Modified protocol to overcome in vitro recalcitrance of premature micro-shoots abscission by silver supplementation in Kinnow mandarin (Citrus nobilis L. × Citrus deliciosa T.)

The present study focused on alleviating the issue of premature abscission of micro-shoots emerged from the cultured nodal segments in Kinnow mandarin. To resolve the problem, explants of different maturity stages, culture media, growth regulator combinations and ethylene inhibitors (silver compounds) were evaluated. Hardwood stage was observed to be the most efficient explant maturity stage to boost the vigorous early shoot emergence on MS (Murashige and Skoog) medium. Addition of GA3 (Gibberellic acid) 10 mg L?1 resulted in delayed abscission. Further reduction in abscission rate and healthy shoot growth were observed when subculturing of micro-shoots was done on MS medium supplemented with BAP 2.5 mg L?1, GA3 10 mg L?1 and silver thiosulfate 5 mg L?1. This modified procedure could be effectively utilized for mass multiplication as well as crop improvement in Kinnow mandarin.

Limonoid detection and profile in callus culture of sweet orange

Plant tissue culture has emerged as an important tool to produce bioactive compounds from various plant species, including the sustainable production of limonoids that are receiving considerable attention due to the benefits associated with human health such as anticancer activities. The purpose of the present study was to evaluate the capacity of limonoids aglycone production from callus culture from sweet orange cv. Pera (Citrus sinensis) seeds and identify the compounds produced in this cell line. Callus induction occurred in Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic (2,4-D), malt extract, agar and coconut water. For the analysis and identification of the limonoids, CG-MS-EI ion-positive mode and UPLC-QTOF-ESI were used operating in positive and negative mode. An intense peak corresponding to limonin appeared in the callus extracts at a retention time of 58.1 min. in CG-MS-EI and four major limonoids aglycone by positive ion mode UPLC-QTOF-ESI: limonin, nomilin, deacetylnomilin, and nomilinic acid. The culture medium was efficient at the bioproduction of limonoids aglycone in callus cultures of C. sinensis seeds. Therefore, data obtained from UPLC-QTOF-ESI proved its importance at identifying new compounds that benefit human health, and may assist future work in the identification of known or new limonoids in Citrus species and related genera.

Germination of Mansonia altissima (A. Chev.) A. Chev. var. altissima: an endangered valuable timber species in Africa

The in vitro seed germination which results in the production of disease-free seedlings and greenhouse germination of the seeds of Mansonia altissima was investigated in order to establish a better way of germination of the timber species. Five levels of GA3 treatment were used in in vitro germination with three replicate and two seeds were inoculated in each of the jam bottle. Whereas, in greenhouse germination, five levels of different treatments were used, replicated three times and each Petri plate contained 15 seeds. The experiment was repeated twice and the data from each experiment was put together and used for the statistical analysis. The results showed that seeds germination occurred eight days after inoculation in in vitro but in the case of greenhouse germination, it took only five days. For in vitro rapid germination of Mansonia altissima, the MS medium should be supplemented with 1.0 μm of GA3. Equally, in greenhouse germination, the seeds need to be soaked in 1.0 mM of GA3 for 24 hours. Alternatively, in the absence of GA3, the seeds can be soaked in water for 24 hours before broadcasting the seeds on the seedbed for germination, as this will help to identify nonviable seeds.

In vitro multiplication of cassava varieties / Multiplicação in vitro de variedades de mandioca / Multiplicación in vitro de variedades de mandioca

The implantation of a model of sustainable development for agriculture can happen with the contribution of the mandiocultura. But for this, culture needs to be strengthened. In vitro propagation is an instrument for this purpose. Micropropagation can provide growers with large quantities of vigorous and healthy cassava seedlings in a short time. The objective of this study was to evaluate the in vitro establishment of four varieties of cassava cultivated in the municipality of Colorado do Oeste, State of Rondônia, popularly known as Arara, Caturra, Cacau Vermelha and Roxinha. For that, an experiment was carried out in the Laboratory of In Vitro Cultivation at the Pole of Technological Innovation of the University of Cruz Alta (UNICRUZ), with a completely randomized design in a 4 x 2 factorial scheme, with 6 replications. The treatments consisted of explants grown in Murashige and Skoog (MS) medium without the presence of growth regulator and MS medium supplemented with of 6-benzylaminopurine (BAP). The results indicate that the mean contamination percentage of the explants was 47.19%, differing among the varieties. The best growth response in culture media, in the multiple comparison of means (Scott-Knott's test, 5%), was obtained with MS medium without BAP addition, with significant difference between varieties. Under the conditions of this experiment, it was evidenced that micropropagation is a viable tool for obtaining varieties of interest, with desired phytosanitary qualities, with varietal and large-scale authenticity.(AU)

