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Molecular characterization of Myb transcription factor (TaMyb) gene that is
expressed in response to hypoxic condition in wheat (Triticum aestivum L.) roots
Tong Geon Lee1, Cheol Seong Jang1, Jae Yoon Kim1, Jae Han Park1, Dae Yeon Kim1, Dong Sub Kim2, and Yong Weon Seo1
(1) Division of Biotechnology and Genetic Engineering, Korea University, Anam-Dong, Seongbuk-Gu, Seoul 136-713, Republic of Korea
(2) Department of Radiation Plant Breeding and Genetics, Korea Atomic Energy Research Institute, Yuseong-Gu, Daejeon 305-600, Republic
of Korea
A
Introduction.………………………………………...
0.180
7.50
oxygen deficiency (DO)
0.160
• Oxygen deficiency associated with waterlogging
in the field is one of the major problems in the crop
growing areas. The appropriate signaling pathway
under oxygen deficiency may help a successful
adaptation to change in O2 concentration.………..
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TaMyb1
Myb1
MybHv1
Zm38
R2R3 Myb protein
Arabidopsis
CRLRWINYLRPDLKRGNFSDEEDELIIKLHSLLGNKWSLIAGRLPGRTDNEIKNYWNTHI
CRLRWINYLRPDLKRGNFSDEEDELIIKLHSLLGNKWSLIAGRLPGRTDNEIKNYWNTHI
CRLRWINYLRPDLKRGNFSHEEDELIIKLHSLLGNKWSLIAGRLPGRTDNEIKNYWNTHI
CRLRWINYLRPDLKRGNFTADEDDLIVKLHSLLGNKWSLIAARLPGRTDNEIKNYWNTHV
CRLRWINYLRPDLKRGNFTADEDDLIIKLHSLLGNKWSLIAARLPGRTDNEIKNYWNTHI
CRLRWLNYLRPDVRRGNITLEEQFMILKLHSLWGNRWSKIAQYLPGRTDNEIKNYWRTRV
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TaMyb1
Myb1
MybHv1
Zm38
R2R3 Myb protein
Arabidopsis
RRKLTSRGIDPVTHRAINSDHAASNITISFEAAQ---RDDKGAVFRRDAEPTKVAAAAAA
RRKLTSRGIDPVTHRAINSDHAASNITISFETAQ---RDDKGAVFRRDAEPTKVAAAAAA
RRKLTSRGIDPVTHRAINSDHAASNITISFESAQ---RDDKGAVFRRDAEPAKAAAAAAA
RRKLLGRGIDPVTHRPIAADAVTVTTVSFQPSPS-------------------------RRKLLGRGIDPVTHRPVNAAAATISFHPQPPP---------------------------QKQAKHLRCDVNSNLFKETMRNVWMPRLVERINAQSLPTTCEQVESMITDPSQPVNEPSP
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TaMyb1
Myb1
MybHv1
Zm38
R2R3 Myb protein
Arabidopsis
ITHVDHHHRS--NPHHQMEWGQGKPLKCPDLNLDLCISPPS-----HEDPMVDTKPVXKR
ITHVDHHHHHRSNPLHQMEWGQGKPLKCPDLNLDLCISPPS-----HEDPMVDTKPVVKR
ISHHVDHHHR---SNPQLDWGQGKPLKCPDLNLDLCISPPI-----HEDPMVDTKPVVKR
---------AAAAAAAEAEATAAKAPRCPDLNLDLCISPPCQQQEEEEVDLKPSAAVVKR
-------------TTKEEQLILSKPPKCPDLNLDLCISPPSCQEEDDDYEAKPAMIVRAP
VEPG---------FVQFSQNHHQQFVPATELSATSSNSPAETFSDVRGGVVNGSGYDPSG
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TaMyb1
Myb1
MybHv1
Zm38
R2R3 Myb protein
Arabidopsis
EA-----------VVGLCFSCSMGLPRSADCSAAAFMGL------------------EA-----------VVGLCFSCSMGLPRSADCKCSSFMGL------------------EAGVGV------GVVGLCFSCSMGLPRSSDCKCSSFMGL------------------EVLLGGRGHGHGHGGALCFGCSLGVQKGAPGCSCSSSNGHRCLGLRGGMLDFRGLKMK
