Cinchona Pubescens Research Articles

Globally Distributed Arbuscular Mycorrhizal Fungi Associated With Invasive Cinchona pubescens on Santa Cruz Island, Galápagos.

The presence of arbuscular mycorrhizal fungi (AMF) is essential for the survival and establishment of most plant species in nature. The reproductive success of invasive plant species in a particular habitat could also depend on these AM fungi. Cinchona pubescens, commonly known as quinine, is highly invasive on Santa Cruz Island, Galápagos, but at the same time severely endangered in its native range on mainland Ecuador due to overexploitation in the past. In this study, we aim at determining the AMF communities associated with C. pubescens at both locations to investigate whether the successful invasion of C. pubescens on Santa Cruz is related to its association with a particular community of AMF. For this, roots of C. pubescens trees were sampled at three sites, one site on Santa Cruz and two sites in the province of Loja, on mainland Ecuador. Communities of AM fungi were determined through the molecular cloning and sequencing of the 18S nrDNA gene and through the delimitation of Operational Taxonomic Units (OTUs), associated with the plant roots. We found 36 AMF OTUs associated with C. pubescens, most of them belonging to the genus Glomus. The highest richness of AMF OTUs was detected in samples from sites located on mainland Ecuador. The AMF communities between Santa Cruz and mainland Ecuador were significantly different, and only five OTUs were shared between both locations. Two dominant OTUs in C. pubescens from Santa Cruz were detected but no dominant OTUs from mainland Ecuador. Almost two thirds of the OTUs associated with C. pubescens had a wide global distribution. Our results suggest that the successful invasion of C. pubescens on Santa Cruz could have been facilitated by local generalist AMF and not by particular AMF. The observed generalist AMF from both locations could be important for conservation plans of restoring the endangered C. pubescens in the native forests on mainland Ecuador.

An undergrad experiment for the at-home study of fluorescence: Extraction of quinine and chlorophyll from Cinchona tree bark

Herein we describe a method for the effective extraction and qualitative detection of quinine and chlorophyll. Both compounds can be successfully extracted from Cinchona tree bark (Cinchona pubescens Vahl) using nothing but household materials, usually cheap and easily acquired items available in any hardware store. During solid-liquid extraction with reflux, using an organic solvent is used to retain both compounds. Afterward, the compounds are partitioned between water and paint solvent. The organic phase containing the chlorophyll is saved, and the aqueous layer is acidified. Finally, an ultraviolet (UV) light is used to provoke fluorescence in both compounds.

Cinchonine: A Versatile Pharmacological Agent Derived from Natural Cinchona Alkaloids.

Cinchonine is one of the Cinchona alkaloids that is commercially extracted from the Peruvian bark of Cinchona officinalis L. (Family: Rubiaceae). It is also obtained in much lower quantities from other species of Cinchona, such as Cinchona calisaya, Cinchona succirubra, and Cinchona pubescens, and in some other plants, such as Remijia peruviana. Cinchonine has been historically used as an anti-malarial agent. It also has a wide range of other biological properties, including anti-cancer, anti-obesity, anti-inflammatory, anti-parasitic, antimicrobial, anti-platelet aggregation, and anti-osteoclast differentiation. This review discusses the pharmacological activity of cinchonine under different experimental conditions, including in silico, in vitro, and in vivo. It also covers the compound's physicochemical properties, toxicological aspects, and pharmacokinetics. A comprehensive literature search was conducted on multiple online databases, such as PubMed, Scopus, and Google Scholar. The aim was to retrieve a wide range of review/research papers and bibliographic sources. The process involved applying exclusion and inclusion criteria to ensure the selection of relevant and high-quality papers. Cinchonine has numerous pharmacological properties, making it a promising compound for various therapeutic applications. It induces anti-cancer activity by activating caspase-3 and PARP-1, and triggers the endoplasmic reticulum stress response. It up-regulates GRP78 and promotes the phosphorylation of PERK and ETIF-2α. Cinchonine also inhibits osteoclastogenesis, inhibiting TAK1 activation and suppressing NFATc1 expression by regulating AP-1 and NF-κB. Its potential anti-inflammatory effects reduce the impact of high-fat diets, making it suitable for targeting obesity-related diseases. However, research on cinchonine is limited, and further studies are needed to fully understand its therapeutic potential. Further investigation is needed to ensure its safety and efficacy in clinical applications. Overall, this review article explains the pharmacological activity of cinchonine, its synthesis, and physicochemical properties, toxicological aspects, and pharmacokinetics.