A implantação de um modelo de desenvolvimento sustentável para a agricultura pode acontecer com a contribuição da mandiocultura. Mas para isso, a cultura precisa ser fortalecida. A propagação in vitro é um instrumento para este fim. A micropropagação pode proporcionar aos produtores grande quantidade de mudas de mandioca vigorosas e sadias em um curto espaço de tempo. O objetivo deste trabalho foi avaliar o estabelecimento in vitro de quatro variedades de mandioca cultivadas no município de Colorado do Oeste, Rondônia, popularmente conhecidas como Arara, Caturra, Cacau Vermelha e Roxinha. Para isso, foi realizado um experimento no Laboratório de Cultivo In Vitro no Polo de Inovação Tecnológica da Universidade de Cruz Alta (UNICRUZ), com delineamento inteiramente casualizado em esquema fatorial 4 x 2, com 6 repetições. Os tratamentos consistiram em explantes cultivados em meio Murashige e Skoog (MS) sem a presença de regulador de crescimento e meio MS suplementado com 6-benzilaminopurina (BAP). Os resultados indicam que a porcentagem média de contaminação dos explantes foi de 47,19%, diferindo entre as variedades. A melhor resposta de crescimento em meios de cultura, na comparação múltipla de médias (teste de Scott-Knott, 5%), foi obtida com meio MS sem adição de BAP, com diferença significativa entre as variedades. Nas condições deste experimento, ficou evidenciado que a micropropagação é uma ferramenta viável para obtenção de variedades de interesse, com qualidades fitossanitárias desejadas, com autenticidade varietal e em larga escala.(AU)

La implantación de un modelo de desarrollo sostenible para la agricultura puede suceder con el aporte de la mandiocultura. Pero, para eso, la cultura necesita ser fortalecida. La propagación in vitro es un instrumento para ese fin. La micropropagación puede proporcionar a los cultivadores grandes cantidades de plántulas de mandioca vigorosas y saludables en poco tiempo. El objetivo de esta investigación ha sido evaluar el establecimiento in vitro de cuatro variedades de mandiocas cultivadas en el municipio de Colorado del Oeste, Rondônia, popularmente conocidas como Arara, Caturra, Cacau Vermelha y Roxinha. Para eso, se realizó un experimento en el Laboratorio de cultivo in vitro en el Polo de Innovación Tecnológica de la Universidad de Cruz Alta (UNICRUZ), con delineamiento completamente casualizado en esquema factorial 4 x 2, con 6 repeticiones. Los tratamientos consistieron en explantes cultivados en medio Murashige y Skoog (MS) sin la presencia de regulador de crecimiento y medio MS suplementado con 6-bencilaminopurina (BAP). Los resultados indican que el porcentaje de contaminación promedio de los explantes fue de 47.19%, diferenciándose entre las variedades. La mejor respuesta de crecimiento en medios de cultivo, en la comparación múltiple de medias (prueba de Scott-Knott, 5%), se obtuvo con medio MS sin adición de BAP, con una diferencia significativa entre las variedades. Bajo las condiciones de este experimento, se evidenció que la micropropagación es una herramienta viable para obtener variedades de interés, con cualidades fitosanitarias deseadas, con autenticidad varietal y de gran escala.(AU)

In Vitro Propagation of Chamomilla Recutita From Capitulum Inflorescence: A Medicinal Plant With Multiple Therapeutic Applications

A protocol has been developed for induction of somatic embryogenesis from whole inflorescence explants of Chamomilla recutita L. (chamomile). Chamomile is a well-known medicinal plant from the Asteraceae family often referred to as the “star among medicinal species.” Nowadays, it is a highly favoured medicinal plant in folk and traditional medicine. Its multitherapeutic, cosmetic and nutritional values have been established through the years of traditional and scientific use and research. Chamomile has an established domestic (Indian) and international market, which is increasing day by day. Among the various major constituents, α-bisabolol and chamazulene have been reported to be more useful than others. Chamazulene occurs in the capitula of the flowers in minute quantities and has been demonstrated to exert antiinflammatory activity in-vivo. Moreover, chamomile is a seasonal 4-5 months winter crop in India but is extensively required in various medicinal applications. Therefore, to increase the overall yield of this plant, its in-vitro propagation is needed. In the present study, somatic embryos were developed from capitulum explants after 2-4 weeks of culture on MS medium supplemented with 26.8 µM NAA and 11.5 µM Kin. The somatic embryos were further subcultured in-vitro, where new plantlets regenerated from embryos. It is concluded that in-vitro propagation is possible in case of chamomile and can be used to increase the overall yield of chamazulene present in the capitula of flowers as well as augment the overall yield of this important plant, which is conventionally propagated by seeds.

In vitro micropropagation of Stevia rebaudiana Bertoni in Malaysia

Stevia rebaudiana Bertoni is a medicinal plants and commercially use as non-caloric sweetener for diabetic patient. In the present study, a protocol was developed for in vitro micropropagation using 6-benzylamino purine (BAP) and Kinetin (Kn) for the formation of multiple shoot proliferation and Indole-3-acetic acid (IAA), Indole-3-butyric acid (IBA) and 1-Naphthaleneacetic acid (NAA) for the induction of roots. Maximum shoot formation (7.82 ± 0.7 shoots per explants) was observed on a Murashige and Skoog (MS) medium supplemented with 0.5 mg L-1 BAP and 0.25 mg L-1 Kn. The maximum number of roots (30.12 ± 2.1 roots per explants) was obtained on a MS medium containing 1.0 mg L-1 IBA. The well rooted plantlets were successfully weaned and acclimatized in plant soil with survival rate of 83.3 %.

