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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (9): 1768-1778.doi: 10.3724/SP.J.1006.2021.04170

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Effects of cold stress on germination in peanut cultivars with normal and high content of oleic acid

XUE Xiao-Meng1(), WU JIE1, WANG Xin1, BAI Dong-Mei2, HU Mei-Ling1, YAN Li-Ying1, CHEN Yu-Ning1, KANG Yan-Ping1, WANG Zhi-Hui1, HUAI Dong-Xin1,*(), LEI Yong1, LIAO Bo-Shou1,*()   

  1. 1Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture and Rural Affairs / Oil Crops Research Institute, Chinese Academy of Agricultural Sciences, Wuhan 430062, Hubei, China
    2Industrial Crops Research Institute, Shanxi Academy of Agricultural Sciences, Fenyang 032200, Shanxi, China
  • Received:2020-07-26 Accepted:2021-01-21 Online:2021-09-12 Published:2021-02-23
  • Contact: HUAI Dong-Xin,LIAO Bo-Shou E-mail:xiaomengxue1991@163.com;dxhuai@caas.cn;lboshou@hotmail.com
  • Supported by:
    National Key Research and Development Program of China “Physiological Basis and Agronomic Management for High-quality and High-yield of Field Cash Crops”(2018YFD1000900);Key Area Research and Development Program of Guangdong Province(2020B020219003)

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Abstract:

High oleate (HO) peanut is highly popular for its improved nutrient value and strengthened storage stability among customers and peanut processing enterprises. In recent years, with the adoption of HO peanut in our country, the cold tolerance of peanut at germination stage in high altitude or high latitude area has become a major concern. To figure out the relationship between the seed oleic acid content and the cold tolerance at germination stage in peanut, the germination rate and germination index of seeds under cold stress were investigated among six peanut cultivars with normal content of oleic acid (NO) and their backcross-derived HO lines, respectively. The results showed that the oleic acid content was not significantly correlated with the cold tolerance at germination stage. The contents of eight main fatty acids under cold stress at germination stage were tracked in Quanhua 551 (Quanhua 551-NO) and its HO offspring line (Quanhua 551-HO), and the germination rate of Quanhua 551-HO was significantly lower than that of Quanhua 551-NO under cold stress. The oleic acid content of Quanhua 551-NO was significantly decreased while the linoleic acid content was significantly increased under cold stress. However, the contents of oleic acid and linoleic acid exhibited the same trend in Quanhua 551-HO, and there was no significant difference. The expression profiles offatty acid desaturase 2 (AhFAD2) genes in both Quanhua 551-NO and Quanhua 551-HO under cold stress revealed that the relative expression level of AhFAD2-1A/B was significantly up-regulated, while that of AhFAD2-4A/B was significantly down-regulated under cold stress in Quanhua 551-NO. Conversely, the relative expression level of AhFAD2-1A/B was significantly decreased, but the relative expression level of AhFAD2-4A/B was significantly increased under cold stress in Quanhua 551-HO. These results implied that the up-regulation of AhFAD2-4A/Bin HO peanut may partly compensate for lost function of AhFAD2-1A/Bin response to cold stress. In conclusion, the oleic acid content in seed was not the main factor to determinate the cold tolerance at germination stage.

Key words: high oleate peanut, cold stress, germination rate, fatty acid desaturase 2 (FAD2)

Table 1

Oleic acid contents of peanut cultivars in this study"

品种
Cultivar 		油酸含量
Oleic acid content (%) 		品种
Cultivars 		油酸含量
Oleic acid content (%)
徐花13号
Xuhua 13 		普通油酸NO 45.58 中花21
Zhonghua 21 		普通油酸NO 38.40
高油酸HO 81.14 高油酸HO 80.83
徐花9号
Xuhua 9 		普通油酸NO 49.73 泉花551
Quanhua 551 		普通油酸NO 50.72
高油酸HO 81.60 高油酸HO 80.62
中花16
Zhonghua 16 		普通油酸NO 50.56 漯花9号
Luohua 9 		普通油酸NO 47.90
高油酸HO 80.70 高油酸HO 80.50

