, Volume 248, Issue 2, pp 369–379 | Cite as

Occurrence and tolerance mechanisms of seed cracking under low temperatures in soybean (Glycine max)

  • Mineo Senda
  • Michio Kawasaki
  • Miho Hiraoka
  • Kazuki Yamashita
  • Hayato Maeda
  • Naoya Yamaguchi
Original Article


Main conclusion

In soybean, occurrence of, or tolerance to, seed cracking under low temperatures may be related to the presence or absence, respectively, of proanthocyanidin accumulation in the seed coat dorsal region.

Soybean seeds sometimes undergo cracking during low temperatures in summer. In this study, we focused on the occurrence and tolerance mechanisms of low-temperature-induced seed cracking in the sensitive yellow soybean cultivar Yukihomare and the tolerant yellow soybean breeding line Toiku 248. Yukihomare exhibited seed cracking when subjected to a 21-day low-temperature treatment from 10 days after flowering. In yellow soybeans, seed coat pigmentation is inhibited, leading to low proanthocyanidin levels in the seed coat. Proanthocyanidins accumulated on the dorsal side of the seed coat in Yukihomare under the 21-day low-temperature treatment. In addition, a straight seed coat split occurred on the dorsal side at the full-sized seed stage, resulting in seed cracking in this cultivar. Conversely, proanthocyanidin accumulation was suppressed throughout the seed coat in low-temperature-treated Toiku 248. We propose the following mechanism of seed cracking: proanthocyanidin accumulation and subsequent lignin deposition under low temperatures affects the physical properties of the seed coat, making it more prone to splitting. Further analyses uncovered differences in the physical properties of the seed coat between Yukihomare and Toiku 248. In particular, seed coat hardness decreased in Yukihomare, but not in Toiku 248, under the low-temperature treatment. Seed coat flexibility was higher in Toiku 248 than in Yukihomare under the low-temperature treatment, suggesting that the seed coat of low-temperature-treated Toiku 248 is more flexible than that of low-temperature-treated Yukihomare. These physical properties of the Toiku 248 seed coat observed under low-temperature conditions may contribute to its seed-cracking tolerance.


4-Dimethylaminocinnamaldehyde Histochemical analysis Physical property Seed coat Texture analysis 



Cold-induced seed coat discoloration


Chalcone synthase




Developing non-SC seeds


Developing SC seeds




Quantitative trait locus


Seed cracking


Seed coat pigmented mutant of cv. Toyohomare


Breeding line Toiku 248


cv. Yukihomare



This work was supported by the Ministry of Agriculture, Forestry, and Fisheries of Japan [Genomics-based Technology for Agricultural Improvement, SFC1006 (to MS, MK, HM, and NY)] and the Japan Society for the Promotion of Science [KAKENHI, grant number 15K07268 (to MS)]. We thank Jennifer Smith, PhD, Emma Tacken, Ph.D., and Barbara Goodson, PhD from Edanz Group ( for editing a draft of this manuscript.

Compliance with ethical standards

Conflict of interest

Authors declare that they have no conflict of interest.

Supplementary material

425_2018_2912_MOESM1_ESM.pptx (10.9 mb)
Supplemental Fig. S1 Cross sections of seed coats of YH and T248 grown under normal conditions (YH-N and T248-N, respectively). Seed coat sections were stained with DMACA to show PA accumulation. a YH-N (hilum region), b YH-N (middle region), c YH-N (dorsal region), d T248-N (hilum region), e T248-N (middle region), f T248-N (dorsal region). Hilum, middle, and dorsal regions are shown in Fig. 2e. pa, parenchyma; pl, palisade layer; tb, tracheid bar. Scale bars 100 μm (a, d) and 20 μm (b, c, e, f) (PPTX 11174 kb)
425_2018_2912_MOESM2_ESM.pptx (43 kb)
Supplemental Fig. S2 Comparison of seed coat physical properties among YH-N, YH-L dNS, and YH-L dSS. Vertical bars represent mean ± SD [n = 51 for YH-N, n = 29 for YH-L (dNS), and n = 22 for YH-L (dSS)]. Different letters indicate significant differences at P < 0.05 by Tukey–Kramer’s test (PPTX 43 kb)


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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Faculty of Agriculture and Life ScienceHirosaki UniversityHirosakiJapan
  2. 2.Hokkaido Research Organization, Tokachi Agricultural Experiment StationKasai-gunJapan

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