ELQR--------RRGGLCFGCSLGLQKECKCS-------------------------QTGFG---------EFNDWGCVGGDNMWTDEESFWFLQDQFCPDTTSYSYN-------
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M
N1A-T1B
N3A-T3D
N4A-T4B
N5A-T5D
N6A-T6D
N7A-T7B
N1B-T1A
N2B-T2D
N3B-T3D
N5B-T5A
N6B-T6D
N7B-T7A
N1D-T1B
N2D-T2A
N3D-T3A
N3D-T3B
N5D-T5B
N6D-T6B
N7D-T7A
Dt3BL
Fig. 1. Amino acid sequence alignment of TaMyb1 with those
of other plant Myb transcription factors. Accession numbers:
TaMyb1 (wheat, DN828996), transcription factor Myb1 (wheat,
AAT37167), MybHv1 (barley, CAA50224), Myb-related protein
Zm38 (maize, P20025), typical P-type R2R3 Myb protein (rice,
A AL 84 62 8 ), A tMY B 2 (A rab id opsis ,
BAA03534).………………………………………………………….
.
.
TaMyb1
Fig. 2. Chromosomal localization of TaMyb1 in aneuploid lines
of Chinese Spring. …………………………………………………
Acknowledgement…………………………………………
This work was financially supported by the grant from the
Proton Engineering Frontier Project, KAERI, Republic of
Korea. This work was partially supported by a grant
(20050301-034-432-006-01-00) from BioGreen 21
Program, Rural Development Administration, Republic of
Korea.………………………………………………………….
* This study has been accepted in Physiologia Plantarum
(PPL-2006-00042.R2).
oxygen deficiency (pH)
1
4
5
0
T
2
1
4
5
6.50
oxygen deficiency+dark (pH)
0.120
0
C
2
TaMyb1
PDC
oxygen deficiency+dark+cutting (pH)
6.00
0.100
5.50
rRNA
pH
O2 concentration (mol m-3)
0.140
DOC (mol m-3):
0.080
0.042
0.037 0.033
0.011
0.011
5.00
0.060
4.50
0.040
0.020
4.00
0.000
3.50
0
1
2
4
5
Time (days)
D
C
0
Time (d)
• We have analyzed and report our results for the
tissue specific transcription of the Myb
transcription factor gene in wheat roots under
oxygen deficiency treatment and abiotic stresses.
--------MGRSPCCEKAHTNKGAWTKEEDDRLTAYIKAHGEGCWRSLPKAAGLLRCGKS
--------MGRSPCCEKAHTNKGAWTKEEDDRLTAYIKAHGEGCWRSLPKAAGLLRCGKS
--------MGRSPCCEKAHTNKGAWTKEEDDRLTAYIKAHGEGCWRSLPKAAGLLRCGKS
--------MGRSPCCEKAHTNRGAWTKEEDERLVAYIRAHGEGCWRSLPKAAGLLRCGKS
--------MGRSPCCEKEHTNKGAWTKEEDERLVAYIRAHGEGCWRSLPKAAGLLRCGKS
MEDYERINSNSPTHEEDSDVRKGPWTEEEDAILVNFVSIHGDARWNHIARSSGLKRTGKS
Time (d)
7.00
oxygen deficiency+dark (DO)
oxygen deficiency+dark+cutting (DO)
• In a previous study, we analyzed 1,344 clones
from the cDNA library of hypoxia-stressed wheat
roots and registered 1,274 ESTs in the GenBank
database (accession nos. DN828708 - DN829789
and DN948989 - DN949180). Several clones
including a clone (accession no. DN828996) that
shared high homology with a Myb transcription
factor gene exhibited various expressions in roots
under hypoxia.…..……………………………………..
