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Scientia Horticulturae 263 (2020) 109108
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Short communication
Willow bark extract and the biostimulant complex Root Nectar® increase propagation efficiency in chrysanthemum and lavender cuttings
Kimber Wisea,b, Harsharn Gilla, Jamie Selby-Phamb,* a b
School of Science, RMIT University, Bundoora, VIC 3083, Australia Nutrifield, Sunshine West, VIC 3020, Australia
Keywords: Horticulture Salicylic acid Kelp extract Aloe vera extract BBCH Rooting
Biostimulants are a powerful tool for optimizing the propagation efficiency of vegetative cuttings, however, their optimal application rates are often species specific. Herein, the root promoting effects of Nutrifield’s biostimulant complex Root Nectar® and the biostimulant willow bark extract were tested on vegetative cuttings from lavender (soft wood) and chrysanthemum (semi-hard wood). Application of Root Nectar® at 1 mL/L achieved consistent improvements in the formation of adventitious roots and root branching in both lavender and chrysanthemum, and 1.06 μL/L willow bark extract achieved best results for both species. The root promoting biostimulant effects of Root Nectar® and willow bark extract can be utilized to speed up propagation of soft and semi-hard wood cuttings, highlighting their applicability to improve process efficiencies in horticultural industries, which are dependent on rapid large-scale vegetative propagation.
1. Introduction Plant propagation is a vital part of both the agricultural and horticultural industries. While there are many forms of plant propagation, vegetative propagation - colloquially known as ‘cuttings’ - remains the preferred method for many species due to the simplicity and continuation of desirable genetic traits (Adam, 2006; Arjana et al., 2015; Caplan et al., 2018; Nakhooda and Jain, 2016; Potter, 2014; Schum and Preil, 1998). Principle methods of propagating by cuttings involve the support of excised tissue with moisture and light until adventitious roots (AR) emerge (Owen and Maynard, 2007). The process of AR formation is a complex process, often requiring particular environmental conditions and hormonal stimuli which are specific to both species and tissue-type (Agbo and Obi, 2007; Hassanein, 2013). Modern methods of propagating cuttings utilise sophisticated technologies such as synthetic plant hormones, misting and fogging systems, aeroponics systems, and advanced hygiene protocols to optimise for AR formation and thereby maximise propagation efficiency (Preece, 2003). Whilst there have been many significant advancements to vegetative propagation, economic loss due to insufficient rooting efficiency remains a burden on the propagation industry, thus additional work is needed to identify biostimulants which promote rooting (Ahkami et al., 2009; Preece, 2003).
Biostimulants are non-fertiliser materials which, when applied to plants in small quantities, have growth promoting/altering effects (Khan et al., 2009). Kelp extracts are well-established biostimulants within the agricultural industry (Arioli et al., 2015; Crouch and Van Staden, 1993), and are associated with a range of improvements to plant growth and yield, including; increased biomass, nutrient uptake, germination, and root growth (Battacharyya et al., 2015; Khan et al., 2009). Similarly, Aloe vera extracts have also been shown to have biostimulating effects, including increases to plant growth, biomass, root growth, and oil production (El Sherif, 2017; Hamouda et al., 2012). Both kelp and Aloe vera extracts are feature active components in the root stimulating biostimulant complex, Root Nectar® (Nutrifield Pty Ltd., Melbourne, VIC). Additionally, willow bark extracts, containing high levels of the phytohormone salicylic acid (SA), have been shown to exhibit biostimulating effects on plant growth, with reported effects on flowering, callus formation, stress mediation, and rooting (AlAmad and Qrunfleh, 2016; Gesto et al., 1977; Hayat and Ahmad, 2007; Kammerer et al., 2005; Kawase, 1964; Raskin, 1992). Salicylic acid and other salicylates have been shown to promote root growth in cuttings, seedling, and mature plants, of several species, including soybean, carrot, and Eranthemum (Basu et al., 1969; Gutiérrez-Coronado et al., 1998; Hayat et al., 2010). Herein, application of Root Nectar® and the biostimulant willow bark extract to vegetative cuttings of
Abbreviations: AR, adventitious root; BBCH, Biologische Bundesanstalt, Bundessortenamt, Chemische Industrie; GLM, general linear model; IAA, indole acetic acid; JA, jasmonic acid; L:D, light:dark; SA, salicylic acid ⁎ Corresponding author at: Research & Development, Nutrifield, Sunshine West, Victoria, 3020, Australia. E-mail address: [email protected] (J. Selby-Pham). Received 9 August 2019; Received in revised form 3 December 2019; Accepted 5 December 2019 0304-4238/ © 2019 Published by Elsevier B.V.
