Particularităţile rizogenezei speciilor Vaccinium vitis-idaea L. şi Vaccinium macrocarpon Aiton în cultura in vitro

This article presents the studies on the process of rhizogenesis of explants of Vaccinium vitisidaea L. and Vaccinium macrocarpon Aiton (Ericaceae family) in vitro. The rhizogenesis was more efficient on Woody Plant Medium, gelled with agar powder and supplemented with IAA (0.2 mg/l) and IBA (0.5 mg/l) and the pH of the growth medium – 5.0.

How Carbon Source and Seedcoat Influence the In Vitro Culture of Peach (Prunus persica L. Batsch) Immature Seeds

The effects of carbon source and concentration and of seedcoat were tested on the in vitro germination of peach seeds derived from crosses performed in the field. Seeds were extracted from the fruit and cultured in Woody Plant Medium (WPM) supplemented with sucrose, glucose, or sorbitol at concentrations of 15, 30, and 45 g·L−1. The percentage of germination as well as the root and hypocotyl lengths were measured after the stratification process and before acclimatization. Seedcoat did not have any influence on seed germination in any tested media and genotype. Glucose at a concentration of 15 g·L−1 and sucrose at 15, 30, and 45 g·L−1 resulted in greater stem seedling growth. The root developed the most when seeds were cultured in media with 15 or 30 g·L−1 of sucrose.

Prácticas del laboratorio de cultivo de células y tejidos vegetales

Este manual está enfocado en la propagación asexual de los vegetales, por ende, las prácticas se orientan a la identificación de tipos de plantas, elaboración de medios de cultivo a partir de sales de Murashige y Skoog, como de Woody Plant Medium (WPM), así como la siembra de semillas in vitro y la organogénesis. Consta de un total de 13 prácticas acompañadas de figuras que ilustran la descripción del procedimiento, además de dos apéndices que guían la creación de medios de cultivo.

Creation and maintanance of in vitro collections of ornamental cultures from the Lomonosov MSU Botanical Garden’s funds

The article contains the data on creating and maintaining the collections in vitro of some ornamental plants – lilac, roses, tree peonies. The analysis of morphogenetic processes during microclonal propagation of lilac cultivars demonstrates well-defined correlation between susceptibility of this culture to mineral mixture of nutrient medium and plant genotype. Rose cultivars from different garden groups keep high growth capacity during 12 subculturing cycles on Murashige Skoog medium (MS) supplemented with 0.5 mg/l BAP. For all culturing tree-peonies cultivars Woody Plant Medium (WPM) with double content of Ca2+ ions and total exception of Cl- ions turns out to be more suitable. The opportunity of 10 lilac cultivars and 4 rose cultivars preservation in slow growth cultures was appreciated. The correlation between long-term cultivation in low temperature and genotype of ornamental plants cultivars was shown. Thus, 18 ºС turns out to be suitable for creation of slow-growth collection of all studied rose cultivars, and 14 ºС – only for 'Nina Weibull'. Maintaining of lilac microshoots at 10 ºС during 5 months practically did not act negatively on their viability.

Comparison of the Effect of Meta-Topolin and Benzyladenine during Daphne mezereum L. Micropropagation

Meta-Topolin (mT) is one of the lesser-known and only recently discovered cytokinins that can be used as an alternative to the commonly used benzyladenine (BA). The experiment aimed to compare the effects of BA and mT on the regeneration from explants of Daphne mezereum, on the levels of several chemical components, the catalase activity, and to assess genetic diversity of microcuttings. The Murashige and Skoog medium (MS) and the woody plant medium (WPM) were used, enhanced with auxin and 1 mg·L−1 mT or 1 mg·L−1 BA. The results suggest that mT has a more positive effect on shoot regeneration. It also positively affected the levels of various compounds which are generally taken as indicative of micro cutting conditions. The levels of chlorophyll and carbohydrates were the highest in microcuttings from the WPM with 0.1 mg·L−1 NAA. Total soluble proteins and amino acids were the highest in plants from the MS medium with 1 mg·L−1 mT. Increased levels of hydrogen peroxide and catalase activity were observed on media with 1 mg·L−1 BA. Screening with 30 randomly amplified polymorphic DNAs (RAPD) and 20 inter-simple sequence repeat (ISSR) primers demonstrated a considerable level of genetic variation among regenerating plants. This variation is attributed to the condition of the starting material (seed propagated); there were no indications that the two cytokinins tested contributed to increased somaclonal variation.

