Expression Profiling of Non-Aflatoxigenic Aspergillus parasiticus Mutants Obtained by 5-Azacytosine Treatment or Serial Mycelial Transfer
Abstract
:1. Introduction
2. Materials and Methods
2.1. Fungal Strains
2.2. Culture Conditions
2.3. Aflatoxin Evaluation
2.4. Microarray Analyses
2.5. Quantitative RT-PCR (qRT-PCR)
NCBI or TIGR Number | Sequence (5' to 3') | Product size (bp) | |
---|---|---|---|
aflJ | TGACTCTCCTTTTGCCGAATGT | 250 | |
GGGAATGGCAACGGTGGGCG | |||
XM_001824706 (TC9112) | TATGAGCCCCAACGCAACAGACAG | 268 | |
CGGGTACGGCTCATTAGAAGGAC | |||
XM_002372755(TC10739) | GCCCCAGCTTACCAAACGAGA | 336 | |
AGGGATGAGGGGGAATGAAGTG | |||
XM_002373878(TC10202) | TGCGGGAAATAATACGACAGGAA | 359 | |
AAAGTAGCGACCCGGGCAGTAGTG | |||
XM_002380627(TC11938) | GACCGCATCGTGGCTTTTCTAC | 334 | |
AAGCATCATTCGTCATTCGTTCTG | |||
XM_002374352(TC9364) | CCCACCGAAGACCTGACCTACAT | 322 | |
CTGCCATCTGCCAAACTCCATTA |
3. Results
3.1. Loss of Aflatoxin Production after 5-Azacytosine Treatment and Serial Mycelial Transfer
3.2. Microarray Transcription Profiles in 5-Azacytosine-Treated and Serial Transfer Isolated Colonies Compared to Wild-Type
Gene | Ratios | Probabilities b | ||
---|---|---|---|---|
Untreated vs. 5-AC-treated | Untreated vs. 5-AC-treated | Untreated vs. Serial transfer | Untreated vs. Serial transfer | |
adhA | 1.22 | 1.10 | 0.4388 | 0.3307 |
aflJ * | 1.76 | 1.59 | 0.0018 | 0.0015 |
aflR | 1.29 | 1.12 | 0.3367 | 0.3465 |
aflT | 1.32 | 1.11 | 0.3321 | 0.2423 |
avfA | 1.31 | 1.17 | 0.2902 | 0.4271 |
avnA | 1.25 | 1.18 | 0.3835 | 0.2162 |
cypX | 1.33 | 1.15 | 0.1235 | 0.0835 |
estA | 1.33 | 1.25 | 0.1929 | 0.0493 |
hexA | 1.37 | 1.12 | 0.0768 | 0.1850 |
hypB1 | 1.24 | 0.95 | 0.2552 | 0.8827 |
hypB2 | 2.49 | 2.16 | 0.4722 | 0.5134 |
moxY | 1.26 | 1.09 | 0.1828 | 0.1875 |
nadA | 1.24 | 1.07 | 0.4882 | 0.4211 |
nor-1 | 1.41 | 1.12 | 0.0570 | 0.0620 |
norA | 1.33 | 1.07 | 0.0553 | 0.2726 |
omtA | 1.35 | 1.28 | 0.0422 | 0.0741 |
omtB | 1.37 | 1.08 | 0.0385 | 0.2716 |
ordA | 1.34 | 1.25 | 0.2363 | 0.1889 |
ordB | 1.35 | 1.31 | 0.1566 | 0.0602 |
pksA | 1.27 | 1.08 | 0.2264 | 0.1826 |
sugR | 1.32 | 1.10 | 0.3869 | 0.3722 |
vbs | 1.26 | 1.05 | 0.1547 | 0.1843 |
ver-1 | 1.23 | 1.10 | 0.7285 | 0.9483 |
verA | 1.16 | 1.08 | 0.5312 | 0.8985 |
verB | 1.24 | 1.15 | 0.1683 | 0.1181 |
TC Number or EST b | Best Match GenBank c | Putative Protein | Ratios d | Probabilities d | ||
---|---|---|---|---|---|---|
Untreated vs. 5-AC-treated | Untreated vs. Serial transfer | Untreated vs. 5-AC-treated | Untreated vs. Serial transfer | |||
TC8344 | XM_001824592 | HET&ankyrin domain protein | 10.02 | 8.53 | 0.0000 | 0.0000 |
TC10739 * | XM_002372755 | Ankyrin repeat-containing protein | 5.69 | 4.52 | 0.0000 | 0.0000 |
TC9364 * | XM_002374352 | FG-GAP (integrin) repeat protein | 4.79 | 3.97 | 0.0000 | 0.0000 |
TC9256 | XM_002375859 | Nitrite reductase (NiiA) | 4.58 | 3.38 | 0.0000 | 0.0000 |
NAGAO52TV | XM_002380681 | Polysaccharide synthase Cps1 | 4.54 | 4.72 | 0.0000 | 0.0000 |
TC10370 | XM_002384463 | Opsin | 4.18 | 3.49 | 0.0000 | 0.0000 |
TC8557 | XM_001827098 | alpha1,3-glucan synthase | 3.71 | 2.62 | 0.0000 | 0.0000 |
TC10767 * | XM_003096248 | Integral membrane protein | 3.54 | 2.49 | 0.0000 | 0.0002 |
TC10268 | XM_002383371 | 12 kda heat shock protein | 3.53 | 3.48 | 0.0000 | 0.0000 |
TC9112 * | XM_001824706 | GPI-anchored protein | 3.45 | 3.00 | 0.0000 | 0.0000 |
TC10523 | XM_002374369 | Integral membrane prot-exocytic transport from Golgi | 3.40 | 3.53 | 0.0000 | 0.0000 |
NAGCY20TV | XM_002379925 | Gal4 transcription factor-AFLA_139560 | 3.28 | 3.20 | 0.0000 | 0.0000 |
NAGAE41TV | XM_002382445 | Calcium binding protein Caleosin | 3.23 | 3.09 | 0.0000 | 0.0000 |
