Abstract:
:Saccharum spontaneum has been used for the development of energy cane a crop aimed to be used for the production of second-generation ethanol, or lignocellulosic ethanol. Lignin is a main challenge in the conversion of cell wall sugars into ethanol. In our studies to isolate the genes the lignin biosynthesis in S. spontaneum we have had great difficulty in RT-PCR reactions. Thus, we evaluated the effectiveness of different additives in the amplification of these genes. While COMT and CCoAOMT genes did not need any additives for other genes there was no amplification (HCT, F5H, 4CL and CCR) or the yield was very low (CAD and C4H). The application of supplementary cDNA was enough to overcome the non-specificity and low yield for C4H and C3H, while the addition of 0.04% BSA + 2% formamide was effective to amplify 4CL, CCR, F5H and CCR. HCT was amplified only by addition of 0.04% BSA + 2% formamide + 0.1 M trehalose and amplification of PAL was possible with addition of 2% of DMSO. Besides optimization of expression assays, the results show that additives can act independently or synergistically.
journal_name
An Acad Bras Ciencjournal_title
Anais da Academia Brasileira de Cienciasauthors
Llerena JPP,Araújo P,Mazzafera Pdoi
10.1590/0001-3765201820170250subject
Has Abstractpub_date
2018-01-01 00:00:00pages
509-519issue
1eissn
0001-3765issn
1678-2690pii
S0001-37652018000100509journal_volume
90pub_type
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