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chrysanthemum and lavender, was explored for their capacities to increase adventitious root formation and root branching, and thereby improve propagation efficiency of vegetative cuttings.
Table 1 Description of the phenological stages of chrysanthemum and lavender cuttings during principal growth stage 0 (rooting), using a 2-digit BBCH scale.
2. Materials and methods 2.1. Stock plants Stock plants of Chrysanthemum sp., and Lavandula x hybrid ‘Frills’ PBR were grown hydroponically in indoor growth rooms. The stock plants were grown under 315 W ceramic metal-halide horticultural lamps, with a light:dark (L:D) photoperiod of 12 h day and 12 h night, day temperature and relative humidity of 21.5 °C, and 70%, respectively, and night temperature and relative humidity of 18 °C, and 51%, respectively. Stock plants were provided with Coco A&B (Nutrifield Pty Ltd., Melbourne, VIC) nutrients weekly, at EC = 1.5, and pH = 5.8 and pruned regularly to prevent flowering.
Description of stage
00 05 06
Cutting, no roots present Adventitious roots present, no root branching Root branching
2.6. Statistical analysis Biostimulant effect on rooting efficiency was assessed by general linear model (GLM), and Tukey’s test 95% confidence grouping analyses, which were performed in the Minitab 18 statistical software package (Minitab Inc., State College, PA). 3. Results
2.2. Cuttings
Chrysanthemum and lavender cuttings were propagated with biostimulants and scored as per the BBCH scale (Fig. 1) to compare devlopemnt rates over time. Application of the biostimulant complex Root Nectar® at 1 mL/L, decreased the time taken for adventitous root formation, and root branching, in vegetative cuttings (Fig. 2). Root Nectar® treatment had a significant reduction in the timing of AR formation (BBCH = 05) for chrysanthemum (P = 0.043), and lavender (P = 0.013). Chrysanthemum cuttings developed AR within 10.4, and 12.3 days, when treated with Root Nectar® or water, respectively (15.5% faster with Root Nectar®). While lavender cuttings treated with Root Nectar® and water developed AR within 12.8, and 19.3 days, respectively (33.8% faster with Root Nectar®). Additionally, Root Nectar® treatment of chrysanthemum (P = 0.003), and lavender (P = 0.001) cuttings had a significant reduction in the timing of root branching initiation (BBCH = 06). Chrysanthemum cuttings began root branching within 14.4, and 18.4 days, when treated with Root Nectar® or water, respectively (22.1% faster with Root Nectar®), while lavender cuttings treated with Root Nectar® and water began root branching in 17.6, and 27.6 days, respectively (36.2% with Root Nectar®). Willow bark extract was applied to chrysanthemum and lavender cuttings at 4 concentrations to assess the biostimulant effect on AR formation and root branching, and to identify the optimal application rate (Fig. 3). AR formation (BBCH = 05) in chrysanthemum cuttings was not significantly different (P = 0.121) between control and willow treatment groups, while root branching (BBCH = 06) was (P = 0.010). The best results for chrysanthemum were observed for 1.06 μL/L willow bark extract treatment, which achieved root branching in 13.9 days, while control cuttings achieved root branching in 18.4 days (12.83% faster with willow bark extract). For lavender, all levels of willow treatment showed similar differences to control for both AR formation (P < 0.001), and root branching (P < 0.001). Lavender cuttings treated with water (control group) achieved AR formation in 19.3 days, while those treated with 1.06 μL/L willow bark extract achieved AR growth in 12.0 days (37.7% faster than control). Additionally, lavender cuttings control group achieved root branching in 27.6 days, while those treated with 1.06 μL/L willow bark extracts achieved root branching in 15.6 days (43.4% faster than control). Of the four concentrations of willow bark extract tested (1.06, 2.12, 4.23, and 10.58 μL/L), 1.06 μL/L overall achieved the best results for vegetative propagation of chrysanthemum and lavender cuttings.