Multiplication and in vitro rooting of Handroanthus impetiginosus (Mart. Ex DC.) Mattos

Este estudo objetivou avaliar a influência das citocininas durante o estágio de multiplicação in vitro e a combinação entre carvão ativado com o ácido indol-3-butírico (AIB) durante o enraizamento in vitro de Handroanthus impetiginosus. Para a multiplicação in vitro, os segmentos nodais dos brotos obtidos a partir do estabelecimento de plântulas in vitro foram inoculados em meio de cultura Woody Plant Medium (WPM), com e sem as seguintes concentrações de 6-benzilaminopurina (BAP), cinetina (KIN) e tidiazuron (TDZ): 0,5; 1,0; 2,0; 4,0 e 8,0 µmol L-1. Para o enraizamento in vitro, brotos foram inoculados em meio WPM contendo combinações entre AIB (1,0; 3,0; 6,0 e 9,0 μmol L-1) e carvão ativado (1,0; 2,0 e 3,0 g L‑1), bem como suas ausências no meio de cultura. O maior número de brotos e gemas foi alcançado após usar 8,0 μmol L-1 de TDZ, enquanto que o melhor enraizamento ocorreu após a adição de 2,0 g L-1 de carvão ativado, independente da concentração de AIB. Nossos resultados mostram que usar TDZ em níveis adequados para o estágio de multiplicação in vitro seguido pelo uso de 2,0 g L-1 de carvão ativado para obter brotos bem enraizados é eficiente para a micropropagação de ipê-roxo de maneira rápida e com baixo custo, além de fornecer conhecimento sobre como manter esta espécie ameaçada de extinção em um ambiente in vitro podendo ajudar outros estudos de conservação.

PERBANYAKAN AKASIA HIBRIDA (Acacia mangium × Acacia auriculiformis) MELALUI SUBKULTUR BERULANG

In vitro culture is a promising technique for mass propagation of high-value species. Study of propagation for Acacia hybrid (A. mangium x A. auriculiformis) through this technique has been conducted using single node stem from seedlings as explants. Growth medium used was modified Murashige and Skoog (MS), basal medium Woody Plant Medium (WPM), and Gamborg (B5) supplemented. The study was conducted in two stages, namely shoot induction and shoot multiplication. The treatment tested was the Benzyl Adenine (BA) supplementation at the concentration of 0.3; 0.7; and 1.0 mgL-1 of. Observation was conducted on the frequency of shoot induction, number of shoot, shoot length and visual performance of the culture. The result showed that treatment of BA 0.7 mgL-1 on modified MS medium is the best for shoot induction, shoot multiplication and visual performace of the culture. The average of number of shoot was 2.6; 5.0 and 7.7 shoots on the first three consecutive subcultures. Changing to different basal medium on the fourth subculture showed that the treatment of BA 0.7 mgl-1 is the best condition for shoot regeneration (12.60 shoots) and shoot length (6.97 cm). The culture from this treatment showed the best visual morphological performance.Keywords:Acacia hybrid; multiplication; subculture; in vitro; BA. ABSTRAKKultur in vitro merupakan suatu teknik yang menjanjikan untuk perbanyakan massal spesies-tanaman bernilai tinggi. Penelitian perbanyakan akasia hibrid (A. mangium x A. auriculiformis) melalui kulturin vitro telah dilakukan dengan menggunakan eksplan berupa batang satu buku yang berasal dari anakan. Media tumbuh yang digunakan adalah media dasar Murashige dan Skoog (MS) yang sudah dimodifikasi, media dasar Woody Plant Medium (WPM), dan Gamborg (B5). Penelitian dilakukan dalam dua tahap yaitu induksi tunas dan perbanyakan tunas. Perlakuan yang diuji adalah penggunaan Benzyl Adenine (BA) dengan konsentrasi 0,3; 0,7 dan 1,0 mg L-1. Pengamatan dilakukan terhadap waktu induksi tunas, jumlah tunas, tinggi tunas dan penampilan biakan secara visual. Hasil penelitian menunjukkan bahwa penggunaan BA 0,7 mg L-1 pada media MS modifikasi merupakan perlakuan terbaik untuk induksi tunas, perbanyakan tunas, tinggi tunas, dan kondisi biakan secara visual. Jumlah rata-rata tunas yang dihasilkan dari perlakuan ini adalah 2,6; 5,0 dan 7,7 tunas pada subkultur pertama, kedua dan ketiga. Pada penggunaan media dasar berbeda pada subkultur keempat menunjukkan bahwa perlakuan BA 0,7 mg L-1 merupakan perlakuan terbaik dengan jumlah tunas sebanyak 12,60 tunas dan rata-rata tinggi tunas 6,97 cm. Biakan yang dihasilkan dari perlakuan tersebut mempunyai penampilan yang baik dan normal.

