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Study of Seed Germination by Soaking Methode of Cacao (Theobroma cacao L.) Sulistyani Pancaningtyas; Teguh Iman Santoso; Sudarsianto .
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 30 No 3 (2014)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v30i3.32

Study of germination methods conduct to get information about seed viability based on germination rate, percentage of germination and vigority. Germination methods was studied to get the efficiency and effectivity of germination, easy to handle, low costs with high vigority. Sand and gunny sack methods for germination, need extensive place and 3-4 days germination period after planting. This research will study the alternative of germination method with soaking. This method can be accelerating germination rate and effectively place usage without decreasing the quality of cacao seedling.The research was done at Kaliwining Experimental Station, Indonesian Coffee and Cocoa Research Institue. This research consist of two experiment was arranged based on factorial completely random design. First experiment will observed to compared germination rate and the second experiment will observed seedling quality between soaking and wet gunny sack germination method.The results showed that length of radicel on soaking method longer than wet gunny sack method. Growth of radicel started from 2 hours after soaking, moreover length of radicel at 4 hours after soaking have significant different value with gunny sack method. On 24 hours after soaking have 3,69 mm and 0,681 mm on wet gunny sack treatment. Except lengt of hipocotyl, there is not different condition between seedling that out came from soaking and wet gunny sack method. Length of hipocotyl on 36 hours after soaking have 9,15 cm and significant different between wet gunny sack germination method that have 5,40 cm. Keywords : seed germination, soaking method, Theobroma cacao L., cocoa seedlings

The effect of 2,4 dichlorophenoxyacetic acid on in vitro callogenesis of cocoa (Theobroma cacao L.) Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 31 No 2 (2015)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v31i2.67

Cocoa (Theobroma cacao L.) development using modern breeding techniques can be facilitated by propagation of planting material through somatic embryogenesis. Various factors that may affect embryogenesis are the composition of culture medium and culture condition. Hormone commonly used to initiate the formation of callus is auxin with type 2.4-D (2.4 Dichlorophenoxy acetic acid). The aim of this study was to determine the effect of the addition of 2.4 -D hormoneson the process of cocoa embryogenesis. The treatments were arragged in factorial combination in completely randomized design, which consisted of two factors. Thefirst factor was the concentration of auxin 2,4-D 25 %, 50 %, 75 %, and 100 %; and the second factor was cocoa clones; Sulawesi 01 and Sulawesi 02. The resultshowed that the addition of 2.4-D hormone up to 100% on somatic embryogenesis of cocoa for Sulawesi 01 clone was not significantly different from Sulawesi 02 clone for all parameters. While on the addition of 2.4-D, there was significant difference between Sulawesi 01 and 02. Cocoa embryogenic callus using the addition of 2.4-D (25%-100%) was significantly different from control. Increased concentrations of 2,4-D hormone which is applied onto media would inhibit the formation of the somatic embryo. Addition of 2.4 D 25%, encouraged towards non-embryogenic callus. Keywords: 2.4 Dichlorophenoxy acetic acid, embryogenic callus, somatic embryos, cocoa, medium culture, hormone

Study on the presence and influence of phenolic compounds in callogenesis and somatic embryo development of cocoa (Theobroma cacao L.). Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 31 No 1 (2015)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v31i1.81

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance

Effectiveness of calcium chloride in reduction of shoot necrosis incocoa (Theobroma cacao L.) in vitro propagation Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 28 No 1 (2012)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v28i1.161

