The effects of 2,4-Dichlorophenoxyacetic Acid on the induction of callus from cotyledon and hypocotyl explants of Butterfly Pea (Clitoria ternatea)

Clitoria ternatea (Butterfly pea) is a tropical medicinal and fodder legume from the Fabaceae family possessing various beneficial phytochemical compounds linked to the mammalian neuroprotective mechanism. Callus and cell suspension cultures are excellent alternatives for harnessing secondary metabolites from medicinal plants. The current study aims to induce callus from cotyledon and hypocotyl explants of C. ternatea for the establishment of cell suspension cultures. Cotyledon and hypocotyl explants from two-weeks-old seedlings were subjected to half-strength MS medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) at different concentrations (0.5 mg/L to 2.5 mg/L) and callus scoring and morphology were assessed at week 8 of culture. Results revealed that the treatment of 0.5 mg/L 2,4-D resulted in the highest percentage of callus induction (100%) and the highest callus scoring for both cotyledon and hypocotyl explants with friable callus morphology. Cotyledon explants exhibited a higher callus scoring with a relative value of 3.03 ± 0.20 compared to hypocotyl explants at 1.80 ± 0.12. This study thereby provides a basis for future studies on callus induction studies and the establishment of cell suspension cultures of C. ternatea for the production of valuable secondary metabolites linked to the memory enhancing properties of the plant

The effects of banana, potato, and coconut water in the regeneration of Ficus carica cv. Japanese BTM 6

Fig (Ficus carica L.) belongs to the family of Moraceae and its fruits are nutritious to the human diet. Organic additives contain beneficial components for plant growth and could also be an alternative carbon source in culture. This study aims to evaluate the effects of organic additives such as banana, potato, and coconut water in the culture media for the regeneration of Ficus carica cv. Japanese BTM 6. In vitro shoots were cultured in MS medium supplemented with 1 mg/L of BAP without sucrose and with different concentrations of organic additives. MS medium supplemented with 1 mg/L BAP and 200 mL/L of coconut water resulted in the highest number of shoots (1.73 ± 0.16) shoot height (2.50 ± 0.34 cm), and several leaves (5.08 ± 0.81). On the other hand, treatments of 10 g/L potato puree and 10 g/L of banana puree showed the highest number of shoots, shoot height, and several leaves for each potato and banana treatments, respectively. The incorporation of coconut water was observed to support the induction of shoots, shoot height, and leaves. The study concludes that coconut water is a suitable source of organic additive for the regeneration of Ficus carica cv. Japanese BTM 6

The incorporation of coconut water and banana homogenate in the regeneration of fig (Ficus carica L.) cv. Violette de Solliès

The common fig (Ficus carica L.) is from the family of Moraceae and is commonly cultivated for its fruits, which are well-known for their exceptional nutritional and medicinal properties. The addition of organic additives functions to supply carbon sources and other essential vitamins, minerals, and natural growth regulators to support the growth of explants. The present study aims to assess the effects of coconut water and banana homogenate in the regeneration of Ficus carica cv. Violette de Solliès (VDS). In vitro shoot, explants were cultured in full-strength MS medium without sucrose but with 1.0 mg/L BAP and different concentrations of coconut water and banana homogenate. Results indicated that MS media with 200 mL/L coconut water resulted in the highest number of induced shoots (3.03 ± 0.122) and shoot height (1.005 ± 0.022 cm) compared to other treatments with coconut water, whereas MS media supplemented with 200 g/L banana homogenate produced the highest number of induced shoots (3.00 ± 0.144) and the highest shoot height (0.958 ± 0.020 cm) of all the banana homogenate treatments. In conclusion, coconut water and banana homogenate are suitable alternatives for carbon sources and other organic growth factors contributing to the regeneration of Ficus carica cv. VDS

Transformasi Gen Proteolisis 6 (PRT6) berperantarakan Agrobacterium tumefaciens ke dalam kotiledon tomato kultivar Micro Tom

