Immunofluorescence localization of ?-amylase in the scutellum, germ aleurone and ?normal? aleurone of germinated barley grains

Pogson, Barry J.; Ashford, A. E.; Gubler, Frank

doi:10.1007/bf01403450

Cited by 17 publications

(5 citation statements)

References 28 publications

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“…Plugs of scutellar tissue, totally devoid of peripheral aleurone tissue, release ␣-amylase 50,58 . Substrate-film tests show that amylase, protease and ribonuclease first appear during germination next to the scutellar epithelium 59 , before enzyme is released from the aleurone layer, results that agree with immunological studies on particular enzymes 58,[60][61][62][63][64] . Hybridisation histochemistry shows that the m-RNA coding for (13, 14)-␤-D-glucanase appears first in the scutellum and then declines while rising in the aleurone layer 65 .…”

Section: LI 6spiw Sj Xli )Qfv]s Erh Xli %Piyvsri 0e]ivsupporting

confidence: 71%

Malt Modification - A Century Of Evolving Views

Briggs

2002

Journal of the Institute of Brewing

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This lecture is the 22nd to commemorate Horace Taberrer Brown, a truly remarkable polymath who made important contributions to brewing practice and science and to 'pure' science. The ideas that Brown and his collaborators developed in the 1890s regarding the origins of the enzymes which catalyse the modification of the endosperm of germinating barley and how modification progresses through the grain are summarized, then the stages by which our understanding has been enhanced are noted. The discovery of gibberellins, their use in malting, the role they play in germinating grain, and how this knowledge has suggested ways to make malts with reduced malting losses are outlined. The discrepant reported patterns of modification and true 'average' patterns are described. It is pointed out that abrasion does not work by inducing two-way modification, as originally proposed. This successful process probably depends on improved access of oxygen to the living grain tissues. Microbes on the grain compete with the embryo and aleurone layer for oxygen, to the detriment of germination and modification. Commercially acceptable ways in which the activities of microbes might be reduced (and malting enhanced) are discussed, including the use of plug-rinses at the end of steeps to reduce the microbial population and the quantity of the microbial nutrients which support growth and respiration and are dissolved in the water around the grains. A newly-proposed method for the bulk-selection of malting quality lines from mixed populations of barley is mentioned. Finally, the dangers to the malting and brewing industries in the U.K. of the continuing reduction in scientific research is indicated.

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Section: LI 6spiw Sj Xli )Qfv]s Erh Xli %Piyvsri 0e]ivsupporting

confidence: 71%

Malt Modification - A Century Of Evolving Views

Briggs

2002

Journal of the Institute of Brewing

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“…In long time exposures we interpret that there is some Ltp2 promoter activity also in barley germ aleurone. In germinating barley grains, m-amylase accumulates in the germ aleurone (Pogson et al, 1989) suggesting that this tissue utilizes a gene programme resembling that of the 'normal' aleurone layer.…”

Section: Discussionmentioning

confidence: 99%

The promoter of the barley aleurone‐specific gene encoding a putative 7 kDa lipid transfer protein confers aleurone cell‐specific expression in transgenic rice

Kalla

Shimamoto²,

Potter³

et al. 1994

The Plant Journal

144

105

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SummaryThis paper describes the aleurone-specific gene Ltp2 from barley, which encodes a putative 7 kDa nonspecific lipid transfer protein. As shown by Northern and in situ hybridization analyses, the Ltp2trenscript is present in barley aleurone cells shortly after the initiation of aleurone cell differentiation. The expression of Ltp2 Increases until grain mid-maturity, but the mRNA is absent from mature grains. The Ltp2 transcript is undstectable in the embryo and vegetative tissues, confirming the aleurone specificity of the Ltp2 gene. The ability of the isolated 801 bp Ltp2 promoter to direct aleurone-specific expression in immature barley grains is demonstrated by perticle bombardment experiments. In these experiments, the activity of the Ltp2 promoter is 5% of the activity of the strong constitutive Actinl promoter from rice, as quantified by GUS activity measurements. In stably transformed rice plants containing the Ltp2 promoter-Gus construct, the specificity of the Ltp2 promoter is confirmed in vivo by the presence of GUS activity exclusively in the aleurone layer. This study demonstrates the conserved nature of the regulatory signals involved in aleurone-specific gene transcription in cereal grains.

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“…To fix water soluble proteins within the tissues the polystyrene films containing the grain sections were immediately after sectioning treated with fixation medium in 2% paraformaldehyde and 0.5% glutaraldehyde, buffered with 0.03 M 1,4-piperazinediethanesulfonic acid (PIPES) buffer, pH 7.5, containing 0.01 M CaCl, at 20°C for 30 s (POGSON et al 1989). The dehydration procedure was left out.…”

Section: Fixation and Immunohistochemistrymentioning

confidence: 99%

Developmental Patterns of Enzymes and Proteins During Mobilization of Endosperm Stores in Germinating Barley Grains

Jensen

2004

Hereditas

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Mobilization of stored reserves in the endosperm of barley grains involves the interaction of enzymes and other proteins synthesized during grain development and those synthesized during germination. In order to obtain information on this interaction, localization of a-amylase, the bifunctional subtilisinla-amylase inhibitor (BASI), the unspecific lipid transfer protein (LTP), and the antimicrobial proteins chitinase and ribosome inactivating protein (RIP) were determined by immunofluorescence microscopy in the different tissues of resting and germinating grains of the Hordeum uulgare L. genotypes Alexis, Klages, and Steptoe. Targeting of a-amylase from the proximal to the distal part of the endosperm progresses with different rates in the three genotypes. LTP is localized in the endosperms, the aleurone layers and in the embryo of the ungerminated grains. LTP is depleted during the first days of germination but then re-synthesized in the embryo and in the aleurone during the following phases of germination and released to the endosperm independent of cell wall hydrolysis. Chitinase is present in the aleurone layers and, depending on variety, in variable amounts in the endosperm of the mature and the germinating grains. Secretion of chitinase from the aleurone and scutellum is indicated by the increasing amount found in the endosperm during germination. Chitinase accumulates in the distal aleurone layers before a-amylase IS detected and thus must be able to penetrate the intact endosperm 0-glucan walls. BASI and RIP are localized in all parts of the mature grain and at later stages of germination the amount in the scutellum and aleurone layer increases. The analysed enzymes and proteins fulfill different functions during germination and are of importance for the industrial use of the barley crop. The efficient immunohistochemical demonstration of the genotypic bility in these characteristics provides a possibility to identify important breeding targets.

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Immunofluorescence localization of ?-amylase in the scutellum, germ aleurone and ?normal? aleurone of germinated barley grains

Cited by 17 publications

References 28 publications

Malt Modification - A Century Of Evolving Views

Malt Modification - A Century Of Evolving Views

The promoter of the barley aleurone‐specific gene encoding a putative 7 kDa lipid transfer protein confers aleurone cell‐specific expression in transgenic rice

Developmental Patterns of Enzymes and Proteins During Mobilization of Endosperm Stores in Germinating Barley Grains

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