Effect of ZnMgO2 nanoparticles used as a nanofertilizer: promoting the growth activities of rice seedlings

Novel ZnMgO2 bimetallic nanoparticles were synthesized using Cinchona succirubra leaf extract and used as a nano fertilizer, promoting the growth activities of rice seedlings, α-amylase and antioxidant activity with average particle sizes of 24.8 nm.

Green nanoarchitectonics of carbon quantum dots from Cinchona Pubescens Vahl as targeted and controlled drug cancer nanocarrier

Carbon quantum dots (CQDs) are a new carbon-based nanomaterial that has attracted tremendous attention due to their excellent fluorescent properties, chemical stability, water solubility, and biocompatibility features. Here, fluorescent CQDs synthesized by a green nanoarchitectonic method using Cinchona Pubescens Vahl extract were evaluated as drug nanocarriers for carboplatin (CBP) delivery. The characterization methods showed CQDs with semispherical shapes and sizes around 5 nm, temperature- and pH-dependent functional groups that interact with the CBP molecule adding specificity to the drug-delivery system. Based on the load efficiency results, it seems that the CQDs can carry almost 100 μg of carboplatin for every 1 mg of CQDs. This is possible due to the self-assembly process that takes place through the interaction between the protonation/deprotonation functional groups of CQDs and the hydrolyzed CBP molecule. Through this process, it is created spherical nanoparticles with an average size of 77.44 nm. The CQDs-CBP nanoparticles release the drug through a diffusion-controlled release mechanism where the acidic media is preferred, and the EPR effect also plays a helpful role. Besides, the viability test shows that the CQDs have almost null cytotoxicity suggesting that they could be used as a promising cancer treatment, improving the efficiency of cell internalization and significantly increasing their drug delivery.

1417 Interest of a cinchona succirubra extract and its association with caffeine in hair loss

1417 Interest of a cinchona succirubra extract and its association with caffeine in hair loss

Endemic Galapagos Tree Daisy Scalesia pedunculata Threatened by Competition with Nonnative Blackberry Rubus niveus

Photo 1. (A) The distribution and population size of the Galapagos Tree Daisy Scalesia pedunculata (1) suffers from competition with nonnative trees such as Cedrela odorata (2), Cinchona pubescens (3) and (B) nonnative undergrowth bushes, most seriously from Rubus niveus (4). (C) A removal experiment and subsequent 8 years of monitoring showed that S. pedunculata can successfully recruit in areas with low undergrowth such as non-native Tradescantia fluminensis (5), but it fails completely to recruit in the presence of R. niveus. Photo credit: B. Riegl (A), H. Jäger (B), M. San José (C). Photo 2. (A) Scalesia pedunculata trees are true trees and follow an asymptotic growth function. Under competition with Rubus niveus, S. pedunculata produces thinner, shorter trees with a reduced life-span. (B) Scalesia pedunculata is a relatively short-lived species and shows strong, pulsed, recruitment dynamics in undisturbed habitat. The shown high density of recruits was only observed in the immediate aftermath of R. niveus removal, but later decreased. However, in the continued presence of R. niveus, no recruitment at all was observed over the 8-year study period. Photo credit: H. Jäger. Photo 3. The endemic Scalesia pedunculata forest is a unique ecosystem that provides an important habitat for many invertebrates and vertebrates, such as this (A) Monarch Butterfly Danaus plexippus megalippe and the (B) endangered Little Vermillion Flycatcher Pyrocephalus nanus. Past control of invasive Rubus niveus has had temporary negative effects on arthropod biomass and bird breeding success of Darwin's finches, but is outweighted by long-term habitat restoration benefits. Photo credit: M. San José. These photographs illustrate the article “Invasive blackberry outcompetes the endemic Galapagos tree daisy Scalesia pedunculata” by Bernhard Riegl, Anna Walentowitz, Christian Sevilla, Rafael Chango, and Heinke Jäger, published in Ecological Applications. https://doi.org/10.1002/eap.2846

A highly contiguous, scaffold-level nuclear genome assembly for the fever tree (Cinchona pubescens Vahl) as a novel resource for Rubiaceae research.

