Dataset Information

Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

ABSTRACT: Premise:Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of Brassica rapa. Methods:FAA (formaldehyde, glacial acetic acid, and ethanol)-fixed samples were stored in ethanol. Without performing any additional clearing or staining, we tested two methods of image acquisition at three locations along the proximal-distal axis of the laminae and estimated the patterns of venation using the program phenoVein. We developed and made available an R script to handle the phenoVein output and then analyzed our data using linear mixed-effects models. Results:Beyond fixation and storage, staining and clearing are not necessary to estimate leaf venation using phenoVein if the images are acquired using a stereomicroscope. All estimates of venation required some manual adjustment. We found a significant effect of location within the laminae for all aspects of venation. Discussion:By removing the clearing and staining steps and utilizing the semi-automated program phenoVein, we quickly and cheaply acquired leaf venation data. Venation may be an important target for crop breeding efforts, particularly if intralaminar variation correlates with variation in physiological processes, which remains an open question.

SUBMITTER: Newsome EL

PROVIDER: S-EPMC7249269 | biostudies-literature | 2020 May

REPOSITORIES: biostudies-literature

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Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

Newsome Eastyn L EL Brock Grace L GL Lutz Jared J Baker Robert L RL

Applications in plant sciences 20200511 5

<h4>Premise</h4>Physiological processes may vary within leaf laminae; however, the accompanying heterogeneity in leaf venation is rarely investigated because its quantification can be time consuming. Here we introduce accelerated protocols using existing software to increase sample throughput and ask whether laminae venation varies among three crop types and four subspecies of <i>Brassica rapa</i>.<h4>Methods</h4>FAA (formaldehyde, glacial acetic acid, and ethanol)-fixed samples were stored in e ...[more]

PMID: 32477842

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Project description:Recent advances in genomics indicate functional significance of a majority of genome sequences and their long range interactions. As a detailed examination of genome organization and function requires very high quality genome sequence, the objective of this study was to improve reference genome assembly of banana (Musa acuminata).We have developed a modular bioinformatics pipeline to improve genome sequence assemblies, which can handle various types of data. The pipeline comprises several semi-automated tools. However, unlike classical automated tools that are based on global parameters, the semi-automated tools proposed an expert mode for a user who can decide on suggested improvements through local compromises. The pipeline was used to improve the draft genome sequence of Musa acuminata. Genotyping by sequencing (GBS) of a segregating population and paired-end sequencing were used to detect and correct scaffold misassemblies. Long insert size paired-end reads identified scaffold junctions and fusions missed by automated assembly methods. GBS markers were used to anchor scaffolds to pseudo-molecules with a new bioinformatics approach that avoids the tedious step of marker ordering during genetic map construction. Furthermore, a genome map was constructed and used to assemble scaffolds into super scaffolds. Finally, a consensus gene annotation was projected on the new assembly from two pre-existing annotations. This approach reduced the total Musa scaffold number from 7513 to 1532 (i.e. by 80%), with an N50 that increased from 1.3 Mb (65 scaffolds) to 3.0 Mb (26 scaffolds). 89.5% of the assembly was anchored to the 11 Musa chromosomes compared to the previous 70%. Unknown sites (N) were reduced from 17.3 to 10.0%.The release of the Musa acuminata reference genome version 2 provides a platform for detailed analysis of banana genome variation, function and evolution. Bioinformatics tools developed in this work can be used to improve genome sequence assemblies in other species.

Dataset Information

Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

Publications

Variation within laminae: Semi-automated methods for quantifying leaf venation using phenoVein.

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