Abstract
In this study, we investigated the roles of the plant hormones indole-3-acetic acid (IAA) and gibberellin (GA) in regulating Pinus massoniana Lamb. plantlet regeneration during long-term in vitro subculture. Six generations (1st, 5th, 10th, 20th, 30th and 40th with 40 days for each transfer cycle) of subcultured shoots derived from mature and juvenile explants were characterised for their different rooting capacities. In the present experiment, shoots were subcultured on rooting medium containing indole-3-acetic acid (IAA), naphthaleneacetic acid (NAA) or gibberellin (GA) inhibitor (paclobutrazol, PBZ). In addition, high-performance liquid chromatography (HPLC) was used to analyse endogenous hormone levels in shoots developed on medium with low concentrations of NAA. The concentration of endogenous IAA was highest in the 20th generation shoots, which also exhibited the strongest rooting ability. However, the highest GA concentration was observed in the 40th generation shoots, which exhibited a poor rooting capacity. For shoots subcultured for 40 generations, incubation on rooting medium containing IAA caused an enhanced root number, shoot length and survival rate. This suggested that the effects of IAA on shoot and root formation were positive. PBZ promoted plantlet regeneration from in vitro cultures but impaired shoot elongation at high levels. Furthermore, PBZ improved performance of in vitro cultures after long-term (3 to 4 y) subculture. These results suggest the inhibitory role of a high endogenous GA level in plant regeneration and the use of appropriate levels of PBZ for plant regeneration of P. massoniana.
Similar content being viewed by others
References
Aloni R, Aloni E, Langhans M, Ullrich CI (2006) Role of cytokinin and auxin in shaping root architecture: regulating vascular differentiation, lateral root initiation, root apical dominance and root gravitropism. Ann Bot 97:883–893
Beltran V, Salvadó N, Butí S, Pradell T (2016) Ageing of resin from Pinus species assessed by infrared spectroscopy. Anal Bioanal Chem 408:4073–4082
Bonga JM (1987) Clonal propagation of mature trees: problems and possible solutions. In: Bonga JM, Durzan DJ (eds) Cell and tissue culture in forestry. Martinus Nijhoff, Dordrecht, pp 249–271
Bonga JM, Von Adrekas P (1993) Rejuvenation of tissues from mature conifers and its implications for propagation in vitro. In: Ahuja MR, Libby WJ (eds) Clonal forestry I. Springer-Verlag, Berlin, pp 182–199
Cano A, Sánche-García AB, Albacete A, González-Bayón R, Justamante MS, Ibáñez S, Acosta M, Pérez-Pérez JM (2018) Enhanced conjugation of auxin by GH3 enzymes leads to poor adventitious rooting in carnation stem cuttings. Front Plant Sci 9:1–17
De Smet I, Signora L, Beeckman T, Inze D, Foyer CH, Zhang HM (2003) An abscisic acid-sensitive check point in lateral root development of Arabidopsis. Plant J 33:543–555
Ding GJ, Zhou ZC, Wang ZR (2006) Cultivation and utilization of pulpwood stand for Pinus massoniana. China Forestry Press, Beijing, pp 1–10
Duncan DB (1955) Multiple range and multiple F test. Biometrics 11:1–42
Eriksson ME, Israelsson M, Olsson O, Moritz T (2000) Increased gibberellin biosynthesis in transgenic trees promotes growth, biomass production and xylem fiber length. Nat Biotech 18:784–788
Fonouni-Farde C, Kisiala A, Brault M, Emery RJN, Diet A, Frugier F (2017) DELLA1-mediated gibberellin signaling regulates cytokinin-dependent symbiotic nodulation. Plant Physiol. https://doi.org/10.1104/pp.17.00919
Fonouni-Farde C, McAdam E, Nichols D, Diet A, Foo E, Frugier F (2018) Cytokinins and the CRE1 receptor influence endogenous gibberellin levels in Medicago truncatula. Plant Signal Behav 13:1–3