The Andean fever tree (Cinchona L.; Rubiaceae) is a source of bioactive quinine alkaloids used to treat malaria. C. pubescens Vahl is a valuable cash crop within its native range in northwestern South America, however, genomic resources are lacking. Here we provide the first highly contiguous and annotated nuclear and plastid genome assemblies using Oxford Nanopore PromethION-derived long-read and Illumina short-read data. Our nuclear genome assembly comprises 603 scaffolds with a total length of 904 Mbp (∼82% of the full genome based on a genome size of 1.1 Gbp/1C). Using a combination of de novo and reference-based transcriptome assemblies we annotated 72,305 coding sequences comprising 83% of the BUSCO gene set and 4.6% fragmented sequences. Using additional plastid and nuclear datasets we place C. pubescens in the Gentianales order. This first genomic resource for C. pubescens opens new research avenues, including the analysis of alkaloid biosynthesis in the fever tree.

A Review: Pharmacological Activities of Quinoline Alkaloid of Cinchona sp.

Cinchona is a plant used in traditional malaria treatment. This review aims to examine pharmacological activities to increase the benefits and uses of the bioactive compounds found in the bark of the Cinchona trees as medicine and cosmetics. A literature search was conducted through the internet database https://pubmed.ncbi.nlm.nih.gov/; https://scholar.google.com/; https://www.sciencedirect.com/; https://www.wiley.com/en-us from 1998-2022. The main phytochemical content of Cinchona bark is quinoline alkaloids (quinine, cinchonidine, cinchonine, and cinchonidine), with total alkaloid concentrations varying between 6 and 15% (Cinchona succirubra ranging from 5-7%, Cinchona calisaya 4-7% and Cinchona ledgeriana 5-14%). Cinchona quinoline alkaloids have the same active site on the nitrogen atom in the quinuclidine ring and the methylene alcohol functional group, which plays an essential role in their pharmacological activity. Besides being used as an antimalarial, Cinchona alkaloids are currently being developed as their anticancer, antioxidant, antidiabetic, antifungal, muscle cramps, hair growth stimulant, antimicrobial, antiobesity, antiplatelet, antiviral, anesthetic, and antipyretic properties. Conclusion: Quinoline alkaloids of Cinchona sp have various pharmacological activities that have the potency to be developed as drugs and cosmetics.

Carbon Dots Synthesized from Cinchona Pubescens Vahl. An Efficient Antibacterial Nanomaterial and Bacterial Detector.

AbstractCarbon dots (CDs) synthesized using Cinchona pubescens Vahl from the hydrothermal method displayed blue emission radiation, pH dependency, homogeneous sizes, colloidal stability in water, and antibacterial features. They consist of graphene oxide and reduced graphene oxide, attributed to the antioxidant species extracted from the Cinchona pubescens Vahl that simultaneously synthesized and reduced graphene oxide. They reach a bacterial minimal inhibitory concentration of 62.5 μg mL−1 against Gram‐positive Staphylococcus aureus and Gram‐negative Pseudomonas aeruginosa strains, which is better than traditional antibiotics and other CDs obtained from similar natural sources and methods. The photoluminescent CDs were also used to detect bacteria, an essential application in microbiology, biotechnology, medical diagnosis, and food safety, showing the capability of distinguishing between gram‐positive and gram‐negative strains and between dead and alive gram‐positive bacteria. Thus, CDs obtained by green resources represent a promising platform to produce functional antibacterial agents and bacteria detectors.

Formulation of Cinchona Extract (Cinchona succirubra) Cream and The Safety as Hair Fertilizer

Background: cinchona extract contains quinoline alkaloids, has telangiectatic activity, is practically insoluble in water so penetration into hair follicles is low and requires a delivery system, namely cream. Objective: to determine the formula of cinchona extract cream with good activity, physical stability and safety. Methodology: cinchona bark extraction (soxhletation method), phytochemical screening, cinchona extract dosage determination, optimization of cream formula using Design Expert software version 11. Evaluation: pH, adhesion, spreadability, viscosity and cycling test stability. Activity test using guinea pigs (Cavia porcellus) for 14 days, parameters of hair length/2 days, hair mass (14th day). Acute dermal irritation test using rabbit (Orycogalus cuniculus) by erythema and edema index parameters. Results: cinchona extract (13.32% yield) contained quinoline alkaloids, cinchona extract dosage was 20% (p<0.05). The optimum formula is F2 with characteristics: pH 6.48±0.07, adhesion 1.16±0.07, spreadability 4.99±0.85 and viscosity 3067±416.45, has good physical stability. Activity test: test area hair length 13.54 – 62.14% longer than normal control and 10.10 – 49.17% than negative control. The hair mass of test area was 122.91% heavier than normal control and 104.48% of negative control (p<0.05). Cinchona extract and excipients were non-irritant. Conclusion: Cinchona extract cream (20%) has hair fertilizer activity, good physical stability and safety.

