EFFECT OF BIOSTIMULANTS ON SOME BIOACTIVE COMPOUNDS AND NITRATE LEVEL IN LETTUCE (Lactuca sativa L.) GROWN UNDER UNHEATED PLASTIC TUNNEL

Based on the regulation (EU) No 2015/1107 in 8 July 2015, Willow (Salix alba) bark extract is a recent approved basic substance in Europe as a safe and effective environmentally friendly fungicide to be used in plant protection. Because of having indole butyric acid content, it is also used as a growth hormone to promote rooting in cutting propagation of wooden part in horticultural plants. This research aimed to evaluate the effect of willow extract and Ferbanat L (Bistep) on lettuce leaves. Bistep is an organic solution of nano fertilizer which is produced in Hungary and allowed as „Bistep plant conditioner”. Furthermore, the experiment evaluated their interaction on some bioactive compounds in different lettuce varieties, too. Three different lettuce varieties (King of May, Kobak and Great Lakes) were evaluated for total polyphenols, vitamin C and nitrate content in fresh leaves. The results indicate that the contents were significantly affected by the used biostimulants rather than varieties. Significantly difference of the total polyphenols was recorded in the plants treated with Willow + Bistep (80.64 mg GAE / 100g) in comparison to the control (55.85 mg GAE / 100g) in the mean of varieties. Similar result was recorded for the vitamin C. However, plants treated with Willow extract resulted in the highest level of nitrate content (692.00 mg/100g), while the lowest amount was measured in the control (488.00 mg/100g). According to our results, Willow bark extract can be a beneficial source of natural growth regulator or fungicide; however, when it is used alone for vegetables, the nutrient content (mainly nitrate level) must be checked before consuming and delivering to the market.

ROLE OF VENTILATION, CALCIUM AND BORON ON SHOOT TIP NECROSIS AND HYPERHYDRICITY REDUCTION OF IN VITRO ROORSTOOCKS OF PISTACIO ‘BADAMI’ ȘI ‘UCB1’

Shoot tip necrosis (STN) is one of the main physiological disorders in the micropropagation of pistachios. In the current study, the effects of CaNO3.4H2O at 196 mg/L and 291 mg/L, H3BO3 at 196 mg/L and 291 mg/L, and CaCl2.2H2O at 2,980 mg/L on STN and hyperhydricity reduction of Pistacia vera L., ‘Badami’ and ‘UCB1’ rootstocks were assessed, compared to the MS standard medium containing 3% sucrose, 0.7% agar supplemented with benzyladenine (BA) (1.5 mg/L), indole butyric acid (IBA) (0.1 mg/L). For ventilation parameter, filter container vessels with a 50-µm microporous polypropylene membrane (Pardis®) were used. Based on the results, an increase in calcium chloride content of the MS standard medium prevented hyperhydricity in the UCB1 rootstock, whereas it increased STN, yellow leaves, decreasing the multiplication of shoots in the ‘Badami’ rootstock. The results also showed that increasing boric acid from 196 mg/L up to 291 mg/L decreased STN in the UCB1 rootstock and increased this disorder by 37% in the ‘Badami’ rootstock. Ventilation showed no significantly reducing effect on the percentage of STN in the regenerated shoots of the ‘Badami’ rootstock, whilst it decreased the STN of the ‘UCB1’ rootstock to the lowest percentage. For the ‘Badami’ rootstock, CaNO3.4H2O at 196 mg/L led to the highest proliferation rate, shoot height, shoot diameter, and leaf number, but for the ‘UCB1’ rootstock, an increase in the concentration of CaNO3.4H2O up to 291 mg/L under ventilated conditions resulted in an increase in proliferation, shoot height, and shoot diameter.

Metal Akümülatör Bitki olan Brassica nigra L.’nin in vitro Kallus İndüksiyonu ve Sürgün Gelişimi

This study established an efficient in vitro callus formation and plant regeneration protocol for a prevıously reported Cu accumulator, Brassica nigra, black mustard collected from Diyarbakir (Station site). Node explants from 10-day old mature plants were used for callus formation and shoot regeneration. The highest callus formation frequency (100%) was observed on Murasige Skoog (MS) medium containing 0.1 mg/L Benzylaminopurine (BAP) + 0.5 mg/L Naphthylacetic acid (NAA) (MS 2), 0.6 mg/L BAP + 0.2 mg/L NAA (MS 7), the highest shoot regeneration frequency (100%) was achieved on MS medium containing 0.6 mg/L BAP + 0.05 mg/L Indole butyric acid (IBA) (MS 8), 0.2 mg/L IBA + 0.2 mg/L NAA (MS 10) and the highest number of shoots per explant (3,25) was obtained on MS medium supplemented 0.6 mg/L BAP + 0.05 mg/L IBA (MS 8). After root, stem, and leaf formation from explants in MS medium, these plants were transferred to soil and grown in the plant growth room for one month. A dependable and effective shoot regeneration procedure was developed, laying the groundwork for genetic transformation in Brassica nigra.

