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Plant and Cell Physiology Advance Access originally published online on April 23, 2009
Plant and Cell Physiology 2009 50(6):1090-1098; doi:10.1093/pcp/pcp061
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

Molecular Mechanism of Seed Coat Discoloration Induced by Low Temperature in Yellow Soybean

Atsushi Kasai1,5, Shizen Ohnishi2,5, Hiroyuki Yamazaki2, Hideyuki Funatsuki3, Tasuku Kurauchi1, Takuro Matsumoto1, Setsuzo Yumoto2,4 and Mineo Senda1,*

1Faculty of Agriculture and Life Science, Hirosaki University, Hirosaki, 036-8561 Japan
2Tokachi Agricultural Experiment Station, S9-2, Shinsei, Memuro-cho, Kasai-gun, Hokkaido, 082-0071 Japan
3National Agricultural Research Center for Hokkaido Region, Hitsujigaoka, Sapporo, 062-8555 Japan

*Corresponding author: E-mail, senda{at}cc.hirosaki-u.ac.jp; Fax, +81-172-39-3757.


  Abstract

Seed coat pigmentation is inhibited in yellow soybean. The I gene inhibits pigmentation over the entire seed coat. In yellow soybean, seed coat discoloration occurs when plants are exposed to low temperatures after the onset of flowering, a phenomenon named ‘cold-induced discoloration (CD)’. Inhibition of seed coat pigmentation results from post-transcriptional gene silencing (PTGS) of the chalcone synthase (CHS) genes. PTGS is a sequence-specific RNA degradation mechanism in plants and occurs via short interfering RNAs (siRNAs). Similar post-transcriptional suppression is called RNAi (RNA inter-ference) in animals. Recently, we identified a candidate of the I gene designated GmIRCHS. In this study, to elucidate the molecular mechanism of CD, CHS mRNA and siRNA levels in the seed coat were compared between CD-sensitive and CD-tolerant cultivars (Toyomusume and Toyoharuka, respectively). In Toyomusume, the CHS siRNA level was reduced markedly by low temperature treatment, and subsequently the CHS mRNA level increased rapidly after treatment. In contrast, low temperature treatment did not result in severe reduction of the CHS siRNA level in Toyoharuka, and the CHS mRNA level did not increase after the treatment. These results suggest that the rapid increase in CHS mRNA level after low temperature treatment may lead to enhanced pigmentation in some of the seed coat cells and finally in seed coat discoloration. Interestingly, we found a Toyoharuka-specific difference in the GmIRCHS region, which may be involved in CD tolerance.

Keywords: CHS genes - Low temperature - PTGS - Seed coat discoloration - Tolerance - Yellow soybean

Abbreviations: ASGS, antisense-mediated gene silencing; CD, cold-induced discoloration; CHS, chalcone synthase; CMV, cucumber mosaic virus; DPT, days post-treatment; dsRNA, double-stranded RNA; ET, end of treatment; GmASCHS, Glycine max antisense CHS pseudogene; GmIRCHS, Glycine max inverted repeat of CHS pseudogene; IR, inverted repeat; miRNA, microRNA; PTGS, post-transcriptional gene silencing; RDR, RNA-dependent RNA polymerase; RFLP, restriction fragment length polymor-phism; RISC, RNA-induced silencing complex; RT–PCR, reverse transcription–PCR; siRNA, short interfering RNA; SMV, soybean mosaic virus; THM, Toyohomare pigmented mutant line; TM, Toyomusume; TR, Toyoharuka.

4Present address: National Agricultural Research Center for Tohoku Region, Kariwano, Daisen, 019-2112 Japan.

5These authors contributed equally to this work.

(Received March 25, 2009; Accepted April 21, 2009)
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