Eight cuttings of each chrysanthemum and lavender (a single propagation event) were taken from stock plants with young shoots and minimal branching. All cuttings were tip pruned and cut at the base 2 mm below the last node. Chrysanthemum cuttings (10 nodes and 54–144 mm in length), were taken from semi-hard wood tissue and all leaves and side shoots were removed except leaves from the top two nodes (two leaves total per cutting). While lavender cuttings (6 nodes and 54–117 mm in length), were taken from softwood tissue and retained leaves on the top most node only (two leaves total per cutting). 2.3. Propagation Excised cuttings were immediately placed into aeroponic mist-propagation units (EzClone Aeroponic Classic Cutting System - 16, EZCLONE Enterprises Inc., Sacramento, CA) and maintained in indoor growth rooms under 600 W metal-halide horticultural lamps, with a L:D photoperiod of 18 h day and 6 h night, day temperature and humidity of 22 °C and 70%, respectively, and night temperature and humidity of 19 °C and 51%, respectively. Lavender cuttings were placed into EZ clone collars with 2 nodes above the top of the collar, while chrysanthemum cuttings were placed into collars with 3 nodes above the top of the collar. 2.4. Biostimulants Propagation solutions (10 L per aeroponic unit containing 16 cuttings) were made using the root stimulating liquid fertiliser, Root Nectar® (Nutrifield Pty Ltd., Melbourne, VIC) containing 4.02 g/L nitrate, 1.74 g/L phosphate, 2.32 g/L potassium, 0.1% Aloe vera extract, and 0.1% Ascophyllym nodosum kelp extract, applied at 0, or 1 mL/L (as per recommended usage instructions), and willow bark extract (Willow bark extract (10:1), Native Extracts Pty Ltd., Alstonville, NSW) applied at 0, 1.06, 2.12, 4.23, and 10.58 μL/L. Propagation solutions were replaced with new solution weekly. Root Nectar® was provided by Nutrifield Pty Ltd. for testing purposes. 2.5. BBCH scoring
4. Discussion
Cuttings were scored daily to measure root development progress. Scoring was done using the extended BBCH (Biologische Bundesanstalt, Bundessortenamt, CHemische Industrie) numerical scale, adapted as presented in Table 1 (Herraiz et al., 2015; Meier et al., 2009; Niemenak et al., 2010).
The root promoting biostimulant complex Root Nectar® was found to decrease the time taken to achieve AR formation and root branching, in chrysanthemum (semi-hard wood) and lavender (soft wood) cuttings 2
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Fig. 1. Principal growth stages of chrysanthemum and lavender cuttings 00–06. Representative photographs (from 8 biological replicates) of the phenological stages of Chrysanthemum sp., and Lavandula x hybrid ‘Frills’ PBR (lavender), categorised according to the BBCH scale. Descriptions of two-digit scale codes presented in Table 1.