Carbon capture in living aerial biomass in Tingo María National Park

The present research work, has the objective of determining the quantity of carbon in the live aerial arboreal biomass; to do so, seven 50m x 50m plots were defined; randomly distributed within the area of the current research; twenty eight families and seventy three arboreal species were identified from a total of 1837 specimens; the family with the most abundance was Rubiaceae, the specie with the most abundance was Cinchona pubescens Vahl (Quina amarillo) with 143 specimens identified. The quantity of stored carbon in the aerial arboreal biomass was an average of 70.08 tC/ac with respect to the plots studied; statistically, the stored carbon has a heterogenous behavior within the area in study; due to the fact that it is found to be influenced by the height and type of vegetation category; at 1066 – 1187 masl the greatest quantity of carbon was found; and the mature trees influence the quantity of carbon at a 50%, followed by the rest of the categories.

Genetic profiles of three Cinchona species in Junghuhn Natural Reserve, Indonesia

Cinchona species were widely used as ancient medicines for different diseases because they contain the active component quinine and its derivatives. However, studies on the molecular aspects of cinchona, including its genetic diversity, have not been reported because most previous works focused on the administration of the antimalarial cinchona alkaloid. Quinine is also being tested as alternative compound for the treatment of Covid-19. The Junghuhn Natural Reserve in Indonesia contains three different types of cinchona plants, namely, Cinchona calisaya, Cinchona pubescens, and Cinchona sp. Given that the genetic diversity and kinship of these species have never been studied, collecting data on the cinchona gene pool has become imperative. This study analyzed the genetic diversity of the cinchona species in the Junghuhn Natural Reserve, Indonesia, by using eight RAPD markers, i.e., OPA-2, OPA-9, OPB-02, OPB-03, OPB-04, OPB-05, OPB-7, and OPJ-07, during 2020 at the University of Jenderal Soedirman, Purwokerto-Indonesia. Polymorphic band data were obtained. Then, phenogram analysis was conducted by using UPGMA and maximum parsimony with MEGA7. The RAPD profiles of Cinchona species (C. calisaya, C. pubescent, and Cinchona sp.) revealed polymorphism with different markers, i.e., OPA-2 (90%), OPB-2 (75%), OPB-5 (75%), OPB-3 (66.66%), OPB-4 (66.66%), OPB-7 (66.66%), OPJ-7 (66.66%), and OPA-9 (58.33%) sequentially with total polymorphism (70.62%). C. calisaya was identified as the most distinctive species. UPGMA yielded a coefficient of 0.200 and two distinctive groups: Group I, which comprised C. pubescens and Cinchona sp. with the p-distance value of 0.333, and Group II, which contained C. calisaya. Ixora sp. was treated as an outgroup plant. The topology of the dendrogram was consistent with that of the UPGMA dendrogram. Results may be used for the further exploration of the genetic diversity of cinchona species.

Potential bioactive compounds of medicinal plants against new Coronavirus (SARS-CoV-2): A review

Nowadays, COVID-19 cases are growing worldwide, and there is no evidence of an effective drug to combat the new SARS-CoV-2 virus. In this sense, medicinal plants as an alternative treatment are being used without control to handle the pandemic situation. Therefore, bioactive compounds against Coronavirus of medicinal plant species that grow in Ecuador are discussed. The plant species described in this review are Azadirachta indica, Cinchona pubescens, Coriandrum sativum, Cúrcuma longa, Eucalyptus spp, Morus alba, and Salvia rosmarinus[A3] ; and compunds are curcumin, coriandrin, deoxynojirimycin, kuwanon G, mulberroside A, oleanolic acid, rosmarinic acid, ursolic acid, quinine, eucalyptol, and jensenone.

Il y a deux siècles : la quinine

On September 11th 1820, Joseph Pelletier and Joseph Bienaimé Caventou communicated about their researches on Cinchona trees. They first described the isolation of Cinchonine from grey cinchona tree. Then they described the process they followed to isolate and identify Quinine from yellow Cinchona trees. They proved that it was a new and an alkaline compound and studied its chemical properties. Then they compared the compositions of the three kinds of Cinchona tree. They showed that the red one, Cinchona succirubra at the time, was richer than the others, so far as Quinine and Cinchonine were concerned. François Magendie performed the first pharmacological studies. Quinine Sulfate was administered by François Clément Maillot in 1834 to soldiers of French Army in Algeria, in order to treat them with success against malaria.

