Kinetin induces chromosomal abnormalities in African Blue Lily (Agapanthus praecox ssp. minimus) grown in In Vitro

Kinetin has been reported to exert inhibitory effect when used in tissue culture and in some cases reverse the action of auxin and cause growth inhibition and retardation of root formations. Kinetin also acts as ‘mitotic poison’, mimicking the effect of pesticides and toxic chemicals and interferes in mitosis mechanism of plants. The effect of kinetin on size of cell and nucleus as well as chromosome behaviour in root tip meristems of Agapanthus praecox ssp. minimus was studied. The results showed that prolong exposure to kinetin caused chromosome abnormalities to occur more frequently. Chromosome breakage yielded fragmented chromosomes, while abnormal spindle fibers caused delay in chromosome movement, termed as laggard chromosomes. Abnormal nucleus was also observed with kinetin treatments, such as micronucleus, binucleated and tripolar cells

Plant Regeneration and Cellular Behaviour Studies in Celosia cristata Grown In Vivo and In Vitro

Tissue culture studies of Celosia cristata were established from various explants and the effects of various hormones on morphogenesis of this species were examined. It was found that complete plant regeneration occurred at highest percentage on MS medium supplemented with 2.0 mg/L NAA and 1.5 mg/L BAP, with the best response showed by shoot explants. In vitro flowering was observed on MS basal medium after six weeks. The occurrence of somaclonal variation and changes in cellular behavior from in vivo and in vitro grown plants were investigated through cytological studies and image analysis. It was observed that Mitotic Index (MI), mean chromosome numbers, and mean nuclear to cell area ratio of in vitro root meristem cells were slightly higher compared to in vivo values. However, in vitro plants produced lower mean cell areas but higher nuclear areas when compared to in vivo plants. Thus, no occurrence of somaclonal variation was detected, and this was supported by morphological features of the in vitro plants

Identification of crocin, crocetin and zeaxanthin in Crocus sativus grown under controlled environment in Malaysia

Purpose – The purpose of the study is to identify the high valuable compounds which are crocin, crocetin and zeaxanthin in the stigmas and stamens of Crocus sativus grown under controlled environment in Malaysia. Design/methodology/approach – Spectrophotometry and high-performance liquid chromatography (HPLC) analysis were used to identify and measure crocin, crocetin and zeaxanthin content qualitatively and quantitatively in the stigmas and stamens of C. sativus grown under controlled environment in Malaysia. Findings – The results of this study showed that crocin, crocetin and zeaxanthin were detected in the stigmas. However, among those three compounds, only crocetin was detected in the stamens. In the stigmas, the detectable level of crocin was high compared to crocetin and zeaxanthin. It was also found that crocetin was higher in the stamens compared to in the stigmas. Originality/value – This is the first attempt in Malaysia that the stigmas and stamens were directly purified from the natural sources by means of no addition of preservatives as C. sativus has never been grown here before. Furthermore, limited reports are available regarding the identification of compounds in saffron stamens

Micropropagation of Bioencapsulation and Ultrastructural Features of Sainfoin ( Onobrychis viciifolia

To explore the potential of in vitro rapid regeneration, three varieties (Golpaygan-181, Orumieh-1763, and Gorgan-1601) of sainfoin (Onobrychis viciifolia Scop. syn. Onobrychis sativa L.) were evaluated. For the first time, an encapsulation protocol was established from somatic embryogenic callus in torpedo and cotyledonary stages to create artificial seeds. Callus derived from different concentrations of Kinetin (0–2.0 mg L−1) and Indole-3-acetic acid (0–2.0 mg L−1) was coated with sodium alginate and subsequently cultured either in Murashige and Skoog (MS) medium or in soil substrate. Adventitious shoots from synthetic beads developed into rooting in full and half strength MS medium supplemented with various concentrations of auxin and cytokinin. Prolonged water conservation of black and red soils (1 : 1) had the highest rate of survival plantlets in the acclimatization process. Diverse resistance techniques in Onobrychis viciifolia were evaluated when the plants were subjected to water deficiency. Higher frequency of epicuticular waxes was observed in in vivo leaves compared to in vitro leaves. Jagged trichomes nonsecreting glands covered by spines were only observed in the lower leaf side. Ultimately, stomata indices were 0.127 (abaxial), 0.188 (adaxial) in in vivo and 0.121 (abaxial), 0.201 (adaxial) in in vitro leaves