Fu X, Harberd NP (2003) Auxin promotes Arabidopsis root growth by modulating gibberellin response. Nature 421:740–743
Galavotti A (2013) The role of auxin in shaping shoot architecture. J Exp Bot 64:2593–2608
Hardtke CS (2003) Gibberellin signaling: GRASs growing roots. Curr Biol 13:366–367
Hedden P (2003) The genes of the green revolution. Trends Genet 19:5–9
Heide OM (2019) Juvenility, maturation and rejuvenation in plants: adventitious bud formation as a novel rejuvenation process. J Hort Sci Biotech 94:2–11
Himanen K, Vuylsteke M, Vanneste S, Vercruysse S, Boucheron E, Alard P, Chriqui D, Van Montagu M, Inze D, Beeckman T (2004) Transcript profiling of early lateral root initiation. Proc Natl Acad Sci U S A 101:5146–5151
Huang Y, Ji KS, Zhai JR (2007) Relationship between rooting ability and endogenous phytohormone changes in successive continuous generation cuttings of Buxus sinica var. parvifolia, an endangered woody species in China. Forest Stud China 9:189–197
Ji KS, Wang ZR, Chen TH, Wang MX (1996) Cyclophysis and effect of rejuvenation with continued cuttage in Pinus massoniana cutting propagation. J Zhejiang Forest Coll 16:341–345
Kamran M, Wennan S, Ahmad I, Xiangping M, Wenwen C, Xudong Z, Siwei M, Khan A, Qingfang H, Tiening L (2018) Application of paclobutrazol affect maize grain yield by regulating root morphological and physiological characteristics under a semi-arid region. Sci Rep 8:1–15
Li QF, Wang JD, Xiong M, Wei K, Zhou P, Huang LC, Zhang CQ, Fan XL, Liu QQ (2018) iTRAQ-based analysis of proteins co-regulated by brassinosteroids and gibberellins in rice embryos during seed germination. Int J Mol Sci 19:3460–3479
Li XY, Lv CQ, Huang BL, Wu QM, Zhang MH (2009) Adventitious roots induction of Pinus massoniana shoots in test tubes and anatomical observation. J Northwest For Univ 24:80–84
Marhavý P, Bielach A, Abas L, Abuzeineh A, Duclercq J, Tanaka H, Pařezová M, Petrášek J, Friml J, Kleine-Vehn J, Benková E (2011) Cytokinin modulates endocytic trafficking of PIN1 auxin efflux carrier to control plant organogenesis. Dev Cell 21:796–804
Marhavý P, Vanstraelen M, De Rybel B, Zhaojun D, Bennett MJ, Beeckman T, Benkova E (2013) Auxin reflux between the endodermis and pericycle promotes lateral root initiation. EMBO J 32:149–158
Mauriat M, Petterle A, Bellini C, Moritz T (2014) Gibberellins inhibit adventitious rooting in hybrid aspen and Arabidopsis by affecting auxin transport. Plant J 78:372–384
Merkle SA, Dean JF (2000) Forest tree biotechnology. Curr Opin Biotech 11:298–302
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Olszewski N, Sun TP, Gubler F (2002) Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell 14:S61–S80
Pullman GS, Zeng XY, Copeland-Kamp B, Crockett J, Lucrezi J, May SW, Bucalo K (2015) Conifer somatic embryogenesis: improvements by supplementation of medium with oxidation–reduction agents. Tree Physiol 35:209–224
Salari H, Baninasab B, Akbari M, Rohani AM (2017) Effect of paclobutrazol on adventitious root formation of IBA-treated cuttings of ‘Zard’ and ‘Dakal’ olive (Olea europaea L.) cultivars. Asian J Appl Sci 5:692–699
Schaller GE, Street IH, Kieber JJ (2014) Cytokinin and the cell cycle. Curr Opin Plant Biol 21:7–15
Shi XX, Du GQ, Wang C, Ma BK, Ge YN (2007) Effects of subculture times on organogenesis characteristics of apple in vitro shoot explants. Acta Hort Sin 34:561–564
Su XC (2000) Study on the differences of the seedling of different generations from successive tissue culture of Chinese fir clone. J Fujian Coll For 20:353–356
Sun TP, Gubler F (2004) Molecular mechanism of gibberellin signaling in plants. Ann Rev Plant Biol 55:197–223
