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作物学报 ›› 2021, Vol. 47 ›› Issue (9): 1768-1778.doi: 10.3724/SP.J.1006.2021.04170

• 研究论文 • 上一篇    下一篇

低温胁迫对普通和高油酸花生种子萌发的影响

薛晓梦1(), 吴洁1, 王欣1, 白冬梅2, 胡美玲1, 晏立英1, 陈玉宁1, 康彦平1, 王志慧1, 淮东欣1,*(), 雷永1, 廖伯寿1,*()   

  1. 1中国农业科学院油料作物研究所 / 农业农村部油料作物生物学与遗传育种重点实验室, 湖北武汉 430062
    2山西省农业科学院经济作物研究所, 山西汾阳 032200
  • 收稿日期:2020-07-26 接受日期:2021-01-21 出版日期:2021-09-12 网络出版日期:2021-02-23
  • 通讯作者: 淮东欣,廖伯寿
  • 作者简介:E-mail: xiaomengxue1991@163.com
  • 基金资助:
    国家重点研发计划项目“大田经济作物优质丰产的生理基础与调控”(2018YFD1000900);广东省重点领域研发计划项目(2020B020219003)

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 Published:2021-09-12 Published online:2021-02-23
  • Contact: HUAI Dong-Xin,LIAO Bo-Shou
  • 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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摘要：

高油酸花生以其营养价值高和耐储藏等特点深受广大消费者和加工企业的喜爱。近年来, 随着高油酸花生品种在我国的推广应用, 高油酸花生在高海拔、高纬度地区的发芽期耐寒性成为关注热点。为探究花生种子的油酸含量与其萌发期耐寒性的相关性, 本研究调查6组不同遗传背景的花生品种及其高油酸回交后代品系(BC4F8)在低温条件下的发芽率和发芽指数发现, 在低温胁迫下花生的萌发期耐寒性与其油酸含量无显著相关性。在泉花551高油酸后代品系(Quanhua 551-HO)低温发芽率显著低于其普通油酸含量亲本(Quanhua 551-NO)的组合中, 追踪分析8种主要脂肪酸在低温胁迫萌发过程中含量的变化发现, 在低温胁迫下Quanhua 551-NO中油酸含量显著减少且亚油酸含量显著增加, 而Quanhua 551-HO中也表现出了油酸含量减少、亚油酸含量增加的趋势, 但是未达到显著水平。进而分析上述2种材料中低温胁迫下各脂肪酸脱氢酶(fatty acid desaturase 2, FAD2)基因的表达模式发现, Quanhua 551-NO中AhFAD2-1A/B受低温诱导显著上调表达, 而AhFAD2-4A/B显著下调表达; 在Quanhua 551-HO中AhFAD2-4A/B受低温诱导持续显著上调表达, 而AhFAD2-1A/B显著下调表达, 推测由于高油酸花生中AhFAD2-1A/B编码蛋白失活, AhFAD2-4A/B在低温诱导下高量表达, 部分弥补了AhFAD2-1A/B缺失的功能。综上所述, 花生种子中油酸含量并不是决定其萌发期耐寒性的关键因素。

关键词: 高油酸花生, 低温胁迫, 发芽率, 脂肪酸脱氢酶(FAD2)

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)

表1

供试材料及其油酸含量"

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

表2

Real time PCR引物及其碱基序列"

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

图1

低温胁迫对不同品种花生的高油酸和普通油酸种子发芽率影响 NO: 普通油酸花生; HO: 高油酸花生。*、**和***分别表示在0.05、0.01和0.001水平显著差异。"

图2

低温胁迫下不同品种花生高油酸和普通油酸种子的发芽指数 CK: 种子在25℃条件下萌发6 d; T: 种子第1~3天在15℃条件下萌发, 第4~6天在25℃条件下萌发; NO: 普通油酸花生; HO: 高油酸花生。经单因素方差分析差异达到显著水平, 标以不同小写字母的柱值表示经单因素方差分析和最小显著差异法(LSD)检验差异达到显著水平(P < 0.05)。 "

图3

泉花551高油酸花生和普通油酸花生在不同温度处理下的发芽情况 25-NO: 泉花551普通油酸花生在25℃条件下萌发6 d; 15-NO: 第1~3天泉花551普通油酸花生在15℃条件下萌发, 第4~6天在25℃条件下萌发; 25-HO: 泉花551高油酸花生在25℃条件下萌发6 d; 15-HO: 第1~3天泉花551高油酸花生在15℃条件下萌发, 第4~6天在25℃条件下萌发。"

图4

低温胁迫下Quanhua 551种子发芽过程中脂肪酸含量的变化情况 C16:0: 棕榈酸; C18:0: 硬脂酸; C18:1: 油酸; C18:2: 亚油酸; C20:0: 花生酸; C20:1: 花生烯酸; C22:0: 山嵛酸; C24:0: 木蜡酸。CK: 种子在25℃条件下萌发6 d; T: 种子第1~3天在15℃条件下萌发, 第4~6天在25℃条件下萌发; NO: 普通油酸花生; HO: 高油酸花生。*表示在0.05水平差异显著。"

图5

低温胁迫下各AhFAD2基因在泉花551高油酸花生和普通油酸花生中的表达模式 a: 在不同温度处理条件下, AhFAD2-1A/B在Quanhua 551-NO中的表达模式; b: 在不同温度处理条件下, AhFAD2-3A/B在Quanhua 551-NO中的表达模式; c: 在不同温度处理条件下, AhFAD2-4A/B在Quanhua 551-NO中的表达模式; d: 在不同温度处理条件下, AhFAD2-1A/B在Quanhua 551-HO花生中的表达模式; e: 在不同温度处理条件下, AhFAD2-3A/B在Quanhua 551-HO中的表达模式; f: 在不同温度处理条件下, AhFAD2-4A/B在Quanhua 551-HO中的表达模式。CK: 种子在25℃条件下萌发6 d; T: 种子第1~3 天在15℃条件下萌发, 第4~6天在25℃条件下萌发; NO: 普通油酸花生; HO: 高油酸花生。*、**和***分别表示在0.05、0.01和0.001水平显著差异。 "

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