Yu Christine K Y, Springob Karin, Schmidt Jürgen, Nicholson Ralph L, Chu Ivan K, Yip Wing Kin, Lo Clive
Department of Botany, University of Hong Kong, Hong Kong, China.
Plant Physiol. 2005 May;138(1):393-401. doi: 10.1104/pp.105.059337. Epub 2005 Apr 8.
A chalcone synthase (CHS)-like gene, SbCHS8, with high expressed sequence tag abundance in a pathogen-induced cDNA library, was identified previously in sorghum (Sorghum bicolor). Genomic Southern analysis revealed that SbCHS8 represents a single-copy gene. SbCHS8 expression was induced in sorghum mesocotyls following inoculation with Cochliobolus heterotrophus and Colletotrichum sublineolum, corresponding to nonhost and host defense responses, respectively. However, the induction was delayed by approximately 24 h when compared to the expression of at least one of the other SbCHS genes. In addition, SbCHS8 expression was not induced by light and did not occur in a tissue-specific manner. SbCHS8, together with SbCHS2, was overexpressed in transgenic Arabidopsis (Arabidopsis thaliana) tt4 (transparent testa) mutants defective in CHS activities. SbCHS2 rescued the ability of these mutants to accumulate flavonoids in seed coats and seedlings. In contrast, SbCHS8 failed to complement the mutation, suggesting that the encoded enzyme does not function as a CHS. To elucidate their biochemical functions, recombinant proteins were assayed with different phenylpropanoid-Coenzyme A esters. Flavanones and stilbenes were detected in the reaction products of SbCHS2 and SbCHS8, respectively. Taken together, our data demonstrated that SbCHS2 encodes a typical CHS that synthesizes naringenin chalcone, which is necessary for the formation of different flavonoid metabolites. On the other hand, SbCHS8, now retermed SbSTS1, encodes an enzyme with stilbene synthase activity, suggesting that sorghum accumulates stilbene-derived defense metabolites in addition to the well-characterized 3-deoxyanthocyanidin phytoalexins.
先前在高粱(Sorghum bicolor)中鉴定出一个查尔酮合酶(CHS)类似基因SbCHS8,其在病原体诱导的cDNA文库中的表达序列标签丰度很高。基因组Southern分析表明,SbCHS8代表一个单拷贝基因。接种异旋孢腔菌(Cochliobolus heterotrophus)和亚线炭疽菌(Colletotrichum sublineolum)后,高粱中胚轴中SbCHS8的表达被诱导,分别对应非寄主和寄主防御反应。然而,与至少一个其他SbCHS基因的表达相比,这种诱导延迟了约24小时。此外,SbCHS8的表达不受光诱导,也不是以组织特异性方式发生。SbCHS8与SbCHS2一起在CHS活性有缺陷的转基因拟南芥(Arabidopsis thaliana)tt4(透明种皮)突变体中过表达。SbCHS2挽救了这些突变体在种皮和幼苗中积累类黄酮的能力。相反,SbCHS8未能弥补该突变,这表明编码的酶不具有CHS的功能。为了阐明它们的生化功能,用不同的苯丙烷类辅酶A酯对重组蛋白进行了检测。在SbCHS2和SbCHS8的反应产物中分别检测到了黄烷酮和芪。综上所述,我们的数据表明,SbCHS2编码一种典型的CHS,其合成柚皮素查尔酮,这是形成不同类黄酮代谢物所必需的。另一方面,现在重新命名为SbSTS1的SbCHS8编码一种具有芪合酶活性的酶,这表明高粱除了积累特征明确的3-脱氧花青素植物抗毒素外,还积累芪衍生的防御代谢物。