Antioxidant and antibacterial activities of ethanolic extracts of Asparagus officinalis cv. Mary Washington: Comparison of in vivo and in vitro grown plant bioactivities

The antioxidant and antibacterial activities of ethanolic extracts of in vivo grown Asparagus officinalis cv. Mary Washington were investigated using superoxide dismutase, erythrocyte haemolysis and 2,2- diphenyl-1-picrylhydrazil free radical scavenging methods. The measured antioxidant and antimicrobial potential were then compared to the activities offered by the ethanolic extracts of in vitro grown A. officinalis as well as ethanolic extract of undifferentiated callus cells of A. officinalis produced on Murashige and Skoog medium containing 1.5 mg/l 6-benzylaminopurine combined with 0.5 mg/l naphthalene acetic acid. The highest antioxidant capacity was obtained from the in vivo grown plant extract followed by in vitro grown plant extract in all three examined assays. Although, no antibacterial activity was detected from both in vivo and in vitro grown plant extracts in the disc diffusion antimicrobial assay, ethanolic extract of A. officinalis offered antibacterial activity against Bacillus cereus.Key words: Antibacterial activity, antioxidant activity, Asparagus officinalis

Germination and Plantlet Regeneration of Encapsulated Microshoots of Aromatic Rice ( Oryza sativa

Plant tissues such as somatic embryos, apical shoot tips, axillary shoot buds, embryogenic calli, and protocom-like bodies are potential micropropagules that have been considered for creating synthetic seeds. In the present study, 3–5 mm microshoots of Oryza sativa L. Cv. MRQ 74 were used as explant sources for obtaining synthetic seeds. Microshoots were induced from stem explants on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L benzylaminopurine (BAP). They were encapsulated in 3% (w/v) sodium alginate, 3% sucrose, 0.1 mg/L BAP, and 0.1 mg/L α-Naphthalene acetic acid (NAA). Germination and plantlet regeneration of the encapsulated seeds were tested by culturing them on various germination media. The effect of storage period (15–30 days) was also investigated. The maximum germination and plantlet regeneration (100.0%) were recorded on MS media containing 3% sucrose and 0.8% agar with and without 0.1 mg/L BAP. However, a low germination rate (6.67%) was obtained using top soil as a sowing substrate. The germination rate of the encapsulated microshoots decreased from 93.33% to 3.33% after 30 days of storage at 4°C in the dark. Therefore, further research is being done to improve the germination rate of the synthetic seeds

Micropropagation of an Exotic Ornamental Plant, Calathea crotalifera

A successful protocol was established for micropropagation in two selected varieties of exotic ornamental plants, Calathea crotalifera. The effects of different sterilization techniques, explant type, and the combination and concentration of plant growth regulators on shoots induction were studied. The axillary shoot buds explants sprouted from rhizomes in soil free conditions showed high induction rate of shoots with lowest contamination percentage when treated with combination of 30% (v/v) NaOCl, 70% (v/v) ethanol, and 0.3% (w/v) HgCl2. In the present study, the highest number of multiple shoots was obtained in MS basal medium supplemented with 3.5 mg/L 6-Benzylaminopurine (BAP), 1.0 mg/L 1-Naphthaleneacetic acid (NAA), 3% sucrose, and 6 g/L plant agar for both varieties and was used as multiplication medium. Microshoots were highly induced when the young shoot bud explants were incised longitudinally prior subculture. Chlorophyll analysis was studied to test the effects of activated charcoal and L-glutamine on reduction of necrosis problem. The maximum roots induction was recorded on MS medium supplemented with 1.0 mg/L 1-Naphthaleneacetic acid (NAA) compared to indolebutyric acid (IBA). The complete regenerated plantlets were successfully acclimatized in the soilless medium under greenhouse condition. This is the first report of rapid mass propagation for C. crotalifera

Production of Gymnemic Acid Depends on Medium, Explants, PGRs, Color Lights, Temperature, Photoperiod, and Sucrose Sources in Batch Culture of Gymnema sylvestre

