Establishment Of in vitro Plantlets Of Artemisia annua L. For The Analysis Of Artemisinin Biosynthetic Gene (CYP71AV1) And Trichome Initiation Gene (GL3)

Artemisia annua L. is an herb known for its secondary metabolite, artemisinin. Artemisinin is used as antimalarial drug but its availability is limited by low yield in plantation. To produce artemisinin using in vitro technique, a high yielding in vitro cultivar must first be selected. Artemisia annua L. ialah sejenis tumbuhan herba yang terkenal untuk metabolit sekundernya iaitu, artemisinin. Artemisinin digunakan sebagai ubat antimalaria tetapi pengunaannya terhad disebabkan oleh hasil artemisinin yang rendah di ladang. Untuk menghasilkan artemisinin menggunakan teknik in vitro, kultivar in vitro yang menghasilkan artemisinin tinggi perlu dipilih terlebih dahul

Effect of Basal Medium on In Vitro Leaf Morphology, Growth and Artemisinin Production of Artemisia annua L.

Artemisia annua L. was classified as one of the important medicinal plants due to its potential in the treatment of malaria. However, the propagation of this plant was limited by environmental and geographical factors. Therefore, in vitro culture technique was an alternative to overcome these limitations. Five different basal media were examined for their effect on the growth and artemisinin content of in vitro plantlets of A. annua. They were found to give different effect on the growth in term of height, fresh biomass and rooting ability of the plantlets. Glandular and non-glandular filamentous trichomes were observed on the adaxial and abaxial surface of A. annua leaf. The five basal media was found to affect the distribution and the number of trichomes and stomata formed on the leaf surfaces. LV medium induced more trichomes formation of both types on both leaf surfaces. Highest number of stomata was found on the leaf surface of the plantlets cultured in MS medium. While B5 medium resulted in non formation of stomata on the abaxial leaf surface of all the studied clones. Artemisinin production was found to greatly affect by the choice of basal medium used for cultivation. Keywords: Artemisia annua, artemisinin, basal medium, growth pattern, stomata, trichome

High quality of bacterial dna extraction from Corbicula fluminea (Müller, 1774) tissue in Kelantan

Corbicula fluminea is a freshwater clam that is served as a popular traditional food in Kelantan, Malaysia. However, there are limited studies that report on high quality bacterial metagenome deoxyribonucleic acid (DNA) from C. fluminea. Therefore, this study compares the effectiveness and efficiency of conventional cetyltrimethylammonium bromide (CTAB) protocol, a commercial kit, and modified CTAB protocol for bacterial DNA extraction from the soft tissue surface of raw C. fluminea. The instruments used to examine the quality of the extracted bacterial DNA were DeNovix DS-11 spectrophotometer, gel electrophoresis machine, and UV transilluminator. The results showed that the bacterial DNA extracted from modified CTAB protocol had the highest purity and integrity with the A260/A280 ratio of 1.92 ± 0.01 and A260/A230 ratio of 1.83 ± 0.06 as well as the DNA band with minimum smear. This concludes that modified CTAB protocol is the best approach for the bacterial extraction from the C. fluminea

Potential of Leucas zeylanica extract to eliminate E. coli and S. aureus in Corbicula fluminea (“Etak”) tissue

“Etak” or Corbicula fluminea, is a freshwater mollusc species regularly consumed as a popular snack among the Kelantanese in Malaysia. The “etak” is usually heated with traditional smoking process which is considered as half cooked and the smoked C. fluminea is commonly known as “etak salai”. This study focuses on the potential of Leucas zeylanica leaves extract to eliminate the bacteria content in “etak salai”. Extraction of bacterial genomic DNA was performed and confirmed the existence of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) in “etak salai”. Antibacterial properties of L. zeylanica leaves extract was identified using disc diffusion assay and the result obtained exhibit that 70 μg/μL of L. zeylanica extract was the optimum concentration to give the effect of 11 mm inhibition zone for E. coli and 15 mm inhibition zone for S. aureus. This finding proof that L. zeylanica leaves could be the ingredients in the paste for “etak salai” preparation

Antimicrobial Activity of Artemisinin and Precursor Derived from In Vitro Plantlets of Artemisia annua L.

Artemisia annua L., a medicinal herb, produces secondary metabolites with antimicrobial property. InMalaysia due to the tropical hot climate, A. annua could not be planted for production of artemisinin, themain bioactive compound. In this study, the leaves of three in vitro A. annua L. clones were, extracted and two bioactive compounds, artemisinin and a precursor, were isolated by thin layer chromatography. These compounds were found to be effective in inhibiting the growth of Gram-positive and Gram-negative bacteria but not Candida albicans.Their antimicrobial activity was similar to that of antibactericidal antibiotic streptomycin. They were found to inhibit the growth of the tested microbes at the minimum inhibition concentration of 0.09mg/mL, and toxicity test using brine shrimp showed that even the low concentration of 0.09mg/mL was very lethal towards the brine shrimps with 100% mortality rate. This study hence indicated that in vitro cultured plantlets of A. annua can be used as the alternative method for production of artemisinin and its precursor with antimicrobial activities

