Identification of Tawd40a, a Wheat WD40 Repeat-Containing Protein Important for Plant Tolerance

Identification of TaWD40AWD40D, a wheat WD40 repeat-containing protein important for plant tolerance to abiotic stresses

Dejing Kong · Mengjun Li · Zhanghui Dong · Hongtao Ji · Xia Li

D. Kong ·M. Li ·Z. Dong ·H. Ji ·X. Li ()

The State Key Laboratory of Plant Cell Chromosome Engineering, Center of Agricultural Resources Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 286 Huaizhong Road, Shijiazhuang, Hebei 050021, China

e-mail:

Dejing Kong

University of Chinese Academy of Sciences, 19A Yuquan Road, Beijing 100049, China

Present Address:

M. Li

Shijiazhuang Academy of Agriculture and Forestry Sciences, 479 Shenglibei Street, Shijiazhuang, Hebei 050021, China

Present Address:

Z. Dong

Shijiazhuang Academy of Agriculture and Forestry Sciences, 479 Shenglibei Street, Shijiazhuang, Hebei 050021, China

Supplementary Material

Fig. S1. Chromosomal location of TaWD40D. NT1A1B to NT7A5D are 21 nulli-tetrasomic (NT) lines of Chinese Spring lacking the specified chromosome. CS is the Chinese Spring (Triticum aestivum L.). Markers are 250 bp DNA ladder containing 6 bands from 100-2000 base pairs. TaWD40D was not amplified by NT4D4B.

Fig.ureS1S2.The three-dimensional structure of TaWD40AWD40D protein. The Seven-bladed (1-7) β-propeller structure was predicted using PyMOL ( and four stranded antiparallel β sheet structure (A-D).

Fig.ure S32.TaWD40AWD40Doverexpression lines are comparable to the wild type at different developmental stages.

The seeds of the TaWD40AWD40Doverexpression lines and the controls were germinated on MS medium and the 6 day-old seedlings after stratification were transplanted to pots. The pictures were taken at 15 days (A), 30 days (B) and 60 days (C) after transplantation; (D) The siliques of the plants. The representive plants were shown. EV, empty vector transgenic line; OE1 and OE2, TaWD40AWD40D overexpression lines; WT, wild type.

Fig.ure S43. Expression of stress-responsive genes in the wild type and TaWD40AWD40D overexpression linesunder normal or stress conditions.

The expression of stress-responsive genes in the Arabidopsis plants overexpressing TaWD40AWD40D. Six day-old seedlings were exposed to either 200 mM NaCl (A-H), 300 mM mannitol (I-P), or 100 µM ABA (Q-X) for 3 h. Gene-specific primers were used to detect the relative transcript levels of stress-responsive genes. The Arabidopsis Actin2 gene was used as an internal control.Under normal conditions (Control), the gene expression in WT was regarded as standard, and EV, OE1, and OE2 values were compared with it. Under the stress conditions, the gene expression in WT was regarded as standard, and EV, OE1, and OE2 values were compared with it. The gene expression of each genotype under normal conditions was compared with the genotype under the stress condition, respectively. Error bars represent the standard deviation among three biological replicates. Significant differences (T-test, *P < 0.05) are marked with asterisks. EV, empty vector transgenic line. OE1 and OE2, TaWD40AWD40D overexpression transgenic lines. WT, wild type.

Table S1.Primers used in this study

Genes / Forward primers / Reverse primers
TaWD40AWD40D-EST / GCACGAGGGGACAACCCCGC / TCGACCCAGTACCAGTAGGT
TaWD40D-locationTaWD40A-genomic / TTCGTGTGAACTGTTGTGGTGGGTAACCACCAGAA / CGTCAACTACACAGACCAGATGTTGAAGGTCCTGTCCC
TaWD40AWD40D-cDNA / ATGGGTAACCACCAGAAGCTCC / TCACGCCGCTGGCACGCGCTTCG
TaWD40AWD40D-RT-PCR / TCCTCCTCCTCCGCCT / GCCACGGTGTTGATGG
TaWD40AWD40D-qRT-PCR / CGGTGTGGCTTCGGCA / GCTTCGTGGCGTCGCT
TaWD40AWD40D-VIGS / AAGGAAGTTTAAGTGGGACAGGACCTTCAA / AACCACCACCACCGTAGCACCTTCTCCAGGACAT
Wheat tubulin / CTCCCTCACAACAACCGC / TACCAGGAACTTCCATACCAAC
TaPDS / AAGGAAGTTTAATTTCTCCAGGAGAAG / AACCACCACCACCGTTCTCCAGTTATTTGAG
Arabidopsis actin2 / AAGCAATGAGCATGAGCAAG / GGAAGACACGACAGGAAACAC
RAB18 / GGAGAAGTTGCCAGGTCATC / ACCGGGAAGCTTTTCCTTGATC
DREB2A / AAACCTGTCAGCAACAACAGCAGG / TTAAGCCTGCAAACACATCGTCGC
ABI2 / GTTCTTGTTCTGGCGACGGAGC / CCATTAGTGACTCGACCATCAAG
SOS2 / GGCTTGAAGAAAGTGAGTCTCG / GCTACATAGTTCGGAGTTCCACA
SOS3 / CATTCACGGTAGAAGAAGTGGA / GCTTGGATGGAAGACACCTAA
ABI1 / AGAGTGTGCCTTTGTATGGTTTTA / CATCCTCTCTCTACAATAGTTCGCT
RD29A / GGAAGTGAAAGGAGGAGGAGGAA / CACCACCAAACCAGCCAGATG
RD29B / GAATCAAAAGCTGGGATGGA / TGCTCTGTGTAGGTGCTTGG
SOS1 / ATTTCTGGAGGAAGCGACC / GCGACAGTGTAAGGAACCCT
ABA1 / GCAATGGTATGCATTTCACG / CATGCTTCATCAAGCTCCAA
DREB1A / CGGTGGCTGATAGGCTAAAG / CCGCCTTATTTTCAACCGTA