Swain SM, Singh DP (2005) Tall tales from sly dwarves: novel functions of gibberellins in plant development. Trends Plant Sci 10:123–129
Ubeda-Tomas S, Swarup R, Coates J, Swarup K, Laplaze L, Beemster GTS, Hedden P, Bhalerao R, Bennett MJ (2008) Root growth in Arabidopsis requires gibberellin/DELLA signalling in the endodermis. Nat Cell Biol 10:625–628
Vaičiukynė M, Žiaukaa J, Žūkienė R, Vertelkaitė L, Kuusienė S (2019) Abscisic acid promotes root system development in birch tissue culture: a comparison to aspen culture and conventional rooting-related growth regulators. Physiol Plant 165:114–122
Von Aderkas P, Bonga JM (2000) Influencing micropropagation and somatic embryogenesis in mature trees by manipulation of phase change, stress and culture environment. Tree Physiol 20:921–928
Waldie T, Leyser OC (2018) Targets auxin transport to promote shoot branching. Plant Physiol 177:803–818
Wang Y, Yao R (2017) Plantlet regeneration of adult Pinus massoniana lamb. Trees using explants collected in march and thidiazuron in culture medium. J For Res 28:1169–1175
Wang Y, Yao RL (2019a) Increased endogenous indole-3-acetic acid:abscisic acid ratio is a reliable marker of Pinus massoniana rejuvenation. Biotech Histochem 94:546–553
Wang Y, Yao RL (2019b) Optimization of rhizogenesis for in vitro shoot culture of Pinus massoniana lamb. J For Res in press:1–7. https://doi.org/10.1007/s11676-019-01076-8
Watson GW (1996) Tree root system enhancement with paclobutrazol. J Arbor 22:211–217
Watson G (2004) Effect of transplanting and paclobutrazol on root growth of ‘green column’ black maple and ‘summit’ green ash. J Environ Hortic 22:209–212
Wendling I, Trueman SJ, Xavier A (2014) Maturation and related aspects in clonal forestry-part II: reinvigoration, rejuvenation and juvenility maintenance. New Forest 45:473–486
Yang JC, Zhang JH, Wang ZQ, Zhu QS, Wang W (2001) Hormonal changes in the grains of rice subjected to water stress during grain filling. Plant Physiol 127:315–323
Yang MH, Zhang DL, Li ZH, Jin XC, Ding GJ (2011a) Somatic embryogenesis with immature embryos of Masson pine (Pinus massoniana lamb.). Plant Physiol J 47:904–912
Yang MH, Zhang DL, Yang Y, Ding GJ, Li ZH (2011b) Micropropagation in immature embryos of Pinus massoniana in vitro. J Central South Univ For Tech 31:90–96
Yao RL, Wang Y (2016) An effective protocol for regenerating mature Pinus massoniana L. trees by tissue culture. Res J Biotech 11:75–80
Yao RL, Wang Y, Wu YM (2016) Key factors affecting rooting of Pinus massoniana by tissue culture. Guihaia 36:1288–1294
Zhu LH, Wu XQ, Qu HY, Ji J, Ye JR (2010) Micropropagation of Pinus massoniana and mycorrhiza formation in vitro. Plant Cell Tiss Org Cult 102:121–128
Funding
This work was supported by the key programme of Guangxi Forestry Bureau under Grant [2016]13, the project of Scientific and Technological Plan from the Department of Science and Technology of Guangxi under Grants 2017GXNSFAA198037, AD17195078, 2018GXNSFDA281020, AA17204087-1, and 2016GXNSFBA380224, and the Natural Science Foundation of China under Grant 31960311 and 31360178.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare that they have no conflicts of interest.
Additional information
Editor: Wenhao Dai
Rights and permissions
About this article
Cite this article
Wang, Y., Yao, R. Increased endogenous gibberellin level inhibits root growth of Pinus massoniana Lamb. plantlets during long-term subculture. In Vitro Cell.Dev.Biol.-Plant 56, 470–479 (2020). https://doi.org/10.1007/s11627-020-10067-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11627-020-10067-y