SECTION I – Introduction generale
A- PHASE ADMINISTRATIVE
5) Pourvoi devant la C.N.R.F
Com os dados apresentados no presente trabalho pode-se concluir que os quatro genótipos cultivados em campo, previamente selecionados com base no teor de lignina, não se distinguem claramente em dois grupos contratantes, de maior e menor teor do polímero. Porém, as análises bioquímicas distinguem claramente os tecidos analisados, ainda que os genótipos tenham sido cultivados em duas localidades. O efeito de localidade também foi claramente notável através do perfil dos oligômeros, sendo possível observar maior diversidade de estruturas quando os genótipos foram cultivados na cidade de PIN.
A complexidade do genoma da cana-de-açúcar, a variação da expressão gênica dos dezenove genes identificados anteriormente, juntamente com a influência de outros fatores que intermeiam o processo de lignificação dificultam a identificação de uma correlação direta entre acúmulo de lignina e a expressão dos genes da sua via de biossíntese. No entanto, apesar da complexidade dos dados apresentados, este trabalho traz conhecimento complementar para futuras manipulações de genes envolvidos nos processos de biossíntese de lignina como forma de otimizar a produção de etanol lignocelulósico.
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APÊNDICE
Figura S1. Esquema ilustrativo dos dados mais relevantes de expressão gênica relativa e a possível correlação entre eles e as análises bioquímicas de quantificação e composição (razão
Tabela S1. Valores obtidos pela análise de expressão gênica relativa dos 19 genes relacionados à biossíntese de lignina para o genótipo IACSP04-028. Resultados expressos como as médias de três replicatas independentes.
Gene Mean Ribeirão Preto Pindorama
P value YR‡ MR YP MP YR MR YP MP
ShPAL1 0,0026 4,6061Aa 4,2085Aa 2,2050Aa 4,0835Aa 6,4920Aa 0,1057Ab 1,4474Ab 1,0095Ab
ShPAL2 0,0024 0,00376Aa 0,00098Aab 0,00286Aab 0,00072Ab 0,00175Aa 0,00079Aa 0,00045Ba 0,00156Aa
ShPAL3 <0,0001 0,01199Aa 0,00178Ab 0,00596Aab 0,00035Ac 0,00311Aa 0,00066Aa 0,00133Aa 0,00012Ab
ShPAL4 0,0002 0,8435Aa 0,4166Aa 0,3610Aa 0,1961Aa 0,7420Aa 0,0080Bb 0,2125Aac 0,0753Abc
ShC4H1 0,0329 0,0041Aab 0,0023Bb 0,0050Aab 0,0074Aa 0,0050Aa 0,0069Aa 0,0066Aa 0,0062Aa
ShC4H2 0,0006 0,00010Ab 0,00041Aa 0,00013Aab 0,00033Aab 0,00041Aab 0,00083Aa 0,00016Ab 0,00048Aab
Sh4CL1 0,0024 0,00780Aa 0,00680Aa 0,00238Aa 0,00432Aa 0,00712Aa 0,00347Aab 0,00181Ab 0,00120Ab
Sh4CL2 0,0026 0,0545Aa 0,0319Aa 0,0334Aa 0,0408Aa 0,0472Aa 0,0137Ab 0,0248Aab 0,0225Aab
ShHCT1 0,0552 0,00008Aa 0,00005Aa 0,00001Aa 0,00001Aa 0,00002Aa 0,00001Aa 0,00001Aa 0,00006Aa
ShHCT-like <0,0001 0,00026Ab 0,00025Ab 0,00072Aa 0,00050Aab 0,00026Aa 0,00008Bb 0,00009Bb 0,00015Bab
ShC3H1 0,0029 0,0678Aa 0,0472Aa 0,0340Aa 0,0339Aa 0,0166Aa 0,0116Aa 0,0119Aa 0,0071Aa
ShC3H2 0,0165 0,0265Aa 0,0246Ba 0,0276Aa 0,0210Aa 0,0231Ab 0,1129Aa 0,0259Ab 0,0324Aab
ShCCoAOMT1 0,0003 0,8533Aa 0,5911Aa 0,5467Aa 1,2545Aa 0,3862Aa 0,2057Aa 0,0972Ba 0,1794Ba
ShCCoAOMT2 <0,0001 0,9345Aa 0,0824Ab 0,2219Ab 0,0903Ab 0,5321Aa 0,0202Bb 0,1921Aa 0,0604Ab
ShCCR1 <0,0001 0,00134Aa 0,00063Aa 0,00066Aa 0,00110Aa 0,00154Aa 0,00008Bb 0,00027Ac 0,00077Aac
ShF5H <0,0001 0,00107Ab 0,00301Aa 0,00042Bc 0,00071Abc 0,00092Aa 0,00141Ba 0,00095Aa 0,00033Bb
ShCOMT1 <0,0001 0,1533Aa 0,3064Aa 0,0224Ab 0,1612Aa 0,0251Ba 0,0171Ba 0,0105Aa 0,0294Ba
ShCAD2 <0,0001 0,0052Aa 0,0052Aa 0,0019Ab 0,0049Aa 0,0032Aac 0,0047Aa 0,0007Bb 0,0025Bc
ShCAD8 0,0006 0,0046Aa 0,0033Aa 0,0033Aa 0,0024Aa 0,0019Aa 0,0012Aa 0,0007Ba 0,0013Aa
† A expressão de cada gene foi normalizada pela expressão do gene GAPDH.
