The effect of 2,4-dichlorophenoxyacetic acid, benzyl amino purin and cupric sulphate on in vitro propagation system from shoot apices of shoot tiller of hybrid Napier grass (Pennisetum purpureum Schum)

An efficient micropropagation method of hybrid Napier grass (Pennisetum purpureumn Schum) for in vitro plant production and material breeding was established from multiple-shoot clumps (MCS) regeneration system. This system was important for forage breeding system. Shoot apices from shoot-tillers produced MSC on Murashige-Skoog (MS) induction medium containing several combinations of BAP and 2,4-D in induction stage. The addition of 5 μM (v/v) and 50 μM (v/v) CuSO4 were added in best medium for inoculation to proliferate the clump in proliferation/multiplication stage. Plant regeneration was achieved by culturing on solid MS with several combination of medium containing NAA and BAP in regeneration stage. The best results for induction were Murashige-Skoog (MS) induction medium containing 2 mgL−1 BAP and 0.1 mgL−1 2,4-D. The proliferation stage on MS medium containing 5 μM CuSÜ4 effective for proliferation (50% multiple shoot formation). The regeneration stage using 0.1 mgL−1 NAA and 2.0 mgL−1 BAP (51.6% number of shoot can regenerate). All plantlets were successfully grown up in an acclimatization stage. Based on the results, the hybrid Napier grass regeneration via MSC was a stable tissue culture system (no albino plats), which could be applied either for further genetic transformation assay or for alternative supply of nursery plant in the future.

Cabe Jawa (Piper retrofractum Vahl.) : PENGGUNAAN TRADISIONAL, FITOKIMIA dan AKTIVITAS FARMAKOLOGI Piper retrofractum Vahl. : Traditional Uses, Phytochemical and Pharmacological Activities

<em><em><em> </em></em></em><p align="center">ABSTRAK</p><p align="center"><strong> </strong></p><p>Cabe jawa<strong><em> (</em></strong><em>Piper</em><em> retrofractum</em> Vahl.) adalah tanaman daerah tropis asli Indonesia yang dijumpai juga di negara Asia Tenggara seperti Thailand dan Malaysia, dan sejak dahulu telah digunakan secara turun-temurun sebagai bahan tambahan makanan ataupun obat tradisional. Secara tradisional di masyarakat, buah cabe jawa<strong><em> </em></strong>dapat digunakan dalam ramuan untuk mengobati demam, perut kembung, mulas, muntah, mengatasi gangguan pencernaan, merangsang nafsu makan, dan lemah syahwat. Akarnya sering digunakan untuk mengobati sakit gigi, luka dan kejang, serta bagian daunnya digunakan juga untuk obat kumur. Beberapa penelitian menyebutkan aktivitas farmakologi cabe Jawa<strong><em> </em></strong>memiliki efek afrodisiaka, antipiretik, antihiperurisemia, antikanker, dan antimikroba. Pengujian klinis terhadap cabe jawa telah dilakukan dan potensial dikembangkan menjadi obat tradisional golongan fitofarmaka. Cabe jawa<strong><em> </em></strong>memiliki aktivitas sebagai imunostimulan, lebih tinggi dibandingkan dengan kelompok imunostimulan fitofarmaka. Cabe jawa<strong><em> </em></strong>mempunyai potensi sebagai anti-<em>photoaging</em>, aktivitas antituberkular, antiproliferasi, aktivitas larvasida, dan aktivitas sitotoksik. Studi fitokimia senyawa metabolit sekunder utama yang terkandung dalam cabe jawa<strong><em> </em></strong>antara lain beberapa jenis alkaloid seperti <em>piperine</em>, <em>pipernonaline</em>, <em>guineensine</em>, <em>piperoctadecalidine,</em> minyak atsiri buah cabe jawa mengandung tiga komponen utama yaitu yaitu <em>β</em>-<em>caryophyllene</em> (17%), <em>pentadecane</em> (17,8%) dan <em>β</em>- <em>bisabollene</em> (11,2%) . Selain senyawa utama tersebut, terdapat senyawa baru pada buah cabe jawa, diantaranya; senyawa amida, amida glikosida, fenilpropanoid glikosida, dan alkaloid. Sebagai afrodisiaka bagian yang digunakan adalah buahnya dan senyawa piperin yang diduga bertanggung jawab terhadap aktivitas tersebut. Piperin merupakan senyawa utama dan zat berkhasiat yang terkandung dalam buah<em> </em>cabe jawa dan berfungsi sebagai penurun demam, mengurangi rasa sakit, antioksidan, mengurangi peradangan, antitumor, dan sebagai imunomodulator. Berdasarkan aktifitas farmakologi yang baik dari cabe jawa maka studi atau penelitian-penelitian pada tanaman ini harus terus dilakukan seperti pengembangan formulasi dan upaya perbanyakan tanaman karena populasi cabe jawa jumlahnya terbatas. Media terbaik dalam induksi kalus tanaman cabe jawa adalah <em>Murrashige Skoog</em> (MS) yang ditambah 6-<em>Benzil Amino Purin</em> (BAP) dan <em>Naphtalene</em> <em>Acetic</em> <em>Acid</em> (NAA).</p><p align="center">ABSTRACT</p><p align="center"><strong><em> </em></strong></p><p><em>Piper retrofractum</em> vahl. is a tropical plant native to Indonesia which is also found in Southeast Asian countries such as Thailand and Malaysia, and has been used for generations as a food additive or traditional medicine. Traditionally in the community, <em>P. retrofractum </em> fruit can be used in potions to treat fever, flatulence, heartburn, vomiting, overcome digestive disorders, stimulate appetite, and impotence. The roots are often used to treat toothaches, wounds, and seizures, and the leaves are also used for mouthwash. Several studies have stated that the pharmacological activity of <em>P. retrofractum </em> has aphrodisiac, antipyretic, anticancer, and antimicrobial effects. Clinical testing on <em>P. retrofractum </em> has been carried out and has the potential to be developed into a traditional medicine of the phytopharmaceutical class. <em>P. retrofractum </em> has activity as an immunostimulant, which is higher than the phytopharmaceutical immunostimulant group. <em>P. retrofractum </em>has potential as anti-photoaging, antitubercular, antiproliferative, larvicidal activity, and cytotoxic activity. Phytochemical studies of the main secondary metabolites contained in P. rectofractum include several types of alkaloids such as piperine, pipernonaline, guineensine, piperoctadecalidine, fruit essential oils. Javanese chili contains three main components, namely-caryophyllene (17%), pentadecane (17.8%) and -bisabollene (11.2%). In addition to these main compounds, there are new compounds in <em>P. retrofractum </em> fruit, including; amide compounds, amide glucosides, phenylpropanoid glucosides, and alkaloids. As an aphrodisiac, the part used is the fruit and the piperine compound which is thought to be responsible for this activity. Piperine is the main compound and efficacious substance contained in <em>P. retrofractum </em> fruit and functions as a fever reducer, pain reliever, antioxidant, reducing inflammation, antitumor, and immunomodulator. Based on the good pharmacological activity of <em>P. retrofractum </em>, studies or researches on this plant must continue to be carried out such as formulation development and plant propagation efforts because the population of <em>P. retrofractum </em> is limited. The best medium for callus induction of cabe jawa was Murashige Skoog (MS) with 6-Benzyl Amino Purine (BAP) and Naphtalene acetic (NAA) added.</p><p> </p>