TC10524 | XM_002374369 | Exocytic transport protein | 3.21 | 3.28 | 0.0000 | 0.0000 |
TC10334 | XM_002379834 | GABA-permease | 3.11 | 2.69 | 0.0000 | 0.0000 |
TC11140 | XM_002383723 | MFS-transporter-related | 3.02 | 2.18 | 0.0000 | 0.0007 |
TC11938 * | XM_002380627 | EsdC | 2.98 | 2.71 | 0.0000 | 0.0000 |
NAFCO41TV | XM_001819214 | Transcriptional regulatory protein pro1 (HLH protein) | 2.88 | 2.35 | 0.0000 | 0.0000 |
TC11262 | XM_002384600 | Helix-loop-helix DNA-binding domain protein | 2.75 | 2.40 | 0.0000 | 0.0001 |
TC10225 | XM_003189329 | SH3 domain kinase binding protein (predicted membrane protein) | 2.70 | 2.47 | 0.0000 | 0.0000 |
TC8802 | XM_002378642 | Ser-thr rich protein AOS28(solid-state culture specific gene) | 2.66 | 2.63 | 0.0000 | 0.0000 |
TC11294 | XM_002384659 | Arrestin domain protein | 2.54 | 1.92 | 0.0000 | 0.0007 |
TC8781 | XM_002375350 | MAPEG superfamily protein pfam01124 | 2.45 | 1.81 | 0.0000 | 0.0003 |
TC10523 | XM_002374369 | Hypothetical protein | 2.40 | 2.54 | 0.0000 | 0.0000 |
TC10950 | XM_002377280 | RTA domain protein (RTA1 superfamily, pfam04479) | 2.15 | 1.82 | 0.0000 | 0.0000 |
TC8911 | XM_002383317 | Aspartic endopeptidase; aspartyl proteinase | 2.13 | 1.82 | 0.0000 | 0.0002 |
TC8680 | XM_002382580 | GPI-anchored glycoprotein (AFLA_004200) | 2.03 | 1.98 | 0.0000 | 0.0000 |
TC9120 | XM_002381180 | bZIP transcription factor ATFB (atf21, AFLA_094010) | 2.02 | 1.96 | 0.0000 | 0.0000 |
TC8511 | XM_002383753 | Hypothetical protein | 1.92 | 2.11 | 0.0000 | 0.0000 |
NAFBR39TV | XM_002378094 | alpha/beta hydrolase domain protein | 1.90 | 1.66 | 0.0000 | 0.0002 |
TC9105 | XM_001824752 | Hypothetical protein | 1.87 | 1.96 | 0.0000 | 0.0000 |
TC10397 | XM_002379378 | PHD finger and SET domain protein | 1.67 | 1.75 | 0.0000 | 0.0000 |
TC9269 | XM_002384586 | Metallophophodiesterase (AFLA_119200) | 1.66 | 1.55 | 0.0000 | 0.0001 |
TC9718 | XM_002372994 | CVNH superfamily (cyanovirin-N family, pfam08881) | 0.53 | 0.61 | 0.0000 | 0.0001 |
NAFEY64TV | XM_002377133 | Asparagine synthase (glutamine-hydrolyzing) | 0.49 | 0.52 | 0.0001 | 0.0003 |
Gene | Ratios | Probabilities | Expressed Sequence Tag (EST) | GenBank Accession Number | AFLA # | |||
---|---|---|---|---|---|---|---|---|
Untreated vs. 5-AC-treated | Untreated vs. Serial transfer | Untreated vs. 5-AC-treated | Untreated vs. Serial transfer | |||||
laeA | 0.75 | 0.77 | 0.8998 | 0.8874 | Unknown | XM_002374798 | 033290 | |
laeA-1 1 | 0.41 | 0.40 | 0.0002 | 0.0002 | TC9243 | XM_002375064 | 035950 | |
veA | 0.76 | 0.68 | 0.0307 | 0.0041 | TC11465 | XM_002380164 | 066460 | |
nsdD | 1.83 | 0.99 | 0.0145 | 0.9512 | TC10891 | XM_002376000 | 020210A |
4. Discussion
5. Conclusions
Acknowledgements
References
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Wilkinson, J.R.; Kale, S.P.; Bhatnagar, D.; Yu, J.; Ehrlich, K.C. Expression Profiling of Non-Aflatoxigenic Aspergillus parasiticus Mutants Obtained by 5-Azacytosine Treatment or Serial Mycelial Transfer. Toxins 2011, 3, 932-948. https://doi.org/10.3390/toxins3080932
Wilkinson JR, Kale SP, Bhatnagar D, Yu J, Ehrlich KC. Expression Profiling of Non-Aflatoxigenic Aspergillus parasiticus Mutants Obtained by 5-Azacytosine Treatment or Serial Mycelial Transfer. Toxins. 2011; 3(8):932-948. https://doi.org/10.3390/toxins3080932
Chicago/Turabian StyleWilkinson, Jeffrey R., Shubha P. Kale, Deepak Bhatnagar, Jiujiang Yu, and Kenneth C. Ehrlich. 2011. "Expression Profiling of Non-Aflatoxigenic Aspergillus parasiticus Mutants Obtained by 5-Azacytosine Treatment or Serial Mycelial Transfer" Toxins 3, no. 8: 932-948. https://doi.org/10.3390/toxins3080932