant mt genomes and species closely related share the most homologous sequences. For species that have both mt and chloroplast genome sequences available, we checked the location of cp-like migration and found several fragments closely linked with mitochondrial genes. Conclusion The G. hirsutum mt genome possesses most of the common characters of higher plant mt genomes. The existence of syntenic gene clusters, as well as the conservation of some intergenic sequences and genic content among the plant mt genomes suggest that evolution of mt genomes is consistent with plant taxonomy but independent among different species.

Dynamic QTL and epistasis analysis on seedling root traits in upland cotton

Roots are involved in acquisition of water and nutrients, as well as in providing structural support to plant. The root system provides a dynamic model for developmental analysis. Here, we investigated quantitative trait loci (QTL), dynamic conditional QTL and epistatic interactions for seedling root traits using an upland cotton F2 population and a constructed genetic map. Totally, 37 QTLs for root traits, 35 dynamic conditional QTLs based on the net increased amount of root traits (root tips, forks, length, surface area and volume) (i) after transplanting 10 days compared to 5 days, and (ii) after transplanting 15 days to 10 days were detected. Obvious dynamic characteristic of QTL and dynamic conditional QTL existed at different developmental stages of root because QTL and dynamic conditional QTL had not been detected simultaneously. We further confirmed that additive and dominance effects of QTL qRSA-chr1-1 in interval time 5 to 10 DAT (days after transplant) offset the effects in 10 to 15 DAT. Lots of two-locus interactions for root traits were identified unconditionally or dynamically, and a few epistatic interactions were only detected simultaneously in interval time of 5–10 DAT and 10–15 DAT, suggesting different interactive genetic mechanisms on root development at different stages. Dynamic conditional QTL and epistasis effects provide new attempts to understand the dynamics of roots and provide clues for root architecture selection in upland cotton.

Partial Dominance, Overdominance, Epistasis and QTL by Environment Interactions Contribute to Heterosis in Two Upland Cotton Hybrids.

Based on two recombinant inbred line (RIL) populations, two corresponding backcross (BC) populations were constructed to elucidate the genetic basis of heterosis in Upland cotton (Gossypium hirsutum L.). The yield, and yield components, of these populations were evaluated in three environments. At the single-locus level, 78 and 66 quantitative trait loci (QTL) were detected using composite interval mapping in RIL and BC populations, respectively, and 29 QTL were identified based on mid-parental heterosis (MPH) data of two hybrids. Considering all traits together, a total of 50 (64.9%) QTL with partial dominance effect, and 27 (35.1%) QTL for overdominance effect were identified in two BC populations. At the two-locus level, 120 and 88 QTL with main effects (M-QTL), and 335 and 99 QTL involved in digenic interactions (E-QTL), were detected by inclusive composite interval mapping in RIL and BC populations, respectively. A large number of QTL by environment interactions (QEs) for M-QTL and E-QTL were detected in three environments. For most traits, average E-QTL explained a larger proportion of phenotypic variation than did M-QTL in two RIL populations and two BC populations. It was concluded that partial dominance, overdominance, epistasis, and QEs all contribute to heterosis in Upland cotton, and that partial dominance resulting from single loci and epistasis play a relatively more important role than other genetic effects in heterosis in Upland cotton.

Seedling root QTLs analysis on dynamic development and upon nitrogen deficiency stress in Upland cotton

Roots are vital to plant anchorage and efficient uptake of water and nutrients, and play an important role in environmental fitness, performance and yield formation in crops. The main objective of this study was to dissect the developmental behavior of seedling root traits at different developmental stages, and try to explore stress effect of nitrogen deficiency at seedling stage by mapping quantitative trait loci (QTLs) using an intraspecific population of recombinant inbred lines in Upland cotton (Gossypium hirsutum L.). Totally, 34 QTLs and 27 conditional QTLs for root length, root surface area, root volume, number of root tips and number of root forks were detected, respectively, in which 18 were congruent QTLs. The performance of five seedling root traits and QTLs showed obvious dynamic characteristics at three developmental stages. Special conditional QTLs were detected by QTL mapping strategy, in which conditional QTLs with different effects were identified at certain stages, demonstrating that the expression of genes showed temporal characteristics during root development. After nitrogen deprivation and deficiency stress for five days, a total of 11 QTLs were identified. Of these, ten QTLs were also identified in previous three stages under normal nutritional condition, and one QTL qNRT-chr19-1 referring number of root tips was newly detected, which suggested that it might be related to stress response of N-deficiency. After nitrogen deprivation for eight days, a total of 5 QTLs for plant height, maximum root length, fresh weight of root, and root/shoot ratio were identified. These QTLs and dynamic QTLs might offer different clues to understand the developmental mechanism and genetic basis of seedling root traits and to select root trait architecture in breeding program of Upland cotton.

Overexpression of Heteromeric GhACCase Subunits Enhanced Oil Accumulation in Upland Cotton

Acetyl-CoA carboxylase (ACCase) catalyzes acetyl-CoA to produce malonyl-CoA, which is the essential first step in the biosynthesis of de novo fatty acids. The heteromeric ACCase of higher plants, such as upland cotton, is composed of four subunits: biotin carboxyl carrier protein (BCCP), biotin carboxylase (BC), α-subunit of carboxyltransferase (CTα), and β-subunit of carboxyltransferase (CTβ). In this study, four subunits encoding heteromeric ACCase were amplified and the structures were analyzed further. The result showed that 7, 16, 10, and 1 exon(s) were identified from GhBCCP1, GhBC1, GhCTα2, and GhCTβ, respectively. The expression pattern analysis showed that the transcripts of these four subunits were ubiquitous in all the tested tissues and may have a positive correlation between the four subunits and oil accumulation. The four subunits coordinate to enhance oil content. The expression patterns and levels varied under ABA, MeJA treatments, and cold stress. Furthermore, under the control of a seed-specific AGP promoter, overexpression of the four subunits separately in Upland cotton demonstrated that GhBCCP1 transgenic plants showed a significant increase of 21.92% in oil content in cotton seeds, while about a 17% increase was noted for GhBC1 and GhCTβ, respectively. So overexpression of each subunit of heteromeric GhACCase may effectively increase seed oil content in Upland cotton.

