Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap

Gaupels, Frank; Buhtz, Anja; Knauer, Torsten; Deshmukh, S. D.; Waller, Frank; Bel, Aart J. E. van; Kogel, Karl‐Heinz; Kehr, Julia

doi:10.1093/jxb/ern181

Cited by 69 publications

(56 citation statements)

References 57 publications

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“…Both b-1,3-glucanase GIII (contig1636_at, contig1639_at) ) and GII (contig1637_s_at) (Hoj et al 1989) are extracellular proteins, and they may be secreted into the phloem. However, they were not found among proteins in phloem sap reported from barley (Gaupels et al 2008). Increased levels of b-1,3-glucanases may be involved in the breakdown of induced callose (Hao et al 2008).…”

Section: Discussionmentioning

confidence: 84%

The constitutive expression and induction of three β-1,3-glucanases by bird cherry-oat aphid in relation to aphid resistance in 15 barley breeding lines

Mehrabi

Åhman

Jonsson

2016

Arthropod-Plant Interactions

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In this study, 15 closely related barley genotypes were analyzed for the abundance of three b-1,3-glucanase transcripts immediately before and during infestation by the bird cherry-oat aphid (Rhopalosiphum padi L.). The barley lines are doubled haploid lines in backcross (BC) generations BC1 and BC2 from a cross between cultivar Lina and a wild barley accession. Previously, they have been characterized as susceptible (S) or resistant (R) to R. padi based on their ability to support nymphal growth. Here we also tested whether resistance was manifested as reduced aphid settling on the plants. Indeed, aphid numbers were lower on R than on S lines in all cases where there were significant differences between R and S lines. The choice of b-1,3-glucanase sequences is based on earlier results comparing two S and two R genotypes, suggesting that at least two of the three studied sequences are susceptibility factors. The comparisons of transcript abundance in plants with aphids showed for two of the b-1,3-glucanase sequences that there were several cases where an S genotype had significantly higher abundance than an R genotype, and in no case did an R line have significantly higher abundance than an S line. Thus, there was some support for the idea that b-1,3-glucanase sequences are susceptibility factors in the interaction between barley and R. padi.

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Section: Discussionmentioning

confidence: 84%

The constitutive expression and induction of three β-1,3-glucanases by bird cherry-oat aphid in relation to aphid resistance in 15 barley breeding lines

Mehrabi

Åhman

Jonsson

2016

Arthropod-Plant Interactions

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“…Phloem sap obtained this way can be used to analyze proteins, small molecules, lipids, and RNAs and has been used successfully in many plants. While a limited amount of experiments has been performed in monocots 11 , the method appears to be more suitable for dicots (Perilla 17,20 , Arabidopsis . Collection of exudates has to occur in a humid environment to avoid loss of exudates through transpiration.…”

Section: Edta Chelates Camentioning

confidence: 99%

Collection and Analysis of <em>Arabidopsis</em> Phloem Exudates Using the EDTA-facilitated Method

Tetyuk

Benning

Hoffmann-Benning

2013

JoVE

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The plant phloem is essential for the long-distance transport of (photo-) assimilates as well as of signals conveying biotic or abiotic stress. It contains sugars, amino acids, proteins, RNA, lipids and other metabolites. While there is a large interest in understanding the composition and function of the phloem, the role of many of these molecules and thus, their importance in plant development and stress response has yet to be determined. One barrier to phloem analysis lies in the fact that the phloem seals itself upon wounding. As a result, the number of plants from which phloem sap can be obtained is limited. One method that allows collection of phloem exudates from several plant species without added equipment is the EDTA-facilitated phloem exudate collection described here. While it is easy to use, it does lead to the wounding of cells and care has to be taken to remove contents of damaged cells. In addition, several controls to prove purity of the exudate are necessary. Because it is an exudation rather than a direct collection of the phloem sap (not possible in many species) only relative quantification of its contents can occur. The advantage of this method over others is that it can be used in many herbaceous or woody plant species (Perilla, Arabidopsis, poplar, etc.) and requires minimal equipment and training. It leads to reasonably large amounts of exudates that can be used for subsequent analysis of proteins, sugars, lipids, RNA, viruses and metabolites. It is simple enough that it can be used in both a research as well as in a teaching laboratory.

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“…This information superhighway has been implicated in regulating development, responding to biotic stress, delivering nutrients, and as a vehicle commandeered by viruses for spreading infections (Lough and Lucas, 2006). Numerous full-length transcripts have been identified in the sieve elements of several plant species (Asano et al, 2002;Vilaine et al, 2003;Omid et al, 2007;Deeken et al, 2008;Gaupels et al, 2008;Kehr and Buhtz, 2008). One of these mobile RNAs is StBEL5, a BEL1-like transcription factor that is expressed in potato (Solanum tuberosum; Banerjee et al, 2006a).…”

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The Impact of the Long-Distance Transport of aBEL1-Like Messenger RNA on Development

et al. 2012

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BEL1-and KNOTTED1-type proteins are transcription factors from the three-amino-loop-extension superclass that interact in a tandem complex to regulate the expression of target genes. In potato (Solanum tuberosum), StBEL5 and its Knox protein partner regulate tuberization by targeting genes that control growth. RNA movement assays demonstrated that StBEL5 transcripts move through the phloem to stolon tips, the site of tuber induction. StBEL5 messenger RNA originates in the leaf, and its movement to stolons is induced by a short-day photoperiod. Here, we report the movement of StBEL5 RNA to roots correlated with increased growth, changes in morphology, and accumulation of GA2-oxidase1, YUCCA1a, and ISOPENTENYL TRANSFERASE transcripts. Transcription of StBEL5 in leaves is induced by light but insensitive to photoperiod, whereas in stolon tips growing in the dark, promoter activity is enhanced by short days. The heterodimer of StBEL5 and POTH1, a KNOTTED1-type transcription factor, binds to a tandem TTGAC-TTGAC motif that is essential for regulating transcription. The discovery of an inverted tandem motif in the StBEL5 promoter with TTGAC motifs on opposite strands may explain the induction of StBEL5 promoter activity in stolon tips under short days. Using transgenic potato lines, deletion of one of the TTGAC motifs from the StBEL5 promoter results in the reduction of GUS activity in new tubers and roots. Gel-shift assays demonstrate BEL5/POTH1 binding specificity to the motifs present in the StBEL5 promoter and a double tandem motif present in the StGA2-oxidase1 promoter. These results suggest that, in addition to tuberization, the movement of StBEL5 messenger RNA regulates other aspects of vegetative development.

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Adaptation of aphid stylectomy for analyses of proteins and mRNAs in barley phloem sap

Cited by 69 publications

References 57 publications

The constitutive expression and induction of three β-1,3-glucanases by bird cherry-oat aphid in relation to aphid resistance in 15 barley breeding lines

The constitutive expression and induction of three β-1,3-glucanases by bird cherry-oat aphid in relation to aphid resistance in 15 barley breeding lines

Collection and Analysis of <em>Arabidopsis</em> Phloem Exudates Using the EDTA-facilitated Method

The Impact of the Long-Distance Transport of aBEL1-Like Messenger RNA on Development

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