Propagación in vitro de Quishuar (buddleja incana ruíz&pav).

Quishuar es una especie forestal apreciada por sus usos. La explotación intensiva y la oferta insatisfecha de plantas han sido limitantes para cumplir con programas de reforestación. La micropropagación es una técnica que ayudaría a erradicar este problema ya que el propósito es producir mayor cantidad de plantas en menor tiempo. El objetivo de esta investigación fue desarrollar protocolos para la desinfección, establecimiento y multiplicación in vitro, para el efecto se realizaron ensayos partiendo de semillas y brotes de plantas. En la primera fase con semillas, utilizando kilol y benomil, junto con NaOCl al 3 % (10 min), los resultados indican que el porcentaje de germinación fue del 100 % en MS (Murashige y Skoog) y porcentaje de contaminación y oxidación 0 %. Por otro lado, los brotes sometidos a fungicidas con adición de antioxidantes y NaOCl al 1 % (10 min) no presentaron contaminación ni oxidación. El porcentaje de brotación fue del 100 % en WPM (Woody Plant Medium). En la segunda fase en medio MS sin adición de hormonas se observó una mayor longitud de brote (1.95 cm), número de nudos (1.94 nudos) e índice de multiplicación (2.47). Basándose en los resultados, se sugiere que los protocolos son efectivos para la propagación in vitro.

Efecto de las citoquininas en la multiplicación in vitro de cuatro variedades de Vaccinium corymbosum, a partir de segmentos nodales Effect of cytokinins on the in vitro multiplication of four varieties of Vaccinium corymbosum

<p>El objetivo principal de este trabajo fue evaluar el efecto de las citoquininas en la multiplicación in vitro de cuatro variedades de arándano (Vaccinium corymbosum) a partir de segmentos nodales. Para lo cual se emplearon cuatro variedades comerciales de arándano (Biloxi, Legacy, Star y Bluecrop), bajo la aplicación de tres citoquininas del tipo Isoprenoides (zeatina, trans-zeatina y cis zeatina). La metodología comprendió la selección de la planta madre y la recolección de tallos jóvenes de 5 a 10cm. libres de plagas y enfermedades, para su desinfección se aplicaron agua desionizada estéril, alcohol al 70%, NaOCl al 3% y Tween 20. Los segmentos desinfectados y seccionados se establecieron en un medio de cultivo líquido Woody Plant Medium (WPM) por un periodo de 45 días, posteriormente los brotes obtenidos se sub cultivaron en un medio de cultivo sólido WPM, suplementado con diferentes concentraciones de zeatina, trans-zeatina y cis-zeatina zeatin (0.0, 1.0, 2.0 y 4.0 mg.L-1), los cuales fueron sometidos a 16 horas luz y 8 horas de oscuridad por un lapso de 50 días. Para el análisis de los datos se empleó la prueba estadística no paramétrica de Kruskal Wallis, observándose que zeatina y trans-zeatina, ambas citoquininas fueron muy efectivas en concentraciones de 2.0 mg.L-1 presentando un mayor número de brotes, hojas, ramificaciones y altura de plántula con promedios de (3.6), (27.5 y 32.5), (2) y (3.5 y 4.5cm.) respectivamente, por otro lado se evidenció que cis-zeatina presentó un comportamiento inactivo en las cuatro variedades de arándano.</p>