Differences in the Abundance of Auxin Homeostasis Proteins Suggest Their Central Roles for In Vitro Tissue Differentiation in Coffea arabica

Coffea arabica is one of the most important crops worldwide. In vitro culture is an alternative for achieving Coffea regeneration, propagation, conservation, genetic improvement, and genome editing. The aim of this work was to identify proteins involved in auxin homeostasis by isobaric tandem mass tag (TMT) and the synchronous precursor selection (SPS)-based MS3 technology on the Orbitrap Fusion™ Tribrid mass spectrometer™ in three types of biological materials corresponding to C. arabica: plantlet leaves, calli, and suspension cultures. Proteins included in the β-oxidation of indole butyric acid and in the signaling, transport, and conjugation of indole-3-acetic acid were identified, such as the indole butyric response (IBR), the auxin binding protein (ABP), the ATP-binding cassette transporters (ABC), the Gretchen-Hagen 3 proteins (GH3), and the indole-3-acetic-leucine-resistant proteins (ILR). A more significant accumulation of proteins involved in auxin homeostasis was found in the suspension cultures vs. the plantlet, followed by callus vs. plantlet and suspension culture vs. callus, suggesting important roles of these proteins in the cell differentiation process.

Response of Growing Media and IBA Application under Different Weather Conditions on Growth of Pomegranate (Punica granatum L.) Cuttings

Pomegranate (Punica granatum L.) is an important commercial horticultural crop. The cultivation of this crop is increasing day by day. Mainly due to its survival nature, particularly in dry or less water available areas. Its propagation is possible by using seed but vegetative propagation methods are very popular. Even commercial propagation of pomegranate is done by cuttings. To fulfill the rising demand for high-quality plant material, easy, fast, reliable, economical multiplications are required, and under such circumstances, cuttings are the best-suited material. This method of propagation can further be improved by the use of optimum growing media and application of various plant growth regulators Indole Butyric acid and its derivations. Moreover, the success of propagation depends on prevailing weather conditions and the time of planting. Perlite + vermiculite and a combination of vermiculite + coco peat in equal proportions, quick dip application of 2500 ppm IBA, and planting in the spring season are considered ideal for fast and economical multiplication of pomegranate cuttings. Humid weather conditions during the spring season exhibit a good possibility of rooting and survival of the same. Therefore, the main aim of this review article is to collect/compile the information regarding the effect of growing media and applications of IBA under different weather regimes over previous years along with recent happenings.

Sprouting and rooting ability of the plus trees of Eucalyptus pellita, E. brassiana and its hybrid in Wanagama, Indonesia

The demand for eucalypts has greatly increased since they are raw materials for timbers, plywoods, pulp and papers, and essential oil production. This study aimed to select the plus trees and determine the sprouting and rooting ability of E. pellita, E. brassiana, and their hybrids in Wanagama. Plus trees selection and girdling were conducted in December 2019. Cuttings were picked from the 3rd-4th nodes of sprouting at 8-10 cm in length. A completely randomized design was applied in a factorial experiment with three levels of Indole Butyric Acid (IBA) (100, 1000, and 4000 ppm) and two types of media (media 1: sand, rice husk charcoal and cocopeat (2:2:1), media 2: sand, topsoil and dung-fertilizer (2:2:1)), with ten replications. Results selected a total of 53 plus trees candidates (29 of E. pellita, 12 of E. brassiana, and 12 of hybrid). E. pellita is the best in sprouting ability (100% survival; 41-60 shoot/trees, 127-161cm shoot length, 0.53-0.57cm shoot diameter, and 194-252 leaves/shoot). Cuttings of E. pellita treated with 100 ppm IBA gained the highest survival and leaves production (96%-100%; 14-16 leaves), followed by E. brassiana (52%-66.67%; 7 leaves) and the hybrid (4% to 8.33%; 3-4 leaves). These results may contribute to arranging better strategies for mass production of selected eucalypts.

