Functional analyses of PtrHBI 1 gene in Populus trichocarpa based on CRISPR/Cas9

WANG Zhuwen, GUO Yanjiao, LI Shuang, ZHOU Chenguang, CHIANG Vincent, LI Wei

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (6) : 31-39.

PDF(4684 KB)
南京林业大学学报（自然科学版）
PDF(4684 KB)
JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2021, Vol. 45 ›› Issue (6) : 31-39. DOI: 10.12302/j.issn.1000-2006.202107030

Functional analyses of PtrHBI 1 gene in Populus trichocarpa based on CRISPR/Cas9

Author information +
History +

Abstract

【Objective】 We used the CRISPR/Cas9 system to generate PtrHBI1 loss of function mutants of Populus trichocarpa in order to investigate the function of the PtrHBI1 transcription factor in wood formation. This investigation provides new clues for creating superior varieties of trees suitable for pulp and papermaking. 【Method】 Based on RNA-seq data of different cell types (cambium, xylem and phloem) of wild-type P. trichocarpa by laser capture microdissection, the bHLH transcription factor PtrHBI1 was identified. Subcellular localization of PtrHBI1 was analyzed using the stem-differentiating xylem (SDX) protoplast system of P. trichocarpa, and tissue-specific expression of PtrHBI1 was analyzed by in situ hybridization. We then used the CRISPR/Cas9 system to generate ptrhbi1 mutants of P. trichocarpa and analyzed the growth traits of ptrhbi1. Meanwhile, we performed a morphological analysis using paraffin sections and analyzed the wood composition of the mutant using the Klason acid hydrolysis method and high-performance liquid chromatography. 【Result】 According to the results, we found that PtrHBI1 was localized in the nucleus of P. trichocarpa SDX cells. The results of in situ hybridization showed that PtrHBI1 was mainly expressed in the cambium and xylem regions. The height of ptrhbi1 mutants created by CRISPR/Cas9 was significantly increased, and the internode number and stem basal diameter were significantly larger than those of the wild type during a certain period. In addition, the ptrhbi1 mutant showed a significant increase in the lumen area of per vessel cells and a significant decrease in the number of fiber cells. However, the number of vessel cells and layers of cambium cells in the ptrhbi1 mutant were similar to those in the wild type. The wood composition analysis showed that the glucose content of the ptrhbi1 mutant increased. Compared with the wild type, although the total lignin content was not significantly changed, the mass ratio of syringly to guaiacy (S/G) ratio of the mutant was significantly decreased. 【Conclusion】 PtrHBI1 was involved in regulating the growth and secondary xylem development of P. trichocarpa, which may play an important role in wood formation.

Key words

Populus trichocarpa / wood formation regulation / CRISPR/Cas9 / PtrHBI1 transcription factor

Cite this article

EndNote

Ris (Procite)

Bibtex

Download Citations
WANG Zhuwen , GUO Yanjiao , LI Shuang , et al . Functional analyses of PtrHBI 1 gene in Populus trichocarpa based on CRISPR/Cas9[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2021, 45(6): 31-39 https://doi.org/10.12302/j.issn.1000-2006.202107030

