The effect of cytokinins on shoot proliferation, biochemical changes and genetic stability of Rhododendron ‘Kazimierz Odnowiciel’ in the in vitro cultures

Shoot proliferation is a very important micropropagation phase, decisive for economic efficiency of this method for a given taxon. To obtain a high multiplication ratio and a good quality of microshoots a detailed propagation protocol must be developed for particular species or even cultivars. Rhododendron ‘Kazimierz Odnowiciel’ is a relatively new cultivar distinguished by large, beautiful flowers and high frost resistance so there is a need to develop an efficient method of its propagation to satisfy a growing demand for this plant. The aim of the experiment was to evaluate effects of cytokinins: meta-Topolin (mT), zeatin (ZEA), 6-benzyladenine (BA), thidiazuron (TDZ), 2-isopentenyladenine (2iP), or the combination of 2iP+ZEA on proliferation of shoots in R. ‘Kazimierz Odnowiciel’ cultured on Anderson’s medium (AN). Biochemical changes in plant material affected by cytokinins during this phase of micropropagation were determined and occurrence of genetical changes was followed using ISSR markers. TDZ, ZEA or the combination of ZEA+2iP resulted in 100% explant regeneration. On the medium with TDZ or ZEA over two new shoots per explant were produced but the highest proliferation was attained on the medium containing ZEA+2iP – over three shoots per explant. Microshoots developed in this treatment had also the highest contents of chlorophyll, carotenoids and soluble sugars as well as the highest catalase activity. Microshoots formed on the medium with zeatin showed the lowest polymorphism (below 4%) relative to a stock plant.

Dynamic versus Concurrent Lighting with Red and Blue Light-emitting Diodes as the Sole Light Source Can Potentially Improve Campanula Stock Plant Morphology for Cutting Production

Short campanula (Campanula portenschlagiana ‘PGM Get MEE’®) stock plants present a difficulty in machine-harvesting of cuttings. Light adjustment may be an effective approach to mediate plant elongation. Two experiments were performed to 1) investigate whether short-term (five weeks) daily 24-h dynamic lighting (DL) with red and blue light-emitting diodes (LEDs) can promote elongation without inducing flowering, and 2) explore whether DL can be used to modify stock plant morphology to improve the cutting quality and rooting success in a controlled environment. Two lighting treatments were used: concurrent lighting (CL) with red (85%) and blue (15%) LEDs (RB) at 100 µmol·m−2·s−1 and DL with red (170 µmol·m−2·s−1), blue (30 µmol·m−2·s−1), and RB (100 µmol·m−2·s−1) LEDs sequentially at three different lighting stages, respectively, in both experiments. In Expt. 1, at final harvest of stock plants, the side branches were longer under DL compared with CL, but the five (= 2 + 2 + 1) weeks of 24-h daily lighting resulted in visible flower buds under both treatments. Based on the results of Expt. 1, a second experiment (Expt. 2) was conducted with the same cultivar and experimental conditions, but with a shorter photoperiod (10 h·d−1) for 11 (= 8 + 2 + 1) weeks. In Expt. 2, at final harvest, DL compared with CL caused more upright side branches, and reduced the dry biomass of side branches with one branching order and leaf chlorophyll content. However, the harvested cutting quality and rooting success were similar between both treatments. In both experiments, side branch number under DL was greater compared with CL at the end of the first lighting stage. Stock plants under DL were taller from the second lighting stage on to final harvest compared with CL, and the final heights of stock plants under DL met the target for machine-harvest in both experiments. Therefore, if the lighting strategy is further optimized, DL can potentially benefit controlled-environment production of campanula cuttings.

Involvement of the auxin–cytokinin homeostasis in adventitious root formation of rose cuttings as affected by their nodal position in the stock plant

Main conclusion Enhanced levels of indole-3-acetic and raised auxin to cytokinin ratios in the stem base contribute to the positive acropetal gradient in rooting capacity of leafy single-node stem cuttings of rose. Abstract Cuttings excised from different nodal positions in stock plants can differ in subsequent adventitious root formation. We investigated the involvement of the auxin–cytokinin balance in position-affected rooting of Rosa hybrida. Leafy single-node stem cuttings of two rose cultivars were excised from top versus bottom positions. Concentrations of IAA and cytokinins were monitored in the bud region and the stem base during 8 days after planting using chromatography–MS/MS technology. The effects of nodal position and external supply of indole-butyric acid on rooting were analyzed. Most cytokinins increased particularly in the bud region and peaked at day two before the bud break was recorded. IAA increased in both tissues between day one and day eight. Top versus bottom cuttings revealed higher levels of isopentenyladenosine (IPR) in both tissues as well as higher concentrations of IAA and a higher ratio of IAA to cytokinins particularly in the stem base. The dynamic of hormones and correlation analysis indicated that the higher IPR contributed to the enhanced IAA in the bud region which served as auxin source for the auxin homeostasis in the stem base, where IAA determined the auxin–cytokinin balance. Bottom versus top cuttings produced lower numbers and lengths of roots, whereas this deficit was counterbalanced by auxin application. Further considering other studies of rose, it is concluded that cytokinin-, sucrose- and zinc-dependent auxin biosynthesis in the outgrowing buds is an important factor that contributes to the enhanced IAA levels and auxin/cytokinin ratios in the stem base of apical cuttings, promoting root induction.

