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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Optimization of RT-PCR reactions in studies with genes of lignin biosynthetic route in Saccharum spontaneum

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Author(s):
Llerena, Juan P. P. [1] ; Araujo, Pedro [2] ; Mazzafera, Paulo [1, 3]
Total Authors: 3
Affiliation:
[1] Univ Estadual Campinas, Dept Biol Vegetal, Lab Fisiol Mol Plantas, Inst Biol, Rua Monteiro Lobato 255, Caixa Postal 6109, BR-13083862 Campinas, SP - Brazil
[2] Univ Estadual Campinas, Inst Biol, Dept Genet Evolucao & Bioagentes, Lab Genoma Func, Rua Monteiro Lobato 255, BR-13083862 Campinas, SP - Brazil
[3] Univ Sao Paulo, Escola Super Agr Luiz Queiroz, Dept Prod, Av Padua Dias 11, Caixa Postal 9, BR-34294100 Piracicaba, SP - Brazil
Total Affiliations: 3
Document type: Journal article
Source: Anais da Academia Brasileira de Ciências; v. 90, n. 1, p. 509-519, MAR 2018.
Web of Science Citations: 2
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. (AU)

FAPESP's process: 14/25994-1 - Lignin in species of the genus Saccharum
Grantee:Juan Pablo Portilla Llerena
Support type: Scholarships in Brazil - Master
FAPESP's process: 08/58035-6 - Control of lignin biosynthesis in sugar cane: many gaps still to be filled
Grantee:Paulo Mazzafera
Support type: Program for Research on Bioenergy (BIOEN) - Thematic Grants