References

List( Publishing order | Descend order by publishing year | Descend order by cited within ) Chart analysis
[1]
KO J H, KIM H T, HAN K H. Biotechnological improvement of lignocellulosic feedstock for enhanced biofuel productivity and processing[J]. Plant Biotechnol Rep, 2011, 5(1):1-7.DOI: 10.1007/s11816-010-0159-7.
https://doi.org/10.1007/s11816-010-0159-7
http://link.springer.com/10.1007/s11816-010-0159-7
Cited in this article [1]
[2]
许会敏, 王莉, 曹德昌, 等. 维管形成层活动周期调控研究进展[J]. 科学通报, 2015, 60(7):619-629.
XU H M, WANG L, CAO D C, et al. Research progress on the regulation of cambium activity periodicity[J]. Chin Sci Bull, 2015, 60(7):619-629.DOI: 10.1360/N972014-01037.
https://doi.org/10.1360/N972014-01037
http://engine.scichina.com/doi/10.1360/N972014-01037
Cited in this article [1]
[3]
文静, 王春涛, 杨永平. 植物木质部次生细胞壁加厚调控的研究进展[J]. 西南林业大学学报, 2021, 41(2):182-188.
WEN J, WANG C T, YANG Y P. Advances in regulation of xylem secondary cell wall thickening in plants[J]. J Southwest For Coll, 2021, 41(2):182-188.DOI: 10.11929/j.swfu.201909077.
https://doi.org/10.11929/j.swfu.201909077
Cited in this article [1]
[4]
LIU C, YU H, RAO X L, et al. Abscisic acid regulates secondary cell-wall formation and lignin deposition in Arabidopsis thaliana through phosphorylation of NST1[J]. PNAS, 2021, 118(23): e2106367118.DOI: 10.1073/pnas.2010911118.
https://doi.org/10.1073/pnas.2010911118
http://www.pnas.org/lookup/doi/10.1073/pnas.2106367118
Cited in this article [1]
[5]
MARRIOTT P E, GóMEZ L D, MCQUEEN-MASON S J,. Unlocking the potential of lignocellulosic biomass through plant science[J]. New Phytol, 2016, 209(4):1366-1381.DOI: 10.1111/nph.13684.
https://doi.org/10.1111/nph.13684
https://onlinelibrary.wiley.com/toc/14698137/209/4
Cited in this article [1]
[6]
ZHONG R, LEE C, ZHOU J, et al. A battery of transcription factors involved in the regulation of secondary cell wall biosynjournal in Arabidopsis[J]. Plant Cell, 2008, 20(10):2763-2782.DOI: 10.1105/tpc.108.061325.
https://doi.org/10.1105/tpc.108.061325
https://academic.oup.com/plcell/article/20/10/2763-2782/6093073
Cited in this article [1]
[7]
ZHANG J, NIEMINEN K, SERRA J A, et al. The formation of wood and its control[J]. Curr Opin Plant Biol, 2014, 17:56-63.DOI: 10.1016/j.pbi.2013.11.003.
https://doi.org/10.1016/j.pbi.2013.11.003
https://linkinghub.elsevier.com/retrieve/pii/S1369526613001660
Cited in this article [1]
[8]
CHEN Y, ZHU P H, WU F, et al. Identification and characterization of the basic helix-loop-helix transcription factor family in Pinus massoniana[J]. Forests, 2020, 11(12):1292.DOI: 10.3390/f11121292.
https://doi.org/10.3390/f11121292
https://www.mdpi.com/1999-4907/11/12/1292
Cited in this article [1]
[9]
ZHANG T, LV W, ZHANG H, et al. Genome-wide analysis of the basic Helix-Loop-Helix (bHLH) transcription factor family in maize[J]. BMC Plant Biol, 2018, 18(1):235.DOI: 10.1186/s12870-018-1441-z.
https://doi.org/10.1186/s12870-018-1441-z
https://doi.org/10.1186/s12870-018-1441-z
Cited in this article [1]
[10]
BAI M Y, FAN M, OH E, et al. A triple helix-loop-helix/basic helix-loop-helix cascade controls cell elongation downstream of multiple hormonal and environmental signaling pathways in Arabidopsis[J]. Plant Cell, 2012, 24(12):4917-4929.DOI: 10.1105/tpc.112.105163.
https://doi.org/10.1105/tpc.112.105163
https://academic.oup.com/plcell/article/24/12/4917/6098050
Cited in this article [1]
[11]
陈儒钢, 巩振辉, 逯明辉, 等. 植物抗逆反应中的转录因子网络研究进展[J]. 农业生物技术学报, 2010, 18(1):126-134.
