cDNA-Amplified fragment length polymorphism analysis reveals differential gene expression induced by exogenous MeJA and GA3 in oilseed rape (Brassica napus L.) flowers
cDNA-Amplified fragment length polymorphism analysis reveals differential gene expression induced by exogenous MeJA and GA3 in oilseed rape (Brassica napus L.) flowers
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cDNA 扩增片段长度多态性分析揭示了外源 MeJA 和 GA3 在油菜 (Brassica napus L.) 花中诱导的差异基因表达
DOI：
10.1016/s2095-3119(16)61407-7
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发表时间：
2017-01-01
影响因子：
4.8
通讯作者：
Jiang Li-xi
中科院分区：
文献类型：
--
作者：
Pak, Haksong;Li Yu-ling;Jiang Li-xi
植物从营养阶段到生殖阶段的转变是由一大群基因控制的，这些基因对环境和内源刺激作出反应。茉莉酸甲酯（MeJA）和赤霉素（GA(3)）对油籽植物甘蓝型油菜（Brassica napus L.）的应用中断了其脆弱的内源平衡，导致各种花器官异常。外源MeJA或GA(3)会影响拟南芥开花初期的转录组，但油菜花组织中转录组的相应变化尚不清楚。本研究通过分析dna扩增片段长度多态性（cDNA-AFLP）来鉴定MeJA和GA(3)对花蕾表达调控的基因。利用64对引物组合共计数了2787个cDNA片段，并比较了水分控制、MeJA （50 pmol L-1）、MeJA （100 pmol L-1）和GA(3) (50 pmol L-1) 4个处理中大于50 bp的条带。总体而言，168个转录衍生片段（TDFs）在不同处理之间存在差异表达。通过半定量RT-PCR分析证实了部分TDFs的表达模式，并克隆了一组106个差异TDFs并对其进行了测序。鉴定了拟南芥基因同源物，并将其划分为12类功能。共有34例、39例和24例tdf分别对GA(3)、MeJA和GA(3)和MeJA均有反应。这一发现表明，这两种激素之间的串扰可能参与调节花的发育。本研究为油菜重要农艺性状开花时间和花器官起始的调控提供了潜在的靶基因。
The transition of a plant from vegetative to reproductive stage is controlled by a large group of genes, which respond to environmental and endogenous stimuli. Application of methyl jasmonate (MeJA) and gibberellins (GA(3)) to oilseed plants (Brassica napus L.) interrupts the delicate endogenous balake and results in various floral organ abnormalities. Exogenous MeJA or GA(3) influences the transcriptome at the initial flowering stage in Arabidopsis, but the corresponding changes of transcriptome in floral tissues of oilseed rape remain unknown. In this study, cDNA-amplified fragment length polymorphism (cDNA-AFLP) was analyzed to identify genes whose expression was modulated by application of MeJA and GA(3) to flower buds. A total of 2787 cDNA fragments were counted using 64 primer pair combinations, and bands larger than 50 bp were compared among four treatments, namely, water control, MeJA (50 pmol L-1), MeJA (100 pmol L-1), and GA(3) (50 pmol L-1). Overall, 168 transcript-derived fragments (TDFs) were differentially expressed among the treatments. The expression pattern of some TDFs was confirmed by semi-quantitative RT-PCR analysis, and a group of 106 differentially displayed TDFs was cloned and sequenced. Homologs of Arabidopsis genes were identified and classified into 12 functional categories. A total of 34, 39, and 24 TDFs were responsive to GA(3), MeJA, and both GA(3) and MeJA, respectively. This finding indicated that cross-talk between these two hormones may be involved in regulating flower development. This study provides potential target genes for manipulation in terms of flowering time and floral organ initiation, important agronomic traits of oilseed rape.