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In order to study the effect of chalcone isomerase on the synthesis of flavonoids during the development of rubber leaves, this study used the CHI functional domain chalcone (PF02431) to identify the HbCHI gene family members of the Ccalcone isomerase, and to analyze the physicochemical properties, conserved domain, conserved motif and phylogenetic tree of the protein sequences. qRT-PCR was used to analyze the expression pattern of HbCHI during the development of rubber leaves. The effect of HbCHI2 on plant flavonoid synthesis was analyzed using Agrobacterium-mediated transient expression technique in Nicotiana benjamina. Nine HbCHI gene family members (HbCHI1-HbCHI9) were identified from the genomic database of H. brasiliensis. The proteins encoded by HbCHI1-HbCHI9 contained 177-474 amino acids with isoelectric points ranging from 5.1 to 9.52. The proteins were mainly hydrophilic proteins. Only HbCHI1 contained 6 conserved motif1-motif6 motifs, and phylogenetic analysis showed that HBCHI1-HBCHI4 was type I CHI. HbCHI6-HbCHI9 showed fatty acid binding sites, which were type III CHI. HbCHI5 was type IV CHI. The relative expression level of HbCHI gene family in rubber trees showed a trend of first decreasing and then increasing from small bronzer to mature stage. The relative expression level was the lowest in light green stage, and the change trend of total flavonoids content was consistent with the relative expression level of CHI gene. Correlation analysis showed that HbCHI2 had the highest significant correlation with flavoniods content. The Pearson coefficient was 0.97. Overexpression of HbCHI2 in N. benjamina promoted the biosynthesis of flavonoids and increased the total flavonoids content in leaves, which was 2.13 times that of control. These results indicate that HbCHI2 gene regulates flavonoid biosynthesis in rubber leaves.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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