Supporting Information
Identification of Autophagy-related Genes ATG4 and ATG8 from Wheat (Triticum aestivum L.) and Profiling of Their Expression Patterns Responding to Biotic and Abiotic Stresses
Dan Pei, Wei Zhang, Hong Sun, Xiaojing Wei, Jieyu Yue, Huazhong Wang
School of Life Sciences, Tianjin Normal University / Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin 300387, China
First author:
Ms. Dan Pei
School of Life Sciences, Tianjin Normal University / Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin, China
393 Binshuixi Road, Tianjin 300387, China
E-mail:
Corresponding author:
Prof. Huazhong Wang
School of Life Sciences, Tianjin Normal University / Tianjin Key Laboratory of Animal and Plant Resistance, Tianjin, China
393 Binshuixi Road, Tianjin 300387, China
Telephone and fax: 86-22-23766539
E-mail:
Journal: Plant Cell Reports
Primer name / Sequence（5' to 3'） / Product size (bp) / ApplicationA8a/eF / GTCGCAAACCCTCGCCCAAATCC / 438 / Amplification of TaATG8a, 8e full-length cDNA by RT-PCR
A8a/eR / CTCGCCCAAATCCGCCGAATCCC
A8b/dF / ACCAAGGTAGATCGATGGCGAAGA / 436 / Amplification of TaATG8b, 8d full-length cDNA by RT-PCR
A8b/dR / TTGATTGGCACGCCTTATTTACA
A8cF / GCGTCGCAAACCCTCGCCCAAAT / 559 / Amplification of TaATG8c full-length cDNA by RT-PCR
A8cR / ATAGGATAAGTCAGGATACGAATAGG
A8fF / CAAGGTAGATCGATGGCGAAGAG / 474 / Amplification of TaATG8f full-length cDNA by RT-PCR
A8fR / TTAAAGATCCTACGACGAGGCAA
A8gF / GAGCGACTTCCAATCCAGTC / 654 / Amplification of TaATG8g full-length cDNA by RT-PCR
A8gR / AATCATCCCAGAGCATGAGG
A8hF / TCTATCCGCGATTCAATCAAATAC / 446 / Amplification of TaATG8h full-length cDNA by RT-PCR
A8hR / CAGTCAGGCAGATGTCACCCAAA
A4a/bF / GGGTGCCTGATGCCCTGATTGAC / 1724(4a)
1727(4b) / Amplification of TaATG4a,4b full-length cDNA by RT-PCR
A4a/bR / TTCCCATCGAACCAACTTGTGCC
NotI-A8a/bF / ATAAGAATGCGGCCGCATGGCGAAGAGCTCGTTCAAGC / 390 / Construction of TaATG8a and 8b yeast or prokaryotic expression vectors
NotI-A8a/bR / ATAAGAATGCGGCCGCCTAGAGCAATCCGAAGGTGT
NotI-A8gF / ATAAGAATGCGGCCGCATGGCCAAGACTTGCTTCAA / 392 / Construction of TaATG8g yeast or prokaryotic expression vectors
NotI-A8gR / ATAAGAATGCGGCCGCTTAGGCGGAGCCGAAAGTGT
NotI-A8hF / ATAAGAATGCGGCCGCATGAAGTCCTTCAAGAAGGA / 383 / Construction of TaATG8h yeast or prokaryotic expression vectors
NotI-A8hR / ATAAGAATGCGGCCGCTCACCCAAATGTCTTCTCGC
NotI-A4aF / ATAAGAATGCGGCCGCATGACGAGCTTGCCTGAGAG / 1470 / Construction of TaATG4a yeast or prokaryotic expression vectors
NotI-A4aR / ATAAGAATGCGGCCGCTCAGAGAATCTGCCACTCAT
NotI-A4bF / ATAAGAATGCGGCCGCATGACGAGCTTGCCTGAGAG / 1473 / Construction of TaATG4b yeast or prokaryotic expression vectors
NotI-A4bR / ATAAGAATGCGGCCGCTCAGAGAATCTGCCACTCGT
SmaI-A8a/bF / TCCCCCGGGAATGGCGAAGAGCTCGTTCAA / 360 / Construction of GFP-TaATG8a or -TaATG8b expression vectors
BamHI-A8a/bR / CGGGATCCCTAGAGCAATCCGAAGGTGT
XbaI-A8gF / GCTCTAGAAGCCAAGACTTGCTTCAAGACC / 358 / Construction of GFP -TaATG8g expression vector
SacI-A8gR / CGAGCTCTTAGGCGGAGCCGAAAGTGT
XbaI -A8hF / GCTCTAGAAATGAAGTCCTTCAAGAAGGA / 351 / Construction of GFP -TaATG8h expression vector
