Melia dubia Cav. of family Meliaceae is a fast growing, high value tree species native to India. Isolating DNA from matured dried leaves of M. dubia was difficult due to accumulation of secondary metabolites, majorly ...Melia dubia Cav. of family Meliaceae is a fast growing, high value tree species native to India. Isolating DNA from matured dried leaves of M. dubia was difficult due to accumulation of secondary metabolites, majorly polyphenolics, which resulted in dark brown to black colour of the pellet. In this study, a modified STE-(Sucrose, Tris-HCl and Ethylene Diamine Tetra Acetic Acid) CTAB (hexadecyltrimethylammonium bromide) method was standardized for removal of polyphenolics. The protocol developed yielded 200 - 1000 ng/μl of quality DNA without any impurities as evident by A260/280 ratio ranging from 1.75 - 2.0. It was also suitable for extracting quality DNA from other members of Meliaceae like Azadirachta indica and Melia azedarach. In downstream applications, the extracted DNA was used for PCR amplification by using ISSR and SSR markers. ISSR PCR conditions were optimized in a reaction volume of 25 μl, consisting of 30 ng of template DNA, 1.5 mM MgCl<sub>2</sub>, 200 μM of each of dNTPs and 2 U of Taq polymerase. The best amplification was observed and the same was applicable for SSR markers.展开更多
This research was carried out on nine provenances from the north Western Ghats to quantify proximate principles,mineral matter,total phenols and other phytochemicals in Melia dubia drupe pulp.Drupe biometric,biomass a...This research was carried out on nine provenances from the north Western Ghats to quantify proximate principles,mineral matter,total phenols and other phytochemicals in Melia dubia drupe pulp.Drupe biometric,biomass attributes,proximate principles,mineral matter and total phenols diff ered signifi cantly among the provenances.The average drupe length and width was 26.5 and 22.3 mm,respectively.Average drupe and pulp fresh weight,pulp dry weight and stone weight were 783.1,552.1,165.1,and 179.8 g/100 drupes,respectively.Pulp,dry matter and moisture per cent were 70.5%,29.9%and 70.0%,respectively.Average crude fi bre,protein,ether extract,total ash and nitrogen-free extract were 8.3%,7.6%,5.1%,6.3%,and 72.6%,respectively.Average Ca,Mg,P and K levels were 0.5%,0.2%,0.2%,and 2.0%,respectively.Total phenols were 0.4%.Cluster analysis showed that the study area may be divided into three distinct provenances,each having diff erent drupe traits and proximate composition,including mineral components.Gas-chromatography mass-spectrometry analysis revealed 27 phytochemicals,many of which possess benefi cial biological attributes.The implications of this provenance variation study should be useful for future improvement in terms of better drupe pulp nutrition for livestock feed.Further study suggests that M.dubia pulp could be categorized as a good feed source with low fi bre,energy,fat and mineral levels.All proximate parameters,mineral matter content and total phenols are within prescribed limits.展开更多
文摘Melia dubia Cav. of family Meliaceae is a fast growing, high value tree species native to India. Isolating DNA from matured dried leaves of M. dubia was difficult due to accumulation of secondary metabolites, majorly polyphenolics, which resulted in dark brown to black colour of the pellet. In this study, a modified STE-(Sucrose, Tris-HCl and Ethylene Diamine Tetra Acetic Acid) CTAB (hexadecyltrimethylammonium bromide) method was standardized for removal of polyphenolics. The protocol developed yielded 200 - 1000 ng/μl of quality DNA without any impurities as evident by A260/280 ratio ranging from 1.75 - 2.0. It was also suitable for extracting quality DNA from other members of Meliaceae like Azadirachta indica and Melia azedarach. In downstream applications, the extracted DNA was used for PCR amplification by using ISSR and SSR markers. ISSR PCR conditions were optimized in a reaction volume of 25 μl, consisting of 30 ng of template DNA, 1.5 mM MgCl<sub>2</sub>, 200 μM of each of dNTPs and 2 U of Taq polymerase. The best amplification was observed and the same was applicable for SSR markers.
基金Master‘s thesis research undertakenunder two projects namely“Strengthening of College of Forestry,Navsari Agricultural University,Navsari”and“Popularizing MeliaComposita based Agroforestry System in Gujarat through Productionof Quality Material”,funded by Government of Gujarat,India.
文摘This research was carried out on nine provenances from the north Western Ghats to quantify proximate principles,mineral matter,total phenols and other phytochemicals in Melia dubia drupe pulp.Drupe biometric,biomass attributes,proximate principles,mineral matter and total phenols diff ered signifi cantly among the provenances.The average drupe length and width was 26.5 and 22.3 mm,respectively.Average drupe and pulp fresh weight,pulp dry weight and stone weight were 783.1,552.1,165.1,and 179.8 g/100 drupes,respectively.Pulp,dry matter and moisture per cent were 70.5%,29.9%and 70.0%,respectively.Average crude fi bre,protein,ether extract,total ash and nitrogen-free extract were 8.3%,7.6%,5.1%,6.3%,and 72.6%,respectively.Average Ca,Mg,P and K levels were 0.5%,0.2%,0.2%,and 2.0%,respectively.Total phenols were 0.4%.Cluster analysis showed that the study area may be divided into three distinct provenances,each having diff erent drupe traits and proximate composition,including mineral components.Gas-chromatography mass-spectrometry analysis revealed 27 phytochemicals,many of which possess benefi cial biological attributes.The implications of this provenance variation study should be useful for future improvement in terms of better drupe pulp nutrition for livestock feed.Further study suggests that M.dubia pulp could be categorized as a good feed source with low fi bre,energy,fat and mineral levels.All proximate parameters,mineral matter content and total phenols are within prescribed limits.