Various efforts have been developed for the optimization of the various stages in vitro micropropagation. The maturation stage and pre-acclimatization plantlets is an important stages that must be considered to produce vigorous plants and ready to be planted in the field. The purpose of this study was to reduced shoot tip necrosis in cocoa planlets to obtaine vigorous planlets from in vitro propagation through the addition of Calcium Chloride (CaCl2). The study used two stages of embryogenic development. The first was embryo maturation stage and the second was the shoot growth development stage. The study was arranged factorially in experiment design of Completely Randomized Design consisting of two factors i.e. concentration of CaCl2 consisted of 0, 50, 100, 150, and 200 mg/l and clones consisted of Sulawesi 1 and Sca 6. Each experiment was repeated three times, so the number of combination trials were 5x2x3=30 experimental units. The parameters observed included shoot growth percentage and vigorous planlets percentage. The results showed that the addition of CaCl2 at a concentration of 150 mg/l during maturition stage increased the embryos performance and percentage of shoot tip. However, it could not prevent the shoot tip necrosis. Whereas, the addition at a concentration of 50 mg/l during the shoot m growth development stage could reduced necrosis, suggested to increase the quality of in vitro planlets.Key words: Cocoa, necrosis, calcium chloride, somatic embryogenesis, embryos, planlets, in vitro.

Evaluation of Quantity and Hyperhydricity of Cocoa Somatic Embryo Obtained from Solid Culture, Liquid Culture, and Sequence Subculture Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 29 No 1 (2013)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v29i1.187

Research aimed to study the effect of solid culture, liquid culture, and sequence subculture on quantity and hyperhydricity of somatic embryo wascarried out at Laboratory of Biotechnology, Indonesian Coffee and Cocoa Research Institute. Materials used in this study were embryogenic callitransferred on somatic embryos expression both in solid and liquid media with the same media composition, namely MS medium with the addition ofAdenine (0.025 mg/L). Gelling agent used in solid media was gelrite (3 g/L). Clones used in this study was Sca 6. This research consisted of two trials, namely1) effect of medium type (solid and liquid), and 2) sequence subculture (four subcultures). This results showed that the production of somatic embryosin liquid medium was higher than in the solid medium. Regeneration of somatic embryos on solid medium culture showed the highest percentage of abnormality embryos due to hyperhydricity at the cotyledonary phase 60%. Meanwhile, the regeneration of somatic embryos in liquid culture showed the highest percentage of abnormality embryos due to hyperhydricity at the globular and cotyledonary phase 37%. Frequent subculture increased abnormal embryos and decreased the number of somatic embryos.Key words: Cacao, hyperhydricity, somatic embryos, solid culture, liquid culture, subculture, in vitro.

Role of Exogenous Salicylic Acid and Benzoic Acid Applications to Vascular Streak Dieback Disease Attack on Cocoa Seedlings Fakhrusy Zakariyya; Agung Wahyu Susilo; Teguh Iman Santoso; Hardian Susilo Addy; Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 34 No 1 (2018)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v34i1.305

Currently, vascular streak dieback (VSD) attacks have widespread in areas ofthe center of cocoa (Theobroma cacaoL.) production, especially inSulawesi. Salicylic acid and benzoic acid are part of phenol compounds reportedlyacts as bioimmunoregulator in increasing of plant resistance to that pathogen. The objec-tiveof this research was to investigate effect of exogenous salicylic acid and benzoic acid applications to VSD disease attack on cocoa seedlings. The research was conducted at Indonesian Coffee and Cocoa Research Institute,Jember, East Java. This experiment designed using randomized completely block design with three replications. The treatments included concentrations of salicylic acid in 0.5 mMol, 1 mMol, 2 mMol; benzoic acid with concentration 0.5 mMol, 1 mMol,2 mMol; and control (water spray). Observations were based on the incidence and severity damage of disease, plant height, stem diameter, number of leaves, stelate trichome, chlorophyll, width and length of stomatal apperture, and number of stomata. Exogenoussalicylic acid and benzoic acid as phytohormone can reduce disease insidence and severity of VSD. In conclusion, our results indicated that exogenous salicylic acid and benzoic promoted stomatal closure in cocoa leaf by decreasing width-stomatal closure. Salicylic and benzoic acid had no effect to plant height, stem diameter, number of leaves, length of stomatal apperture, number of stomata, and number of stelate trichome.

