Expressão gênica durante a germinação de sementes de café

Silva, Edvaldo Aparecido Amaral da; Acencio, Marcio Luis; Bovolenta, Luiz Augusto; Lemke, Ney; Varani, Alessandro de Mello; Bravo, Juliana Pereira; Hoshino-Bezerra, Andrea Akemi; Lemos, Eliana Gertrudes Macedo

doi:10.1590/2317-1545v41n2208263

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ORIGINAL ARTICLE • J. Seed Sci. 41 (2) • Apr-Jun 2019 • https://doi.org/10.1590/2317-1545v41n2208263 copiar

Expressão gênica durante a germinação de sementes de café

Autoria SCIMAGO INSTITUTIONS RANKINGS

Resumo:

A germinação da semente do café (Coffea arabica L.) é resultado de eventos que ocorrem simultaneamente no embrião e endosperma. Para entender os mecanismos moleculares responsáveis por estes eventos, nós realizamos uma análise transcricional do embrião e dos endospermas micropilar e lateral de sementes embebidas por 10 dias. O sequenciamento obteve contigs que codificam 16.813 proteínas. Destas, 14.005 (~ 83%) foram altamente similares a pelo menos uma sequência de proteína na base de dados nr. 162 genes foram significativamente expressos em embrião, 36 em endosperma micropilar e 72 em endosperma lateral. A análise de tecido-especificidade dos genes significativamente expressos mostrou que a maior proporção de genes específicos está no embrião (113/162, ~70%), enquanto 11 genes foram expressos nos endospermas lateral e micropilar. No embrião, os genes foram associados principalmente com estresse abiótico, crescimento celular e comunicação intercelular. Nos endospermas micropilar e lateral, eles foram associados com estresse abiótico e degradação da parede celular. A acurácia da análise de RNA-seq foi confirmada por RT-qPCR. Este trabalho acrescenta novas informações sobre os mecanismos moleculares envolvidos na germinação de sementes de café.

Termos de indexação:
RNA-seq; embrião; endosperma micropilar; endosperma lateral; Coffea arabica

ABRATES - Associação Brasileira de Tecnologia de Sementes Av. Juscelino Kubitschek, 1400 - 3° Andar, sala 31 - Centro,, CEP 86020-000 Londrina/PR - Londrina - PR - Brazil
E-mail: jss@abrates.org.br

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[1] *Corresponding author: <amaral.silva@unesp.br>

Gene ID	Gene name	Primer Forward (5’-3’)	Primer Reverse (5’-3’)
2890	Indole-3-acetic acid induced protein 4	AACTAGTCACCGTTGCTTCAT	ATTACACCTTAGGCTTCATGGA
8606	Indole-3-acetic acid induced protein 9	ATTGTCTGAGTGTTCATCGGT	GGCAAAGTTCTGAATCCCTC
3078	α-Tubulin	AGAGGGCATTATACAGTCGG	CCAATCCAGAACCAGTACCA
898	Actin 7	TGCCATCAAGAATCACCCTG	TGACTCACACCATCTCCAGA
396	Glyceraldehyde 3-phosfate dehydrogenase	TTTGGAATCAGGAATCCCGAA	AGCCTTGTCCTTGTCTGTAAA
475	Fructose bisphosfate aldolase	TTGTGGATGTGTTGAAGGAGG	TCTCACCATTAGTGCCAGGA
698	Heat Shock Protein Hsp40	TGCCATCAAGAATCACCCTG	CCACCCATTCCTTCCTTCAAT
1023	Catalase 3	AAGGTACGATCCTGTTCGTC	GTTGCTTGAAGTTGTTCTCCTT
104	Endo-β-mannanase	AGGATTTGTTAGGACTCAGGG	GACACTTTATGCCTCTCGCTA
3111	β-mannosidase	GACGAGTACCACAGATACAAGA	TTCCAATTTACTTTCCCAGTTCC

