Overexpression of glycine-rich RNA-binding protein in tomato renders fruits with higher protein content after cold storage

Abstract

Glycine-rich RNA-binding proteins (GR-RBPs) are involved in RNA processing and also some of them are output signals of the circadian clock. In tomato, one GR-RBP gene family (LeGRP1) is composed by three highly homologous genes (LeGRP1a-c); each one rendering three transcriptional products: the un-spliced pre-RNA (preLegrp1a-c), the mature mRNA (mLegrp1a-c) and the alternatively spliced mRNA (asLegrp1a-c). To get insight into their regulation and impact on RNA metabolism in fruits, Solanum lycopersicum cv. Micro-Tom was transformed with preLeGRP1a fused to the polygalacturonase promoter, which drives expression to fruits from the mature green stage. Our results demonstrated a complex positive regulation of LeGRPs, in which LeGRP1a overexpression led to the induction of the others LeGRP1 members. Even though the LeGRP1 transcription and the content of three LeGRPs proteins were affected, the overall LeGRP protein circadian rhythm profile was similar in transgenic and wild type (WT) fruits. However, when the fruits were kept at a chilling temperature after harvest, total protein content was significantly higher in transgenic than in WT fruits, and the content of some free amino acids was modified. The results obtained suggest a probable role of LeGRP1s: structural rearrangements and/or stabilization of mRNA to allow efficient processing of fruits under cold conditions.

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Abbreviations

CI:

chilling injury

CSD:

cold shock domain

DAA:

days after anthesis

GR-RBPs:

glycine-rich RNA-binding proteins

IG:

immature green

MG:

mature green

PG:

polygalacturonase

PITC:

phenylisothiocyanate

RT-qPCR:

reverse

transcription:

quantitative polymerase chain reaction

RBP:

RNA binding protein

RR:

red ripe

RRM:

RNA recognition motif

TEA:

triethylamine

WT:

wild-type

Y:

green yellowish

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Correspondence to G. L. Müller.

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Acknowledgments: This work was funded by the Argentine National Research Council and the National Agency for Science Promotion and Technology (PICT 2012 Nº19). The first two authors contributed equally to this work.

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Ruggieri, G.M., Triassi, A., Alvarez, C.E. et al. Overexpression of glycine-rich RNA-binding protein in tomato renders fruits with higher protein content after cold storage. Biol Plant 62, 501–510 (2018). https://doi.org/10.1007/s10535-018-0794-3

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Additional key words

  • amino acids
  • circadian rhythm
  • polygalacturonase promoter
  • RNA stabilization