TaMyb1
Myb1
MybHv1
Zm38
R2R3 Myb protein
Arabidopsis
B
1
2
4
E
Time (d)
TaMyb1
TaMyb1
rRNA
rRNA
DOC (mol m-3): 0.047
0
1
2
4
light DOC
L
0
12
18
21 (h)
N
rRNA
0.016
D
N
rRNA
Fig. 3. Oxygen deficiency treatment and Northern blot hybridization of TaMyb1. (A) Changes in O2 concentration and pH in the
solution as treatment time progresses. O2 is presented as a measure of the concentrations of dissolved O2 (mol m-3). The
values are means of three replicates ±s.e. (B) Northern blot hybridization of the TaMyb1 and PDC under oxygen deficiency
treatment. (C) Northern blot hybridization of the TaMyb1 under oxygen deficient condition combined with dark treatment. (D)
Northern blot hybridization of the TaMyb1 under oxygen deficiency, darkness, together with cutting treatment in the roots. (E)
Northern blot hybridization of the TaMyb1 under dark condition (normoxia, DOC: > 0.06 mol m-3) in the roots. h, hour(s); d,
day(s); C, control; T, treatment; D, dark conditions; L, light conditions; N, normoxia; DOC, dissolved oxygen concentration;
PDC, pyruvate decarboxylase. ………………………………………………………………………………………………………………..
T
C
MX
A
E
C
B
MX
AE
E
PC
C
D
PX
Time (h)
0
18
24
36
48
0
18
24
36
48
ABA
PC
E
AE
D
MX
MX
PC
D
PEG
AE
P
PX
Fig. 4. In situ hybridization of the TaMyb1 mRNA in radicle of
control and hypoxia-stressed wheat. Radicles from the
control (A) and anaerobic stressed (B, C, and D) plants were
collected under hypoxia (DOC: 0.011 mol m -3 ). Solid
arrowheads indicate a high positive signal. Magnifications
are ×400 (A and B) and ×1000 (C and D). AE, aerenchyma;
C, cortex; D, epidermis; E, endodermis; MX, metaxylem; P,
phloem; PC, pericycle; PX, protoxylem. Bars = 100 ㎛. …...
NaCl
Citric acid
rRNA
Fig. 5. Northern blot hybridization of TaMyb1 in seminal roots
treated with citric acid (10 mM), NaCl (250 mM),
polyethylene glycol (PEG, 25%) or ABA (100 µM). h, hours;
C, control; T, treatment. …………………………………………
Results and Discussion.…………………………………………………………………………….…………
• A clone (accession no. DN828996) contained a 750-bp open reading frame (ORF) encoding for the
putative Myb transcription factor with 250 amino acids. This clone was designated as TaMyb1 (Triticum
aestivum Myb transcription factor 1).……………………………………………………………………………..
• Presence of TaMyb1 on the 3BL was confirmed by using Chinese Spring aneuploid accessions
including nullisomic-tetrasomic and ditelosomic lines.….............……………………………………………...
• Dramatic increases in the transcripts of a TaMyb1 occurred under hypoxia. The transcriptional
expression of TaMyb1 was continued until approximate anoxia, being enhanced by light under hypoxia,
but little expression during anoxia could be shown by Northern blot hybridization. Under normoxic
condition, the presence or absence of light did not affect expression of TaMyb1.………………………....
• In situ hybridization of the hypoxic stressed radicle showed that TaMyb1 expression was occurred in
the epidermis, endodermis, and cortex which was in contact with the endodermis...………………………
• TaMyb1 transcription levels in roots gradually increased as the result of treatment with NaCl.………..
• The results of our Northern blot analysis and tissue specific hybridization revealed that TaMyb1
expression was strongly related to the oxygen concentration in root environment and the responses to
the abiotic stresses in the wheat roots. …………………………………………………………………………