directly after excision (Santos et al., 2009). This is consistent with the observation that Root Nectar® treatment imparted a greater effect size for root branching than AR formation, which may be attributed to plant nutrients, biostimulants, or some combination thereof. Root Nectar® treatment achieved a consistent improvement in rooting efficiency of both chrysanthemum and lavender cuttings, and should be considered as an improvement for large scale propagation of soft, and semi-hard wood cuttings. Unlike Root Nectar®, willow bark extract did not improve root development efficiency for all developmental stages for both plants. Whilst the lowest dose of willow extract (1.06 μL/L) significantly reduced the time to achieve root branching for chrysanthemum, higher doses were ineffective, and willow extract did not impart significant effects to chrysanthemum AR formation. By contrast, willow extract significantly reduced the time to achieve both AR formation and root branching for lavender, irrespective of the applied willow extract concentration. Consistent with the results presented herein for lavender and chrysanthemum, previous studies exploring application of willow extract to cuttings of other species, such as olive and bean, identified that stimulation of root development by willow extract has speciesspecific efficacies (Al-Amad and Qrunfleh, 2016; Gesto et al., 1977; Kawase, 1964). While the root stimulating effects of willow bark extract have been attributed to the phytochemical catechol (Gesto et al., 1977; Kling and Meyer, 1983), this effect is now more commonly associated with the phytohormone SA, which is a major component of willow bark extracts (Hayat et al., 2010; Hayat and Ahmad, 2007). Salicylic acid is a phenolic signalling molecule, and is associated with the regulation of flowering processes (D’Maris Amick Dempsey et al., 2011), biotic and abiotic stress responses (Horváth et al., 2007), and improved root growth and development (Basu et al., 1969; Gutiérrez-Coronado et al., 1998). While stress mediation after excision may contribute indirectly to improved root growth and development in cuttings, Salicylic acid is likely having more direct effects on rooting through interactions with rooting-related hormones. Exogenous application of SA with the prorooting hormone IAA is observed to enhance the hormone’s effects and improve AR formation compared with IAA application alone (Basu et al., 1969; Raskin, 1992). SA has also been shown to negatively
when applied at 1 mL/L (Fig. 1). Root Nectar® contains several biostimulants including extracts from kelp and Aloe vera that are known to contain many biological compounds. Kelp extracts contain plant nutrients, simple and complex sugars, vitamins, amino acids, and plant hormones (Battacharyya et al., 2015; du Jardin, 2015; Hamza and Suggars, 2001), and have been shown to have beneficial effects on rooting, germination, early plant growth, and the reduction of plant stress (Battacharyya et al., 2015; Carvalho et al., 2013; Rao and Chatterjee, 2014; Sangha et al., 2014). Similarly, Aloe vera extracts contain nutrients, vitamins, enzymes, amino acid, and sugars, as well as plants sterols, gibberellins, and salicylic acid (SA), and are associated with improvements in vegetative growth (particularly root growth), plant mineral composition, and oil production (Chatterjee et al., 2013; El Sherif, 2017; Hamouda et al., 2012). Whilst the exact modes of action of these plant extracts are as yet unknown, many of their constituents have been shown to affect root growth or aspects of nutrient uptake which may impact root development in cuttings. Sugars have been shown to promote rooting in cuttings when applied to the rooting medium, by providing the plant with a carbon source (Ahkami et al., 2009), while amino acids are associated with increases to nutrient uptake via chelation (Callahan et al., 2007; Halpern et al., 2015). Although plant hormones have been identified in kelp and Aloe vera extracts (include the rooting hormone indole acetic acid (IAA), and SA, respectively), these are often in small and varied quantities (Buggeln and Craigie, 1971; El Sherif, 2017; Kingman and Moore, 1982; Sharma et al., 2011; Surjushe et al., 2008) and more research is needed to confirm their activity and effects. Additionally, Kelp and Aloe vera extracts contain hormone-like compounds which may also contribute to these effects (Davis and Maro, 1989; Stirk and Van Staden, 1996). Therefore, either indirectly by stress mediation or chelation, or directly by hormones or carbon supply, the kelp and Aloe vera extracts in Root Nectar® confer root stimulating effects when applied to soft and semihard wood cuttings. In addition to biostimulants, Root Nectar® also contains plant nutrients (4.02 g/L nitrate, 1.74 g/L phosphate, 2.32 g/L potassium), which likely contribute to the beneficial effects observed. Cuttings have been shown to take up nutrients by their stem, and are observed to develop faster (after AR emergence) when treated with nutrients 3
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Fig. 2. Effect of Root Nectar® on chrysanthemum and lavender cuttings. Days for chrysanthemum (a) and lavender (b) vegetative cuttings to achieve BBCH = 05, and BBCH = 06, were compared between cuttings propagated with vs without the addition of Root Nectar®. Data presented as mean ± standard deviation of 8 biological replicates. Significant differences are indicated by * for P < 0.05, ** for P < 0.01, *** for P ≤ 0.001 by Student’s ttest performed between treatments.