‡ Tecidos: YR, córtex jovem; YP, medula jovem; MR, córtex maduro; MP, medula madura.
§Estatística entre o mesmo tecido entre as localidades. Médias seguidas pela mesma letra não são significativamente diferentes de acordo com
¶Estatística entre tecidos de uma localidade. Médias seguidas pela mesma letra não são significativamente diferentes de acordo com teste de Tukey (P ≤ 0,05) a partir da análise de uma via da variância (ANOVA).
Tabela S2. Valores obtidos pela análise de expressão gênica relativa dos 19 genes relacionados à biossíntese de lignina para o genótipo IACSP04-053. Resultados expressos como as médias de três replicatas independentes.
Gene Mean Ribeirão Preto Pindorama
P value YR‡ MR YP MP YR MR YP MP
ShPAL1 <0.0001 10.7328A§a¶ 6.8575Aab 5.6643Abc 2.2998Ac 6.4602Ba 1.7163Bb 7.5976Aa 1.7425Ab
ShPAL2 <0.0001 0.00332Aa 0.00023Ab 0.00245Aa 0.00020Ab 0.00260Aab 0.00093Ab 0.00680Aa 0.00090Ab
ShPAL3 <0.0001 0.02238Aa 0.00674Aa 0.01149Aa 0.00082Ab 0.01336Aa 0.00561Aab 0.01616Aa 0.00210Ab
ShPAL4 <0.0001 0.8068Aa 0.5129Aa 0.4201Aa 0.0588Ab 0.7810Aa 0.1360Ab 0.6815Aa 0.0414Ab
ShC4H1 <0.0001 0.0030Ba 0.0036Ba 0.0058Ba 0.0061Aa 0.0087Abc 0.0112Aac 0.0228Aa 0.0047Ab
ShC4H2 0.0030 0.00021Ab 0.00068Aa 0.00019Ab 0.00060Aa 0.00035Aa 0.00058Aa 0.00039Aa 0.00030Aa
Sh4CL1 <0.0001 0.0174Ab 0.0469Aa 0.0072Ac 0.0097Abc 0.0108Aa 0.0200Ba 0.0049Ab 0.0148Aa
Sh4CL2 <0.0001 0.0622Aa 0.0176Ab 0.0526Aa 0.0120Ab 0.0511Aa 0.0127Ab 0.0526Aa 0.0202Ab
ShHCT1 <0.0001 0.00133Aa 0.00018Aab 0.00012Bab 0.00003Ab 0.00014Ac 0.00001Bb 0.00218Aa 0.00002Ab
ShHCT-like <0.0001 0.00005Ab 0.00026Aa 0.00010Ac 0.00013Ac 0.00006Ab 0.00022Aa 0.00012Aab 0.00006Bb
ShC3H1 0.0004 0.0346Aa 0.0954Aa 0.0298Aa 0.0296Aa 0.0263Aab 0.0473Aa 0.0081Bb 0.0321Aa
ShC3H2 0.0051 0.0284Aab 0.0340Aa 0.0151Ab 0.0163Bab 0.0271Aa 0.0356Aa 0.0258Aa 0.0424Aa
ShCCoAOMT1 0.0001 0.4916Ab 2.6958Aa 0.3347Ab 0.5647Ab 0.3124Aab 1.1077Aa 0.1721Ab 0.6934Aab
ShCCoAOMT2 0.0002 0.6903Aa 0.1890Aa 0.4079Aa 0.0242Bb 0.4581Aa 0.1143Aa 0.4121Aa 0.1139Aa
ShCCR1 0.0012 0.00091Aab 0.00357Aa 0.00055Ab 0.00054Ab 0.00060Aab 0.00124Aab 0.00029Ab 0.00166Aa
ShF5H <0.0001 0.00118Abc 0.00591Aa 0.00063Bb 0.00235Ac 0.00148Ab 0.00228Bab 0.00295Aa 0.00220Aab
