Tissue culture-induced flower-color changes in Saintpaulia caused by excision of the transposon inserted in the flavonoid 3′, 5′ hydroxylase (F3′5′H) promoter
Tissue culture-induced flower-color changes in Saintpaulia caused by excision of the transposon inserted in the flavonoid 3′, 5′ hydroxylase (F3′5′H) promoter
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DOI：
10.1007/s00299-011-1016-z
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发表时间：
2011-05-01
影响因子：
6.2
通讯作者：
Doi, Motoaki
中科院分区：
文献类型：
--
作者：
Sato, Mitsuru;Kawabe, Takashi;Doi, Motoaki
杂色的Saintpaulia栽培品种Thamires（Saintpaulia sp.），粉红色的花瓣上有蓝色的斑点，通常是通过剪叶来维持的。与此相反，组织培养衍生的后代的品种不仅表现出高比例的突变体与纯蓝色的花瓣，但也有其他纯色的变种，这还没有观察到从叶插。纯色表型在组织培养后代中遗传稳定。从每个纯色变种的花瓣进行了分析，通过高效液相色谱法，并显示含有不同比例的三个主要的花青素衍生物：锦葵色素，牡丹色素，和天竺葵色素。类黄酮3 '，5'-羟化酶（F3 '5'H）序列分析表明，变异体和杂色个体的编码区没有差异。然而，在杂色个体的启动子区发现了属于hAT超家族的转座子，并且转座子相关插入或缺失的存在与观察到的花色表型相关。纯蓝色花突变体含有8个碱基对（bp）插入（转座子切除足迹），而纯粉红色突变体有58- 70-bp插入，紫色和深紫色突变体分别有21-和24-bp缺失。实时逆转录聚合酶链反应（RT-PCR）分析表明，F3 '5'H的表达水平与插入和缺失（indels）所造成的hAT切除，导致花的颜色差异。我们的研究结果表明，组织培养的圣保罗'Thamires' eliminate转座子切除，这反过来又改变F3 '5'H的表达水平和花色。
The variegated Saintpaulia cultivar Thamires (Saintpaulia sp.), which has pink petals with blue splotches, is generally maintained by leaf cuttings. In contrast, tissue culture-derived progeny of the cultivar showed not only a high percentage of mutants with solid-blue petals but also other solid-color variants, which have not been observed from leaf cuttings. Solid-color phenotypes were inherited stably by their progeny from tissue culture. Petals from each solid-color variant were analyzed by high-performance liquid chromatography and shown to contain different proportions of three main anthocyanin derivatives: malvidin, peonidin, and pelargonidin. Analysis of flavonoid 3', 5'-hydroxylase (F3'5'H) sequences showed no differences in the coding region among the variants and variegated individuals. However, a transposon belonging to the hAT superfamily was found in the promoter region of variegated individuals, and the presence of transposon-related insertions or deletions correlated with the observed flower-color phenotypes. Solid-blue flower mutants contained 8-base pair (bp) insertions (transposon excision footprints), while solid-pink mutants had 58- to 70-bp insertions, and purple- and deep-purple mutants had 21- and 24-bp deletions, respectively. Real-time reverse transcription polymerase chain reaction (RT-PCR) analysis showed that F3'5'H expression levels correlated with insertions and deletions (indels) caused by hAT excision, resulting in flower-color differences. Our results showed that tissue culture of Saintpaulia 'Thamires' elicits transposon excision, which in turn alters F3'5'H expression levels and flower colors.