Efectele genetice implicate în răspunsul grăului comun la filtratul de cultură Drechslera sorokiniana (SACC.) subram

Helminthosporiosis caused by the fungus Drechslera sorokiniana (Sacc.) causes significant crop and quality losses to Triticum aestivum L. in agroecological conditions with extreme humidity. Increasing the resistance is considered the most cost-effective and sustainable approach to disease control. The aim of this study was to determine the genetic effects involved in the inheritance of resistance, using the ge-netic model of character reproduction in descendants of wheat. Generations F1, F2, BCP1 and BCP2, de-scended from the mutual crossing of the parents Basarabeanca / Moldova 30 and Moldova 30 / Moldova 3 (P1 and P2) were evaluated for the response of callus characters to the action of D. sorokiniana culture filtrate on the medium Murashige Skoog. Fungal metabolites have decreased the effects of gene actions and epistatic interactions, but also their variance. The phenomenon corresponds to the decrease of callus indices. A great importance for the heredity of the character of the surface of the callus manifested the epistatic effects of additive-dominant (ad) type. In the case of callus biomass comparable to the mean val-ues were the a actions, but also the ad and dd epistatic effects. The predominant involvement of epistatic effects indicates the need for resistance selections to helminthosporiosis in late generations of wheat.

In Vitro Regeneration of The Endangered Cactus Turbincarpus Mombergeri Riha, A Hybrid of T. Laui x T. Pseudopectinatus

Turbinicarpus mombergeri is a cacti species formed by a hybridization process between Turbinicarpus laui and Turbinicarpus pseudopectinatus. Under natural conditions, it is very difficult for two species be genetically compatible for hybridization, and to produce flowers at the same time. Thus, T. mombergeri is a very interesting and a rare species. Unfortunately, the current populations are decreasing and now it is considered critically endangered. The aim of this research was to develop a successful protocol for propagating T. mombergeri using the in vitro culture techniques. Seed disinfection was performed with Plant Preservative Mixture, and 80% of germination occurred at day 45 in Murashige-Skoog medium. The shoots were cut longitudinally, and the segments were transferred to media containing 2.22 or 4.44 µM benzyladenine to induce shooting. The generated shoots were highly hydrated, and presented abundant callus. The hyperhydricity was controlled by reducing salt medium concentration, by increasing calcium levels and by using polyethylenglycol. The reduction of callus was attained by adding tri-iodo benzoic acid. Vigorous and thick shoots were generated in medium containing urea, and rooting improved in the presence of 0.5 µM indoleacetic acid. Plantlets with normal morphology were obtained, and the survival rate of the plants in soil was 80%. The methodology developed represents an alternative for propagation of T. mombergeri under controlled conditions for commercial or conservation purposes.

Analisis Peran Air Kelapa Terhadap Pertumbuhan Eksplan Kultur Vanili (Vanilla planifolia)

Prospek vanili (Vanilla planifolia) sebagai komoditas penghasil devisa negara masih sangat berpeluang untuk dikembangkan. Salah satu teknik dalam pengembangan vanili melalui metode budidaya kultur jaringan tanaman dengan penambahan air kelapa sebagai sumber zat pengatur tumbuh alami agar eksplan asal ruas batang vanili dapat tumbuh dan berkembang secara optimal. Air kelapa dapat diperoleh dengan mudah sehingga lebih efisien pada segi biaya dan waktu. Oleh karena itu, tujuan dari penelitian ini adalah menganalisis peran air kelapa terhadap pertumbuhan eksplan kultur vanili. Pelaksanaan penelitian pada bulan Februari - Juni 2021 di Laboratorium Kultur Jaringan Politeknik Negeri Jember. Metode analisis T test digunakan untuk membandingkan perlakuan tanpa penambahan air kelapa (P0) dengan perlakuan penambahan air kelapa 15% (P1) pada media dasar Murashige-Skoog. Hasil penelitian memperlihatkan bahwa 1) Penambahan air kelapa 15% mempengaruhi tinggi tunas eksplan vanili dengan rerata 4,45 cm/eksplan dan jumlah akar dengan rerata 2,03 akar/eksplan pada umur eksplan 56 hsi, 2) Penambahan air kelapa 15% memiliki pengaruh yang sama dengan tanpa penambahan air kelapa terhadap kedinian bertunas, jumlah akar pada 14, 28, 42 hsi dan panjang akar eksplan vanili, 3) Penambahan air kelapa 15% mampu mendukung pertumbuhan panjang tunas dan akar tetapi tidak mampu menggandakan tunas eksplan vanili.

Problems of Managing Potato Plant Growth Under Micropropagation for Primary Seeds

This study shows the possibility of controlling the growth of potato microplants when adding the Murashige-Skoog (MS) nutrient medium chlorocholine chloride (CCC) to the composition at a concentration of 0.05-0.25 mg ⋅ L −1. Varietal specificity of the reaction of potato plants to the preparation was revealed. At optimal concentrations of CCC, plants with a stem length of 7-10 cm in 250 ml flasks were grown from single-node microcuttings for 1.5 months. Microplants were removed from the flasks using 20 cm scissors and unsterilized tweezers. The roots were shortened to 2-3 mm and the basal part of the stems was treated with a chalk-based paste containing 0.04% indolylbutyric acid (IBA). Plants were planted on 36 mm diameter Jiffy 7 peat pellets directly in the laboratory. The trays with the material were transferred to the greenhouse at the end of April at a temperature of 20-25 °C and placed in the conditions of the upper fine irrigation. After 3-4 days, mass root formation began in the plants, and another week later the seedlings were ready for use. By then the height of the plants was 10-15 cm, and the roots were clearly visible on the surface of the peat pellets. Planting these seedlings in a technological greenhouse to obtain minitubers was convenient and inexpensive. Keywords: potato, microcuttings, microtubers, minitubers, chlorocholine chloride, peat pellets

