A functional study of bHLH106 transcription factor based on CRISPR/Cas9 in Populus trichocarpa

SUN Jiatong, GUO Yanjiao, LI Shuang, ZHOU Chenguang, CHIANG Vincent, LI Wei

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (6) : 15-23.

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南京林业大学学报（自然科学版）
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JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (6) : 15-23. DOI: 10.12302/j.issn.1000-2006.202107031

A functional study of bHLH106 transcription factor based on CRISPR/Cas9 in Populus trichocarpa

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Abstract

【Objective】 We generated CRISPR-based mutants of PtrbHLH106 to explore its function in the wood formation of Populus trichocarpa. 【Method】 Based on the previous RNA-seq data of various cell types (cambium, xylem and phloem cells) of P. trichocarpa, we cloned a bHLH transcription factor, PtrbHLH106, which is highly expressed in the cambium and xylem. The CRISPR/Cas9 gene editing system was used to generate ptrbhlh106 mutants. Phenotype observations of ptrbhlh106 and wild-type (WT) plants grown for 60, 90 and 120 d were carried out. Paraffin sections of stem internodes of ptrbhlh106 and WT plants grown for 120 days were created, and toluidine blue staining was used for observation and statistical analyses of different wood cells. 【Result】 Phenotype observation showed that there were no significant differences in plant height and ground diameter. Compared to that in WT plants, the internodes length of 8th stem reduced, total number of stem internodes increased, and the layer of cambium cells increased in mutants. However, the differences in the above indexes were not significant. In addition, the lumen area of per vessel increased, and the number of fiber cells decreased significantly in ptrbhlh106. 【Conclusion】 The phenotypic differences between ptrbhlh106 mutants and WT plants showed that PtrbHLH106 is involved in the regulation of secondary xylem development in P. trichocarpa.

Key words

Populus trichocarpa / secondary xylem development / CRISPR/Cas9 gene editing system / PtrbHLH106 transcription factor

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SUN Jiatong , GUO Yanjiao , LI Shuang , et al . A functional study of bHLH106 transcription factor based on CRISPR/Cas9 in Populus trichocarpa[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2021, 45(6): 15-23 https://doi.org/10.12302/j.issn.1000-2006.202107031

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