Chin Bull Bot ›› 2019, Vol. 54 ›› Issue (4): 509-514.doi: 10.11983/CBB19101


• TECHNIQUES AND METHODS • Previous Articles     Next Articles

Identification of Chilling Tolerance of Rice Seedlings by Cold Water Bath

Liu Dongfeng,Tang Yongyan,Luo Shengtao,Luo Wei,Li Zhitao,Chong Kang,Xu Yunyuan()   

  1. Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China
  • Received:2019-05-29 Accepted:2019-06-11 Online:2020-01-08 Published:2019-07-01
  • Contact: Xu Yunyuan


The premise of studies on rice chilling tolerance is to find an efficient and accurate way to evaluate chilling tolerance of rice seedling. In this study, we developed an efficient technology to evaluate the cold tolerance at seedling stage by using a constant temperature water bath, based on the characteristics of excellent temperature uniformity for circulation of water. In this method, the setting temperature of environment temperature and water bath was 20°C and 4°C, respectively. From the results of two subspecies (indica/xian and japonica/geng), we summarized the reference treatment time for different survival rate of cultivars. Some attention to the cold treatment procedure was also discussed.

Key words: rice, seedling, cold tolerance, low temperature water bath

Figure 1

Germination and sowing of rice seeds, and the way of seedling culture (A) Germination of rice seeds; (B) Placement of germination seeds in 96-well plates without bottom; (C) Trefoil stage seedlings (Bar=2 cm)"

Figure 2

The diagram of low temperature water bath treatment of rice seedlings (A) The temperature of water bath within 24 h; (B) Seedlings under chilling treatment in 4°C water bath; (C) Partially enlarged detail of the seedlings in the cold water bath tank; (D) A diagram of the fixation pattern of seedlings in cold water bath"

Figure 3

Effect of submerged depth of cold water on low temperature tolerance of rice Red arrow indicated the site of shoot apical meristem under different water depth in chilling treatment I, II and III. Dotted lines represent the water surface. The survival rates were indicated on the top of the seedlings recoveried for 7 days. Treatment with 28oC water was used as control (left). Bars=6 cm"

Figure 4

The evaluation of cold tolerance for different rice varieties at seedling stage (A) Phenotypic response to chilling in DJ, NIP and MH86, the survival rate was determined after cold-water treatment at 4°C for 96 h and subsequent recovery at 28°C for 7 days (means±SD, n≥30, three replicates); (B) The phenotype for seedlings of ZH10 and OX8 after the chilling treatment at 4°C for 120 h, subsequently at 28°C for 50 days. Bars=2 cm"

Table 2

The recommended treatment time for different survival rates of cultivars of indica/xian and japonica/geng"

Cultivars Survival rate (>50%) Survival rate (<10%)
KY131, Kitaake 60-80 h 96-120 h
DJ, HY 72-84 h 120-144 h
ZH10, NIP 72-96 h 120-144 h
YD 96-144 h 168-192 h
ZF802, GLA4, IR64 12-28 h 40-60 h
93-11, MH86 24-40 h 48-60 h
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