Studies on the effect of age of in vitro grown seedlings on regenerative response of explants in tomato (Solanum lycopersicum).

Studies on the effectiveness of age of in vitro grown seedlings on regenerative response of explants derived from them were conducted in tomato (Solanum lycopersicum L.) cv. PKM-1. The results revealed that cotyledons and hypocotyls of 10 days old seedlings were found to be superior compared to explants collected from 8, 12, 14 days old seedlings when MS medium is supplemented with BAP 1.5 mg/L + Kinetin 1.0 mg/L. These findings could be well exploited for further development of quick regenerative and transformation protocols for the tomato cv.PKM-1.

Studies on the effect of various sterilants and culture conditions on in-vitro seed germination in tomato (solanum lycopersicum).

Studies on the effectiveness of various sterilants and culture conditions on in-vitro seed germination in tomato (Solanum lycopersicum L.) cv. PKM-1 revealed that among three sterilants used, surface sterilization of seeds with 5 % NaOCl for 20 minutes was found to be more effective resulting in high germination rate and contamination free cultures. Similarly among the different media and culture conditions considered in the present experiment, MS medium without sucrose with dark incubation for three days recorded lower no. of days (3.3 days) for initiation of germination with 96 % germination. These findings could be well exploited for further development of quick regenerative and transformation protocols for the tomato cv.PKM-1.

In vitro propagation of a rare medicinal fern of Western Ghats – Diplazium esculentum (Reytz.).

Present study aimed for in vitro culture of circinate part of young leaves of D. esculentum which is amongst the leafy vegetables consumed as vegetable by Paniya and Chetti tribes of Western Ghats. The circinate part of young leaves (crosiers), excised before the beginning of foliar expansion, was inoculated on half strength Murashige and Skoog (MS) medium supplemented with auxins indole-3-butyric acid (IBA) or α-napthalene acetic acid (NAA) or 2,4-Dichlorophenoxyacetic acid (2,4-D) and cytokinin 6- benzylaminopurine (BA) in a range 0.5 to 2.5 mg L-1. Combinations of different concentrations of 2,4 D + BA, IBA + BA as well as of NAA+ BA were also tested in half strength MS medium with 3% sucrose and with pH 5.8. The best morphogenic response was obtained with half strength MS medium supplemented with 2,4-D 0.5 mg L-1 and BA 2.5 mg L-1, 3% sucrose, at pH 5.8. For rooting of the microshoots, half strength MS medium supplemented with 2,4-D ( 2 and 1 mg L-1 ) exhibited best results. Present study reports the successful in vitro culturing of D. esculentum.

Effects of growth regulator and culture methods on rooting of Ajuga lobata and content of β-ecdysone / 中草药

Objective: To study the effects of indole butyric acid (IBA) and NAA at different concentrations and culture methods on the growth of Ajuga lobata root and content of β-ecdysone. Methods: Taking MS as basic medium supplemented with IBA (0.5, 1.0, and 2.0 mg/L) or NAA (0.3, 0.6, and 0.9 mg/L), the tissue culture was carried out. HPLC was used to determinate the content of β-ecdysterone. Results: The results showed that the roots could grew well on MS as basic medium supplemented with IBA 0.5-1 mg/L. The best culture condition for the highest β-ecdysone accumulation was MS medium supplemented with IBA 2 mg/L. The content of β-ecdysterone in the whole plant and the roots of A. lobata with tissue culture was twice times of hydroponics and about three times of cultivation. Conclusion: MS medium supplemented with IBA at the proper concentration could promote the root growth of A. lobata. The content of β-ecdysterone in the root of A. lobata is the highest in tissue cultures.

Resposta à regeneração e crescimento de brotos in vitro de Kielmeyera coriacea quando influenciado por diferentes concentrações dos sais e de sacarose / In vitro regeneration and growth response of Kielmeyera coriacea shoots when affected by salts and sucrose concentrations

Avaliaram-se diferentes concentrações de sacarose e sais do MS (MURASHIGE & SKOOG, J 962) na regeneração e crescimento de brotos de Kielmeyera coriacea in vitro. A redução e o aumento da quantidade de sacarose partindo de 30 e 45g/L, respectivamente, causou um decréscimo no número total de brotos. O uso integral ou da metade das concentrações dos sais do MS proporcionou maiores taxas de multiplicação. A maior porcentagem de brotos com mais de 1,0cm de comprimento pôde ser obtida empregando-se 30 g/l de sacarose associado a concentração 1/1 do MS. A porcentagem média de brotos decresceu linearmente com a diluição salina.

Trials were carried out to test sucrose and salt concentrations added to growth medium on regeneration and growth response of Kielmeyera coriacea shoots. The reduction and increase of sucrose of 30 and 45g/l, rerspectively, caused a decrease in total number of shoots. A higher shoots percentage of with more than 1.0cm, occurred on media with 30g/L of sucrose. The use of total force (1/1) and half strength (1/2) of MS salt proportioned a higher shoot ratio and growth.