Regulation of LBD12 transcription factor on wood formation in Populus trichocarpa

GAO Yuan, SUN Jiatong, ZHOU Chenguang, CHIANG Vincent, LI Wei, LI Shuang

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2024, Vol. 48 ›› Issue (1) : 29-38.

PDF(4417 KB)
PDF(4417 KB)
JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2024, Vol. 48 ›› Issue (1) : 29-38. DOI: 10.12302/j.issn.1000-2006.202301002

Regulation of LBD12 transcription factor on wood formation in Populus trichocarpa

Author information +
History +

Abstract

【Objective】The formation of wood, one of the most important raw materials for pulp and energy, depends on a complex and precise transcriptional regulation process in which transcription factors figure prominently. In Populus trichocarpa, PtrLBD12 (lateral organ boundaries domain 12) is a transcription factor lying downstream of PtrbHLH186, a key regulator of wood formation. Hence, PtrLBD12 was studied in depth here to investigate its function in tree growth and wood formation.【Method】 To determine the role of the PtrLBD12 transcription factor in the growth and wood formation of poplar trees, we analyzed its expression characteristics, generated PtrLBD12 overexpressing plants of P. trichocarpa, and assessed the growth and wood formation traits of transgenic plants. (1) Xylem, phloem, and terminal bud and leaf samples of wild-type P. trichocarpa plants, cultivated in a greenhouse, were collected to extract their respective RNA. Next, transcriptome sequencing was done to analyze the patterns of differentially expressed genes in those distinct tissues, as well as their expression levels of PtrLBD12. (2) To clarify the expression pattern of the PtrLBD12 protein, its subcellular localization was investigated by using the transient transformation system of stem-differentiating xylem protoplasts of P. trichocarpa. (3) To create PtrLBD12-overexpressing plants, the Agrobacterium tumefaciens-mediated transformation system of P. trichocarpa was used, with transgenic plants identified at both the DNA and RNA level. (4) Plant stem height, ground (basal) stem diameter, number of stem nodes, and length of the 8th stem node of transgenic and wild-type plants were measured at 30, 60 and 90 days after planting. (5) The 2nd, 4th, 6th and 8th stem segments of overexpressing and wild-type greenhouse plants cultivated for 4 months were paraffin-sectioned. Their stem characteristics were then respectively observed via Safranin O/Fast Green and Toluidine Blue staining. LAS X V2.0 software was used to calculate the number of vessel and fiber cells, as well as average lumen area of vessel. (6) The relative expression levels of 22 monolignol biosynthetic pathway genes in the PtrLBD12-overexpressing plants were determined by using quantitative PCR and applying the 2△△Ct calculation.【Result】 (1) Transcriptome analysis of the xylem, phloem, and terminal bud and leaf tissues of P. trichocarpa revealed a higher expression level of PtrLBD12 in both the xylem and phloem. (2) Subcellular localization showed that PtrLBD12 was expressed in the nucleus, where transcription factors in general were found. (3) Three PtrLBD12 overexpressing transgenic lines of P. trichocarpa were obtained, whose relative expression levels were 40.91, 79.51 and 102.19. (4) Overexpressing PtrLBD12 adversely affected the normal growth and development of P. trichocarpa: plant height, ground stem diameter, number of stem nodes, and length of the 8th stem node all decreased significantly in transgenic plants vis-à-vis the wild type. (5) However, overexpression of PtrLBD12 did lead to significantly more vessel and fiber cells per unit of area in the stems of transgenic plants, but these vessels had smaller lumen area. Furthermore, the augmented expression of PtrLBD12 enhanced lignification of the plant stem. (6) Finally, PtrLBD12’s overexpression bolstered the expression levels of multiple genes in the monolignol biosynthetic pathway, namely PtrPAL1, PtrC4H1, PtrC4H2, PtrHCT6, PtrCSE, PtrCSE2, PtrCCoAOMT1, PtrCCoAOMT2, PtrCCoAOMT3, PtrCCR2 and PtrCAld5H1.【Conclusion】As the gene downstream of PtrbHLH186, a key regulator of wood formation in poplar, the LBD12 transcription factor is able to govern the expression of monolignol biosynthetic genes, change the mode of lignin deposition, alter the morphology of xylem cells, and thereby affect plant growth and development.

