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

In vitro regeneration and acclimatization protocols of selected ornamental plants (agapanthus praecox, justicia betonica and celosia cristata)

This paper discussed on the effectiveness of BAP and NAA growth hormones on establishment of plant regeneration for selected ornamentals; Agapanthus praecox, Justicia betonica and Celosia cristata. Various explants (leaf, stem, shoot tip and bulb) derived from one-month-old aseptic seedlings of A. praecox and C. cristata, as well as explants from intact plants of J. betonica were utilized to achieve complete plant regeneration of these species. MS medium supplemented with various hormones, with an emphasis on BAP and NAA was tested to obtain direct and indirect regeneration. Both A. praecox (bulbs) and C. cristata (shoots) formed complete plantlets on MS added with 0.5-2.0 mg/L BAP and NAA, while direct regeneration was achieved for J. betonica on MS media containing BAP. Several methods were attempted to acclimatize the regenerants, with A. praecox gave the highest percentage of survival rates (96.67%), followed by J. betonica (80.00%) and C. cristata (75.00%)

Effects of NAA and BAP, double-layered media, and light distance on in vitro regeneration of nelumbo nucifera Gaertn. (Lotus), an aquatic edible plant

In vitro direct regeneration of Nelumbo nucifera Gaertn. was successfully achieved from immature explants (yellow plumule) cultured on a solid MS media supplemented with combinations of 0.5 mg/L BAP and 1.5 mg/L NAA which resulted in 16.00 ± 0.30 number of shoots per explant and exhibited a new characteristic of layered multiple shoots, while normal roots formed on the solid MS basal media. The double-layered media gave the highest number of shoots per explant with a ratio of 2 : 1 (liquid to solid) with a mean number of 16.67 ± 0.23 shoots per explant with the formation of primary and secondary roots from immature explants. In the study involving light distance, the tallest shoot (16.67 ± 0.23 mm) obtained from the immature explants was at a light distance of 200 mm from the source of inflorescent light (1000 lux). The plantlets were successfully acclimatized in clay loam soil after 8 months being maintained under in vitro conditions

Tissue culture and cellular behaviour studies of rice (Oryza sativa L. CV. MRQ 74) / Azani Saleh

Tissue culture or in vitro studies of rice (Oryza sativa L. cv. MRQ 74) locally known as “padi Mas Wangi” has been successfully investigated in this project. Callus induction was obtained on MS media supplemented with various concentrations of 2,4-D, applied singly and in combinations with BAP. Stem was identified as the most responsive explant, followed by root, while leaf explants failed to produce any callus. The highest means of callus dry weight of stem (71.60 ± 6.40 mg) and root (66.70 ± 10.90 mg) explants were recorded on MS media supplemented with 0.5 mg/L 2,4-D and 0.5 mg/L 2,4-D in combination with 0.5 mg/L BAP, respectively. Stem explants produced either creamy white, globular and compact or creamy white, globular and friable callus. On the other hand, creamy white, globular and sticky or mucilageneous callus was observed from root explants. Somatic embryos were induced by transferring the obtained callus from MS media supplemented with 2.0 mg/L BAP in combination with 1.0 mg/L 2,4-D onto MS media containing various concentrations of ABA, kinetin and L-Proline. MS media supplemented with 1.0 mg/L ABA in combination with 1.0 mg/L kinetin showed the highest mean number of somatic embryos (14.33 ± 0.27). The addition of 400 mg/L L-Proline had significantly (P<0.05) increased the mean number of somatic embryos (17.37 ± 0.66). Stem was found to be the only responsive explant for in vitro regeneration of this species. The best hormone for shoot induction was BAP at the concentration of 1.5 mg/L with mean number of shoots per explant of 4.03 ± 0.31. The highest mean number of roots produced (25.33 ± 1.89) was achieved when stem explants were cultured on MS media supplemented with 0.1 mg/L BAP in combination with 0.1 mg/L NAA. The addition of TDZ at the concentration of 0.1 mg/L had significantly increased the mean number of shoots per explant (8.23 ± 1.09). Synthetic seeds were created from microshoots of stem explants that were cultured on MS media containing 1.5 mg/L BAP. The best encapsulation matrix was Ca-free MS supplemented with 30 g/L sucrose with survival rate of 100 %, after 30 days of culture. The survival rate of plantlets (100 %) were best achieved on MS basal and MS media supplemented with 0.1 mg/L BAP. It was found that the viability of seeds decreased from 93.33 % to 3.33 % after one month of storage at 4 oC. Regenerated plantlets from stem explants cultured on MS media containing 0.5 mg/L 2,4-D were successfully acclimatized on all types of growing substrates with different survival rates of plantlets. A combination of black soil and red soil at a ratio of 1:1, showed the highest survival rate after 4 and 8 weeks of acclimatization, 90.00 ± 1.53 % and 83.33 ± 1.20 %, respectively. Cytological studies revealed that Mitotic Index (MI) values of root tip meristem cells was significantly lower in MS media supplemented with NAA, kinetin and 2,4-D as compared with hormone-free MS. The obvious effect of 2,4-D was observed on nuclear DNA content, mean cell and nuclear areas

