Sequencing and Transcriptional Analysis of the Biosynthesis Gene Cluster of Abscisic Acid-Producing Botrytis cinerea
Abstract
:1. Introduction
2. Results
2.1. ABA Production of B. cinerea TB-3-H8
2.2. Sequence Analysis of the ABA Cluster of B. cinerea TB-3-H8
Primers | Forward (5'-3') | Reverse (5'-3') | Sequence Length (bp) | |
---|---|---|---|---|
In B05.10 Supercontig 41 | In TB-3-H8 Genome | |||
1 | GAGACGGAGAAGCTAAGGAGG | CATGATGGCAATACACCAGTT | 1046 (71,207–72,252) | 897 |
2 | CCATTTGGTTTGAGCTGCTTTG | CCTCCTTAGCTTCTCCGTCTC | 1193 (72,232–73,424) | 1193 |
3 | AATTATGCGAAGTCATGTCGA | ACTACCAAAGCAGCTCAAACC | 1240 (73,398–74,637) | 1240 |
4 | GGGTTTCACTGCGGAGTCTTG | GACTGATCCCATGCGGCAGAT | 1221 (74,359–75,579) | 1221 |
5 | ACTGTAGCCGAGATAGTAGGG | AGGCTTACGATTGGGCTGGAA | 1371 (75,467–76,837) | 1386 |
6 | AATGTGGAACTTGCCTTTGGT | CGTTACAACCCTACTATCTCG | 933 (76,809–77,741) | 933 |
7 | TCGATTCAATCCATATTATGACAAC | AAGTTCCACATTGCGCTCCATCTC | 1177 (77,730–78,906) | 1177 |
8 | CATAATGTGGGCTAACTACTCTGA | ACCGACCTCCAAGTCAGGCATAT | 1034 (78,786–79,819) | 1034 |
9 | ATTCGTCTGTGCGTAACCGTGCA | TCAGGTGGTGACGAATACAAGAT | 1554 (79,689–81,242) | No data |
10 | GCAGAAAGGGTGCTCAAAGTGTT | TGCACGGTTACGCACAGACGAAT | 986 (81,220–82,205) | No data |
11 | TTCCTGTGGTCGAGTTTATTG | AACACTTTGAGCACCCTTTCT | 4500 (82,183–866,682) | No data |
12 | TCAGTATGCCTCCTACACGAACA | GCAGCGTACTACAATACGAAACGT | 1005 (86,564–87,568) | No data |
13 | AGGCGGTTCCCTCTTAC | ACGGATACACTCGGCACAACG | 2299 (87,402–89,700) | No data |
14 | ACGATGTTTGTAGAAAGCCACTG | ATCCTCAAGAGTCGCAGTTCATA | 1226 (89,625–90,850) | 1226 |
15 | CGTCGCTGGGACAACATGCTCAA | AAGGCGAGTGAAAGACGAGATGG | 1303 (90,617–91,919) | 1303 |
16 | CGCAATGTCAGGAGGGTAGTA | TTATCGCAGAGAATCAGGCAG | 10,943 (79,374–90,316) | 3762 |
Gene Name | Strains Names | GenBank Accession No. | DNA Length (bp) | cDNA Length (bp) | Protein Size (Amino Acids) | No. of Exons | No. of Introns |
---|---|---|---|---|---|---|---|
bcaba1 | TB-3-H8 | / | 1769 | 1530 | 509 | 5 | 4 |
B05.10 | XM_001553921 | 1925 | 1482 | 493 | 5 | 4 | |
T4 | FQ790338.1 | 1769 | 1530 | 509 | 5 | 4 | |
bcaba2 | TB-3-H8 | / | 1810 | 1584 | 527 | 5 | 4 |
B05.10 | XM_001553920 | 1507 | 1332 | 443 | 4 | 3 | |
T4 | FQ790338.1 | 1810 | 1587 | 528 | 5 | 4 | |
bcaba3 | TB-3-H8 | / | 1323 | 1323 | 440 | 1 | 0 |
B05.10 | XM_001553924 | 1323 | 1269 | 422 | 2 | 1 | |
T4 | FQ790338.1 | 1323 | 1323 | 440 | 1 | 0 | |
bcaba4 | TB-3-H8 | / | 842 | 777 | 258 | 2 | 1 |
B05.10 | XM_001553919 | 842 | 777 | 258 | 2 | 1 | |
T4 | FQ790338.1 | 842 | 777 | 258 | 2 | 1 |
2.3. Transcriptional Analysis of the bcaba Cluster
3. Discussions
4. Experimental Section
4.1. Strains, Plasmids and Culture Conditions
4.2. Fed-Batch Fermentation and Determination of ABA
4.3. DNA Manipulation and PCR Amplification
4.4. DNA Sequencing
4.5. RNA Extraction
4.6. Coding Sequence and Intron Identification by Reverse Transcriptase (RT)-PCR
Genes | Forward Primers (5'-3') | Reverse Primers (5'-3') |
---|---|---|
bcababa1 | 5'-ATGTCTAATTCTATATTGAAC-3' | 5'-CTATTTGTATTCTGTTCCC-3' |
bcababa2 | 5'- ATGCTGCTTAGCATTAAAGA-3' | 5'-CTATCTAGGAACCTCTTTTA-3' |
bcababa3 | 5'- ATGCAGCAAGTTATTACTCA-3' | 5'-CTAGGTACTTTCTCCACGAT-3' |
bcababa4 | 5'-ATGTCCTCTCAACCATTCAC-3' | 5'-CTAACATCTCCATCCGCCAT-3' |
4.7. Sequence Analysis
4.8. Analysis of Gene Expression Profiling by SYBR Green Real-Time PCR Assays
Genes | Forward Primers (5'-3') | Reverse Primers (5'-3') | Product Size (bp) |
---|---|---|---|
bcaba1 | 5'-GCCCAAAGCCTACTGATAAA-3' | 5'-TGTCGAATGAATGACCCAAG-3' | 124 |
bcaba2 | 5'-CTTATTACTTCCCGTTTACTC-3' | 5'-CTTATTACTTCCCGTTTACTC-3' | 101 |
bcaba3 | 5'-CAAGGAACTCAGCAAGCCC-3' | 5'-AGTCGATGCCAACAAAAGG-3' | 102 |
bcaba4 | 5'-CTTGGACGAGTGGGAGTT-3' | 5'-GCCGTTGTTAGCCATTAC-3' | 91 |
18S rRNA | 5'-GAAACTCACCAGGTCCAGA-3' | 5'-CAAATCACTCCACCAACTAAG-3' | 104 |
5. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Gong, T.; Shu, D.; Yang, J.; Ding, Z.-T.; Tan, H. Sequencing and Transcriptional Analysis of the Biosynthesis Gene Cluster of Abscisic Acid-Producing Botrytis cinerea. Int. J. Mol. Sci. 2014, 15, 17396-17410. https://doi.org/10.3390/ijms151017396
Gong T, Shu D, Yang J, Ding Z-T, Tan H. Sequencing and Transcriptional Analysis of the Biosynthesis Gene Cluster of Abscisic Acid-Producing Botrytis cinerea. International Journal of Molecular Sciences. 2014; 15(10):17396-17410. https://doi.org/10.3390/ijms151017396
Chicago/Turabian StyleGong, Tao, Dan Shu, Jie Yang, Zhong-Tao Ding, and Hong Tan. 2014. "Sequencing and Transcriptional Analysis of the Biosynthesis Gene Cluster of Abscisic Acid-Producing Botrytis cinerea" International Journal of Molecular Sciences 15, no. 10: 17396-17410. https://doi.org/10.3390/ijms151017396