Gen Proteolisis 6 (PRT6) merupakan gen yang memainkan peranan penting dalam tapak jalan N-end rule dan berfungsi sebagai enzim E3 ligase. PRT6 berperanan dalam pengenalan protein sasaran bagi proses degradasi. Objektif utama kajian ini adalah untuk mentransformasi konstruk RNAi PRT6 ke dalam tomato berperantarakan Agrobacterium tumefaciens. Ini bertujuan untuk memahami peranan tapak jalan N-end rule semasa proses pemasakan buah. Beberapa faktor yang memberi kesan kepada transformasi seperti masa ko-penanaman dan juga kepekatan antibiotik yang digunakan telah dioptimumkan. Keputusan kajian menunjukkan pengeraman kotiledon selama 48 jam pada medium ko-penanaman dapat meningkatkan penghasilan kalus sebanyak 61% manakala penggunaan 500 mg/L antibiotik karbenisilin dalam medium regenerasi pucuk dapat mengurangkan kontaminasi A. tumefaciens sehingga 5.2%. Selain itu, strain A. tumefaciens C58 merupakan strain A. tumefaciens yang paling sesuai digunakan sebagai perantara dalam kajian ini. Tindak balas berantai polimerase (PCR) telah dijalankan pada pucuk yang terhasil untuk mengesahkan integrasi fragmen PRT6 ke dalam genom tomato. Berdasarkan analisis PCR, kesemua tujuh pucuk putatif transgenik adalah merupakan transforman positif

The effect of adenosine monophosphate deaminase overexpression on the accumulation of umami-related metabolites in tomatoes

Taste is perceived as one of a combination of five sensations, sweet, sour, bitter, salty, and umami. The umami taste is best known as a savoury sensation and plays a central role in food flavour, palatability, and eating satisfaction. Umami flavour can be imparted by the presence of glutamate and is greatly enhanced by the addition of ribonucleotides, such as inosine monophosphate (IMP) and guanosine monophosphate (GMP). The production of IMP is regulated by the enzyme adenosine monophosphate (AMP) deaminase which functions to convert AMP into IMP. We have generated transgenic tomato (Solanum lycopersicum) lines over expressing AMP deaminase under the control of a fruit-specific promoter. The transgenic lines showed substantially enhanced levels of AMP deaminase expression in comparison to the wild-type control. Elevated AMP deaminase levels resulted in the reduced accumulation of glutamate and increased levels of the umami nucleotide GMP. AMP concentrations were unchanged. The effects on the levels of glutamate and GMP were unexpected and are discussed in relation to the metabolite flux within this pathway

Effects of light illumination and subculture frequency on biomass production in cell suspension cultures of Clinacanthus nutans

Clinacanthus nutans (Burm. f.) Lindau is a medicinal plant rich in polyphenols and possesses multiple pharmacological properties. It is widely used as a side treatment for cancer and to prevent oxidative stress-related diseases. The present study aims to investigate the effects of light illumination and subculture frequency on the growth and proliferation of cells in the suspension culture of C. nutans. Friable callus induced from leaf explants C. nutans were cultured in liquid Murashige & Skoog medium supplemented with 0.25 mg/L 2, 4-dichlorophenoxyacetic acid, and 0.25 mg/L 6-benzylaminopurine to initiate the cell suspension culture. The suspension cultures were incubated under different conditions (16 hr photoperiod and continuous darkness). Subcultures were conducted every two weeks for ten subculture cycles. Results revealed that higher cell biomass was obtained under the exposure of 16 hr photoperiod with the highest biomass production at the fifth subculture cycle (growth index of 11.8) exhibiting stable growth from the eighth to the tenth subculture cycle. The suspension cells showed stable growth rates up to the 10th subculture cycle producing small aggregates and single cells. The present study indicated that light illumination significantly impacts the growth of suspension cultures for C. nutans in the production of valuable secondary metabolites

The establishment of aseptic cultures and multiple shoot induction of olive (Olea europaea) cv.1

Olea europaea or the olive plant is from the family of Oleaceae and has been cultivated for its fruits that are highly nutritious, capable of reducing risks of cardiovascular diseases and to prevent cancer. The current propagation technique of olive plants in Malaysia relies on conventional methods that are less efficient in producing good quality plant stocks. Plant tissue culture offers an alternative to multiply plants from novel mother plants and is commonly applied in the propagation of various crops. This study aims to establish sterile cultures and to induce multiple shoots from Olea europaea cv.1 as a preliminary study to micropropagate olive plants for commercial farms in Malaysia. Nodal explants were surfaced sterilised with ethanol and Clorox© at different durations followed by treatments in MS media supplemented with different concentrations of Zeatin and BAP. Nodal explants surface sterilised with 70% (v/v) ethanol for 2 min and 30% (v/v) Clorox© for 8 min produced sterile explants with the survival rate of 85%. MS media supplemented 5.0 mg/L BAP was optimal for shoot induction (2.10 ± 0.31 shoots per explant) and shoot elongation (6.50 ± 1.17 mm). The current study serves as preliminary assessment for the establishment of local olive cultures

