A modified method for enzymatic isolation of and subsequent wax extraction from Arabidopsis thaliana leaf cuticle

Abstract Background The plant cuticle represents one of the major adaptations of vascular plants to terrestrial life. Cuticular permeability and chemical composition differ among species. Arabidopsis thaliana is a widely used model for biochemical and molecular genetic studies in plants. However, attempts to isolate the intact cuticle from fresh leaves of Arabidopsis have failed so far. The goal of this study was to optimise an enzymatic method for cuticle isolation of species with a thin cuticle and to test it on several A. thaliana wild types and mutants. Results We developed a method for isolation of thin cuticles that allows reducing the isolation time, the separation of abaxial and adaxial cuticles, and avoids formation of wrinkles. Optical microscopy was used for studying cuticle intactness and scanning electron microscopy for visualisation of external and internal cuticle structures after isolation. Wax extracts were analysed by GC–MS. Isolation of intact cuticle was successful for all tested plants. The wax compositions (very-long-chained fatty acids, alcohols and alkanes) of intact leaves and isolated cuticles of wild type Col-0 were compared. Conclusions We conclude that the optimised enzymatic method is suitable for the isolation of A. thaliana adaxial and abaxial cuticles. The isolated cuticles are suitable for microscopic observation. Analysis of wax composition revealed some discrepancies between isolated cuticles and intact leaves with a higher yield of wax in isolated cuticles.

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Then There Were Plenty-Ring Meristems Giving Rise to Many Stamen Whorls

Plants ◽

10.3390/plants10061140 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1140

Author(s):

Doudou Kong ◽

Annette Becker

Keyword(s):

Arabidopsis Thaliana ◽

Dynamic Systems ◽

Molecular Genetic ◽

Floral Organ ◽

Fixed Number ◽

Floral Meristem ◽

Ecological Value ◽

Genetic Studies ◽

Stamen Primordia ◽

Floral Meristems

Floral meristems are dynamic systems that generate floral organ primordia at their flanks and, in most species, terminate while giving rise to the gynoecium primordia. However, we find species with floral meristems that generate additional ring meristems repeatedly throughout angiosperm history. Ring meristems produce only stamen primordia, resulting in polystemous flowers (having stamen numbers more than double that of petals or sepals), and act independently of the floral meristem activity. Most of our knowledge on floral meristem regulation is derived from molecular genetic studies of Arabidopsis thaliana, a species with a fixed number of floral organs and, as such of only limited value for understanding ring meristem function, regulation, and ecological value. This review provides an overview of the main molecular players regulating floral meristem activity in A. thaliana and summarizes our knowledge of ring primordia morphology and occurrence in dicots. Our work provides a first step toward understanding the significance and molecular genetics of ring meristem regulation and evolution.

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Genomic biotechnology of assessment and selection of linseed breeding material

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v21.802 ◽

1970 ◽

Vol 21 ◽

pp. 33-36

Author(s):

V. A. Lemesh ◽

M. V. Bogdanova ◽

E. L. Andronyk ◽

I. A. Golub

Keyword(s):

Fatty Acids ◽

Linolenic Acid ◽

Linseed Oil ◽

Molecular Genetic ◽

Fatty Acid Desaturase ◽

Wild Type ◽

Genetic Studies ◽

Breeding Material ◽

Linum Usitatissimum L ◽

Selection Of

Aim. The aim of this study was to develop of genomic biotechnology for the assessment and selection of the linssed breeding material by a complex of genes controlling the ratio of fatty acids in seeds oil to the creation of a new linseed variety. Methods. Breeding studies were combined with molecular-genetic studies. Results. We developed the genomic biotechnology to detect the mutant alleles of linseed fad3A and fad3B genes responsible for reduced α-linolenic acid levels in linseed oil. Using this biotechnology, it was possible to classify plants as homozygous mutant, homozygous wild type, or heterozygous at fad3A and fad3B loci, that can be used to breed new linseed varieties of food or industrial quality. Conclusions. By results of 3-year molecular-genetic and breeding studies the variety "Dar" was created with using the developed genomic biotechnology of an assessment and selection of linseed breeding material by a complex of the genes controlling the ratio of fatty acids in seeds oil.Keywords: linseed (Linum usitatissimum L.), α-linolenic acid, fatty acid desaturase, fad3 genes.

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Quantification of Water Stress-Induced Osmotic Adjustment and Proline Accumulation for Arabidopsis thaliana Molecular Genetic Studies

Methods in Molecular Biology - Plant Stress Tolerance ◽

10.1007/978-1-60761-702-0_19 ◽

2010 ◽

pp. 301-315 ◽

Cited By ~ 9

Author(s):

Paul E. Verslues

Keyword(s):

Arabidopsis Thaliana ◽

Water Stress ◽

Osmotic Adjustment ◽

Molecular Genetic ◽

Proline Accumulation ◽

Genetic Studies

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γ-tubulin gene intron length polymorphism of Arabidopsis thaliana

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v26.1253 ◽

2020 ◽

Vol 26 ◽

pp. 120-124

Author(s):

A. M. Rabokon ◽

Yu. O. Bilonozhko ◽

A. S. PostovoitovA ◽

L. O. Kalafat ◽

Ya. V. Pirko ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Length Polymorphism ◽

Molecular Genetic ◽

Intron Length ◽

Degenerate Primers ◽

Tubulin Gene ◽

Specific Primers ◽

Intron Length Polymorphism ◽

Genetic Studies ◽

Dna Profile

Aims. Verification of the possibility of using the γ-tubulin gene intron length polymorphism method in genetic studies of plants on the example of Arabidopsis thaliana. Methods. The γ-tubulin gene intron length polymorphism evaluating method was used. Amplified fragments DNA were fractionated by electrophoresis in non-denaturing polyacrylamide gel. DNA bands were detected using silver nitrate staining. Results. Arabidopsis was first time analyzed using the γ-tubulin gene intron length polymorphism method. During amplification with degenerate primers 2 amplicons (520 bp and 555 bp) were formed in all samples. However, using selected arabidopsis-specific primers for the second intron of the γ-tubulin genes, it was possible to find several samples that differ in their DNA profile. Conclusions. It is established that the proposed method can be used in molecular genetic studies of plants. Moreover, the developed specific primers for γ-tubulin gene introns can probably be used both for the study of Arabidopsis and related species. The use of degenerate primers can be useful in the study of plants for which there is no information about their genome. Keywords: molecular-genetic markers, intron length polymorphism, γ-tubulin, A. thaliana.

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