The effects of different strength of MS media in solid and liquid media on in vitro growth of Typhonium flagelliforme

Objective: To determine the effects of different strength of Murashige and Skoog (MS) media (full, ½ and ¼) in solid and liquid media on in vitro growth of Typhonium flagelliforme (T. flagelliforme), whereby an optimum media composition can be provided for mass propagation of T. flagelliforme. Methods: Rhizome bud of T. flagelliforme was obtained from the axenic in vitro established T. flagelliforme plantlets in Plant Tissue Culture Laboratory, Universiti Teknologi MARA, Shah Alam. Rhizome bud was used as explant and cultured onto shoot proliferation medium under different strength of MS media (full, ½, ¼) in solid and liquid culture media. Results: After 6 weeks of culture, the number of shoot, number of leaf, number of root, height of shoot, fresh weight, dry weight and chlorophyll content of T. flagelliforme were analyzed. A comparison was made between liquid and solid culture media. The results revealed that the liquid culture media were more effective for all the growth parameters (shoot height, shoot number, leaf number, root number, fresh weight, dry weight, chlorophyll a and chlorophyll b content) compared to solid culture media. Apart from that, this study revealed the positive relationship between strength of MS media and type of culture media (solid and liquid media) to the growth of T. flagelliforme. Growth of T. flagelliforme was improved when MS strength was increased in liquid media. In contrast, growth of T. flagelliforme was improved when MS strength was decreased in solid media. Conclusions: Through this study, an optimum media composition for mass propagation of T. flagelliforme had been established by observing effects of MS media strength and type of culture media (solid and liquid media) on the growth of T. flagelliforme

Detection and quantification of natural pigments extracted from callus of Echinocereus cinerascens

Purpose: This paper aims to study the effect of different organic solvents on the extraction of pigments present in callus cultures of E. cinerascens. Design/methodology/approach: Attempts have been made to extract pigments from callus cultures through tissue culture system as an alternative replacement for conventional plant cultivation as tissue culture provides unlimited supplies of plant samples. Callus of E. cinerascens was induced from stem explant cultured in Murashige and Skoog medium supplemented with combination of 0.5 mg/L 6-benzylaminopurine and 0.5 mg/L α-naphthaleneacetic acid maintained under photoperiod of 16 h light and 8 h dark. Fresh samples of the callus were harvested and dissolved in various types and concentrations of solvents such as 100 per cent acetone, 80 per cent acetone, 95 per cent ethanol, 100 per cent methanol and 90 per cent methanol. Each of the mixtures was directly centrifuged to get clear supernatant containing pigments of interest. The pigments were detected and subsequently quantified via two simple techniques, ultraviolet-visible (UV-Vis) spectrophotometer and thin layer chromatography (TLC). Findings: UV-Vis spectrophotometer detected two families of pigments present in the callus cultures, namely, carotenoids (carotene and xanthophyll) and tetrapyrroles (chlorophyll a and b). Pigment contents in various solvent extractions were estimated using spectroscopic quantification equations established. Through TLC, spots were seen on the plates, and Rf values of each spots were assessed to indicate the possible existence of carotenoids and tetrapyrroles. Originality/value: This preliminary study offers significant finding for further advance research related on natural pigments extracted from E. cinerascens that would provide profits in the future applications, especially in food industry, medicine, agriculture, etc

Effects of NAA and BAP, Double-Layered Media, and Light Distance on In Vitro Regeneration of Nelumbo nucifera Gaertn. (Lotus), an Aquatic Edible Plant

In vitro direct regeneration of Nelumbo nucifera Gaertn. was successfully achieved from immature explants (yellow plumule) cultured on a solid MS media supplemented with combinations of 0.5 mg/L BAP and 1.5 mg/L NAA which resulted in 16.00 ± 0.30 number of shoots per explant and exhibited a new characteristic of layered multiple shoots, while normal roots formed on the solid MS basal media. The double-layered media gave the highest number of shoots per explant with a ratio of 2 : 1 (liquid to solid) with a mean number of 16.67 ± 0.23 shoots per explant with the formation of primary and secondary roots from immature explants. In the study involving light distance, the tallest shoot (16.67 ± 0.23 mm) obtained from the immature explants was at a light distance of 200 mm from the source of inflorescent light (1000 lux). The plantlets were successfully acclimatized in clay loam soil after 8 months being maintained under in vitro conditions