Regulating the regulators: responses of four plant growth regulators during clonal propagation of Lachenalia montana

Abstract

Lachenalia species are endemic southern African plants with narrow geographical distribution, and are well-traded as ornamental plants in the international floriculture industry. In an attempt to have a better understanding of their growth and hormonal physiology, we evaluated the effects of different plant growth regulators (PGRs) during the clonal regeneration of Lachenalia montana. An auxin (α-naphthaleneacetic acid = NAA) and three cytokinin (CK) types (benzyladenine = BA, meta-topolin riboside = mTR and isopentenyladenine = iP), each at three concentrations (1, 5 and 10 µM), were tested and the effect of these PGRs on the accumulation of endogenous CK metabolites was evaluated to provide clues on the observed morphological responses. As the most efficient PGR, 10 µM mTR treatment produced the highest number of shoots (approximately five shoots per explant) while 1 µM BA-treated plants had more bulbs (approximately three bulbs per plantlet). Rooting was generally lower with increasing concentration of PGRs especially with the aromatic-type CKs. Based on the concentrations of endogenous CKs, 10 µM mTR regenerants also had the highest CKs (40 142.5 pmol g−1 DW) which were mainly of the aromatic-type (98%). In terms of the functional role of the CKs, O-glucosides (which are reversible CK storage forms) were the most dominant CK-type in the regenerants from 10 µM mTR treatment. On the other hand, the poor rooting, mostly prominent in regenerants from BA treatments was closely related to the high accumulation of N 9-glucosides (well-known CK metabolites directly involved in rooting inhibition) when compared to regenerants from other treatments. Overall, the current findings provide evidence on the interrelationship existing among the exogenous PGRs, phenotypic responses and the endogenous CKs in the in vitro regenerants.

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Abbreviations

BA:

N 6-Benzyladenine

BA9G:

N 6-Benzyladenine-9-glucoside

BAR:

N 6-Benzyladenine-9-riboside

BAR5′MP:

N 6-Benzyladenine-9-riboside-5′-monophosphate

CK:

Cytokinin

cZ:

cis-Zeatin

cZOG:

cis-Zeatin-O-glucoside

cZR:

cis-Zeatin-9-riboside

cZR5′MP:

cis-Zeatin-9-riboside-5′-monophosphate

cZROG:

cis-Zeatin-O-glucoside riboside

DHZ:

Dihydrozeatin

DHZOG:

Dihydrozeatin-O-glucoside

DHZR:

Dihydrozeatin-9-riboside

DHZROG:

Dihydrozeatin-O-glucoside riboside

ESI:

Electro-spray interface

iP:

N 6-Isopentenyladenine

iPR:

N 6-Isopentenyladenine-9-riboside

iPR5′MP:

N 6-Isopentenyladenine-9-riboside-5′-monophosphate

MS:

Murashige and Skoog medium

mT:

meta-Topolin

mT9G:

meta-Topolin-9-glucoside

mTOG:

meta-Topolin-O-glucoside

mTR:

meta-Topolin-9-riboside

mTR5′MP:

meta-Topolin-9-riboside-5′-monophosphate

mTROG:

meta-Topolin-O-glucoside riboside

NAA:

α-Naphthaleneacetic acid

oT:

ortho-Topolin

oTR:

ortho-Topolin-9-riboside

PPF:

Photosynthetic photon flux density

pT:

para-Topolin

pTR:

para-Topolin-9-riboside

pTROG:

para-Topolin-O-glucoside riboside

tZ:

trans-Zeatin

tZOG:

trans-Zeatin-O-glucoside

tZR:

trans-Zeatin-9-riboside

tZR5′MP:

trans-Zeatin-9-riboside-5′-monophosphate

tZROG:

trans-Zeatin-O-glucoside riboside

UHPLC® :

Ultra-high performance liquid chromatography

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Acknowledgements

We thank Prof G. D. Duncan for the generous gift of bulbs used for the study. This work was financially supported by the University of KwaZulu-Natal and National Research Foundation (Green Economy Fellowship—U98028), South Africa. Additional finance was provided by the Ministry of Education, Youth and Sport of the Czech Republic (the Program “Návrat” for Research, Development, and Innovations, no. LK21306), National Program for Sustainability (Grant LO1204) and the Czech Science Foundation (Grant 14-34792S). We thank Mrs Alison Young (UKZN Botanical Garden, Pietermaritzburg, South Africa) and her staff for maintaining the mother plants in the greenhouse. We acknowledge the contribution of the Southern African Systems Analysis Centre, the National Research Foundation and the Department of Science and Technology in South Africa as well as the International Institute of Applied Systems Analysis in Austria.

Author contributions

AOA conceived the research idea and designed the experiments. Micropropagation experiments and data collection were conducted by AOA, NAM, MM and SOA. LP and ON conducted and (together with KD) analysed the CK data. AOA prepared the draft manuscript with help of all the other authors. KD prepared mTR, KD and JVS contributed research facilities/reagents/materials and supervised the research. All authors read and edited the final manuscript.

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Correspondence to Johannes Van Staden.

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Aremu, A.O., Plačková, L., Masondo, N.A. et al. Regulating the regulators: responses of four plant growth regulators during clonal propagation of Lachenalia montana . Plant Growth Regul 82, 305–315 (2017). https://doi.org/10.1007/s10725-017-0260-9

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Keywords

  • Asparagaceae
  • Floriculture
  • Phytohormones
  • Physiological disorders
  • meta-Topolin
  • Ornamentals