Plant Mitochondrial Genome Evolution and Cytoplasmic Male Sterility

ABSTRACT Mitochondria are responsible for providing energy currency to life processes in the molecular form of ATP and are therefore typically referred to as the power factories of cells. Plant mitochondria are also relevant to the common phenomenon of cytoplasmic male sterility, which is agronomically important in various crop species. Cytoplasmic male sterility (CMS) is a complex trait that may be influenced by patterns of mitochondrial genome evolution, and by intergenomic gene transfer among the organellar and nuclear compartments of plant cells. Here, we review patterns and processes that shape plant mitochondrial genomes, some relevant interactions between organelles, and the general features shared by the majority of cytoplasmic male-sterile genes in plants to further the goal of understanding CMS.

Chloroplast DNA Structural Variation, Phylogeny, and Age of Divergence among Diploid Cotton Species

The cotton genus (Gossypium spp.) contains 8 monophyletic diploid genome groups (A, B, C, D, E, F, G, K) and a single allotetraploid clade (AD). To gain insight into the phylogeny of Gossypium and molecular evolution of the chloroplast genome in this group, we performed a comparative analysis of 19 Gossypium chloroplast genomes, six reported here for the first time. Nucleotide distance in non-coding regions was about three times that of coding regions. As expected, distances were smaller within than among genome groups. Phylogenetic topologies based on nucleotide and indel data support for the resolution of the 8 genome groups into 6 clades. Phylogenetic analysis of indel distribution among the 19 genomes demonstrates contrasting evolutionary dynamics in different clades, with a parallel genome downsizing in two genome groups and a biased accumulation of insertions in the clade containing the cultivated cottons leading to large (for Gossypium) chloroplast genomes. Divergence time estimates derived from the cpDNA sequence suggest that the major diploid clades had diverged approximately 10 to 11 million years ago. The complete nucleotide sequences of 6 cpDNA genomes are provided, offering a resource for cytonuclear studies in Gossypium.

Molecular evolution of the plastid genome during diversification of the cotton genus

Cotton (Gossypium spp.) is commonly grouped into eight diploid genomic groups, designated A-G and K, and one tetraploid genomic group, namely AD. To gain insight into the phylogeny of Gossypium and molecular evolution of the chloroplast genome duringdiversification, chloroplast genomes (cpDNA) from 6 D-genome and 2 G-genome species of Gossypium (G. armourianum D2-1, G. harknessii D2-2, G. davidsonii D3-d, G. klotzschianum D3-k, G. aridum D4, G. trilobum D8, and G. australe G2, G. nelsonii G3) were newly reported here. In combination with the 26 previously released cpDNA sequences, we performed comparative phylogenetic analyses of 34 Gossypium chloroplast genomes that collectively represent most of the diversity in the genus. Gossypium chloroplasts span a small range in size that is mostly attributable to indels that occur in the large single copy (LSC) region of the genome. Phylogenetic analysis using a concatenation of all genes provides robust support for six major Gossypium clades, largely supporting earlier inferences but also revealing new information on intrageneric relationships. Using Theobroma cacao as an outgroup, diversification of the genus was dated, yielding results that are in accord with previous estimates of divergence times, but also offering new perspectives on the basal, early radiation of all major clades within the genus as well as gaps in the record indicative of extinctions. Like most higher-plant chloroplast genomes, all cotton species exhibit a conserved quadripartite structure, i.e., two large inverted repeats (IR) containing most of the ribosomal RNA genes, and two unique regions, LSC (large single sequence) and SSC (small single sequence). Within Gossypium, the IR-single copy region junctions are both variable and homoplasious among species. Two genes, accD and psaJ, exhibited greater rates of synonymous and non-synonymous substitutions than did other genes. Most genes exhibited Ka/Ks ratios suggestive of neutral evolution, with 8 exceptions distributed among one to several species. This research provides an overview of the molecular evolution of a single, large non-recombining molecular during the diversification of this important genus.

Overexpression of GhDof1 improved salt and cold tolerance and seed oil content in Gossypium hirsutum

A homologous GhDof1, which belongs to a large family of plant-specific transcription factor DOF, was isolated from Upland cotton (Gossypium hirsutum L.). GhDof1 protein was located in the nucleus of onion epidermal cells, the core domain of transcriptional activity existed in the C-terminal, and the activity elements of GhDof1 promoter existed in the regions of -645∼ -469bp and -286∼ -132bp of transcriptional start codon. GhDof1 constitutively expressed in leaves, roots and stems, accumulated highest in leaves. The salinity and cold treatments induced GhDof1 transcript accumulation. The GhDof1-overexpressed cotton showed significantly higher salt and cold tolerance over the wild-type plants. Under salt stress, the root growth of overexpressed GhDof1 lines was promoted. The expression levels of stress-responsive genes, GhP5CS, GhSOD and GhMYB, were differently up-regulated in transgenic lines. Oil contents increased in some transgenic plants, and protein contents reduced compared with transformed receptor. These results suggested that GhDof1 was a functional transcription factor for improving the abiotic tolerance and seed oil content in Upland cotton.