Fig. 3. Effect of willow bark extract on chrysanthemum and lavender cuttings. Days for chrysanthemum (a) and lavender (b) vegetative cuttings to achieve BBCH = 05, and BBCH = 06, were compared between cuttings propagated with vs without the addition of willow bark extract. Data presented as mean ± standard deviation of 8 biological replicates. Data followed by the same letter do not differ significantly based on Tukey’s test 95% confidence grouping analyses.
regulate the wound response hormone jasmonic acid (JA), which is associated both positively and negatively with AR formation in cuttings (Lee et al., 2004; Lischweski et al., 2015). Although IAA and JA are present in the base and stem of cuttings (Ahkami et al., 2009; Garrido et al., 2002), it is unclear if exogenous SA has the capacity to be taken up and interact with these molecular mechanisms. Whilst the exact mode/s of willow bark extract efficacy is yet to be definitively identified, treatment achieved improved rooting efficiency of chrysanthemum and lavender cuttings, suggesting the need for further studies to elucidate molecular mechanisms involved.
improvement of AR formation and root branching in both soft and semihard wood cuttings, whilst application of willow bark extract improved AR formation and root branching in soft wood cuttings but only improved root branching in semi-hard wood cuttings. These results support the use of Root Nectar® and willow bark extract for improving the propagation efficiency of soft and semi-hard wood cuttings, and should be considered for large scale propagation within the horticulture industry. CRediT authorship contribution statement
5. Conclusion Kimber Wise: Conceptualization, Methodology, Formal analysis, Investigation, Writing original draft. Harsharn Gill: Conceptualization, Resources, Writing - review & editing, Supervision, Funding acquisition. Jamie Selby-Pham: Conceptualization, Methodology, Formal analysis, Resources, Writing - review & editing,
Herein the root promoting effects of Nutrifield’s biostimulant complex Root Nectar® and the biostimulant willow bark extract were tested on vegetative cuttings of lavender (soft wood) and chrysanthemum (semi-hard wood). Application of Root Nectar® achieved consistent 4
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Supervision, Project administration.
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Declaration of Competing Interest The authors wish to confirm that there are no conflicts of interest associated with this publication and there has been no significant financial support for this work that could have influenced its outcome. Acknowledgements The author’s would like to thank Nutrifield Pty Ltd. for supporting this work and providing the required resources. The authors would like to thank Abigail Jabines for her mentoring and guidance throughout propagation experiments. References Adam, K.L., 2006. Lavender Production, Products, Markets, and Entertainment Farms. A Publication of ATTRA: National Sustainable Agriculture Information Service, USA. Agbo, C., Obi, I., 2007. Variability in Propagation Potentials of Stem Cuttings of Different Physiological Ages of Gongronema Latifolia Benth. Ahkami, A.H., Lischewski, S., Haensch, K.T., Porfirova, S., Hofmann, J., Rolletschek, H., Melzer, M., Franken, P., Hause, B., Druege, U., 2009. Molecular physiology of adventitious root formation in Petunia hybrida cuttings: involvement of wound response and primary metabolism. New Phytol. 181, 613–625. Al-Amad, I., Qrunfleh, M., 2016. Effect of Babylon Weeping Willow (Salix Babylonica L.) Extracts on Rooting of Stem Cuttings of Olive (Olea Europaea L.)’ Nabali’, 1130th ed. International Society for Horticultural Science (ISHS), Leuven, Belgium, pp. 391–396. Arioli, T., Mattner, S.W., Winberg, P.C., 2015. Applications of seaweed extracts in Australian agriculture: past, present and future. J. Appl. Phycol. 