2个玉米光敏色素C基因的转录丰度对多种光质处理的响应

doi:10.3864/j.issn.0578-1752.2017.12.002

中国农业科学 ›› 2017, Vol. 50 ›› Issue (12): 2209-2219.doi: 10.3864/j.issn.0578-1752.2017.12.002

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

2个玉米光敏色素C基因的转录丰度对多种光质处理的响应

牛骧^1,2，郭林²，杨宗举^2,3，孙蕾^2,3，李红丹^2,3，游光霞²，徐宏⁴，孟凡华²，佘跃辉¹，杨建平²

¹四川农业大学农学院，成都 611130；²中国农业科学院作物科学研究所，北京 100081；³中国农业科学院研究生院，北京100081；⁴河南省潢川县农业科学研究所，河南潢川 465150

收稿日期:2016-12-16 出版日期:2017-06-16 发布日期:2017-06-16
通讯作者: 杨建平，Tel/Fax：010-82105859；E-mail：yangjianping02@caas.cn。佘跃辉，Tel：13880283512；E-mail：syuehui@sina.com
作者简介:牛骧，Tel：010-82105851；E-mail：cnau323_niux@163.com
基金资助:
国家转基因生物新品种培育科技重大专项（2016ZX08010002-003-002）、国家重点研究发展计划（2016YFD0101002）、北京市自然科学基金（重点）（6151002）、中国农业科学院科技创新工程

Transcription Abundances of Two Phytochrome C in Response to Different Light Treatments in Zea mays

NIU Xiang ^1,2, GUO Lin ², YANG ZongJu^2,3, SUN Lei ^2,3, LI HongDan^2,3, YOU GuangXia ², XU Hong ⁴, MENG FanHua ², SHE YueHui ¹, YANGJianPing ²

¹College of Agronomy, Sichuan Agricultural University, Chengdu 611130; ²Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081; ³Graduate School, Chinese Academy of Agricultural Sciences, Beijing 100081; ⁴Huangchuan Institute of Agricultural Sciences of Henan Province, Huangchuan 465150, Henan

Received:2016-12-16 Online:2017-06-16 Published:2017-06-16

摘要/Abstract

摘要： 【目的】通过NCBI数据库获得2个玉米PHYC及相关数据，并进行相关生物信息学分析。利用实时荧光定量PCR（qRT-PCR）分析2个玉米PHYC在玉米各器官的转录丰度，以及其转录丰度对多种光质处理、黑暗到各种光质转换和光周期处理（长日照和短日照）的响应，为研究玉米PHYC在玉米幼苗去黄化与开花期的调控机制奠定基础。【方法】采用玉米B73自交系为研究材料，通过RT-PCR分别对ZmPHYC1和ZmPHYC2的全长ORF序列进行克隆；借助相关软件对其进行生物信息学分析，利用qRT-PCR分析这两个基因在玉米各器官中的转录丰度，及其转录丰度对各种光照处理的响应。【结果】ZmPHYC1和ZmPHYC2的全长ORF均为3 408 bp，编码1 135个氨基酸基序，分子量分别为126.14和126.07 kD。生物信息学分析表明，玉米phyC蛋白可以分为6个功能区段：节奏周期蛋白—Ah核转运接受蛋白—专一蛋白区段（Per-Arnt-Sim，PAS）、cGMP受激磷酸二酯酶区段（GAF）、色素区段（PHY）和PAS相关区段（PRD，包含2个PAS区段）、组氨酸激酶A区段和组氨酸激酶ATP酶区段，但是ZmphyC2在PRD区段仅有一个PAS区段。氨基酸水平的系统发育树分析表明，ZmphyC1和ZmphyC2与禾本科物种phyC有很高的一致性，且与甘蔗和高粱phyC的亲缘关系较近。qRT-PCR分析表明，ZmPHYC1和ZmPHYC2的表达在根和叶中的转录丰度均较高，同时对持续蓝光和白光响应强烈；在黑暗到各种光质转换处理中，这两个PHYC的表达模式相似。在黑暗转到远红光、红光、蓝光和白光的0.5 h，ZmPHYC1和ZmPHYC2的转录表达均急剧上升，随后迅速下降到自身起始黑暗时的水平以下，并上下波动。这两个基因对长日照和短日照的光周期处理也能积极响应，在长日照条件下，2个ZmPHYC出现了极其相似的表达模式，均在光照和黑暗阶段各出现1个峰值；在短日照条件下，这两个基因的表达模式差异较大，ZmPHYC1的峰值出现在进入黑暗后6 h，而ZmPHYC2的峰值出现在进入光照阶段2 h。【结论】玉米phyC蛋白可以分成6个功能区段，但是ZmphyC2在PRD区段仅具有一个PAS相关区段。2个玉米PHYC转录丰度具有组织特异性。在各种光质处理中，ZmPHYC1和ZmPHYC2的表达模式相近，可能二者存在功能冗余，在转录水平上前者的丰度高于后者，推测ZmPHYC1在玉米中起更重要的作用，并且可能二者在功能上存在分工。ZmPHYC1和ZmPHYC2对各种光质和光周期处理均有较强的响应，推测二者在调控玉米光形态建成和开花中具有重要作用。

