谷子<i>SiPRR73</i>基因的光温调控模式及非生物胁迫响应特性

doi:10.7668/hbnxb.20193022

摘要/Abstract

摘要：

利用RT-PCR技术从延谷11号谷子中克隆SiPRR73基因,通过系统分析SiPRR73组织表达特异性,对不同光周期处理、不同光温组合处理以及5种非生物胁迫处理的响应特点,揭示光周期、温度对SiPRR73的调控模式及SiPRR73对非生物胁迫的应答模式。结果表明,从延谷11号克隆得到SiPRR73基因2 928 bp的cDNA序列,包含2 283 bp的CDS区,编码760个氨基酸;SiPRR73蛋白与C4家族作物糜子、哈氏黍、高粱、玉米PRR73蛋白具有较近的进化关系。 SiPRR73在顶端第2片叶中表达水平最高,在根、茎、穗部表达水平较低。短日照、长日照条件下该基因均表现光照期表达水平高于黑暗期的特点,且整个营养生长期短日照条件下表达水平明显低于长日照,SiPRR73受光诱导表达和光周期调控。温度决定SiPRR73表达峰的数目,光周期决定SiPRR73表达峰出现的时间,光周期和温度共同参与了SiPRR73的表达调控。PEG和低温胁迫总体上诱导SiPRR73表达,NaCl在胁迫早期诱导该基因表达,后期总体表现为抑制状态,Fe胁迫则早期抑制该基因表达,后期总体诱导其表达,SiPRR73对ABA胁迫的响应特点接近NaCl。以上研究表明,谷子SiPRR73表达具有光依赖性,光周期和温度以互作模式调控SiPRR73表达,推测SiPRR73参与了谷子对不同光温环境的适应性调节过程,并且在应对干旱、低温、ABA、NaCl、Fe胁迫过程中发挥了一定作用。

关键词: 谷子, PRR73, 光周期, 温度, 非生物胁迫

Abstract:

The SiPRR73 gene was cloned from Yangu 11 using RT-PCR technology,and through analyzing tissue-specific expression,responsive features of SiPRR73 to different photoperiods,photo-thermal combinational treatments and five abiotic stress treatments,the regulation mode of photoperiod and temperature on SiPRR73,and the responsive pattern of SiPRR73 to abiotic stresses in foxtail millet were explored. The results showed that totally 2 928 bp cDNA sequence of SiPRR73 was obtained from Yangu 11,which included 2 283 bp CDS region,encoding 760 amino acids. The SiPRR73 proteins of C4 crops including Panicum miliaceum,Panicum hallii,Sorghum bicolor and Zea mays showed relatively close relationship with SiPRR73. The second parietal leaf was the highest expression tissue of SiPRR73,but the expression level at root,stem and panicle tissues was relatively lower. The expression level of SiPRR73 was higher at light period than that at dark period under both short-day and long-day conditions,and during the whole vegetative growth phase,SiPRR73 showed higher expression level under long-day compared to short-day,which indicated that the expression of SiPRR73 was induced by light and controlled by photoperiod. The temperature determined expression peak number of SiPRR73 and the photoperiod determined occurrence time of expression peaks,so temperature and photoperiod participated in regulating of SiPRR73 expression mutually. PEG and low temperature stresses induced SiPRR73 expression totally,NaCl induced SiPRR73 expression at early stress stage,but inhibited it at later stress stage. Fe stress inhibited SiPRR73 expression at early stage,but induced it at later stage. ABA stress caused the close responsive feature of SiPRR73 to NaCl. This study indicated that SiPRR73 showed light-dependent expression feature,and photoperiod and temperature regulated SiPRR73 by interaction pattern,suggesting that SiPRR73 participated in adaptability regulation process to different photo-thermal conditions and might play a certain role in coping with drought,low temperature,ABA,NaCl and Fe stresses in foxtail millet.

