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The CUT&Tag technology is a new method for protein-DNA interactions, which uses a novel pG-Tn5 transposase with ultra-high activity, to precisely target and cleave DNA sequences near the target protein under antibody guidance, thus enabling cDNA library construction and sequencing analysis. This technology is widely used in human and animal research, but due to the unique structure of plant cells, its application in plant research is relatively limited. In previous studies, we found histone acetylation can participate in the regulation of jasmonic acid induced secondary laticifer differentiation, but the molecular mechanism of histone acetylation modification regulating secondary laticifer differentiation has not been elucidated in rubber tree. In this article, we used the experimental system of secondary laticifer differentiation induced by cornetin (COR) in the vascular cambium by the CUT&Tag technology, and the high quality protoplasts of cambium cells were obtained by enzymolysis, and the Histone H3 acetylation modified antibody was used to identify in situ the region of histone acetylation modification during secondary laticifer differentiation. A cDNA library of cambium cells treatment by COR was successfully constructed using CUT&Tag technology in rubber tree bark. Quality inspection and sequencing analysis were conducted on the construction of cDNA libraries, and found that the quality of libraries were good. And GO and KEGG enrichment analysis of differential genes found the genes related to auxin, flavonoid metabolism, and protein ubiquitination were enriched. The results would provide an operational method for constructing cDNA libraries of plant tissues using CUT&Tag technology, and provide a theoretical basis for elucidating the molecular mechanism of histone acetylation modification regulating secondary laticifer differentiation in rubber tree.
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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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