CHEN R G, GONG Z H, LU M H, et al. Research advance of the transcription factors networks related to plant adverse environmental stress[J]. J Agric Biotechnol, 2010, 18(1):126-134.DOI: 10.3969/j.issn.1674-7968.2010.01.020.
https://doi.org/10.3969/j.issn.1674-7968.2010.01.020
Cited in this article [1]
[12]
TOLEDO-ORTIZ G, HUQ E, QUAIL P H. The Arabidopsis basic/helix-loop-helix transcription factor family[J]. Plant Cell, 2003, 15(8):1749-1770.DOI: 10.1105/tpc.013839.
https://doi.org/10.1105/tpc.013839
https://academic.oup.com/plcell/article/15/8/1749/6010092
Cited in this article [1]
[13]
HEIM M A, JAKOBY M, WERBER M, et al. The basic helix-loop-helix transcription factor family in plants:a genome-wide study of protein structure and functional diversity[J]. Mol Biol Evol, 2003, 20(5):735-747.DOI: 10.1093/molbev/msg088.
https://doi.org/10.1093/molbev/msg088
https://academic.oup.com/mbe/article-lookup/doi/10.1093/molbev/msg088
Cited in this article [1]
[14]
LEDENT V, VERVOORT M. The basic helix-loop-helix protein family:comparative genomics and phylogenetic analysis[J]. Genome Res, 2001, 11(5):754-770.DOI: 10.1101/gr.177001.
https://doi.org/10.1101/gr.177001
http://genome.cshlp.org/lookup/doi/10.1101/gr.177001
Cited in this article [1]
[15]
CHU X, LI M, ZHANG S, et al. HBI1-TCP20 interaction positively regulates the CEPs-mediated systemic nitrate acquisition[J]. J Integr Plant Biol, 2021, 63(5):902-912.DOI: 10.1111/jipb.13035.
https://doi.org/10.1111/jipb.13035
https://onlinelibrary.wiley.com/toc/17447909/63/5
Cited in this article [1]
[16]
CAI H, CHAI M, CHEN F, et al. HBI1 Acts downstream of ERECTA and SWR1 in regulating inflorescence architecture through the activation of the brassinosteroid and auxin signaling pathways[J]. New Phytol, 2021, 229(1):414-428.DOI: 10.1111/nph.16840.
https://doi.org/10.1111/nph.16840
https://onlinelibrary.wiley.com/toc/14698137/229/1
Cited in this article [1]
[17]
FERRERO V, VIOLA I L, ARIEL F D, et al. Class I TCP transcription factors target the gibberellin biosynjournal gene GA20ox1 and the growth-promoting genes HBI1 and PRE6 during thermomorphogenic growth in Arabidopsis[J]. Plant Cell Physiol, 2019, 60(8):1633-1645.DOI: 10.1093/pcp/pcz137.
https://doi.org/10.1093/pcp/pcz137
https://academic.oup.com/pcp/article/60/8/1633/5530963
Cited in this article [1]
[18]
WANG S, LI L, XU P, et al. CRY1 interacts directly with HBI1 to regulate its transcriptional activity and photomorphogenesis in Arabidopsis[J]. J Exp Bot, 2018, 69(16):3867-3881.DOI: 10.1093/jxb/ery209.
https://doi.org/10.1093/jxb/ery209
https://academic.oup.com/jxb/article/69/16/3867/5026618
Cited in this article [1]
[19]
FAN M, BAI M Y, KIM J G, et al. The bHLH transcription factor HBI1 mediates the trade-off between growth and pathogen-associated molecular pattern-triggered immunity in Arabidopsis[J]. Plant Cell, 2014, 26(2):828-841.DOI: 10.1105/tpc.113.121111.
https://doi.org/10.1105/tpc.113.121111
https://academic.oup.com/plcell/article/26/2/828/6097975
Cited in this article [2]
[20]
LI Z, LIU Z B, XING A, et al. Cas9-guide RNA directed genome editing in soybean[J]. Plant Physiol, 2015, 169(2):960-970.DOI: 10.1104/pp.15.00783.
https://doi.org/10.1104/pp.15.00783
https://academic.oup.com/plphys/article/169/2/960-970/6114113
Cited in this article [1]
[21]
HSU P D, LANDER E S, ZHANG F. Development and applications of CRISPR-Cas9 for genome engineering[J]. Cell, 2014, 157(6):1262-1278.DOI: 10.1016/j.cell.2014.05.010.