Key words

Populus trichocarpa / wood formation / genetic transformation / transcription factor / PtrLBD12

Cite this article

Download Citations
GAO Yuan , SUN Jiatong , ZHOU Chenguang , et al . Regulation of LBD12 transcription factor on wood formation in Populus trichocarpa[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2024, 48(1): 29-38 https://doi.org/10.12302/j.issn.1000-2006.202301002

References

[1]
ZHANG J, NIEMINEN K, SERRA J A, et al. The formation of wood and its control[J]. Current Opinion Plant Biology, 2014, 17: 56-63. DOI: 10.1016/j.pbi.2013.11.003.
[2]
SIEDLECKA A, WIKLUND S, PÉRONNE MA, et al. Pectin methyl esterase inhibits intrusive and symplastic cell growth in developing wood cells of Populus[J]. Plant Physiology, 2008, 146(2): 554-565. DOI: 10.1104/pp.107.111963.
[3]
孙滨, 占小登, 曹立勇, 等. 水稻AP2/ERF转录因子的研究进展[J]. 农业生物技术学报, 2017, 25(11): 1860-1869.
SUN B, ZHAN X D, CAO L Y, et al. Research progress of AP2/ERF transcription factor in rice (Oryza sativa)[J]. Journal of Agricultural Biotechnology, 2017, 25(11): 1860-1869. DOI: 10.3969/j.issn.1674-7968.2017.11.013.
[4]
姜秀明. 番茄HD-Zip转录因子家族生物信息学分析及HD-ZipⅠ亚族抗逆相关基因鉴定分析[D]. 哈尔滨: 东北农业大学, 2017.
JIANG X M. Bioinformatics analysis of HD-zip gene family and identification of HD-zip ⅠGenes related to stress resistance in tomato[D]. Harbin:Northeast Agricultural University, 2017.
[5]
毛昱力. 毛果杨PtrDLTPtrWOX4基因在维管形成层发育中的功能解析[D]. 哈尔滨: 东北林业大学, 2020. DOI: 10.27009/d.cnki.gdblu.2020.000763.
MAO Y L. Functional analysis of PtrDLT and PtrWOX4 genes in the development of vascular cambium in Populus trichocarpa[D]. Harbin:Northeast Forestry University, 2020.
[6]
CHEN H, WANG J P, LIU H Z, et al. Hierarchical transcription factor and chromatin binding network for wood formation in black cottonwood (Populus trichocarpa)[J]. Plant Cell, 2019, 31(3): 602-626. DOI: 10.1105/tpc.18.00620.
[7]
MCCARTHY R L, ZHONG R Q, YE Z H. MYB83 is a direct target of SND1 and acts redundantly with MYB46 in the regulation of secondary cell wall biosynthesis in Arabidopsis[J]. Plant Cell Physiology, 2009, 50(11): 1950-1964. DOI: 10.1093/pcp/pcp139.
[8]
WANG Z F, MAO Y L, GUO Y J, et al. MYB transcription Factor161 mediates feedback regulation of Secondary wall-associated NAC-Domain1 family genes for wood formation[J]. Plant Physiology, 2020, 184(3): 1389-1406. DOI: 10.1104/pp.20.01033.
[9]
ZHONG R Q, LEE C H, YE Z H. Functional characterization of poplar wood-associated NAC domain transcription factors[J]. Plant Physiology, 2010, 152(2): 1044-1055. DOI: 10.1104/pp.109.148270.
[10]
ZHONG R Q, MCCARTHY R L, LEE C H, et al. Dissection of the transcriptional program regulating secondary wall biosynthesis during wood formation in poplar[J]. Plant Physiology, 2011, 157(3): 1452-1468. DOI: 10.1104/pp.111.181354.
[11]
OHTANI M, NISHIKUBO N, XU B, et al. A NAC domain protein family contributing to the regulation of wood formation in poplar[J]. Plant J, 2011, 67(3): 499-512. DOI: 10.1111/j.1365-313X.2011.04614.x.
[12]
LI Q Z, LIN Y C, SUN Y H, et al. Splice variant of the SND1 transcription factor is a dominant negative of SND1 members and their regulation in Populus trichocarpa[J]. PNAS, 2012, 109(36): 14699-14704. DOI: 10.1073/pnas.1212977109.
[13]
LIN Y C, LI W, SUN Y H, et al. SND1 transcription factor-directed quantitative functional hierarchical genetic regulatory network in wood formation in Populus trichocarpa[J]. Plant Cell, 2013, 25(11): 4324-4341. DOI: 10.1105/tpc.113.117697.
[14]
LU S F, LI Q Z, WEI H R, et al. Ptr-miR397a is a negative regulator of laccase genes affecting lignin content in Populus trichocarpa[J]. PNAS, 2013, 110(26): 10848-10853. DOI: 10.1073/pnas.1308936110.
[15]
YEH C S, WANG Z F, MIAO F, et al. A novel synthetic-genetic-array-based yeast one-hybrid system for high discovery rate and short processing time[J]. Genome Research, 2019, 29(8): 1343-1351. DOI: 10.1101/gr.245951.118.
[16]
