Phytochemical and mutagenesis studies of neolamarckia cadamba and leucaena leucocephala

Mohamed Zaky Abdel-Latif, Zayed Sadawe (2015) Phytochemical and mutagenesis studies of neolamarckia cadamba and leucaena leucocephala. PhD thesis, Universiti Malaysia Sarawak.

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Abstract

(This study was conducted to determine the effects of soaking duration in 100°C water and ethyl methanesulphonate (EMS) on seed germination, the nutritive value and mimosine content, and the genetic variation of the EMS-induced Neolamarckia cadamba and Leucaena leucocephala seedlings by using ISSR markers. The phytochemical constituents ofN. cadamba leaf extracts were aJ 0 investigaied. The effects of soaking duration in 100°C water on seed gennination were determined for N. cadamba and L. leucocephala by using the following treatments: (I) untreated (control); (2) soaking in 100°C water for 20 seconds followed 48 hours, respectively at room temperature; Results showed that soaking in 100°C water for 20 econds and subsequently in water at room temperature for 48 hours had the highest seed germination rate, higher cumulative gennination (CGP) and shortened the period of complete donnancy (CDP) over soaking duration of 24 hours or untreated seeds before planting. For EMSinduced mutation study, 120 seeds for each treatment of N. cadamba and L. leucocephala and three different concentrations (0.1, 0.3 and 0.6%) of EMS were used to treat the N. cadamba and L. leucocephala seeds in detennining the seed gennination percentage, lethality, seedling height, survival percentage, mimosine content, nutritive value and genetic variation of the mutagenized N. cadamba and L. leucocephala seedlings after 7 months of planting. The results showed that the germination percentage, survivability and seedling height were decreased; whereas lethality increased with increasing mutagenic doses. The mimosine content, crude protein and crude fiber decreased with increasing mutagenic doses for L. leucocephala. However, the mimosine content was not significantly affected by the concentration of EMS for N. cadamba. The mimosine content was highest in the control of L. leucocephala (1.6%) and lowest in N. cadamba (0.03%). Crude protein content was 23.48% and 20.90% for L. leucaena and N. cadamba, respectively. L. leucocephala was lower in nitrogen free extract, crude fiber and ash compared to N. cadamba. Results from this study indicate that N. cadamba has high forage quality and comparable to the traditional L. leLicocephala as forage for ruminants. Mimosine content was reduced from 1.6% to 0.2% or 87.46% reductions and therefore new L. leucocephala clone with reduced mimosine content and high crude protein (18.69%) was successfully produced. To identify the phytochemical constituents of N. cadamba leaf extracts using gas chromatography mass spectrometry (GC-MS). Solvents with increasing polarities viz. hexane, petroleum ether, chlorofonn, ethyl acetate and methanol were used in this study. The solvent extracts were analyzed using GC-MS and the mass spectra of the compounds found in the respective extract were matched with the National Institute of Standards and Technology (NIST) library. A total of 26 compounds were identified and the major chemical constituents were n-hexadecanoic acid '(44.88%), hexadecanoic acid ethyl ester (1 7.96%) and octadecanoic acid ethyl ester (11.71%). Some of the identified compounds have been reported to possess various biological activities such as antioxidant, antimicrobial, anesthetic, antiseptic, antidiabetic, hypocholesterolemic and etc. The results thus concluded that N. cadamba leaves possess various potent bioactive compounds and is recommended as a plant of phytophannaceutical importance. For the determination of genetic variation among the N. cadamba and L. leucocephala seedlings, 0.6% EMS treated samples exhibited the highest level of variability in comparison to 0.1 % EMS treated samples as revealed by using ISSR markers. This indicates that 0.6% EMS treatment is much more beneficial as compared to other EMS treatments. Further, EMS has been successfully used to produce a range of novel traits and broaden the genetic diversity of N. cadamba and L. leucocephala as observed in the present study.

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