Key words: Foxtail millet, PRR73, Photoperiod, Temperature, Abiotic stress

闫留延, 李剑峰, 张世文, 张博, 王永芳, 张小梅, 祖超凡, 王振山, 桑璐曼, 何占祥, 贾小平, 董志平. 谷子SiPRR73基因的光温调控模式及非生物胁迫响应特性[J]. 华北农学报, 2022, 37(4): 11-19. doi: 10.7668/hbnxb.20193022.

YAN Liuyan, LI Jianfeng, ZHANG Shiwen, ZHANG Bo, WANG Yongfang, ZHANG Xiaomei, ZU Chaofan, WANG Zhenshan, SANG Luman, HE Zhanxiang, JIA Xiaoping, DONG Zhiping. Regulation Pattern of Photoperiod,Temperature and Abiotic Stress on SiPRR73 in Foxtail Millet[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 11-19. doi: 10.7668/hbnxb.20193022.

http://www.hbnxb.net/CN/Y2022/V37/I4/11

图/表 8

表1 基因扩增及荧光定量PCR引物

Tab.1 Primers used for gene amplification and fluorescence quantitative PCR

引物名称 Primer name	引物序列(5'-3') Primer sequence(5'-3')	预期片段大小/bp Expected fragment size	目的 Purpose
SiPRR73-G1	F-GCTCTGCTCGTCCTCGTT	2 398	克隆SiPRR73
	R-GCCACCTCCAGCCACAT
SiPRR73-G2	F-CGGGTAACAGGGAGAGAGTAA	804
	R-CAACCACAGAAACGATAACTGG
SiActin-FQ	F-GGCAAACAGGGAGAAGATGA	229	内参基因荧光定量
	R-GAGGTTGTCGGTAAGGTCACG
SiPRR73-FQ	F-ACCATCTGTGAACTCCCTCT	198	SiPRR73荧光定量
	R-ATCTTCCTTTTCCCTCCATT

图1 电泳检测谷子总RNA及SiPRR73基因扩增产物 A.谷子总RNA电泳:1,2.两管RNA。B.SiPRR73基因的RT-PCR产物电泳:1.DL2000 Marker;2,3.分段扩增产物SiPRR73-G1、SiPRR73-G2。

Fig.1 Total RNA and SiPRR73 gene amplified products in foxtail millet detected by electrophoresis A.Electrophoresis of total RNA of foxtail millet:1,2.Two tubes of RNA.B. Electrophoresis of SiPRR73 gene RT-PCR product:1.DL2000 Marker;2,3.Segmental amplification products,SiPRR73-G1 and SiPRR73-G2.

图2 SiPRR73蛋白所包含的结构域

Fig.2 The domains contained in SiPRR73 protein

图3 PRR73蛋白的分子系统进化树

Fig.3 Molecular phylogenetic tree of PRR73 proteins

图4 谷子不同组织部位SiPRR73的表达特性不同小写字母表示处理间差异显著(P<0.05)。图5-7同。

Fig.4 Expression profile of SiPRR73 in different tissue parts of foxtail millet Different lowercase letters indicate significant differences between treatments at P<0.05. The same as Fig.5-7.

图5 SiPRR73基因的光周期响应模式 A.昼夜表达规律;B.不同叶期表达规律。

Fig.5 Photoperiod responsive pattern of SiPRR73 gene A. Diurnal expression pattern;B. Expression pattern at different leaf stages.

图6 光温互作对SiPRR73基因昼夜表达模式的影响

Fig.6 The effect of photo-thermal interaction on diurnal expression pattern of SiPRR73

图7 非生物胁迫条件下SiPRR73基因的表达模式

Fig.7 Expression pattern of SiPRR73 under abiotic stress conditions

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谷子SiPRR73基因的光温调控模式及非生物胁迫响应特性

Regulation Pattern of Photoperiod,Temperature and Abiotic Stress on SiPRR73 in Foxtail Millet

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Regulation Pattern of Photoperiod,Temperature and Abiotic Stress on SiPRR73 in Foxtail Millet

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