关键词: 玉米, ZmPHYC, 光信号转导, 表达模式, 光质处理

Abstract: 【Objective】To study the functions of phytochrome C genes in seedling de-etiolation and flowering regulation in maize (Zea mays L.), two phytochrome C genes of maize (ZmPHYC1 and ZmPHYC2) were selected from the NCBI database and analyzed by bioinformatic methods. The transcription abundances of two ZmPHYC genes was analyzed in different tissues and in response to light qualities, transitions from the dark to different light conditions, photoperiod treatment (long day and short day) by quantitative RT-PCR (qRT-PCR).【Method】B73 inbred line was used in this study, the full length ORFs of two ZmPHYC genes were cloned by RT-PCR. The proper clones were sequenced and analyzed by bioinformatics software. The transcription abundances of two ZmPHYC genes in different tissues and in response to light treatments was detected using qRT-PCR.【Result】Both the full length ORFs of ZmPHYC1 and ZmPHYC2 contained 3408 nucleotides and encoded two polypeptides with 1135 amino acid motifs, and their molecular weight was 126.14 kD and 126.07 kD, respectively. Two ZmphyC proteins were able to be further divided into six domains: Per (period circadian protein)-Arnt (Ah receptor nuclear translocator protein)-Sim (single-minded protein) (PAS), cGMP-stimulated phosphodiesterase (GAF), phytochrome (PHY), PAS-related domain (PRD) containing two PAS, His Kinase A domain (HisKA), Histidine kinase-like ATPases (HATPase_c), while ZmphyC2 lacked a PAS in PRD domain. Phylogenetic analysis indicated that the two ZmphyC proteins belonged to the same branch with phyC proteins from other graminaceous species, especially with the phyC proteins from sugarcane and sorghum. qRT-PCR assays showed that both ZmPHYC1 and ZmPHYC2 belonged to tissue-specific genes and highly expressed in roots and leaves. The transcription abundances of the both genes were very high under blue and white light conditions. Both ZmPHYC1 and ZmPHYC2 displayed similar expression patterns during transitions from the dark to different light conditions. The transcription abundances of the both genes went dramatically up at 0.5 h after transitions from the dark to far-red, red, blue, or white light. And then they quickly dropped and waved below their own levels in the dark. Both ZmPHYC1 and ZmPHYC2 were also able to respond to long-day or short-day treatments. During long-day treatment, they likely had one peak in either light or dark period. During short-day treatment, they showed different expression patterns, the peak of ZmPHYC1 happened at 6 h after conversion to dark period, while ZmPHYC2 occurred at 2 h after conversion to light period.【Conclusion】 ZmphyC1 protein kept six domains, while ZmphyC2 lacks a PAS in PRD domain. The transcription abundances of the both PHYC genes were tissue-specific in maize. Similar expression patterns of ZmPHYC1 and ZmPHYC2 genes in response to different light treatments suggest that they both might keep redundant functions. Since the transcription abundances of ZmphyC1 were higher than these of ZmphyC2, ZmphyC1 might have more important role in seedling responding to light than ZmphyC2, which may be due to the existence of different functions in maize. Both ZmPHYC1 and ZmPHYC2 respond to light and photoperiod treatments, suggesting that they are involved in seedling de-etiolation and flowering time control in maize. Thus their roles in crop improvement are worthy of more exploration in the future.

Key words: Zea mays, ZmPHYC, light signal transduction, expression pattern, light treatment

牛骧，郭林，杨宗举，孙蕾，李红丹，游光霞，徐宏，孟凡华，佘跃辉，杨建平. 2个玉米光敏色素C基因的转录丰度对多种光质处理的响应[J]. 中国农业科学, 2017, 50(12): 2209-2219.

NIU Xiang, GUO Lin, YANG ZongJu, SUN Lei, LI HongDan, YOU GuangXia, XU Hong, MENG FanHua, SHE YueHui, YANGJianPing. Transcription Abundances of Two Phytochrome C in Response to Different Light Treatments in Zea mays[J]. Scientia Agricultura Sinica, 2017, 50(12): 2209-2219.

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2个玉米光敏色素C基因的转录丰度对多种光质处理的响应

Transcription Abundances of Two Phytochrome C in Response to Different Light Treatments in Zea mays

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