Plant Growth Regulator Impacts on Vegetative Cutting Production of Moroccan Pincushion (Pterocephalus depressus) Plants

Moroccan pincushion (Pterocephalus depressus) is a drought-tolerant perennial that is being used in landscapes throughout arid areas of the western United States. This paper describes two experiments researching vegetative cutting production from stock plants. Moroccan pincushion stock plants received foliar applications of gibberellic acid (GA3), benzyladenine, ethephon, or auxin [indole-3-butyric acid (IBA)] plant growth regulators (PGR). Plant growth regulators were applied singularly and in combination with GA3 to determine efficacy on stock plant growth. A propagation study was conducted simultaneously to determine effects of these different PGR treatments applied to stock plants on the rooting of moroccan pincushion cuttings. The stock plant study showed GA3 + benzyladenine application increased cutting production over other PGR treatments. Fresh weight of moroccan pincushion cuttings did not differ among treatments. While cuttings did not differ in dry weight in experiment 1, statistical differences were observed in experiment 2. However, these differences in dry weight did not affect the quality of the cuttings. Cuttings from stock plants treated with GA3 + IBA treatment had the highest numerical growth index [(height + width + width)/3]. Cuttings from stock plants treated with GA3 alone or in combination with another PGR were all greater in average growth index and statistically differed from those without GA3 being applied. PGR treatments did not affect rooting percentages of the cuttings with nontreated stock plant cuttings successfully rooting at an average rate of 95%. However, GA3 + IBA was the only treatment where cuttings had 100% rooting for both experiments, indicating potential rooting benefits. Index words: Plant growth regulator, propagation, Pterocephalus depressus, vegetative cuttings. Species used in this study: Moroccan pincushion [Pterocephalus depressus Archibald]. Chemicals used in this study: gibberellic acid (GA3), benzyladenine, ethephon, indole-3-butyric acid (IBA).

Influencing Stock Production of Mojave Sage and Cape Daisy with the Application of Plant Growth Regulators

This study aimed to assess the effects of plant growth regulators (PGRs) on stock plant production of mojave sage (Salvia pachyphylla) and ‘Avalanche’ cape daisy (Osteospermum hybrid) that received foliar sprays of the following three PGRs: 200 and 400 ppm ethephon; 250 and 500 ppm benzyladenine; and 50 and 100 ppm gibberellic acid 4 and 7 (GA4+7) plus benzyladenine. Vegetative growth [height and width growth index (GI)], the number of vegetative cuttings, and fresh weight (FW) and dry weight (DW) of the harvested vegetative cuttings data were collected. A propagation study was conducted concurrently to determine the effects of the PGR treatments on rooting vegetative cuttings. GA4+7 plus benzyladenine (50 and 100 ppm) increased the production of both mojave sage and ‘Avalanche’ cape daisy cuttings by ≥18% more than the other treatments. The GI, FW, and DW results showed similar trends across experiments 1 and 2 for each perennial. In the propagation study, the rooting percentage did not differ after 4 weeks, indicating that the use of GA4+7 plus benzyladenine in production protocols could benefit producers of both perennials.

Vegetative propagation of mate from shoot cuttings induced by coppicing of selected plants

Mate (Ilex paraguariensis A. St.-Hil) is an arboreal species of great economic and socio-environmental importance, which presents difficulties in the adventitious rooting process. The rhizogenic competence of vegetative propagules of mate can be attained by coppicing, but the factors that influence the success of this technique have not yet been elucidated. Therefore, the aim of this study was to evaluate the effects of the stock plant, time of shoot collection, treatment with indolebutyric acid (IBA) and diameter of vegetative propagules in the adventitious rooting of cuttings from shoots induced by coppicing of selected adult plants of mate. Shoots produced from coppiced 13SM01 and 13SM05 stock plants were collected at different times of the year to prepare single-bud cuttings. The cuttings were measured as their diameter in the central portion and treated with 0 and 3000 mg L-1 of IBA. After 60 days, cuttings were evaluated to determine survival and rooting percentages, number of roots, and length of the three largest roots. A completely randomized design was used in a 4 × 2 × 2 × 13 (collection times x stock plants x IBA treatment x diameter classes) factorial scheme with 15 replications. Competence for adventitious rooting was highest for stock plant 13SM05. Cuttings collected in November and February had the highest rooting percentages, without the need of IBA treatment. Cuttings from 13SM01 and 13SM05 with diameters smaller than 0.636 cm (classes 1 to 6) and smaller than 0.712 cm (classes 1 to 7), respectively, had the highest rooting percentages and differed significantly from cuttings with larger diameters. Thus, the results of the present study indicate that stock plants, time of shoot collection, and diameter of vegetative propagules influence adventitious rooting competence of mate cuttings from coppicing adult plants. Rooting of mate cuttings with lower rhizogenic competence is influenced by the application of 3000 mg L-1 of IBA.

Effects of Growth Substrate and Container Size on Cutting Production from Mojave Sage Stock Plants

Stock plant productivity is an important concern for growers of mojave sage (Salvia pachyphylla) because this species produces more woody growth as the plant ages. The objective of the study was to determine the best growth substrate and container size combination to maximize stock plant productivity. A secondary objective was to determine whether the stock plant treatments influenced the rooting of vegetative cuttings. Three different container sizes (3, 12, and 15.5 qt) and four soilless substrates composed primarily of bark, peat, and perlite (substrate 1); bark, peat, and vermiculite (substrate 2); bark, peat, and coarse perlite (substrate 3); and peat (substrate 4) were used. The stock plant experiment was conducted using 12 treatment combinations, and a subset of those stock plants was selected randomly for the rooting study that immediately followed the stock plant experiment. Stock plants responded to substrate treatments differently. The most successful stock plants, which produced more cuttings per plant and per square foot, as well as larger cuttings, were those grown in substrate 3. Regardless of substrate, the highest number of cuttings per square foot was obtained from stock plants grown in 3-qt containers, indicating that the smaller containers allow for the most efficient use of space when growing mojave sage stock plants for 4 to 6 months. The rooting of vegetative cuttings was successful (88% to 100% of cuttings rooted after 4 weeks under mist) for all treatment combinations.