Dataset Information

Integrated mapping and characterization of the gene underlying the okra leaf trait in Gossypium hirsutum L.

ABSTRACT: Diverse leaf morphology has been observed among accessions of Gossypium hirsutum, including okra leaf, which has advantages and disadvantages in cotton production. The okra leaf locus has been mapped to chromosome 15 of the Dt subgenome, but the underlying gene has yet to be identified. In this study, we used a combination of targeted association analysis, F2 population-based fine mapping, and comparative sequencing of orthologues to identify a candidate gene underlying the okra leaf trait in G. hirsutum. The okra leaf gene identified, GhOKRA, encoded a homeodomain leucine-zipper class I protein, whose closely related genes in several other plant species have been shown to be involved in regulating leaf morphology. The transcript levels of GhOKRA in shoot apices were positively correlated with the phenotypic expression of the okra leaf trait. Of the multiple sequence variations observed in the coding region among GrOKRA of Gossypium raimondii and GhOKRA-Dt of normal and okra/superokra leaf G. hirsutum accessions, a non-synonymous substitution near the N terminus and the variable protein sequences at the C terminus may be related to the leaf shape difference. Our results suggest that both transcription and protein activity of GhOKRA may be involved in regulating leaf shape. Furthermore, we found that non-reciprocal homoeologous recombination, or gene conversion, may have played a role in the origin of the okra leaf allele. Our results provided tools for further investigating and understanding the fundamental biological processes that are responsible for the cotton leaf shape variation and will help in the design of cotton plants with an ideal leaf shape for enhanced cotton production.

SUBMITTER: Zhu QH

PROVIDER: S-EPMC4737076 | biostudies-literature | 2016 Feb

REPOSITORIES: biostudies-literature

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Integrated mapping and characterization of the gene underlying the okra leaf trait in Gossypium hirsutum L.

Zhu Qian-Hao QH Zhang Jian J Liu Dexin D Stiller Warwick W Liu Dajun D Zhang Zhengsheng Z Llewellyn Danny D Wilson Iain I

Journal of experimental botany 20151112 3

Diverse leaf morphology has been observed among accessions of Gossypium hirsutum, including okra leaf, which has advantages and disadvantages in cotton production. The okra leaf locus has been mapped to chromosome 15 of the Dt subgenome, but the underlying gene has yet to be identified. In this study, we used a combination of targeted association analysis, F2 population-based fine mapping, and comparative sequencing of orthologues to identify a candidate gene underlying the okra leaf trait in G. ...[more]

PMID: 26567355

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Project description:Leaf shape is a vital agronomic trait that affects plant and canopy architecture, yield, and other production attributes of upland cotton. Compared with normal leaves, lobed leaves have potential advantages in improving canopy structure and increasing cotton yield. A chromosomal introgression segment from Gossypium barbadense L. conferring sub-okra leaf shape to Gossypium hirsutum L. was identified on chromosome D01. To determine the effects of this transferred sub-okra leaf shape on the leaf anatomical characteristics, photosynthesis-related traits, and yield of short-season cotton, we performed a field experiment with three sets of near-isogenic lines carrying okra, sub-okra, and normal leaf shape in Lu54 (L54) and Shizao 2 (SZ2) backgrounds. Compared with normal leaves, sub-okra leaves exhibited reduced leaf thickness and smaller leaf mass per area; moreover, the deeper lobes of sub-okra leaves improved the plant canopy structure by decreasing leaf area index by 11.24%-22.84%. Similarly, the intercepted PAR rate of lines with sub-okra leaf shape was also reduced. The chlorophyll content of sub-okra leaves was lower than that of okra and normal leaf shapes; however, the net photosynthetic rate of sub-okra leaves was 8.17%-29.81% higher than that of other leaf shapes at most growth stages. Although the biomass of lines with sub-okra leaf shape was less than that of lines with normal leaves, the average first harvest yield and total yield of lines with the sub-okra leaf shape increased by 6.36% and 5.72%, respectively, compared with those with normal leaves. Thus, improvements in the canopy structure and photosynthetic and physiological characteristics contributed to optimizing the light environment, thereby increasing the yield of lines with sub-okra leaf shape. Our results suggest that the sub-okra leaf trait from G. barbadense L. may have practical applications for cultivating short-season varieties with high photosynthetic efficiency, and improving yield, which will be advantageous for short-season varieties.