Table 2

Primer sequences of qRT-PCR"

基因
Gene 		引物名称
Primer name 		引物序列
Primer sequence (5°-3°)
AhFAD2-1A, AhFAD2-1B RTAhFAD2-1-F ATCTGCTATATCACATAGCAACTCT
RTAhFAD2-1-R ACTGTTGCCAATGCTCCTCT
AhFAD2-3A, AhFAD2-3B RTAhFAD2-3-F GGTCTTATCCGTCTTGTCATGG
RTAhFAD2-3-R AGATGAATCGTAATGTGGCAATG
AhFAD2-4A, AhFAD2-4B RTAhFAD2-4-F TCATTCTGCCGGGAAGAGG
RTAhFAD2-4-R ATGGCGACATAGGCGAAAAT
AhActin Actin-F TAAGAACAATGTTGCCATACAGA
Actin-R GTTGCCTTGGATTATGAGC

Fig. 1

Effects of cold stress on germination rate and its corresponding normal oleate seeds in high oleate seeds NO: normal oleate peanut; HO: high oleate peanut. *, **, and *** mean significant difference at the 0.05, 0.01, and 0.001 probability levels, respectively."

Fig. 2

Germination index of high oleate seeds and normal oleate seeds in different peanut varieties under cold stress CK: control group, germination of seed at 25℃ for six days; T: treatment group, germination of seed at 15℃ for 1-3 days, and then 25℃ for 4-6 days; NO: normal oleate peanut; HO: high oleate peanut. Bars superscripted by different lowercase letters are significantly different at P < 0.05 by one-way ANOVA and least significant difference (LSD) test. "

Fig. 3

Seed germination of peanut with high oleate and normal oleate in Quanhua 551 at different temperatures 25-NO: the normal oleate peanut of Quanhua 551 germinated at 25℃ for six days; 15-NO: the normal oleate peanut of Quanhua 551 germinated at 15℃ from the 1st to 3rd day then at 25℃ for the last three days; 25-HO: the high oleate peanut of Quanhua 551 germinated at 25℃ for six days; 15-HO: the high oleate peanut of Quanhua 551 germinated at 15℃ from the 1st to 3rd day then at 25℃ from the 4th to 6th day."

Fig. 4

Changes of fatty acid contents at seed germination stage of Quanhua 551 under cold stress C16:0: palmitic acid; C18:0: stearic acid; C18:1: oleic acid; C18:2: linoleic acid; C20:0: arachidic acid; C20:1: gadoleic acid; C22:0: behenic acid; C24:0: lignoceric acid. CK: control group, germination of seed at 25℃ for six days; T: treatment group, germination of seed at 15℃ from the 1st to 3rd day, and then 25℃ from 4th to 6th day; NO: normal oleate peanut; HO: high oleate peanut. * means significant difference at the 0.05 probability levels."

Fig. 5

Relative expression levels of AhFAD2 genes in high oleate and normal oleate of Quanhua 551 under cold stress a: the expression pattern of AhFAD2-1A/B in Quanhua 551-NO at different temperatures; b: the expression pattern of AhFAD2-3A/B in Quanhua 551-NO at different temperatures; c: the expression pattern of AhFAD2-4A/B in Quanhua 551-NO at different temperatures; d: the expression pattern ofAhFAD2-1A/B in Quanhua 551-HO at different temperatures; e: the expression pattern ofAhFAD2-3A/B in Quanhua 551-HO at different temperatures; f: the expression pattern of AhFAD2-4A/B in Quanhua 551-HO at different temperatures. CK: control group, germination of seed at 25℃ for six days; T: treatment group, germination of seed at 15℃ from 1st to 3rd day, and then 25℃ from the 4th to 6th day; NO: normal oleate peanut; HO: high oleate peanut. *, **, and *** mean significant difference at the 0.05, 0.01, and 0.001 probability levels, respectively. "

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