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Outline
To explore the regulatory effect of novel miRNA phe-novel-miR276 on lignin synthesis in moso bamboo, so as to provide theoretical reference for material quality improvement and molecular breeding of moso bamboo.
Using bioinformatics methods, we analyzed the novel differentially expressed miRNA (phe-novel-miR276) screened in the preliminary study of moso bamboo. qRT-PCR was applied to detect its expression patterns in different moso bamboo tissues, followed by calculation of relative expression levels and differential analysis. The phe-novel-miR276 was cloned to construct an overexpression plant vector, which was transformed into rice via Agrobacterium mediation for screening positive transgenic lines. Lignin content as well as POD and SOD activities of transgenic and wild-type rice were determined, and intergroup differences of each index were compared and analyzed.
1) Sequence analysis revealed that phe-novel-miR276 shares high homology with the miR398 family, suggesting it is a new member of the miR398 family; 2) phe-novel-miR276 is expressed in the roots, stems, and leaves of P. edulis, with the highest relative expression level in the roots; 3) Phloroglucinol staining showed that the lignin staining depth in the stems and leaves of transgenic lines was lighter; meanwhile, the results of the microdetermination method indicated that the total lignin content in the stems and leaves of transgenic lines was significantly reduced; 4) qRT-PCR detection demonstrated that, compared with wild-type rice, the expression level of Os10g36848-the rice homologous target gene of phe-novel-miR276 (encoding ferulate 5-hydroxylase, F5H)-was significantly downregulated in transgenic lines. Additionally, the POD and SOD activities in transgenic lines were significantly higher than those in wild-type rice, respectively.
In P. edulis, phe-novel-miR276 exerts its regulatory role by downregulating the expression of lignin biosynthesis-related target genes, and indirectly affects the activity of the antioxidant enzyme system in plants.
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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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