Response of in vitro propagated fig (Ficus carica L.) shoots to the concentrations of benzyl amino purine and coconut water

In this study, several concentrations of benzyl amino purine (BAP) and coconut water (CW) were investigated along with the interaction between two factors to the growth of in vitro propagated fig shoots. The investigated factors consisted of BAP concentration: 0, 1, 3, 5 mg L−1 and coconut water concentration: 0, 100, 200, 300 ml L−1. A total of 16 treatment combinations with 6 replications resulting in 96 experimental units consisting of a single fig shoot explant per culture medium. The observed parameters including living explant rate, contamination rate, browning rate, day of first shoot emergence, shoot formation rate, explant height addition, number of leaves, callus formation rate, and number of roots were conducted every week from 1 to 8 weeks after proliferation (WAP). The result indicated that in 8 WAP, the living explant rate reached 23.95%. The combination of concentration 200 ml L−1 CW and 3 mg L−1 BAP + 200 ml L−1 CW-induced early emergence of new shoots at 7 days after proliferation (DAP). The highest shoot formation rate (100%) was observed at a concentration of 300 mL L−1CW. The highest explant height addition (7.10 cm) was observed at a concentration of 200 mL L−1 CW. The highest number of leaves (5.80) was observed at a concentration of 1 mg L−1 BAP + 200 mL L−1 CW. The highest callus formation rate (50%) was observed at a concentration of 100 ml L−1CW and 300 ml L−1 CW. The highest number of roots (17) was observed in the control.

Identification Secondary Metabolites From Callus Piper retrofractum Vahl

Javanese chili (Piper retrofractum Vahl) is a traditional medicinal plant originating from Indonesia and has many pharmacological activities, one of which is often used as a base for aphrodisiac herbal medicine. The population of P. retrofractum is limited, so it is necessary to design secondary metabolite production and propagation efforts using plant tissue culture techniques. The materials used in this study were explants of P. retrofractum leaves that were induced in Murashige and Skoog (MS) media and the ratio of growth regulators 2.4-dichlorophenoxyacetis acid (2,4-D): Benzyl Amino Purine (BAP) 0.5: 0.5. The results showed that the callus of P. retrofractum was formed in the growth regulator 2.4D: BAP (0.5: 0.5). TLC and spectrophotometry identified the secondary metabolite content of callus. Secondary metabolite analysis using the thin layer chromatography (TLC) method using the mobile phase ethyl acetate: n-hexane (7: 3) showed a terpenoid compound indicated by purple spots on the visual appearance after spraying 10% spotting vanillin. Identification using infrared spectrophotometry shows functional groups -CH, C = O, C = C, -CH2, and -CH3, characteristic of terpenoid compounds. Based on TLC data and spectrophotometry, callus P. retrofractum is thought to contain terpenoid compounds. This study's results are expected to be the basis for developing secondary metabolite production in P. retrofractum with cell suspension culture and P. retrofractum propagation by micropropagation.Abstrak: Cabai Jawa (Piper retrofractum Vahl) merupakan tanaman obat tradisional yang berasal dari Indonesia dan banyak memiliki aktivitas farmakologis salah satunya sering digunakan sebagai bahan dasar jamu afrodisiaka. Populasi tanaman cabai Jawa terbatas maka perlu dirancang upaya produksi metabolit sekunder dan upaya perbanyakan tanaman cabai Jawa salah satunya menggunakan teknik kultur jaringan tanaman. Bahan yang digunakan pada penelitian ini adalah eksplan daun tanaman cabai Jawa yang diinduksi pada media Murashige and Skoog (MS) dan perbandingan zat pengatur tumbuh 2.4-dichlorophenoxyacetis acid (2,4-D) : Benzyl Amino Purine (BAP) 0,5 : 0,5. Hasil penelitian menunjukkan bahwa kalus cabai Jawa terbentuk dalam zat pengatur tumbuh 2,4D: BAP (0,5: 0,5). Kandungan metabolit sekunder dari kalus diidentifikasi dengan KLT dan spektrofotometri. Analisis metabolit sekunder menggunakan metode kromatografi lapis tipis (KLT) menggunakan fasa gerak etil asetat: n-heksana (7: 3) menunjukkan adanya senyawa terpenoid yang ditunjukkan dengan adanya bercak ungu pada penampakan visual setelah disemprotkan spotting vanilin 10%. Hasil Identifikasi menggunakan spektrofotometri UV-Vis menunjukkan isolat mempunyai panjang gelombang maksimum 272,6 nm. Identifikasi menggunakan spektrofotometri inframerah menunjukkan adanya gugus fungsi -CH, C = O, C = C, -CH2, dan -CH3 yang merupakan ciri khas senyawa terpenoid . Berdasarkan data KLT dan spektrofotometri kalus cabai Jawa diduga mengandung senyawa terpenoid. Hasil penelitian ini diharapkan dapat menjadi dasar pemgembangan produksi metabolit sekunder dalam tanaman cabai Jawa dengan kultur suspensi sel dan perbanyakan tanaman cabai Jawa dengan mikropropagasi.

