Transformation of Campanula carpatica to alter flower colour and ethylene sensitivity
Transformation of Campanula carpatica to alter flower colour and ethylene sensitivity
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DOI：
10.1080/14620316.2015.11668708
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发表时间：
2015-09-01
影响因子：
1.9
通讯作者：
Mibus, H.
中科院分区：
文献类型：
--
作者：
Karimiani, Z. Ghayoor;Serek, M.;Mibus, H.
本研究首次报道了风铃草属植物通过基因操作改变花色。该实验使用风铃草“改良的蓝色统一”（IBU）设计，具有两个主要目的：通过类黄酮3 '，5'羟化酶（F3 '5'H）基因的RNAi沉默下调飞燕草素的产生来产生红色的花颜色，以及通过使用乙烯抗性1-1（etr 1 -1）基因诱导对乙烯的不敏感性来改善花瓶寿命。经农杆菌GV 3101转化获得了3个独立的转基因株系。进行Southern印迹分析以确定整合的T-DNA拷贝数，并进行逆转录-聚合酶链反应（RT-PCR）以评估F3 '5'H和类黄酮3羟化酶（F3 'H）基因的表达。进行乙烯敏感性试验以评价转化植物对暴露于乙烯的耐受性。Southern印迹分析显示，在三个转化系的每一个中整合了两个或三个T-DNA拷贝。转化系的花色没有视觉上改变，并且通过RTPCR证实了F3 ′ 5 ′ H和F3 ′ H基因在所有三个转化系和非转化系中的表达。在未转化风铃草植株上，乙烯处理6 d后，所有的花都衰老，但转基因株系11-213和21-1的存活率分别为63.3%和86.4%。乙烯敏感性试验表明，与非转化植物相比，暴露于乙烯的转化植物显示出衰老的显著延迟。
This study is the first report of gene manipulation to alter flower colour in the genus Campanula. The experiment was designed using Campanula carpatica 'Improved Blue Uniform' (IBU) with two main purposes: to generate a red flower colour by down-regulation of delphinidin production through RNAi silencing of the flavonoid 3', 5' hydroxylase (F3'5'H) gene, and to improve vase-life by inducing insensitivity to ethylene using the ethylene-resistant 1-1 (etr1-1) gene. Three independent transgenic lines were obtained after genetic transformation via Agrobacterium tumefaciens strain GV3101. Southern blot analysis was performed to determine the integrated T-DNA copy number, and reverse transcription-polymerase chain reactions (RT-PCR) were performed to evaluate expression of the F3'5'H and flavonoid 3 hydroxylase (F3'H) genes. An ethylene sensitivity test was conducted to evaluate the tolerance of the transformed plants to exposure to ethylene. Southern blot analysis revealed the integration of two or three T-DNA copies in each of the three transformed lines. The flower colour of the transformed lines was not visually altered, and expression of the F3'5'H and F3'H genes in all three transformed and in non-transformed lines was confirmed by RTPCR. Although all flowers on non-transformed Campanula plants were senescent 6 d after exposure to ethylene, 63.3% of the flowers on transgenic line 11-213 and 86.4% of the flowers on transgenic line 21-1 survived. The ethylene sensitivity test showed that transformed plants exposed to ethylene showed a significant delay in senescence compared to non-transformed plants.