| Anthurium andraeanum L.belongs to the Araceae Anthurium.It is a kind of important flowers in the Chinese flower market.The colorful spathe is one of its important viewing characters.Anthocyanin plays an important role in the formation of its color.Chalcone isomerase?CHI?and cinnamate-4-hydroxylase?C4H?are two kinds of important enzymes in the anthocyanin synthesis pathway.At present,the main varieties of Anthurium were imported from abroad,and the common phenomenon of green lodging in the process of cultivation affected the ornamental value of Anthurium.The study compared the expression of AnCHI and AnC4H in different colors of spathe,analyzed the expression of AnCHI and An C4H treated by ethephon,and used the transgenic technology to explore the mechanism of AnCHI and AnC4H in the synthesis of anthocyanins,and further analyzed the biological mechanism of anthocyanin biosynthesis pathway.The use of modern molecular biology techniques to study the mechanism of An CHI and An C4H provides an effective method for cultivating Anthurium with independent intellectual property rights in China and improving the color saturation of Anthurium buds.Therefore,the research provides an important theoretical support and reference for improving the value of Anthurium.The results of this study are as follows:1.Anthocyanidin content in Anthurium is the key factor for the color of the spathe.AnCHI and AnC4H play an important role in the synthesis of anthocyanin.The contents of total anthocyanin in flower buds and the expression of AnCHI and AnC4H in the red'Dakota',pink'powder champion'and white'white king'were determined.The results showed that the anthocyanin content of Anthurium in red spathe,pink spathe and white spathe decreased successively,and the expression regularity of AnCHI and AnC4H was consistent with that of anthocyanin.The result further verified that AnCHI and AnC4H are two key genes in the synthesis of anthocyanins.2.Ethephon increased the content of anthocyanin in the bud and full period spathes of the Anthurium andraeanum.After treatment with different concentrations of ethephon,the content of anthocyanin in the spathe rose first and then decreased with the increase of ethephon concentration.At budding stage,the content of anthocyanin increased most significantly after treated with ethephon at the concentration of 750 mg·L-1.At full-opening stage,the most significant increase of anthocyanin content was observed at 900 mg·L-11 ethephon.Therefore,the spathes should be treated with a suitable concentration of ethephon.3.Ethephon treating buds of Anthurium spathe,the expression of An CHI and An C4H increased.At the budding stage,the highest expression of An CHI and AnC4H occurred at the concentration of 750 mg·L-11 ethephon 3 h after spraying.In the full-opening spathe,900 mg·L-11 ethephon showed the highest expression of AnC4H at 2h after spraying and AnCHI expression reached the maximum at 3h after spraying.Therefore,the duration of ethephon treatment should be controlled strictly in order to improve the regulation efficiency of ethephon.Meanwhile,the result further confirmed that AnCHI and AnC4H are two key genes in the synthesis of anthocyanins.4.The p CAMBIA3301-AnCHI and pCAMBIA3301-AnC4H fusion expression vectors were constructed through double enzyme digestion,and the recombinant expression vectors were transformed into tobaccos with Agrobacterium GV3101.The transgenic tobaccos with AnCHI gene were obtained.The content of anthocyanin in the transgenic tobaccos with AnCHI was higher than that in wild tobaccos.The result further confirmed that An CHI is a key gene in the process of anthocyanin synthesis in Anthurium.The differentiation culture medium used in the system was:4.43 g·L-11 MS salt+20 g·L-1sucrose+6.5 g·L-11 agar+2 g·L-11 6-BA+0.2mg·L-11 NAA+500 m g·L-11 Cef?cefotaxime?+5 mg·L-11 PPT?Basta 10%?;Rooting medium:2.2 g·L-11 MS salt+20 g·L-11 sucrose+6.5 g·L-11 Agar+500mg·L-11 Cef?cefotaxime?. |
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