Development of an embryo germination protocol for shy-seeded grape (Vitis vinifera L.)

Acquisition and germination of seeds are the most desired targets for the improvement of vegetatively propagated crops. In the present study, we developed a potential embryo germination protocol for the Red Globe grape cultivar having a low seed germination rate. Three grape berries at different developmental stages, viz. 50, 60 and 70 days after flowering (DAF), were selected for in-vitro embryo germination. Three growth media, namely Emershad and Ramming (ER), Nitsch and Nitsch (NN) and Murashige and Skoog (MS), and plant growth regulators (benzyl amino purine (BA), 0.5, 0.7 and 0.9 mg/l; indole butyric acid (IBA), 1.0, 1.5 and 2.0 mg/l; and gibberellic acid (GA), 0.1, 0.3 and 0.9 mg/l) were screened individually in different combinations with three amino acids, namely cysteine, glutamine and proline (2.0 μmol/l each). The maximum embryos germination percentage recorded at 70 DAF was 63.33, 47.78 and 45.56% in ER, NN and MS media, respectively, supplemented with 0.9 mg/l BA, 2.0 mg/l IBA, 0.9 mg/l GA and 2.0 μmol glutamine. Glutamine was found to have the most significant impact, and it improved the rescued embryos germination. The present study provides a potential recipe for a medium that can facilitate efficient germination of grape embryos.

Sterilisasi dan Pertumbuhan In Vitro Tunas Aksilar Pepaya Kultivar Callina dan Caliso

Pepaya tipe sedang dan kecil sudah digemari di Indonesia sejak satu dekade lalu. Sebagian besar tanaman pepaya yang diperbanyak dari biji menghasilkan profit yang rendah karena heterogenitas dan variasi genetik yang diakibatkan oleh penyerbukan silang, sehingga diperlukan kegiatan secara kultur in vitro. Penelitian dilakukan di Laboratorium Kultur Jaringan Pusat Kajian Hortikultura Tropika (PKHT) IPB, Bogor, dari November 2018 hingga Januari 2020. Eksplan merupakan tunas aksilar pepaya kultivar Callina dan Caliso umur 2 minggu. Penelitian dirancang secara faktorial dalam lingkungan Rancangan Acak Kelompok (RAK). Penelitian ini dibagi menjadi dua percobaan: a) Percobaan pertama bertujuan untuk mengetahui pengaruh metode sterilisasi dengan faktor sumber eksplan (rumah kaca dan lapangan) dan metode sterilisasi (M1, M2, M3, dan M4), dan b) Percobaan kedua bertujuan untuk optimalisasi konsentrasi BAP dan GA3 pada pertumbuhan tunas dengan konsentrasi BAP 0.0, 0.5, 1.0, 1.5, dan 2.0 mg L-1, dan konsentrasi GA3 0.0, 0.3, dan 0.4 mg L-1. Hasil penelitian menunjukkan bahwa eksplan dari rumah kaca pada sterilisasi metode M4 menghasilkan tingkat kontaminasi terendah yaitu 19.9% untuk Caliso dan 29.9% untuk Callina. Media terbaik untuk menghasilkan tinggi tunas optimum adalah BAP 0.5 mg L-1 dan GA3 0.3 mg L-1 untuk Caliso dan BAP 0.5 mg L-1 dan GA3 0.4 mg L-1 untuk Callina dengan tinggi rata-rata 1.6 dan 1.8 cm, sehingga tunas dapat dipindahkan ke tahap pengakaran. Kata kunci: Asam giberelat, Benzyl amino purine, Carica papaya, kontaminasi, sterilisasi