INDUCTION OF MICROTUBING OF NEW PROMISING POTATO VARIETIES IN ASEPTIC CULTURE

The studies were carried out with the aim of optimizing microtubing of promising potato varieties in aseptic culture. The experiments studied such factors as the use of vessels of various sizes (test tubes with a diameter of 25 mm and plastic containers 18x18 cm, into which 10 ml of agar and 400 ml of liquid nutrient medium, respectively, were poured, respectively), the density of planting plants in containers (40, 60 and 80 stem explants), the composition of the nutrient medium during ontogenesis, cultivation of mini-tubers from in vitro microplants (control) and microtubers (≥ 0.9 cm and 0.5 ... 0.9 cm in size). When studying the possibility of modifying the nutrient medium based on the Murashige-Skoog recipe to induce tuberization in one variant, the sucrose concentration during growth was changed from 2% before the formation of four internodes by 8% after this phase, the kinetin content during the entire observation period was 0.5 mg/l. In the second variant, microplants were kept on a medium with 6% sucrose and 0.25 mg/l kinetin throughout ontogenesis. The highest yield of microtubers of the standard fraction (27 ... 94%) with a multiplication factor of 0.8 ... 2.7 pcs/plant was noted in the variant with 60 cuttings placed in a container. When grown in test tubes with a change of medium, 1.0 ... 1.5 microtubers were collected per plant with a standard fraction yield of 64 ... 78%. The use of container technology with a similar alternation of nutrient media increased the yield of the standard fraction in most of the studied varieties to 75 ... 86%. In variants with a constant sucrose content in the nutrient medium (6%), a very low multiplication factor was noted, which did not compensate for a sufficiently high yield of the standard fraction, regardless of the laboratory vessel used. The multiplication factor of test tube microplants during planting in the ground was higher than when planting microtubers, with a high yield of the standard fraction

Influence of climatic conditions of the introduction period and varietal characteristics of black currant (Ribes nigrum L.) on the effectiveness of culture initiation in vitro

The article presents data from the effectiveness studies of the initial introduction stage of black currant (Ribes nigrum L.) into in vitro culture depending on the introduction period and the corresponding climatic conditions. The research objects were varieties of black currants selected by the Russian Research Institute of Fruit Crop Breeding: Azhurnaya, Orlovskaya serenada, Ocharovanie, Chudnoye mgnovenye. The introduction into in vitro culture was carried out in several periods characterized by different physiological states of the explants: the period of dormancy release (mid-March), the period of active growth (June), and the period of growth decay (mid-September). The source material in the spring and autumn periods were the buds of one-year stiffened shoots, in the summer introduction period - the buds of growing green shoots. The cultivation was carried out on Murashige-Skoog medium supplemented with 6-BAP (0.5 mg/l). It was noted that the survival rate of explants is determined by the physiological state of the source material due to the corresponding agro-climatic conditions during the introduction period, as well as the genotypic characteristics of the varieties. Thus, explants isolated during the active growing season are characterized by a higher and more stable survival rate. When explants were cultivated in spring and autumn, the physiological state of the explants and their survival rate were influenced by the genotypic response of varieties to the corresponding agroclimatic conditions.

Influence of bacterial metabolites from permafrost on morphophysiological parameters of potato material in vitro*

The effect of secondary metabolites of bacteria from permafrost rocks on the in vitro morphophysiological parameters of potato microplants has been studied. Microplants of Zhukovsky ranniy potato variety were used as the object of the study. The experimental results indicate that when potato microplants are grown in vitro together in the Murashige-Skoog nutrient medium with the bacteria metabolites of the Bacillus cereus 9-08-CH9 and Achromobacter spanius 10-50TS2 strains introduced at the grafting time at a dose of 250 μl, they have the highest inhibitory effect. Secondary metabolites of these bacterial strains have a significant increase in the number of internodes at all stages of plant cultivation, which can accelerate the in vitro replication of material for original potato seed production. Metabolites of the bacterial strain Bacillus cereus 875TS in the indicated concentrations cause growth inhibition and delay in the development of microplants’ root system at the initial cultivation stages.

The effectiveness of the use of the silicon-containing preparation Siliplant in the case of clonal micropropagation of the rose Reine Sammut

Clonal micropropagation of plants is a modern popular method of vegetative reproduction, which allows to obtain high-quality planting material in a large volume in a short time. The research is devoted to the technology of clonal micropropagation of the rose Reine Sammut, in particular, optimization of the nutrient medium composition. To increase the efficiency of micropropagation, the silicon-containing preparation Siliplant was added to the composition of the nutrient medium according to the Murashige-Skoog (MS) recipe as an alternative to the complex of trace elements. At the stage of the actual micropropagation with the combined use of 6-benzylaminopurine (6-BAP) at doses of 1.0; 2.0 and 3.0 mg/l and Siliplant at doses of 1.0; 2.0 and 3.0 ml/l, no large differences were found. Nevertheless, the inclusion of Siliplant (1.0 ml/l) and 6-BAP (3.0 mg/l) in the medium contributed to an increase in the reproduction factor by 3.1 pcs./cuttings at LSD05 = 1.0 pcs. In addition, Siliplant at a dose of 1.0 ml/l as part of a hormone-free nutrient medium MS contributed to a significant increase in shoot growth (by 4.2 mm at LSD05 = 3.6), and at a dose of 3.0 ml/l stimulated the development of standard micro-plants of roses.