ShCOMT1 <0.0001 0.1210Ab 0.7172Aa 0.0768Ab 0.1689Ab 0.0445Ab 0.3971Aa 0.0315Ab 0.0968Aab
ShCAD2 <0.0001 0.0063Abc 0.0215Aa 0.0037Ab 0.0092Ac 0.0035Ab 0.0151Aa 0.0014Bc 0.0053Ab
ShCAD8 <0.0001 0.0072Aa 0.0058Aa 0.0051Aa 0.0039Aa 0.0028Aab 0.0012Bab 0.0040Aa 0.0010Bb
7
† A expressão de cada gene foi normalizada pela expressão do gene GAPDH.
‡ Tecidos: YR, córtex jovem; YP, medula jovem; MR, córtex maduro; MP, medula madura.
§Estatística entre o mesmo tecido entre as localidades. Médias seguidas pela mesma letra não são significativamente diferentes de acordo com teste de Tukey (P ≤ 0,05) a partir da análise de uma via da variância (ANOVA).
¶Estatística entre tecidos de uma localidade. Médias seguidas pela mesma letra não são significativamente diferentes de acordo com teste de Tukey (P ≤ 0,05) a partir da análise de uma via da variância (ANOVA).
Tabela S3. Valores obtidos pela análise de expressão gênica relativa dos 19 genes relacionados à biossíntese de lignina para o genótipo IACSP04-691. Resultados expressos como as médias de três replicatas independentes.
Gene Mean Ribeirão Preto Pindorama
P value YR‡ MR YP MP YR MR YP MP
ShPAL1 0.0178 16.2829A§a¶ 6.8603Ab 7.1768Ab 7.4943Aab 8.1943Aa 9.9294Aa 5.5659Aa 6.5822Aa
ShPAL2 <0.0001 0.0118Aa 0.0013Bb 0.0206Aa 0.0013Bb 0.0113Aa 0.0186Aa 0.0191Aa 0.0096Aa
ShPAL3 <0.0001 0.0409Aa 0.0022Ab 0.0239Aa 0.0019Ab 0.0068Ba 0.0042Ab 0.0090Aa 0.0027Ab
ShPAL4 <0.0001 1.5057Aa 0.0663Bb 0.8902Aa 0.0510Bb 0.7552Aa 0.8220Aa 0.7332Aa 0.4861Aa
ShC4H1 0.0016 0.0042Aa 0.0053Aa 0.0068Aa 0.0026Ba 0.0106Aab 0.0036Ab 0.0147Aa 0.0093Aab
ShC4H2 <0.0001 0.00070Ab 0.00392Aa 0.00109Ab 0.00778Aa 0.00106Aab 0.00218Aa 0.00060Ab 0.00129Bab
Sh4CL1 0.0009 0.0117Aab 0.0127Aa 0.0049Aab 0.0044Bb 0.0235Aa 0.0161Aa 0.0109Aa 0.0175Aa
Sh4CL2 0.0005 0.0354Aa 0.0061Ab 0.0265Aab 0.0100Ab 0.0376Aa 0.0190Aab 0.0324Aab 0.0138Ab
ShHCT1 <0.0001 0.00194Aa 0.00035Ab 0.00032Ab 0.00002Bc 0.00076Aa 0.00049Aa 0.00027Aa 0.00033Aa
ShHCT-like 0.0008 0.00007Ba 0.00020Aa 0.00013Aa 0.00011Aa 0.00058Aa 0.00008Ab 0.00011Ab 0.00003Ab
ShC3H1 0.0099 0.0356Aa 0.0226Aab 0.0268Aab 0.0085Bb 0.0267Aa 0.0300Aa 0.0318Aa 0.0345Aa