Improved in vitro propagation and direct acclimatization of Cryptocoryne wendtii in aquarium in the presence of aquarium fish Puntius tetrazona (Bleeker)

330-337Cryptocoryne wendtii is an important ornamental aquatic plant and has difficulties in conventional cultivation. In the present study, we propose an efficient in vitro propagation and acclimatization protocols for C. wendtii. The highest number of shoots was achieved on Murashige and Skoog (MS) medium containing 4.0 mg/L BA+1.0 mg/L IBA. In the second subculture, the number of shoots per explant increased from 7.2 to 51.8 in this medium. The shoot length per explant, the percentage of shoots rooted, and the number of roots per shoot were maximum in MS medium supplemented with 1.0 mg/L IBA. The in vitro rootless and rooted shoots were acclimatized in aquariums containing different substrates [Calcite (Ca), River Sand (RS), Zeolite (Ze) and Shell Grit (SG)] and tetrazona fish Puntius tetrazona (Bleeker). One third of the water in the aquarium was replaced with fresh tap water every two weeks and the water analyses were carried out before replacement of aquarium water every time. All shoots were successfully acclimatized for a period of 3 months. The RS and rooted shoot explants were found to be more effective than the other substrates, and rootless shoots in terms of all parameters that were tested

Improved in vitro propagation and direct acclimatization of Cryptocoryne wendtii in aquarium in the presence of aquarium fish Puntius tetrazona (Bleeker)

###EgeUn###Cryptocoryne wendtii is an important ornamental aquatic plant and has difficulties in conventional cultivation. In the present study, we propose an efficient in vitro propagation and acclimatization protocols for C. wendtii. The highest number of shoots was achieved on Murashige and Skoog (MS) medium containing 4.0 mg/L BA+1.0 mg/L IBA. In the second subculture, the number of shoots per explant increased from 7.2 to 51.8 in this medium. The shoot length per explant, the percentage of shoots rooted, and the number of roots per shoot were maximum in MS medium supplemented with 1.0 mg/L IBA. The in vitro rootless and rooted shoots were acclimatized in aquariums containing different substrates [Calcite (Ca), River Sand (RS), Zeolite (Ze) and Shell Grit (SG)] and tetrazona fish Puntius tetrazona (Bleeker). One third of the water in the aquariums was replaced with fresh tap water every two weeks and the water analyses were carried out before replacement of aquarium water everytime. All shoots were successfully acclimatized for a period of 3 months. The RS and rooted shoot explants were found to be more effective than the other substrates, and rootless shoots in terms of all parameters that were tested

Immobilization of Rubia tinctorum L. suspension cultures and its effects on alizarin and purpurin accumulation and biomass production

WOS: 000312587800014The present study is investigating the immobilization of Rubia tinctorum L. suspension cultures. The effects of three inoculation volumes and three immobilization materials (loofa, sisal and jute) on fresh and dry weights of biomass as well as on alizarin and purpurin production were determined in this study. Two grams of four-week old callus tissue were transferred to liquid medium to establish suspension cultures. After four weeks, suspension cultures of R. tinctorum at concentration of 8 x 10(5) living cells/ml were immobilized with lignocellulosic materials and the cells were attached to all immobilization materials at the end of the first week and started to form aggregates on them. At the fourth week of these batch systems, biomass was measured approximately three times higher than the starting suspension cultures. The highest fresh weight was obtained (339.40 g/l) from sisal with A1/2 inoculation ratio. Immobilization materials and inoculation volumes had an effect on dry weights, and accordingly, the most effective combinations were jute with A1/4 (J3) and A1/2 (J1) inoculation volumes with 7.86 and 7.82 g/l dry weights, respectively. Alizarin and purpurin contents of immobilized cells, analyzed with U-HPLC method, were 6.05 and 22.91 times higher than inoculated cells. All immobilization materials used in this study had no negative effect on to cells and biomass accumulation was enhanced. Concomitantly with rapid biomass increase, alizarin and purpurin production was ascended

High-frequency protocorm-like bodies and shoot regeneration through a combination of thin cell layer and RITA((R)) temporary immersion bioreactor in Cattleya forbesii Lindl.