Gymnema sylvestre (R.Br.) is an important diabetic medicinal plant which yields pharmaceutically active compounds called gymnemic acid (GA). The present study describes callus induction and the subsequent batch culture optimization and GA quantification determined by linearity, precision, accuracy, and recovery. Best callus induction of GA was noticed in MS medium combined with 2,4-D (1.5 mg/L) and KN (0.5 mg/L). Evaluation and isolation of GA from the calluses derived from different plant parts, namely, leaf, stem and petioles have been done in the present case for the first time. Factors such as light, temperature, sucrose, and photoperiod were studied to observe their effect on GA production. Temperature conditions completely inhibited GA production. Out of the different sucrose concentrations tested, the highest yield (35.4 mg/g d.w) was found at 5% sucrose followed by 12 h photoperiod (26.86 mg/g d.w). Maximum GA production (58.28 mg/g d.w) was observed in blue light. The results showed that physical and chemical factors greatly influence the production of GA in callus cultures of G. sylvestre. The factors optimized for in vitro production of GA during the present study can successfully be employed for their large-scale production in bioreactors

Estudio de organogénesis, morfología y anatomía en el Baybean (Canavalia rosea (Sw.) DC.)

Morphological and anatomical studies comprised leaf venation, histological analysis and epidermal peeling were carried out on Canavalia rosea (Sw.) DC.). Anatomical studies of the leaf and root determined the presence of cuticle and oil glands, arrangement of cells and the structure of vascular system. The dorsiventral leaf with pinnate venation and amphistomatic leaf with paracytic type stomata were embedded below the epidermis. Trichomes and oil glands were also observed on both adaxial and abaxial surfaces of leaf. Longitudinal sections of the callus showed the existence of meristematic cells, which could give rise to plant regeneration. In the process of organogenesis, adventitious shoots emerged from leaf and stem explants within 2 months after culturing on MS media fortified with 6-Benzylaminopurine (1.0-2.0 mg/l). Ultimately, in vitro germination was successfully established for the propagation and future conservation programs of this species.Se realizaron estudios morfológicos y anatómicos de venación foliar, análisis histológico y peeling epidérmico en Canavalia rosea (Sw.) DC). Los estudios anatómicos de la hoja y de la raíz determinaron la presencia de la cutícula y de las glándulas de aceite, la disposición de células y la estructura del sistema vascular. La hoja dorsiventral con venación pinnada y la hoja anfistomática con estomas paracíticos estaban incrustados debajo de la epidermis. También se observaron tricomas y glándulas oleosas en las superficies adaxial y abaxial de la hoja. Las secciones longitudinales del callo mostraron la existencia de células meristemáticas que podrían dar lugar a la regeneración de las plantas. En el proceso de organogénesis, brotes adventicios surgieron de los explantes de hoja y tallo en los 2 meses después del cultivo en medios de MS fortificados con 6-bencilaminopurina (1,0-2,0 mg / l). En última instancia, la germinación in vitro se estableció con éxito para la propagación y los futuros programas de conservación de esta especie

Stimulatory Effects of Gamma Irradiation on Phytochemical Properties, Mitotic Behaviour, and Nutritional Composition of Sainfoin ( Onobrychis viciifolia

Sainfoin (Onobrychis viciifolia Scop. Syn. Onobrychis sativa L.) is a bloat-safe forage crop with high levels of tannins, which is renowned for its medicinal qualities in grazing animals. Mutagenesis technique was applied to investigate the influence of gamma irradiation at 30, 60, 90, and 120 Gy on mitotic behavior, in vitro growth factors, phytochemical and nutritional constituents of sainfoin. Although a percentage of plant necrosis and non-growing seed were enhanced by irradiation increment, the germination speed was significantly decreased. It was observed that gamma irradiated seeds had higher value of crude protein and dry matter digestibility compared to control seeds. Toxicity of copper was reduced in sainfoin irradiated seeds at different doses of gamma rays. Anthocyanin content also decreased in inverse proportion to irradiation intensity. Accumulation of phenolic and flavonoid compounds was enhanced by gamma irradiation exposure in leaf cells. HPLC profiles differed in peak areas of the two important alkaloids, Berberine and Sanguinarine, in 120 Gy irradiated seeds compared to control seeds. There were positive correlations between irradiation dose and some abnormality divisions such as laggard chromosome, micronucleus, binucleated cells, chromosome bridge, and cytomixis. In reality, radiocytological evaluation was proven to be essential in deducing the effectiveness of gamma irradiation to induce somaclonal variation in sainfoin