Rapid In Vitro Propagation of Fig (<i>Ficus carica</i> L.) ‘Violette de Solliès’ Supported by Molecular and Microscopy Analyses

Ficus carica L. is a common fig that is an incredibly nutritional fruit, well-known for its medicinal and economic values. This study aims to establish an efficient protocol for the mass propagation of fig plantlets (Ficus carica L.) for the cultivar ‘Violette de Solliès’. Surface-sterilized shoot-tip explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of cytokinins (6-benzylaminopurine, BAP; thidiazuron, TDZ; kinetin, Kn; and zeatin, Zea). Induced shoots were rooted on Woody Plant Medium (WPM) with different concentrations of auxins (naphthalene-acetic acid, NAA; indole-3-acetic acid, IAA; and indole-3-butyric acid, IBA). Rooted explants were acclimatized in eight different soil substrates prior to cultivation in a commercial plot. The propagated plantlets were analyzed for genetic stability and clonal fidelity using RAPD and SCoT molecular markers, whereas scanning electron microscopy (SEM) was performed to observe the stomata morphology of post-acclimatized plants. MS media supplemented with 5.0 mg/L BAP was the optimal treatment for multiple shoot induction (15.20 ± 1.03 shoots), whereas the highest percentage of rooting (93.33%) was achieved in WPM supplemented with 3.0 mg/L IBA. Plantlets were successfully acclimatized in biochar soil with a survival rate of 100%. RAPD and SCoT analysis showed no polymorphism occurrences across six subculture cycles, whereas observations via SEM indicated normal stomata structures on the leaves of acclimatized plantlets. This study documents an efficient micropropagation protocol for Ficus carica cv. ‘Violette de Solliès’ for the production of uniformed and true-to-type plant stocks suitable for commercial propagation

Effect of Cryopreservation Method Supported with Biochemical Analyses in the Axillary Bud of Jewel Orchid, <i>Ludisia discolor</i>

This study investigated conserving an endangered terrestrial jewel orchid Ludisia discolor, using in vitro grown axillary buds. Excised segments of axillary buds (4–5 mm in length) were precultured on a modified Murashige and Skoog (MS) medium supplemented with 0.2 M sucrose for 24 h and osmoprotected in a loading solution for 20 min. Then, axillary buds were dehydrated in plant vitrification solution 2 (PVS2) for 10 min at 0 °C and incubated in liquid nitrogen for 1 h. Subsequently, axillary buds were rewarmed rapidly by dilution solution and transferred to a growth recovery medium supplemented with 0.05 µM melatonin, which led to an improved survival chance (16.67%) for cryopreserved L. discolor. The osmotic stress and the overproduction of reactive oxygen species (ROS) during cryopreservation stages may result in cryoinjuries and poor survival as increased levels of proline (5.51 µmol/g), catalase (85.64 U/g), peroxidase (565.37 U/g), and ascorbate peroxidase activities (12.19 U/g) were detected after dehydration, preculture, rewarming, and loading stage, respectively. Results obtained from this study indicate that further experimental designs which apply different PVS and exogenous antioxidants are needed for improved survival and regrowth of L. discolor.</i

The effect of MS media strength and cytokinin in the induction of shoots from shoot tip explants of Australian Finger Lime (Citrus australasica cv. Tasty Green)

The Australian Finger Lime (Citrus australasica) is a type of citrus from the Rutaceae family, endemic to the east coast of Australia. The finger lime, loaded with numerous vitamins and renders a unique taste, has also been backed by science to contain essential amounts of antioxidants that are beneficial for cell protection, immune response, cancer prevention, ageing, arthritis and prevention of kidney stones. Current propagation attempts still rely on conventional methods that are less efficient and resulted in the slow establishment of farms for finger lime especially for commercialization purposes. This study focuses on the induction of shoots from shoot tip explants using 6-Benzylaminopurine (BAP) and Kinetin. Aseptic explants were inoculated into Murashige and Skoog (MS) medium of full-strength and half-strength followed by full-strength MS media supplemented with different concentrations of BAP and Kinetin. Results obtained in this study showed no significant differences in terms of the number of axillary shoots produced between explants cultured in full and half-strength MS media. However, the highest number of shoots and increment in shoot length were obtained from MS media supplemented with 2.0 mg/L BAP with the values 1.80 ± 0.27 and 2.56 ± 0.36 cm, respectively. In conclusion, MS media supplemented with 2.0 mg/L BAP was found optimal in the induction of shoots and shoot elongation of C. australasica cv. Tasty Green