In Vitro Adventitious Regeneration of Artemisia annua L. Influencing Artemisinin Metabolism

Artemisia annua L. is a herbaceous plant belonging to the Asteraceae family, known for producing, although at low levels, the sesquiterpene lactone artemisinin (AN), which is highly effective against malaria. In this study, an in vitro regeneration process of A. annua L. using ‘Artemis’ progeny was established and the potential of tissue culture for inducing new variability in terms of AN metabolism of in vitro regenerated plants was investigated. Among the plant growth regulators tested, the cytokinin 6-benzyladenine (BA) at 4.4 μM in combination with the auxin indole-butyric acid (IBA) at 0.35 μM yielded the greatest frequency of shoot induction. The optimal multiplication medium contained BA at 0.9 μM and naphthaleneacetic acid (NAA) at 0.05 μM. Regenerated plants (RPs), after transferring to the greenhouse and subsequently to the field, were analyzed during the growth cycle at different sampling times, showing a peak of AN content 20 days before blossom. Variability among different RPs and sampling times, in terms of AN and its precursors dihydroartemisinic acid (DHAA) and artemisinic acid (AA) was observed. This suggests that adventitious shoot induction could provide a useful strategy to induce variability influencing artemisinin metabolism as a consequence of in vitro manipulation.

Optimization of Callogenesis/Caulogenesis Induction Protocol in Saffron Plant (Crocus sativus L.) Using Response Surface Methodology

The Crocus sativus, an endangered medicinal and aromatic plant in Morocco, has a low propagation rate in natural conditions and, therefore, an efficient method for in vitro propagation is required. This study investigated the effects of various hormones on the induction of callogenesis and callogenesis in C. sativus corms using the Box-Behnken experimental design. The best shoot formation was obtained with Murashige and Skoog fortified with 3 mg/L 6-Benzylaminopurine. On the other hand, callus formation was obtained with 3 mg/L 1-Naphthaleneacetic Acid or 3 mg/L 2,4-Dichlorophenoxyacetic Acid. However, a combination of 3 mg/L 6-Benzylaminopurine, 1.056 mg/L Indole Butyric Acid, and 3 mg/L 2,4-Dichlorophenoxyacetic Acid allows 50% caulogenesis and 60% callogenesis. The in vitro regeneration system could be utilized for both conservation and largescale multiplication of Crocus sativus corms.

Clonal Propagation of Coffea arabica with Indole Butyric Acid and Acclimatization Conditions in Amazonas, Peru

The use of hormones is important in improving the propagation process. The objective of the experiment was to identify the best dose of indole butyric acid (IBA) hormone for the rooting of coffee cuttings of the typical variety through clonal propagation in the Amazon region. The experiment was carried out in two stages. Both stages were carried out under a Completely Randomized Design (CRD). The percentage of rooting of cuttings and the number of roots were evaluated. The coffee tree clones were collected from rust tolerant elite plants. In the laboratory, the cuttings were immersed for 10 minutes in fungicide and then in five doses of IBA hormone (1000, 1500, 2000, 2500, and 3000 mg L−1) of IBA. They were then placed in a microtunnel. With the best dose obtained in stage 1 of IBA (T4: 2000 mg L−1), which achieved a rooting percentage of 52.50% and 5.8 roots on average, stage 2 was installed for the evaluation of acclimatization conditions (greenhouse and nursery). Thirty plants of the Típica variety were evaluated per treatment. Regarding the results after 90 days, the greenhouse obtained the highest values with 72.22% survival, a plant height of 9.48 cm, 4.32 mm stem diameter, 6.53 leaves per plant, 9.65 cm leaf length, and 3.83 cm leaf width. The IBA hormone in a controlled environment such as the greenhouse achieves good vegetative propagation for the Típica variety.

Micropropagation and Cryopreservation of Yukon Draba (Draba yukonensis), a Special Concern Plant Species Endemic to Yukon Territory, Canada

Yukon Draba (Draba yukonensis) is a small, short-lived perennial mustard species that is endemic to southwestern Yukon in Canada. This plant has been categorized as a species of Special Concern. It faces the threat of habitat loss due to natural and man-made causes and a population that is unevenly distributed to a few large and several small subpopulations in the area. It will therefore be judicious to undertake investigations on the conservation of this species to save it from further deterioration which may lead to its extinction. In this study, a protocol was developed for in vitro propagation and cryopreservation of Yukon Draba. The micropropagation protocol was optimized using shoot tips which enabled clonal propagation and in vitro storage of the species. Shoots grew best in the medium containing MS basal salts and had the highest multiplication with the addition of 2 µM 6-benzylaminopurine or 5 µM Kinetin with 3% sucrose. The addition of 10 µM Indole Butyric Acid (IBA) produced the highest number of adventitious roots on the shoots and the longest root length was observed at 2 µM IBA. The rooted plantlets were transferred to greenhouse and the highest survival (87.5%) was observed for the plantlets treated with a lower concentration of IBA (2 µM). Cryopreservation protocol was developed using the droplet-vitrification method for in vitro shoot tips. Two-week-old shoots had the highest survival and regrowth following exposure to plant vitrification solution 3 (PVS3) for 30 min, prior to direct immersion of the droplets into the liquid nitrogen. The optimized protocols for the micropropagation and cryopreservation may be useful for the long-term germplasm conservation and reintroduction of this species in its natural habitat.