ZHONG R Q, LEE C H, ZHOU J L, et al. A battery of transcription factors involved in the regulation of secondary cell wall biosynthesis in Arabidopsis[J]. Plant Cell, 2008, 20(10): 2763-2782. DOI: 10.1105/tpc.108.061325.
[17]
ZHOU J L, LEE C H, ZHONG R Q, et al. MYB58 and MYB63 are transcriptional activators of the lignin biosynthetic pathway during secondary cell wall formation in Arabidopsis[J]. Plant Cell, 2009, 21(1): 248-266. DOI: 10.1105/tpc.108.063321.
[18]
KUBO M, UDAGAWA M, NISHIKUBO N, et al. Transcription switches for protoxylem and metaxylem vessel formation[J]. Genes and Development, 2005, 19(16): 1855-1860. DOI: 10.1101/gad.1331305.
[19]
ZHONG R Q, RICHARDSON E A, YE Z H. Two NAC domain transcription factors, SND1 and NST1, function redundantly in regulation of secondary wall synthesis in fibers of Arabidopsis[J]. Planta, 2007, 225(6): 1603-1611. DOI: 10.1007/s00425-007-0498-y.
[20]
LIN Y C J, CHEN H, LI Q Z, et al. Reciprocal cross-regulation of VND and SND multigene TF families for wood formation in Populus trichocarpa[J]. PNAS, 2017, 114(45): E9722-E9729. DOI: 10.1073/pnas.1714422114.
[21]
LIU H Z, GAO J H, SUN J T, et al. Dimerization of PtrMYB074 and PtrWRKY19 mediates transcriptional activation of PtrbHLH186 for secondary xylem development in Populus trichocarpa[J]. New Phytololgist, 2022, 234(3): 918-933. DOI: 10.1111/nph.18028.
[22]
SHUAI B, REYNAGA-PEÑA C G, SPRINGER P S. The lateral organ boundaries gene defines a novel, plant-specific gene family[J]. Plant Physiology, 2002, 129(2): 747-761. DOI: 10.1104/pp.010926.
[23]
YAMAGUCHI M, OHTANI M, MITSUDA N, et al. VND-INTERACTING2, a NAC domain transcription factor, negatively regulates xylem vessel formation in Arabidopsis[J]. Plant Cell, 2010, 22(4): 1249-63. DOI: 10.1105/tpc.108.064048.
[24]
YORDANOV Y S, REGAN S, BUSOV V. Members of the lateral organ boundaries domain transcription factor family are involved in the regulation of secondary growth in Populus[J]. Plant Cell, 2010, 22(11): 3662-3677. DOI: 10.1105/tpc.110.078634.
[25]
YU J, ZHOU C G, LI D N, et al. A PtrLBD39-mediated transcriptional network regulates tension wood formation in Populus trichocarpa[J]. Plant Communications, 2021, 3(1): 100250. DOI: 10.1016/j.xplc.2021.100250.
[26]
殷小雨, 胡凤荣, 杨盼盼. LBD转录因子调控植物生长发育的研究进展[J/OL]. 分子植物育种, 2021: 1-11. https: //kns.cnki.net/kcms/detail/46.1068.s.20210708.1419.013.html.
YIN X Y, HU F R, YANG P P. Research progress of LBD transcription factors regulating plant growth and development[J/OL]. Mol Plant Breed, 2021: 1-11.
[27]
LI S J, ZHEN C, XU W J, et al. Simple, rapid and efficient transformation of genotype Nisqually-1: a basic tool for the first sequenced model tree[J]. Scientific Reports, 2017, 7(1): 2638. DOI: 10.1038/s41598-017-02651-x.
[28]
LIN Y C, LI W, CHEN H, et al. A simple improved-throughput xylem protoplast system for studying wood formation[J]. Nature Protocols, 2014, 9(9): 2194-205. DOI: 10.1038/nprot.2014.147.
[29]
WANG J P, MATTHEWS M L, WILLIAMS C M, et al. Improving wood properties for wood utilization through multi-omics integration in lignin biosynthesis[J]. Nature Communications, 2018, 9(1): 1579. DOI: 10.1038/s41467-018-03863-z.
[30]
LI S, LIN Y C J, WANG P Y, et al. The AREB1 transcription factor influences histone acetylation to regulate drought responses and tolerance in Populus trichocarpa[J]. Plant Cell, 2019, 31(3): 663-686. DOI: 10.1105/tpc.18.00437.
[31]
KASUGA M, LIU Q, MIURA S, et al. Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor[J]. Nature Biotechnology, 1999, 17(3): 287-291. DOI: 10.1038/7036.
[32]
YAN Y, WANG P, LU Y, et al. MeRAV5 promotes drought stress resistance in cassava by modulating hydrogen peroxide and lignin accumulation[J]. Plant Journal, 2021, 107(3): 847-860. DOI: 10.1111/tpj.15350.
[33]
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]. Journal of Nanjing Forestry University (Natural Sciences Editon), 2020, 44(3): 1-10. DOI: 10.3969/j.issn.1000-2006.202002033.
PDF(4417 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.

/