Growth Evaluation of In-Vitro Propagated Embryo of Morinda Citrifolia L. Seeds

The dormant nature of Morinda citrifolia seeds is a limitation to its efficient in-vitro plantlet multiplication. Hence, the use of embryo culture for successful in-vitro culture initiation. Matured embryo of freshly collected noni seeds were cultured on Murashige and Skoog basal medium supplemented with kinetin (Kn) and Benzyl amino purine (BAP) in the range of A: control (no addition); B: 0.5 mg/l Kn+1.0 mg/l BAP; C: 1.0 mg/l Kn+2.0 mg/l Bap; D: 1.5 mg/l Kn+3.0 mg/l BAP and E: 2.0 mg/l Kn+4.0 mg/l BAP. The results at 4 weeks after inoculation (WAI) showed that germination was faster from medium A without hormone whereas highest percentage germination was obtained from both medium D and E with 80 %. Medium B and C had 65 % each while medium A gave the least (40%). The development of the plantlets showed that longest shoot (3.9 cm) from medium A was closely related to 3.58 cm from Medium B while root lengths (2.28 cm) and number of adventitious roots (26) from medium A were significantly higher than other media at 12 WAI. Highest number of nodes (2.25) obtained from medium D was comparable to Media C and B while medium A had the least at 12 WAI. Number of leaves obtained was similar between the media at 12 WAI. These results indicated that using embryo is reliable for fast in-vitro propagation and shoot development of noni plant with optimum cytokinins (0.5/1.0 mg/l Kn/BAP) application. Keywords: Culture initiation, Cytokinins, Embryo culture, Plantlet, Shoot development

Efficient Protocol for In vitro Regeneration of Ocimum sanctum using Nodal Segments as Explants

Ocimum sanctum commonly called (holy basil) an herb containing medicinal, ornamental values, is often used in culinary applications. This research focuses on the improved and efficient protocol for the direct regeneration and acclimatisation of Ocimum sanctum using nodal segments. Organogenesis and multiplication from explants were observed to a maximum on Murashige and Skoog (MS) medium supplemented with 0.1 mg/L of 6-Benzyl amino purine (BAP) and 0.025 mg/L of Indole-3-acetic acid (IAA). Furthermore, same medium was found effective for the induction of roots, in the in-vitro grown plantlets. A series of experiments were conducted to optimise the acclimatisation of in-vitro grown rooted plantlets of Ocimum sanctum. For this study different types of potting mix in assorted ratios were used to obtain best supporting media for the acclimatisation, A7 media containing soil : farmyard manure (75:25) and A1 media containing (100%) sand were found best supporting medium for the acclimatisation and hardening of Ocimum sanctum.

Induction Shoots From Callus of Cucumber Apple (Cucumis sp.) Using a Combination of Benzil Amino Purine and Naphtalene Acetic Acid Concentrations In Vitro

Research on indirect organogenesis of cucumber apple (Cucumis sp.) was conducted from March to May 2020. This study aims to find the best combination between Benzyl Amino Purine (BAP) and Naphthalene Acetic Acid (NAA) on the growth of cucumber apple (Cucumis) sp.) on Gamborg media (B5) by organogenesis. The callus used came fromcucumber seeds explants with a concentration of NAA 2,0 mg/l + BAP 0,2 mg/l using Murashige and Skoog media. The research method used is an experimental method with nonparametric statistics with 24 treatments repeated three times and analyzed descriptively using the Kruskal Wallis test. The results showed that the best treatment of cucumber apple shoots was the combination of NAA and BAP, namely the B4N2 treatment (2,5 ppm BAP + 0,20 ppm NAA) with shoot height (0,45 cm). At a single concentration, a concentration of 1 ppm BAP was able to provide a faster effect on shoot formation time, namely at 24 days after initiation (hsi) and the best response to shooting height (0,5cm).