A thorough evaluation of matrix-free laser desorption ionization on structurally diverse alkaloids and their direct detection in plant extracts.

Alkaloids represent a major group of natural products (NPs), derived from highly diverse organisms. These structurally varied specialized metabolites are widely used for medicinal purposes and also known as toxic contaminants in agriculture and dietary supplements. While the detection of alkaloids is generally facilitated by GC- or LC-MS, these techniques do require considerable efforts in sample preparation and method optimization. Bypassing these limitations and also reducing experimental time, matrix-free laser desorption ionization (LDI) and related methods may provide an interesting alternative. As many alkaloids show close structural similarities to matrices used in matrix-assisted laser desorption ionization (MALDI), they should ionize upon simple laser irradiation without matrix support. With this in mind, the current work presents a systematic evaluation of LDI properties of a wide range of structurally diverse alkaloids. Facilitating a direct comparison between LDI and ESI-MS fragmentation, all tested compounds were further studied by electrospray ionization (ESI). Moreover, crude plant extracts of Atropa belladonna, Cinchona succirubra, and Colchicum autumnale were analyzed by LDI in order to evaluate direct alkaloid detection and dereplication from complex mixtures. Finally, dose-dependent evaluation of MALDI and LDI detection using an extract of Rosmarinus officinalis spiked with atropine, colchicine, or quinine was conducted. Overall, present results suggest that LDI provides a versatile analytical tool for analyzing structurally diverse alkaloids as single compounds and from complex mixtures. It may further serve various potential applications ranging from quality control to the screening for toxic compounds as well as the build up of MS databases. Graphical abstract.

Efficacy Profile of the Homeopathic Combination for Influenza and Acute Respiratory Viral Diseases Treatment and Prevention

Background. Acute respiratory viral infection (ARVI) is accompanied by a general intoxication syndrome and respiratory mucosa predominant damage. A significant number of viral nature pathogens from various nosological groups (influenza viruses, parainfluenza, adenoviruses, respiratory syncytial viruses, coronaviruses, picornaviruses, rhinoviruses, enteroviruses, herpesviruses, etc.) could cause development of ARVI at human respiratory tract. Methods of ethnoscience and non-traditional medicine can be used as part of integrated treatment and prevention of influenza and ARVI. With the help of anti-cold homeopathic remedies, people can be treated from the very beginning of the disease when an elevated temperature is observed and there are all signs of intoxication and inflammation. Objective. Scientific justification for the safety and efficacy of a complex homeopathic formulation in tablet form ( Aconitum napellus D6, Ammonium bromatum D4, Atropa belladonna D6, Bryonia D6, Cinchona pubescens D6, Echinacea D3, Hydrargyrum bicyanatum D8, Rhustoxicodendron D6) intended to enhance the natural immunity in influenza and respiratory diseases, to normalize the functional state of the immune system and upper respiratory tract. Results. Most patients have acute febrile respiratory syndrome, and some population groups have an increased risk of complications in the form of severe illness or death. These groups include elderly people, very young people, and people with concomitant illnesses. Annually, every third inhabitant of the planet suffers from acute respiratory infections. We have conducted a scientific substantiation of the safety and efficacy of a complex homeopathic tablet formulation ( Aconitum napellus D6, Ammonium bromatum D4, Atropa belladonna D6, Bryonia D6, Cinchona pubescens D6, Echinacea D3, Hydrargyrum bicyanatum D8, Rhustoxico ­dendron D6), which can be used to treat and prevention of these diseases. Conclusions. Components of developed homeopathic preparation are used in medical practice as anti-cold and immunomodulating mono- and complex homeopathic preparations for more than 100 years. There are no restrictions on the use of the drug in the context of its safety profile, since the active ingredient concentrations used are completely non-toxic. The preparation enhances the body’s protective responses and promotes the relief of symptoms of acute respiratory infections and flu (headache, sneezing, runny nose, sore throat, body aches, fever) and rapid recovery.

Identifying potential distributions of 10 invasive alien trees: implications for conservation management of protected areas.