Project description:Background:Genetically modified cotton accounts for 64% of the world's cotton growing area (22.3 million hectares). The genome sequencing of the diploid cotton progenitors Gossypium raimondii and Gossypium arboreum as well as the cultivated Gossypium hirsutum has provided a wealth of genetic information that could be exploited for crop improvement. Unfortunately, gene functional characterization in cotton is lagging behind other economically important crops due to the low efficiency, lengthiness and technical complexity of the available stable transformation methods. We present here a simple, fast and efficient method for the transient transformation of G. hirsutum that can be used for gene characterization studies. Results:We developed a transient transformation system for gene characterization in upland cotton. Using ?-glucuronidase as a reporter for Agrobacterium-mediated transformation assays, we evaluated multiple transformation parameters such as Agrobacterium strain, bacterial density, length of co-cultivation, chemicals and surfactants, which can affect transformation efficiency. After the initial characterization, the Agrobacterium EHA105 strain was selected and a number of binary constructs used to perform gene characterization studies. 7-days-old cotton seedlings were co-cultivated with Agrobacterium and transient gene expression was observed 5 days after infection of the plants. Transcript levels of two different transgenes under the control of the cauliflower mosaic virus (CaMV) 35S promoter were quantified by real-time reverse transcription PCR (qRT-PCR) showing a 3-10 times increase over the levels observed in non-infected controls. The expression patterns driven by the promoters of two G. hirsutum genes as well as the subcellular localization of their corresponding proteins were studied using the new transient expression system and our observations were consistent with previously published results using Arabidopsis as a heterologous system. Conclusions:The Agrobacterium-mediated transient transformation method is a fast and easy transient expression system enabling high transient expression and transformation efficiency in upland cotton seedlings. Our method can be used for gene functional studies such as promoter characterization and protein subcellular localization in cotton, obviating the need to perform such studies in a heterologous system such as Arabidopsis.

Project description:BackgroundF-box proteins are substrate-recognition components of the Skp1-Rbx1-Cul1-F-box protein (SCF) ubiquitin ligases. By selectively targeting the key regulatory proteins or enzymes for ubiquitination and 26S proteasome mediated degradation, F-box proteins play diverse roles in plant growth/development and in the responses of plants to both environmental and endogenous signals. Studies of F-box proteins from the model plant Arabidopsis and from many additional plant species have demonstrated that they belong to a super gene family, and function across almost all aspects of the plant life cycle. However, systematic exploration of F-box family genes in the important fiber crop cotton (Gossypium hirsutum) has not been previously performed. The genome-wide analysis of the cotton F-box gene family is now possible thanks to the completion of several cotton genome sequencing projects.ResultsIn current study, we first conducted a genome-wide investigation of cotton F-box family genes by reference to the published F-box protein sequences from other plant species. 592 F-box protein encoding genes were identified in the Gossypium hirsutume acc.TM-1 genome and, subsequently, we were able to present their gene structures, chromosomal locations, syntenic relationships with their parent species. In addition, duplication modes analysis showed that cotton F-box genes were distributed to 26 chromosomes, with the maximum number of genes being detected on chromosome 5. Although the WGD (whole-genome duplication) mode seems play a dominant role during cotton F-box gene expansion process, other duplication modes including TD (tandem duplication), PD (proximal duplication), and TRD (transposed duplication) also contribute significantly to the evolutionary expansion of cotton F-box genes. Collectively, these bioinformatic analysis suggest possible evolutionary forces underlying F-box gene diversification. Additionally, we also conducted analyses of gene ontology, and expression profiles in silico, allowing identification of F-box gene members potentially involved in hormone signal transduction.ConclusionThe results of this study provide first insights into the Gossypium hirsutum F-box gene family, which lays the foundation for future studies of functionality, particularly those involving F-box protein family members that play a role in hormone signal transduction.

Dataset Information

Integrated mapping and characterization of the gene underlying the okra leaf trait in Gossypium hirsutum L.

Publications

Integrated mapping and characterization of the gene underlying the okra leaf trait in Gossypium hirsutum L.

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