WOS: 000459394000004An efficient in vitro mass propagation through protocorm-like bodies (PLBs) was established in Cattleya forbesii Lindl., a commercially important orchid. Whole PLBs (W-PLB) and transverse thin cell layers of PLB (tTCL-PLB) explants were cultured in RITA((R)) bioreactors based on temporary immersion system. Explants were transferred in semi-solid or RITA((R)) bioreactor for protocorm production or shoot regeneration. The effect of different immersion frequencies, medium volumes and inoculum densities were studied and optimized. RITA((R)) bioreactor cultures were found to be superior compared with semi-solid cultures regarding PLB production and shoot regeneration. tTCL-PLB explant types cultured in the RITA((R)) bioreactor with immersion for 1min/4h, 250mL of medium and 20 explants showed the highest number of PLBs per RITA((R)) (2237 PLBs) and per explant (111.9 PLBs). The highest number of PLBs per explant was 21 times higher than those from semi-solid culture. The highest number of shoots per RITA((R)) (3998 shoots) and per explant (199.9 shoots) were observed on tTCL-PLB cultured in RITA((R)) bioreactor (1min/4h; 150mL of medium and 20 explants). The highest number of shoots per explant was 95 times higher than those grown on semi-solid culture. Mass propagation of PLBs and shoots of C. forbesii Lindl. using combined thin cell layer and RITA((R)) temporary immersion has been adapted in commercial practice.Republic of Turkey Ministry of Science, Industry, and TechnologyMinistry of Science, Industry & Technology - Turkey [1217. STZ.2012-1]; Ege University Scientific Research Projects Coordination UnitEge University [13-BIL-022]Authors are grateful to Republic of Turkey Ministry of Science, Industry, and Technology (Project No: 1217. STZ. 2012-1), Ege University Scientific Research Projects Coordination Unit (Project No: 13-BIL-022) and On Danismanlik Tourism Commitment and Trade Limited Company. In addition, we would like to thank Didem Okmen, Biomedicine and Genome Center, Bioinformatics Department PhD student for her contributions to statistical analyzes and interpretations

High-frequency protocorm-like bodies and shoot regeneration through a combination of thin cell layer and RITA (R) temporary immersion bioreactor in Cattleya forbesii Lindl (vol 136, pg 451, 2019)

WOS: 000459394000005

Micropropagation of centennial tertiary relict trees of Liquidambar orientalis Miller through meristematic nodules produced by cultures of primordial shoots

WOS: 000356876600004This paper provides an important micropropagation method that might be used for conservation and commercial production of tertiary relict tree Liquidambar orientalis. The Liquidambar orientalis Miller is an important tree as a tertiary relict endemic species in terms of plant biodiversity, and has economic value due to the balsam it produces. In the present study, an efficient micropropagation method was developed for the L. orientalis. Initially, primordial shoot explants isolated from axillary buds were cultured on Murashige and Skoog and Woody Plant Medium (WPM) containing different plant growth regulators (PGRs). The highest number of shoots per explant was obtained on WPM supplemented with 1.0 mg/L 6-benzylaminopurine (BAP) and 1.0 mg/L indole-3-butyric acid (IBA) (shoot proliferation medium: SPM). These shoots were then subcultured continuously on SPM for a period of 18 months. Meristematic nodule clusters were formed at the base of the cultured shoots in SPM, where multiple shoots developed from them. The number of shoots per explant was increased approximately 2.8-fold by applying various strategies such as different explant type (single shoots and shoot clusters) and culture vessels (Glass Tube, 210-cc Glass culture jar, 400-cc Glass culture jar, and Vitro Vent(A (R))). Shoot clusters cultured in ventilated Vitro Vent(A (R)) vessels on SPM gave best result for shoot propagation. In vitro shoots rooted best on WPM containing 30 g/L sucrose, 120 mg/L ethylenediamine di-2-hydroxyphenyl acetate ferric (Fe-EDDHA) and 4.0 mg/L IBA. All plantlets were successfully acclimatized in a glasshouse and then plants were transferred to the field. This methodology has been adapted by a commercial producer