Callogenesis and Embryogenic Potential of New Superior Cocoa (Theobroma cacao L.) Genotypes Treated with Ascorbic Acid Sulistyani Pancaningtyas
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 37 No 3 (2021)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v37i3.472

Clonal propagation is an alternative method of maintaining the genetic purity of superior cocoa plants (Theobroma cacao L.). The somatic embryogenesis technique is likely the most effective and efficient method for plant propagation. This study aimed at investigating the callogenesis and embryogenic potential of new superior cocoa clones on culture media added with various concentrations of antioxidants and variation in the concentration of auxin hormone combined with ascorbic acid. The superior clones used in this study were MCC 01, MCC 02, Sulawesi 3, ICCRI 07, and ICCRI 09. The parameters observed in this study were percentage of embryogenic callus formation, number of secondary somatic embryos, and percentage of embryo somatic cells that develop into planlets. The addition of 2,4,5-T combined with ascorbic acid affected the initiation of cocoa secondary somatic embryogenesis. Treatment combination of ascorbic acid 100 mg.L-1 and 2,4,5-T 1 mg.L-1 had a significant effect on the initiation of secondary somatic embryogenesis of cocoa in the embryogenic callus phase (4.73%), globular embryos (2.53%), torpedo embryos (4.67%) and sprouts (0.47%). Three clones i.e ICCRI 07, Sulawesi 3 and ICCRI 09 clones showed a higher percentage (3.0%, 2.10%, and 1.80%, respectively) of embryogenic callus growth than the other clones. Meanwhile, on MCC 01 clone, all treatments did not affect the regeneration of embryogenic callus. The combination treatment of organic acid and adenine showed a low embryogenic callus response in MCC 01 and MCC 02 clones. However, MCC 02 clone also did not show a response in form of globular, torpedo, and germination phase somatic embryo regenerations. This indicates that different plant cultivars show different responses to the addition of organic acids. Treatment combinations of adenine 0.0125 mg.L-1 + ascorbic acid 100 mg.L-1 gave the best response to the regeneration of somatic embryos for the globular, torpedo, and g

Analysis of Cocoa Clonal Seedlings Purity Through Deoxyribonucleic Acid (DNA) Barcoding and Random Amplification of Polymorphic DNA (RAPD) Fingerprinting Sulistyani Pancaningtyas; Agung Wahyu Susilo
Pelita Perkebunan (a Coffee and Cocoa Research Journal) Vol 38 No 1 (2022)
Publisher : Indonesian Coffee and Cocoa Research Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iccri.jur.pelitaperkebunan.v38i1.490

The genetic purity of a plant indicates the similarity properties between seedlings in the field and the description of the plant in the database. Identifying plant purity through a morphological approach has several drawbacks, including time efficiency and environmental factors, and the diversity is limited and inconsistent. This condition encourages the development of detection methods using DNA molecular markers. Plant identification through fingerprinting or the use of molecular markers has not been widely carried out on a commercial scale, considering the investment costs for this analysis are still quite expensive. Another method used for plant identification through DNA Barcoding by comparing variations between DNA sequences. The primers used to identify barcodes on cocoa were derived from the chloroplast genome, including rbcL and matK. This research aims to see the consistency of the rbcL primer when applied to other cocoa clones, and the analysis of the polymorphic diversity of each cocoa clone using DNA fingerprinting RAPD. This method will be tested on clones of Sulawesi 1, Sulawesi 2, ICCRI 03, and ICCRI 09, which were propagated by SE and mother plants in the field using cocoa leaf samples. The stages include DNA extraction, sequencing, and analysis of the sequencing results. The results of seedlings uniformity analysis using DNA barcoding on cocoa plants produced from in vitro propagation showed that the multiplied seeds did not show any difference in sequence with the parent plant (DR2, Sulawesi 1, Sulawesi 2, and ICCRI 09). The analysis of the diversity of cocoa clones DR 2, MCC 2, Sulawesi 1, Sulawesi 2, and ICCRI 09 through DNA Fingerprinting RAPD showed that the OPA 15 primer produced a more apparent polymorphic band than the other three primers (OPP 08, OPW 11, and M 29).

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