Gene ID	Gene name	Gene ID	Gene name
Specifically expressed in embryos
2	60S ribosomal protein l31-like	2280	60S ribosomal protein l30
20	Adenosylhomocysteinase	2281	Polyadenylate-binding protein 2-like
80	60S ribosomal protein l28-1-like isoform 1	2300	Methionine aminopeptidase 2b
85	Polyadenylate-binding protein 2-like	2360	60S ribosomal protein l5-like
137	Eukaryotic initiation factor 4a-2-like	2397	Pectin methylesterase
174	S-adenosylmethionine synthase 1-like isoform 1	2432	Pyrophosphate--fructose 6-phosphate 1-phosphotransferase subunit alpha-like
205	Elongation factor 2-like	2575	40S ribosomal protein
229	Elongation factor 1-	2649	Heat shock protein 70
232	Bag family molecular chaperone regulator 7-like	2660	Serine arginine rich splicing isoform 1
345	Small nuclear ribonucleoprotein sm d3-like	2702	60S ribosomal protein l15-like
369	Ubiquitin-like protein	2743	Glucose-1-phosphate adenylyltransferase large subunit 1-like
371	Polyadenylate-binding protein 2-like	2890	Indole-3-acetic acid induced protein 4
453	Desacetoxyvindoline 4-	2947	RNA-binding protein 39-like
502	60S acidic ribosomal protein p0	2996	Pollen-specific protein c13-like
535	Burp domain protein	3038	60S ribosomal protein l23a-like
565	60S ribosomal protein l3-like	3078	Α-tubulin
568	40S ribosomal protein s5	3267	Ubiquitin-like partial
604	Probable phosphatidylinositol 4-kinase type 2-beta at1g26270-like	3296	Cysteine protease 1 precursor
610	40S ribosomal protein sa-like	3323	Alba DNA RNA-binding protein
639	Ubiquitin-like protein	3510	Vesicle-associated membrane protein 726-like
673	60S ribosomal protein l21	3542	Elongation factor 1-delta-like
690	U1 small nuclear ribonucleoprotein 70 kda-like	3726	40S ribosomal protein S9-2-like
691	Polyadenylate-binding protein rbp47-like	4054	Cathepsin b-like
784	Atp binding	4096	40S ribosomal protein
820	Hydroxyproline-rich glycoprotein	4131	Tonoplast intrinsic protein
826	Calreticulin	4445	40S ribosomal protein S15-like isoform 1
903	60S ribosomal protein l10a-1-like	4522	Pyruvate cytosolic isozyme-like
914	Nep1-interacting protein 2-like	4734	Actin-depolymerizing factor 2-like
949	Signal recognition particle 68 kda	4778	40S ribosomal protein s5-1-like
976	Calmodulin	4863	60S ribosomal protein l44-like
995	Dehydrin	4941	Gtp-binding protein sar1a-like
1015	Elongation factor 1- partial	5256	Polyadenylate-binding protein rbp45-like
1131	Enolase	5513	Serine mitochondrial-like
1151	Cbs domain protein	5840	Mlp-like protein 423-like
1176	Heat shock	6138	60s ribosomal protein l27a-3-like
1217	40S ribosomal protein s8	6674	Transcription factor bhlh63-like
1252	Pyruvate cytosolic isozyme-like	7048	Oleosin kda-like
1312	40S ribosomal protein S13-like	7450	Burp domain-containing protein
1319	Ac012654_15est gb	7525	Cation exchanger 1
1346	40S ribosomal protein S23-like	7549	Serine acetyltransferase 5-like
1427	40S ribosomal protein S11	7583	60S ribosomal protein l18a-like
1434	Cyclin-dependent kinase g-2-like	7823	P-loop containing nucleoside triphosphate hydrolases superfamily protein
1528	Pointed first leaf	7912	Dehydrin-like protein
1562	40S ribosomal protein S4-like	8033	60S ribosomal protein l34-like
1578	Carrier protein mitochondrial-like	8579	Pyrophosphate--fructose 6-phosphate 1-phosphotransferase subunit alpha-like
1596	Protein-protein interaction regulator family	8606	Indole-3-acetic acid induced protein 9
1693	Inositol transporter 1	8891	Galactose oxidase kelch repeat superfamily protein isoform partial
1711	Probable aquaporin pip-type 7a-like	10672	Chaperone protein dnaj 20
1726	Protein disulfide-isomerase-like	10711	Rop4 small gtp binding protein
1787	Heat shock protein 90	12711	Ent-kaurene oxidase
1819	Aspartate aminotransferase	14259	Ferritin 2
1834	Ubiquitin-conjugating enzyme e2 7-like	14595	Serine carboxypeptidase-like 18-like
1945	RNA-binding protein 39-like	14935	40S ribosomal protein S14
2037	Ubiquitin-like partial	16395	Osmotin-like protein
2038	Ubiquitin-like partial	19893	Thaumation-like
2074	Hyaluronan mrna binding family isoform 1	42855	Protodermal factor
2266	Tubulin alpha chain-like
Expressed in micropylar and lateral endosperms
29	BURP domain-containing protein	292	Thaumation-like protein
32	Nonspecific lipid-transfer protein	376	Glutathione s-transferase t1-like
70	β-glucosidase 44-like	1238	MLP-like protein 28
104	Endo-β-mannanase	1318	Dormancy auxin associated protein
110	Cysteine proteinase inhibitor	3111	β-mannosidase
273	Cysteine proteinase
Expressed in embryo, micropylar and lateral endosperms
156	Protein translation factor sui1 homolog isoform 1	636	Thiazole biosynthetic enzyme
158	Elongation factor-1 alpha	698	Heat Shock Protein Hsp40
374	Catalase	765	Ascorbate peroxidase
396	Glyceraldehyde 3-Phosfate dehydrogenase	898	Actin 7
408	Ubiquitin-conjugating enzyme	1022	Catalase
423	Translationally controlled tumor protein	1379	High mobility group b3 protein
475	Fructose bisphosfate aldolase	2508	Mlp-like protein 423-like
516	Polyadenylate-binding protein-interacting p 2	2709	Catalase
608	Ubiquitin-conjugating enzyme e2 10-like	2780	Non-symbiotic hemoglobin class 1