https://doi.org/10.1016/j.cell.2014.05.010
https://linkinghub.elsevier.com/retrieve/pii/S0092867414006047
Cited in this article [1]
[22]
ALI Z, ABUL-FARAJ A, LI L, et al. Efficient virus-mediated genome editing in plants using the CRISPR/Cas9 system[J]. Mol Plant, 2015, 8(8):1288-1291.DOI: 10.1016/j.molp.2015.02.011.
https://doi.org/10.1016/j.molp.2015.02.011
https://linkinghub.elsevier.com/retrieve/pii/S1674205215001628
Cited in this article [1]
[23]
DOUDNA J A, CHARPENTIER E. The new frontier of genome engineering with CRISPR-Cas9[J]. Science, 2014, 346(6213):1258096.DOI: 10.1126/science.1258096.
https://doi.org/10.1126/science.1258096
https://www.science.org/doi/10.1126/science.1258096
Cited in this article [1]
[24]
FAN D, LIU T, LI C, et al. Efficient CRISPR/Cas9-mediated targeted mutagenesis in Populus in the first generation[J]. Sci Rep, 2015, 5:12217.DOI: 10.1038/srep12217.
https://doi.org/10.1038/srep12217
https://doi.org/10.1038/srep12217
Cited in this article [1]
[25]
ZHOU X H, JACOBS T B, XUE L J, et al. Exploiting SNPs for biallelic CRISPR mutations in the outcrossing woody perennial Populus reveals 4-coumarate:CoA ligase specificity and redundancy[J]. New Phytol, 2015, 208(2):298-301.DOI: 10.1111/nph.13470.
https://doi.org/10.1111/nph.13470
https://onlinelibrary.wiley.com/toc/14698137/208/2
Cited in this article [1]
[26]
LIN Y C, LI W, CHEN H, et al. A simple improved-throughput xylem protoplast system for studying wood formation[J]. Nat Protoc, 2014, 9(9):2194-2205.DOI: 10.1038/nprot.2014.147.
https://doi.org/10.1038/nprot.2014.147
https://doi.org/10.1038/nprot.2014.147
Cited in this article [2]
[27]
JAVELLE M, MARCO C F, TIMMERMANS M. In situ hybridization for the precise localization of transcripts in plants[J]. J Vis Exp, 2011(57):e3328.DOI: 10.3791/3328.
https://doi.org/10.3791/3328
Cited in this article [1]
[28]
WANG Z, MAO Y, GUO Y, et al. MYB transcription Factor161 mediates feedback regulation of Secondary wall-associated NAC-Domain1 family genes for wood formation[J]. Plant Physiol, 2020, 184(3):1389-1406.DOI: 10.1104/pp.20.01033.
https://doi.org/10.1104/pp.20.01033
https://academic.oup.com/plphys/article/184/3/1389-1406/6118265
Cited in this article [3]
[29]
LI S, ZHEN C, XU W, et al. Simple,rapid and efficient transformation of genotype Nisqually-1:a basic tool for the first sequenced model tree[J]. Sci Rep, 2017, 7(1):2638.DOI: 10.1038/s41598-017-02651-x.
https://doi.org/10.1038/s41598-017-02651-x
http://www.nature.com/articles/s41598-017-02651-x
Cited in this article [1]
[30]
HEIM M A, JAKOBY M, WERBER M, et al. The basic helix-loop-helix transcription factor family in plants:a genome-wide study of protein structure and functional diversity[J]. Mol Biol Evol, 2003, 20(5):735-747.DOI: 10.1093/molbev/msg088.
https://doi.org/10.1093/molbev/msg088
https://academic.oup.com/mbe/article-lookup/doi/10.1093/molbev/msg088
Cited in this article [1]
[31]
LI L, ZHOU Y, CHENG X, et al. Combinatorial modification of multiple lignin traits in trees through multigene cotransformation[J]. PNAS, 2003, 100(8):4939-4944.DOI: 10.1073/pnas.0831166100.
https://doi.org/10.1073/pnas.0831166100
http://www.pnas.org/cgi/doi/10.1073/pnas.0831166100
Cited in this article [3]
[32]
SAITO K, WATANABE Y, SHIRAKAWA M, et al. Direct mapping of morphological distribution of syringyl and guaiacyl lignin in the xylem of maple by time-of-flight secondary ion mass spectrometry[J]. Plant J, 2012, 69(3):542-552.DOI: 10.1111/j.1365-313x.2011.04811.x.