Astragaloside IV and Cycloastragenol Production Capacity of Astragalus trojanus Calli

WOS: 000350973300004Astragalus species are medicinal plants which produce valuable secondary metabolites, especially cycloartane-type glycosides. In this study, stem and leaf explants of Astragalus trojanus were subjected to different plant growth regulators, environmental conditions and media compositions to identify their callus responses. Stem and leaf explants were cultured in Murashige and Skoog (MS) and woody plant (WPM) media supplemented with different concentrations of kinetin, naphthalene acetic acid, 2,4-dichlorophenoxyacetic acid, thidiazurone and indol acetic acid under two light intensities (1000 and 4000 lux) and also in dark conditions. Both MS and WPM media triggered callus regeneration. Although, callus regeneration was observed on both stem and leaf explants, callus biomass accumulation on stem explants were higher. Addition of 100 mu g/L selenium and doubled concentration of WPM vitamins enhanced callus biomass on stem explants under dark conditions. Stem explants also regenerated shoots at high frequencies (up to 93%), especially in kinetin added media. Astragaloside IV and cycloastragenol accumulation efficiencies were determined in calli tissues. The highest astragaloside IV production (3.5 mu g/mg) was found in callus tissue regenerated from stem explants in D1 medium, whereas the highest cycloastragenol accumulation (4.8 mu g/mg) was detected in callus tissue regenerated from stem explants in N2 medium.Scientific and Technical Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [TOVAG-109O62]The financial support of The Scientific and Technical Research Council of Turkey (TUBITAK) (Project No. TOVAG-109O627) is gratefully acknowledged

Plant Growth Regulator Effects on In Vitro Propagation and Stevioside Production in Stevia rebaudiana Bertoni

WOS: 000392148800022Stevia rebaudiana is of great importance due to its steviol glycosides (SGs) which are natural sweeteners used by the food industry as well as having medicinal purposes. In the present study, the effect of plant growth regulators (PGRs) and explant types on in vitro propagation and shoot growth of S. rebaudiana were studied, the effect of PGRs on SGs production was determined. For this purpose, nodal explants and shoot tip explants were cultured on woody plant medium (WPM) supplemented with cytokinins [6-benzyladenine (BA), kinetin (Kn), or thidiazuron (TDZ)] or cytokinins + auxins combinations [BA + indoleacetic acid (IAA); BA + naphthaleneacetic acid (NAA); Kn + IAA; Kn + NAA]. Although, the best shoot proliferationwas obtained onWPMsupplementedwith BA+ NAA combinations, shoots grown on PGR-containing media produced callus at the base of the shoots and showed chlorosis and necrosis. Additionally, shoots showed at all concentrations of TDZ, and at higher concentrations of BA, morphological changes such as malformed leaves and poor shoot growth. In contrast to PGR-containing media, on the PGR-free control medium, the development of shoots and roots occurred simultaneously and healthy and well-developed plantlets were obtained. Thus, we developed an economical viable means of in vitro propagation byminimizing themicropropagation steps and removing the requirement of PGRs. According to the high-performance liquid chromatography (HPLC) results, PGR-free control medium (WPM) led to considerably higher stevioside content in the leaves compared with the PGR(s)-containing media and the highest stevioside [34 mg.g(-1) dry weight (DW)] and rebaudioside A content was only detected on the control medium without PGRs. Steviolbioside, rubusoside, and dulcoside A were detected qualitatively in the leaves of shoots grown on WPM supplemented with 2.27 mu M TDZ, 4.54 mu M TDZ, 2.22 mu M BA + 2.69 mu M NAA, 2.22 mu M BA + 5.37 mu M NAA, 2.32 mu M Kn + 5.71 mu M IAA, or 2.32 mu M Kn + 2.69 mu M NAA.Celal Bayar University Scientific Research Projects CommissionCelal Bayar University [ALS 2008-069]This research was supported by Celal Bayar University Scientific Research Projects Commission (ALS 2008-069)