SacI -A8hR / CGAGCTCTCACCCAAATGTCTTCTCGC
qA8aF / TTCCTGTGATCGTTGAGAAGGCTG / 80 / Quantitative RT-PCR of TaATG8a transcripts
qA8aR / AAGGTCGGCAGGGACAAGGT
qA8gF / TTGTGAATAGCACCTTGCCACC / 106 / Quantitative RT-PCR of TaATG8g transcripts and amplification in Chinese spring derived aneuploid nulli-tetrasomic(NT) lines for chromosomal localization of TaATG8g
qA8gR / GTGTTCTCGCCACTGTAAGTCATG
qA8hF / GGAGGAGAGGGCGAATGAGTC / 134 / Quantitative RT-PCR of TaATG8h transcripts
qA8hR / GCATGTCACACGGAACCAGGTAC
qA4aF / TATCACTGCAGCACTGTCCG / 245 / Quantitative RT-PCR of TaATG4a transcripts
qA4aR / GGTGTCGATGTTGTCGGAGG
qA4bF / TCAGTTCAGCCGTCGAAACA / 101 / Quantitative RT-PCR of TaATG4b transcripts
qA4bR / GAGCCATCAAGGTCCTCCG
qTubulin-F / GTGGAACTGGCTCTGGC / 234 / Quantitative RT-PCR of the wheat β-tubulin gene transcripts
qTubulin-R / CGCTCAATGTCAAGGGA
Figure S1. Multiple alignment of wheat ATG8s and homologues from other species. The alignment was produced in ClustalX 2.1 software. Sequences in the alignment include the nine wheat ATG8s here identified, AER27507 (TdATG8) from Triticum dicoccoides, XP_003581707 (BdATG8a) and XP_003571383 (BdATG8b) from Brachypodium distachyon, Os07g0512200 (OsATG8a), Os04g0624000 (OsATG8b), Os08g0191600 (OsATG8c) and Os02g0529150 (OsATG8d) from Oryza sativa, ACJ73920 (ZmATG8a), ACJ73921 (ZmATG8b), ACJ73922 (ZmATG8c), ACJ73923 (ZmATG8d), and ACJ73925 (ZmATG8e) from Zea mays, At4g21980 (AtATG8a), At4g04620 (AtATG8b), At1g62040 (AtATG8c), At2g05630 (AtATG8d), At2G45170 (AtATG8e), At4g16520 (AtATG8f), At3g60640 (AtATG8g), At3g06420 (AtATG8h) and At3g15580 (AtATG8i) from Arabidopsis thaliana, ACU13796 (GmATG8a), ACU14633 (GmATG8b), ACU17086 (GmATG8c), BAH22449 (GmATG8d), ACU15101 (GmATG8e), ACU19559 (GmATG8f), ACU13862 (GmATG8g), ACU16419 (GmATG8h), BAH22448 (GmATG8i) and ACU19611 (GmATG8k) from Glycine max, NP_009216 (HsATG8/GATE16/GABARAPL2) from Human and NP_009475 (ScATG8) from Yeast. Names of wheat ATG8s are boxed. One hundred percent similarity, black; 80 to 100% similarity, dark gray; 60 to 80% similarity, light gray; <60% similarity, white. Hashes and asterisks respectively indicate conserved essential amino acids at the N-terminal microtubule binding site and at the ATG7 binding site. The C-terminal conservative glycine for PE-conjugation is indicated by an arrowhead.
Figure S2. Multiple alignment of wheat ATG4s and homologues from other species. The alignment was produced in ClustalX 2.1 software. Sequences in the alignment include the two wheat ATG4s here identified, Os03g0391000 (OsATG4a) and Os04g0682000 (OsATG4b) from Oryza sativa, ACJ73912 (ZmATG4a) and ACJ73914 (ZmATG4b) from Zea mays, At2g44140 (AtATG4a) and At3g59950 (AtATG4b) from Arabidopsis thaliana, NP_443168 (HsATG4a) from Human and NP_014176 (ScATG4) from Yeast. One hundred percent similarity, black; 80 to 100% similarity, dark gray; 60 to 80% similarity, light gray; <60% similarity, white. The conserved Peptidase_C54 domain is under thick line；Arrowheads mask the canonical catalytic triad of cysteine protease: Cys, Asp and His.