Cloning and Expression Pattern Analysis of Rice OsJMJ718 Alternative Polyadenylation Sequences During Reproductive Developmental Stage

doi:10.11983/CBB17114

Abstract

Abstract: Alternative polyadenylation (APA) is one of the important regulatory pathways of eukaryotic gene expression. By forming different lengths of 3' untranslated regions, APA affects the stability, localization and translation efficiency of mRNA and increases the complexity of transcripts. The expression of the Arabidopsis gene increased polyadenylation of BONSAI methylation 1 (IBM1) is regulated by chromatin regulatory factor enhanced downy mildew 2 (EDM2), which can further affect the CHG methylation level of the Arabidopsis thaliana genome. However, whether such regulatory mechanisms exist in other species is unknown. To answer this question, we selected OsJMJ718, an IBM1 homologous gene of rice, for research. By using bioinformatics analysis and 3'RACE experiments, we found that homologous genes of IBM1 also had APA modification. Among them, the OsJMJ718 gene may have 9 alternative polyadenylation sequences. Further sequence alignment analysis revealed that the 3' terminal sequence of OsJMJ718 in the existing Japonica genome in the NCBI database may be different from that of the other ecotype genome sequences, such as 9522 and Minghui 63. Quantitative real-time PCR showed that the 9 transcripts of OsJMJ718 present diverse dynamic expression patterns in different stages of rice reproductive development. The expression of OsJMJ718-TVX5 was higher than that of other transcripts. In summary, this work provides information for APA sequences of OsJMJ718, and the expression pattern analysis of these transcripts would also help further study of the mechanism of APA and biological function of OsJMJ718.

Key words: rice, OsJMJ718, alternative polyadenylation, 3'RACE, expression pattern

Lu Dan, Wang Li, Song Fan, Tao Juhong, Zhang Dabing, Yuan Zheng. Cloning and Expression Pattern Analysis of Rice OsJMJ718 Alternative Polyadenylation Sequences During Reproductive Developmental Stage[J]. Chinese Bulletin of Botany, 2018, 53(5): 594-602.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB17114

https://www.chinbullbotany.com/EN/Y2018/V53/I5/594

Figures/Tables 4

References 10

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Primer name	Primer sequence (5′-3′)	Type
qRT-F-TVX1	GAGCTTGGATAGCCCGCCTC	qRT PCR
qRT-R-TVX1	TCTTTTCTTCCCGGGAGTGC	qRT PCR
qRT-F-TVX2	CAATAATTGAACTCTAGGTC	qRT PCR
qRT-R-TVX2	TAAGGAAATACAATCAGATC	qRT PCR
qRT-F-TVX3	ACGGACGCTGGATCGGCGAG	qRT PCR
qRT-R-TVX3	TAACAAGAGCAGTAGAGCAC	qRT PCR
qRT-F-TVX4	ACGGACGCTGGATCGGCGAG	qRT PCR
qRT-R-TVX4	AAGGACGGGGATGCGGCGT	qRT PCR
qRT-F-TVX5	AACGACAACTTTAGGGTTCG	qRT PCR
qRT-R-TVX5	TCGTTACAAGAAAGATGAAC	qRT PCR
qRT-F-TVX6	ATCGAATTGCCACGTAAGCG	qRT PCR
qRT-R-TVX6	TCATCCTCACTCTCTTCTTC	qRT PCR
qRT-F-TVX7	GAACCACAGGGCCAAAGAAG	qRT PCR
qRT-R-TVX7	TAATCCAATTAAAAGTGTTG	qRT PCR
qRT-F-TVX9	AACTCTTCACCACGCGTATG	qRT PCR
qRT-R-TVX9	TAACCGGCGATGGCTGCATC	qRT PCR
qRT-F-TVX11	GAATAAGATGATAATCTATG	qRT PCR
qRT-R-TVX11	ATATCTCTAACTCTACATGC	qRT PCR
3'RACE-F-OsJMJ718	GATTACGCCAAGCTTAGTGAGACCAACAAGGGAGGTGCT	3'RACE

Primer name	Primer sequence (5′-3′)	Type
qRT-F-TVX1	GAGCTTGGATAGCCCGCCTC	qRT PCR
qRT-R-TVX1	TCTTTTCTTCCCGGGAGTGC	qRT PCR
qRT-F-TVX2	CAATAATTGAACTCTAGGTC	qRT PCR
qRT-R-TVX2	TAAGGAAATACAATCAGATC	qRT PCR
qRT-F-TVX3	ACGGACGCTGGATCGGCGAG	qRT PCR
qRT-R-TVX3	TAACAAGAGCAGTAGAGCAC	qRT PCR
qRT-F-TVX4	ACGGACGCTGGATCGGCGAG	qRT PCR
qRT-R-TVX4	AAGGACGGGGATGCGGCGT	qRT PCR
qRT-F-TVX5	AACGACAACTTTAGGGTTCG	qRT PCR
qRT-R-TVX5	TCGTTACAAGAAAGATGAAC	qRT PCR
qRT-F-TVX6	ATCGAATTGCCACGTAAGCG	qRT PCR
qRT-R-TVX6	TCATCCTCACTCTCTTCTTC	qRT PCR
qRT-F-TVX7	GAACCACAGGGCCAAAGAAG	qRT PCR
qRT-R-TVX7	TAATCCAATTAAAAGTGTTG	qRT PCR
qRT-F-TVX9	AACTCTTCACCACGCGTATG	qRT PCR
qRT-R-TVX9	TAACCGGCGATGGCTGCATC	qRT PCR
qRT-F-TVX11	GAATAAGATGATAATCTATG	qRT PCR
qRT-R-TVX11	ATATCTCTAACTCTACATGC	qRT PCR
3'RACE-F-OsJMJ718	GATTACGCCAAGCTTAGTGAGACCAACAAGGGAGGTGCT	3'RACE

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