van Tunen A J, Koes R E, Spelt C E, van der Krol A R, Stuitje A R, Mol J N
Department of Applied Genetics, Free University, Amsterdam, The Netherlands.
EMBO J. 1988 May;7(5):1257-63. doi: 10.1002/j.1460-2075.1988.tb02939.x.
In this paper we report the isolation of cDNA clones encoding the flavonoid-biosynthetic enzyme chalcone flavanone isomerase (CHI) from Petunia hybrida. A nearly full size cDNA clone, isolated from a corolla-specific expression library, was characterized by sequence analysis. Using this CHI cDNA and the previously cloned flavonoid-specific chalcone synthase (CHS) cDNA we show that CHI and CHS genes are coordinately and tissue-specifically expressed in a developmental and light-regulated manner. Furthermore, coordinate induction of both mRNAs is observed after continuous irradiation of Petunia plantlets with UV light, probably as part of the plants UV defence mechanism. The two CHI genes, denoted A and B, were isolated from a genomic library of the Petunia inbred line V30. Both genes are transcriptionally active: gene A is transcribed in corolla, tube and UV-irradiated plantlets (1.0 kb mRNA), whereas gene B is only transcribed in immature anthers (1.0 kb mRNA). In combination with Southern blot analysis these data implicate the presence of two distinct non-allelic CHI genes in the genome of the P. hybrida line V30. Unexpectedly, mature anthers accumulate a 0.3 kb larger CHI RNA. This RNA is transcribed from CHI gene A and has a 0.3 kb 5' extension relative to the gene A transcript found in corolla tissue. Furthermore it is neither coordinately expressed with CHS mRNA nor UV inducible. Its biological function is still obscure, since no active CHI enzyme could be demonstrated in the same tissue.
在本文中,我们报道了从矮牵牛中分离出编码类黄酮生物合成酶查尔酮黄烷酮异构酶(CHI)的cDNA克隆。从花冠特异性表达文库中分离出一个几乎全长的cDNA克隆,并通过序列分析对其进行了表征。利用这个CHI cDNA和先前克隆的类黄酮特异性查尔酮合酶(CHS)cDNA,我们表明CHI和CHS基因以发育和光调节的方式协调且组织特异性地表达。此外,在用紫外线持续照射矮牵牛幼苗后,观察到两种mRNA的协同诱导,这可能是植物紫外线防御机制的一部分。从矮牵牛自交系V30的基因组文库中分离出两个CHI基因,分别命名为A和B。这两个基因都具有转录活性:基因A在花冠、花管和经紫外线照射的幼苗中转录(1.0 kb mRNA),而基因B仅在未成熟花药中转录(1.0 kb mRNA)。结合Southern印迹分析,这些数据表明在矮牵牛V30品系的基因组中存在两个不同的非等位CHI基因。出乎意料的是,成熟花药积累了一种比CHI RNA大0.3 kb的RNA。这种RNA是从CHI基因A转录而来的,相对于在花冠组织中发现的基因A转录本,其5'端有一个0.3 kb的延伸。此外,它既不与CHS mRNA协同表达,也不是紫外线诱导型的。其生物学功能仍然不清楚,因为在同一组织中未检测到有活性的CHI酶。