Effect of bean sprout on in vitro multiplication of Musa acuminata

Musa acuminata or Pisang Berangan is popular in Kelantan, but due to a lack of knowledge on modern farming technologies, many Musa acuminata are affected by diseases and are of low quality. Plant tissue culture has numerous advantages, including a rapid rate of multiplication, and the prevention of disease. Plant growth regulator such as Benzyl Amino Purine (BAP) is commonly used in media to grow banana explants, however, Murashige and Skoog (MS) media production with BAP hormone is expensive for low-income farmers. Therefore, this research had been done by substituting the BAP hormone by using bean sprout extract as an organic supplement for banana culture. The additives that were used in this project were non-centrifuged and centrifuged bean sprout extracts at different concentrations, which were 5%, 10%, 15% and 20% applied in the MS media. Data were collected based on number of leaves, length of leaves and length of roots. In this study, the positive control treatment with MS + 5 mg/L BAP media recorded the highest mean value in the number of leaves, length of leaves, and length of roots with values of 12.5 ± 0.59, 5.69 ± 0.13cm, and 6.45 ± 0.36cm respectively followed by MS media with centrifuged bean sprout extract. From the observation, the bean sprout has the potential to use as an additive for Musa acuminata media to substitute synthetic hormones such as BAP

A review of durian plant-bat pollinator interactions

Durian (Durio zibethinus) brings in princely revenue for the fruit economy in Southeast Asia, ushering the current trend of clearing forests for durian plantations. Despite the thorny fruit’s popularity and increasing bat-durian papers, not many associate their vital plant-pollinator relationship. This unfamiliarity has led to the persisting negative connotations of bats as agricultural pests and worse, a disease carrier amplified by the Covid-19 pandemic. This review focuses on the bat-durian relationship comprising botanical insights and pollination ecology in relevance to the wider pteropodid-plant interactions. The majority of the studies compiled have concluded that bats are the most effective pollinator for durian than insects. Six fruit bat species (Chiroptera: Pteropodidae) have been recorded pollinating durian flowers, with several other pteropodid species speculated to pollinate durian, including in non-native countries. Lastly, we address the research gaps for the batdurian relationship, which can also be applied to other chiropterophilous plants

Heavy metals concentration in “etak” tissue at different processing stages

Corbicula fluminea, known as “etak” in Kelantan, Malaysia is a freshwater bivalve. This clam is widely consumed as traditional snack by the locals. However, lately there are numerous reports in local newspapers that claim “etak” to cause health effects due to eating contaminated “etak”. Hence, this study aims to determine the heavy metals concentration in fresh, smoked and exposed “etak” sold at the stall as a baseline study in order to develop a method for removing the heavy metals content in its tissues. This study involves sample collection in the stalls around Kelantan, sample preparations via acid digestion and heavy metal determination using Perkin Elmer PinAAcle 900F Atomic Absorption Spectrometer. The results showed the heavy metals (Cr, Zn, Mn and Cu) concentrations (μg/g) in all “etak” tissue were at different level for fresh (Cr: 1.02±0.35ppm, Zn: 74.57±2.76ppm, Mn: 40.22±9.96ppm and Cu: 15.27±1.41ppm), smoked (Cr: 0.42±0.02ppm, Zn: 54.62±17.83ppm, Mn: 50.13±2.31ppm and Cu: 20.94±8.81ppm) and exposed (Cr: 0.53±0.08ppm, Zn: 63.07±8.44ppm, Mn: 50.41±6.92ppm and Cu: 12.80±0.40ppm) samples. The results obtained were compared with the permissible limits set by Malaysian Food Regulations 1985 (Cu: 30.0ppm and Zn: 100.0ppm), FAO/WHO 1984 (Mn: 5.4ppm) and IAEA – 407 (Cr: 0.75ppm). For fresh “etak”, Zn and Cu concentration were below the standards, while Mn and Cr were exceeded the permissible limits. For smoked “etak” and exposed “etak” at stall, all heavy metals concentration were within the permissible limits, except Mn contents. This study successfully determine the baseline concentration of the heavy metals in “etak”

Heavy metals concentration in “etak” tissue at different processing stages

Corbicula fluminea, known as “etak” in Kelantan, Malaysia is a freshwater bivalve. This clam is widely consumed as traditional snack by the locals. However, lately there are numerous reports in local newspapers that claim “etak” to cause health effects due to eating contaminated “etak”. Hence, this study aims to determine the heavy metals concentration in fresh, smoked and exposed “etak” sold at the stall as a baseline study in order to develop a method for removing the heavy metals content in its tissues. This study involves sample collection in the stalls around Kelantan, sample preparations via acid digestion and heavy metal determination using Perkin Elmer PinAAcle 900F Atomic Absorption Spectrometer. The results showed the heavy metals (Cr, Zn, Mn and Cu) concentrations (μg/g) in all “etak” tissue were at different level for fresh (Cr: 1.02±0.35ppm, Zn: 74.57±2.76ppm, Mn: 40.22±9.96ppm and Cu: 15.27±1.41ppm), smoked (Cr: 0.42±0.02ppm, Zn: 54.62±17.83ppm, Mn: 50.13±2.31ppm and Cu: 20.94±8.81ppm) and exposed (Cr: 0.53±0.08ppm, Zn: 63.07±8.44ppm, Mn: 50.41±6.92ppm and Cu: 12.80±0.40ppm) samples. The results obtained were compared with the permissible limits set by Malaysian Food Regulations 1985 (Cu: 30.0ppm and Zn: 100.0ppm), FAO/WHO 1984 (Mn: 5.4ppm) and IAEA – 407 (Cr: 0.75ppm). For fresh “etak”, Zn and Cu concentration were below the standards, while Mn and Cr were exceeded the permissible limits. For smoked “etak” and exposed “etak” at stall, all heavy metals concentration were within the permissible limits, except Mn contents. This study successfully determine the baseline concentration of the heavy metals in “etak”