Tree invasion has the potential to negatively affect biodiversity and ecosystems, with invasive alien trees (IATs) expanding widely in protected areas (PAs) across different habitats. Thus, the effectiveness of PAs might be reduced. Investigation of the distributions of IAT is urgently required to improve the effective conservation management of PAs. We projected the potential distributions of 10 IATs, which included Acacia mearnsii, Ardisia elliptica, Cecropia peltata, Cinchona pubescens, Leucaena leucocephala, Melaleuca quinquenervia, Miconia calvescens, Morella faya, Prosopis glandulosa, and Spathodea campanulata, that have a serious influence on global biodiversity and assessed the distribution possibilities of these IATs in PAs based on the PA categories of the International Union for Conservation of Nature (IUCN). The overall potential distributions of these 10 IATs included Latin America, central and southern Africa, southeastern Asia, eastern Australia and New Zealand, and western Europe. Annual mean temperature, temperature seasonality, annual precipitation, and soil bulk density were found to be important environmental variables for the potential distributions of these IATs. Overall, A. mearnsii, A. elliptica, C. peltata, L. leucocephala, M. quinquenervia, M. calvescens, and S. campanulata were distributed mainly in the IUCN PA categories of national parks and PAs with sustainable use of natural resources. We proposed the following for conservation management of PAs: (1) completion of species inventories for PAs, (2) better understanding of factors driving invasions in PAs, (3) assessment of the efficiency of management within particular PAs, and (4) evaluation of changes in trends regarding plant invasions in PAs under climate change conditions.

Presumption of allelopathic compound(s) released in the leaf litter decomposition process of invasive plants: Calliandra calothyrsus and Cinchona pubescens

Mutaqien Z. 2017. Presumption of allelopathic compound(s) released in the leaf litter decomposition process of invasive plants: Calliandra calothyrsus and Cinchona pubescens. Pros Sem Nas Masy Biodiv Indon 3: 334-338. Invasive alien species is one of the main cause of ecological systems dramatically changes and also native species extinction worldwide. However, more studies are needed to discover its mechanism, including role of allelopathy compound in invasive plants competition. Cinchona pubescens Vahl. and Calliandra calothyrsus Meissn. are recorded as invasive species in some regions. Moreover, C. pubescens is stated as one of 100 of the world’s worst invasive alien species by IUCN. This study aimed to confirm indication of the existence of allelopathic mechanism in invasion process of these two invasive alien species by releasing allelopathic compound(s) from its leaves fall over decomposition process. To clarify this hypothesis, a preliminary study had been conducted by testing the effect of the compounds released from decomposition process over the times (7-180 days) to germination of Vigna radiata (L.) R. Wilczek in the laboratory. Completly Randomize Design was used by applying liquid produced by decomposition process of these two invasive species and control (three repetitions) to test its effects. Radicles and hypocotyls length were measured, compared and Anova analyzed by using R-statistic 3.1.3. Germination of V. radiata only significantly inhibited by compound(s) released by decomposition processes of C. callisaya’s leaves. Its inhibition effect was reduced over the times

Micrografting technique in cinchona (Cinchona ledgeriana Moens) plants propagation

In Indonesia has been found 11 cinchona species but only two species recognized as an economically important Cinchona succirubra known as resistant plant to root disease while Cinchona ledgeriana quinine high content. To put together those two characters from mostly through grafting it took twelve months. Micrografting is one of the propagation techniques through tissue culture. Micrografting has already been used in wide spread for plants propagation either only for research or as mass product. Hopefully, the use of micrografting to obtain cinchona propagation would be short more or less 10 months and provide plants as mass production economically. The result showed that micro grafting at V shape could be used as the propagation method on cinchona species besides bud multiplication through tissue culture. MS medium is the best medium for cinchona micrografting. If there is no IBA on rooting media, there is no root formation from the plants. The increasing of IBA from 1 up to 4 mg/l in media, will increase also the number of rooting plants.different plant material as scion and stock and different concentrarions (0,1,2,3,4 mg/l) of IBA gives the significantly different respons on rooting formation. The increasing of IBA on media will increase the number of rooting plants also. The highest rooting formation are found on medium with 3 dan 4 mg/L IBA. The combination between C. ledgeriana-C. succirubra, C. ledgeriana-C.ledgeriana, C. succrirubra-C.succirubra gives significantly difference on the growth dan development of root. The use of C. ledgeriana as a root stock doesn’t showed good rooting formation. The combination between CSA/QRC205 gives the highest number of leaves result compare wth others combination on as much as 9,8. The first stage of acclimatization on culture room with temperature 25-270C and followed by acclimatization on plastics house covered with paranet 70%. In general in vitro planlet cinchona from micrografting growing well. By the end of observation (8 weeks) growing percentage reach 90%. Â