https://doi.org/10.1111/j.1365-313x.2011.04811.x
http://doi.wiley.com/10.1111/tpj.2012.69.issue-3
Cited in this article [2]
[33]
GUI J, LAM P Y, TOBIMATSU Y, et al. Fibre-specific regulation of lignin biosynjournal improves biomass quality in Populus[J]. New Phytol, 2020, 226(4):1074-1087.DOI: 10.1111/nph.16411.
https://doi.org/10.1111/nph.16411
https://onlinelibrary.wiley.com/toc/14698137/226/4
Cited in this article [3]
[34]
ZHAO K, LI S, YAO W, et al. Characterization of the basic helix-loop-helix gene family and its tissue-differential expression in response to salt stress in poplar[J]. Peer J, 2018, 6:e4502.DOI: 10.7717/peerj.4502.
https://doi.org/10.7717/peerj.4502
https://peerj.com/articles/4502
Cited in this article [3]
[35]
CHEN F, DIXON R A. Lignin modification improves fermentable sugar yields for biofuel production[J]. Nat Biotechnol, 2007, 25(7):759-761.DOI: 10.1038/nbt1316.
https://doi.org/10.1038/nbt1316
https://doi.org/10.1038/nbt1316
Cited in this article [1]
[36]
FREUDENBERG K. Lignin:its constitution and formation from p-Hydroxycinnamyl alcohols:lignin is duplicated by dehydrogenation of these alcohols;intermediates explain formation and structure[J]. Science, 1965, 148(3670):595-600.DOI: 10.1126/science.148.3670.595.
https://doi.org/10.1126/science.148.3670.595
https://www.science.org/doi/10.1126/science.148.3670.595
Cited in this article [1]
[37]
SARKANEN K V. Renewable resources for the production of fuels and chemicals[J]. Science, 1976, 191(4228):773-776.
https://doi.org/10.1126/science.191.4228.773
https://www.science.org/doi/10.1126/science.191.4228.773
Cited in this article [2]
[38]
LI L, CHENG X F, LESHKEVICH J, et al. The last step of syringyl monolignol biosynjournal in angiosperms is regulated by a novel gene encoding sinapyl alcohol dehydrogenase[J]. Plant Cell, 2001, 13(7):1567. DOI: 10.1105/tpc.13.7.1567.
https://doi.org/10.1105/tpc.13.7.1567
https://academic.oup.com/plcell/article/13/7/1567-1586/6009549
Cited in this article [2]
[39]
CHIANG V L. From rags to riches[J]. Nat Biotech, 2002, 20(6):557. DOI: 10.1038/nbt0602-557.
https://doi.org/10.1038/nbt0602-557
https://doi.org/10.1038/nbt0602-557
Cited in this article [1]
[40]
STEWART J J, AKIYAMA T, CHAPPIE C, et al. The effects on lignin structure of overexpression of ferulate 5-hydroxylase in hybrid poplar1[J]. Plant Physio, 2009, 150(2):621-635. DOI: 10.1104/pp.109.137059.
https://doi.org/10.1104/pp.109.137059
https://academic.oup.com/plphys/article/150/2/621/6107898
Cited in this article [1]
[41]
康向阳. 林木遗传育种研究进展[J]. 南京林业大学学报(自然科学版), 2020, 44(3):1-10.
KANG X Y. Research progress of forest genetics and tree breeding[J]. J Nanjing For Univ (Nat Sci Ed), 2020, 44(3):1-10.DOI: 10.3969/j.issn.1000-2006.202002033.
https://doi.org/10.3969/j.issn.1000-2006.202002033

RIGHTS & PERMISSIONS

Copyright reserved © 2021.
PDF(4684 KB)

Accesses

Citation

Detail
Sections
Recommended
The full text is translated into English by AI, aiming to facilitate reading and comprehension. The core content is subject to the explanation in Chinese.

Author

Reader

Reviewer

www.nldxb.njfu.edu.cn

Edited ＆ Published by Editorial Department of Nanjing Forestry University(Natural Sciences Edition)
Address： No.159 Longpan Road,Nanjing 210037 Jiangsu,P.R.China
E-mail ：xuebao@njfu.edu.cn , xuebao@njfu.com.cn
Telephone +86-25-85428247,85427076
Distributed by China International Book Trading Corporation P.O. Box 399, Beijing ,P.R. China

/

〈 〉