An easy and robust method for the isolation of high quality RNA from coconut tissues

BACKGROUND: Coconut tissues consist of a complex network of polysaccharides, proteins, polyphenols, and lipids that can bind to nucleic acids and pose difficulty in isolation. Certainly, a vigorous method is required to isolate high quality and quantity of RNA from such tissues for the purpose of downstream experiments. In this paper, we discuss a newly developed method for the Isolation of RNA from Complex Matrices (IRCM) method from coconut tissues. RESULTS: The method is robust, cheap, and efficient for the extraction of quality RNA in high quantities from the solid endosperm of stored and fresh coconut (150 µg/g FW with A260/280 = 1.89 and 247.5 µg/g FW with A260/280 = 1.91), coconut apple (263.8 µg/g FW with A260/280 = 1.97), and coconut bud (1052.5 µg/g FW with A260/280 = 2.00). The other well established methods, such as Method of RNA Isolation from Palm (MRIP), Cetyl Trimethyl Ammonium Bromide (CTAB), TRIZOL, and RNA plant kit failed to isolate quality RNA in appreciable quantities from the coconut tissues. Furthermore, the resultant RNA performed well in the downstream experiment, that is, RT-PCR for the production and amplification of cDNA. CONCLUSIONS: From the study, we concluded that the present method will play a vital role in the extraction of high quality RNA from complex matrices in a short time.

Estimación de la variabilidad genética entre ecotipos de cocoteros presentes en Cuba por ISTR / Genetics variability estimation among coconut ecotypes in Cuba by ISTR

Se realizó la caracterización molecular entre 16 ecotipos de cocoteros pertenecientes a una población del municipio de Baracoa, provincia Guantánamo, empleando la técnica Inverse Sequence Tagged Repeat (ISTR). El análisis molecular ISTR detectó un polimorfismo del 81,8 por ciento, demostrando la potencialidad de este marcadorpara realizar estudios de diversidad molecular en el cocotero. El porcentaje de identificación fue alto (Pi=91,2 por ciento), lo cual sugiere que las combinaciones de oligonucleótidos empleadas pudieran ser utilizada para estudios de identificación de ecotipos en poblaciones de cocotero de la zona de Baracoa mientras, que la heterocigocidad esperada fue baja (He=0,30). La evaluación de la diversidad en los diferentes ecotipos de cocoteros mediante marcadores ISTR mostró que se cuenta con un nivel de variabilidad, atendiendo a los grupos formados, lo que se corresponde con la gran variación morfológica observada en la poblaciónin situ. Además, estos resultados sugieren que la hibridación natural ha sido un factor determinante en la generación de la variabilidad encontrada entre los ecotipos, característica de las variedades de cocotero.

New biotechnological applications of coconuts

Coconut is a major crop of many poor nations. The present paper shows with inexpensive fluorescence microscopy of the enhanced green fluorescent protein (EGFP), that coconut water has the capability of synthesizing proteins from recombinant DNA vectors. After only 4 hrs of 100 pM application of the EGFP plasmid (pEGFP), a strong fluorescent signal was detected with an off-the-shelf, low-tech CCD webcam. Since natural coconut water is sterile, this heat-stable delivery system may be a good option for the dissemination of food supplements and pharmaceuticals in poor nations. As expensive substances could be expressed in coconut water, through bioreactor technology or micropropagation, the concepts presented here may prove invaluable to the economic and social advancement of many developing countries.