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GrainGenes Reference Report: PPS-130-1335

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Reference	PPS-130-1335
Title	Rubisco small subunit, chlorophyll a/b-binding protein and sucrose:fructan-6-fructosyl transferase gene expression and sugar status in single barley leaf cells in situ. Cell type specificity and induction by light
Journal	Plant Physiology
Year	2002
Volume	130
Pages	1335-1348
Author	Lu C [ Show all 6 ]
Abstract	Summary: We describe a highly efficient two-step single-cell reverse transcriptase-polymerase chain reaction technique for analyzing gene expression at the single-cell level. Good reproducibility and a linear dose response indicated that the technique has high specificity and sensitivity for detection and quantification of rare RNA. Actin could be used as an internal standard. The expression of message for Rubisco small subunit (RbcS), chlorophyll a/b-binding protein (Cab), sucrose (Suc):fructan-6-fructosyl transferase (6-SFT), and Actin were measured in individual photosynthetic cells of the barley (Hordeum vulgare) leaf. Only Actin was found in the non-photosynthetic epidermal cells. Cab, RbcS, and 6-SFT genes were expressed at a low level in mesophyll and parenchymatous bundle sheath (BS) cells when sampled from plants held in dark for 40 h. Expression increased considerably after illumination. The amount of 6-SFT, Cab, and RbcS transcript increased more in mesophyll cells than in the parenchymatous BS cells. The difference may be caused by different chloroplast structure and posttranscriptional control in mesophyll and BS cells. When similar single-cell samples were assayed for Suc, glucose, and fructan, there was high correlation between 6-SFT gene expression and Suc and glucose concentrations. This is consistent with Suc concentration being the trigger for transcription. Together with earlier demonstrations that the mesophyll cells have a higher sugar threshold for fructan polymerization, our data may indicate separate control of transcription and enzyme activity. Values for the sugar concentrations of the individual cell types are reported
Keyword	[ Hide all but 1 of 54 ] actin barley barley leaves barley protein biochemical pathways biosynthesis bundle sheath cells cell chain chlorophyll chloroplast concentration control dark detection difference dose response efficient enzyme epidermal cells epidermis (plant) food crops fructan fructans gene gene expression gene expression regulation gene induction glucose hexosyltransferases hordeum in situ induction light light intensity mesophyll messenger rna nucleotide sequence plant extracts polymerization protein quantification quantitative analysis reverse transcriptase polymerase chain reaction ribulose bisphosphate carboxylase rubisco sensitivity small subunit subunit sucrose sugar sugars transcript transcription

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