27, 2007–2015. Arjana, I.G.M., Situmeang, Y.P., Suaria, I.N., Mudra, N.K.S., 2015. Effect of plant material and variety for production and quality Chrysanthemum. Int. J. Adv. Sci. Eng. Inf. Technol. 5, 407–409. Basu, R., Bose, T., Roy, B., Mukhopadhyay, A., 1969. Auxin synergists in rooting of cuttings. Physiol. Plant. 22, 649–652. Battacharyya, D., Babgohari, M.Z., Rathor, P., Prithiviraj, B., 2015. Seaweed extracts as biostimulants in horticulture. Sci. Hortic. 196, 39–48. Buggeln, R., Craigie, J., 1971. Evaluation of evidence for the presence of indole-3-acetic acid in marine algae. Planta 97, 173–178. Callahan, D.L., Kolev, S.D., Richard, A., Salt, D.E., Baker, A.J., 2007. Relationships of nicotianamine and other amino acids with nickel, zinc and iron in Thlaspi hyperaccumulators. New Phytol. 176, 836–848. Caplan, D., Stemeroff, J., Dixon, M., Zheng, Y., 2018. Vegetative propagation of cannabis by stem cuttings: effects of leaf number, cutting position, rooting hormone, and leaf tip removal. Can. J. Plant Sci. 98, 1126–1132. Carvalho, M.E.A., Castro, P.R.C., Novembre, A.D.C., Chamma, H.M.C.P., 2013. Seaweed extract improves the vigor and provides the rapid emergence of dry bean seeds. Am. J. Agric. Environ. Sci. 13, 1104–1107. Chatterjee, P., Chakraborty, B., Nandy, S., 2013. Aloe vera plant: review with significant pharmacological activities. Mintage J. Pharm. Med. Sci. 2, 21–24. Crouch, I.J., Van Staden, J., 1993. Evidence for the presence of plant growth regulators in commercial seaweed products. Plant Growth Regul. 13, 21–29. D’Maris Amick Dempsey, A.C., Vlot, M.C.W., Daniel, F.K., 2011. Salicylic acid biosynthesis and metabolism. The Arabidopsis Book, vol. 9 American Society of Plant Biologists. Davis, R.H., Maro, N.P., 1989. Aloe vera and gibberellin. Anti-inflammatory activity in diabetes. J. Am. Podiatr. Med. Assoc. 79, 24–26. du Jardin, P., 2015. Plant biostimulants: definition, concept, main categories and regulation. Sci. Hortic. 196, 3–14. El Sherif, F., 2017. Aloe vera leaf extract as a potential growth enhancer for Populus trees grown under in vitro conditions. Am. J. Plant Biol. 2, 101–105. Garrido, G., Ramón Guerrero, J., Angel Cano, E., Acosta, M., Sánchez-Bravo, J., 2002. Origin and basipetal transport of the IAA responsible for rooting of carnation cuttings. Physiol. Plant. 114, 303–312. Gesto, M., Vazquez, A., Vieitez, E., 1977. Rooting substances in water extracts of Castanea sativa and Salix viminalis. Physiol. Plant. 40, 265–268. Gutiérrez-Coronado, M.A., Trejo-López, C., Larqué-Saavedra, A., 1998. Effects of salicylic acid on the growth of roots and shoots in soybean. Plant Physiol. Biochem. 36,

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One of the Best Works of Kevin Henkes. published in multiple languages including English, consists of 32 pages and is available in Paperback format for offline reading.

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Chrysanthemum PDF Details

Author: Kevin Henkes
Book Format: Paperback
Original Title: Chrysanthemum
Number Of Pages: 32 pages
First Published in: September 16th 1991
Latest Edition: December 30th 2008
Series: Mouse Books
Language: English
Awards: California Young Readers Medal Nominee for Primary (1994)
Generes: Childrens, Picture Books, Childrens,
Formats: audible mp3, ePUB(Android), kindle, and audiobook.

The book can be easily translated to readable Russian, English, Hindi, Spanish, Chinese, Bengali, Malaysian, French, Portuguese, Indonesian, German, Arabic, Japanese and many others.

Please note that the characters, names or techniques listed in Chrysanthemum is a work of fiction and is meant for entertainment purposes only, except for biography and other cases. we do not intend to hurt the sentiments of any community, individual, sect or religion

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Great book, nicely written and thank you BooksVooks for uploading

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