Abstract
【Objective】The study established the foundation to understand the role of RAR1 gene in wheat resistance to diseases, especially powdery mildew, as RAR1 is a required component in some signaling pathways mediated by disease resistance (R) genes in many plant species. 【Method】 RT-PCR and RACE methods were used to isolate the full-length cDNA sequence of the RAR1 gene from Thinopyrum intermedium,a wheat wild-relative with multi-resistance to various pathogens. The gene expression vector pUBI∷RAR1 was constructed and then transformed into a wheat variety ‘Yangmai 12’ by the biolistic particle method. The regenerated transgenic plants were detected through PCR, RT-PCR and test of the resistance to powdery mildew.【Result】The full-length cDNA of the RAR1 gene was isolated from Thinopyrum intermedium, named TiRAR1, which encodes the RAR1 protein possessing 2 CHORD domains and 1 CCCH domain. The sequence of the TiRAR1 protein shows high homology with the sequences of RAR1 proteins in barley and rice. The expression vector of TiRAR1 gene, pUBI::RAR1, was correctly constructed, and transformed into the wheat variety ‘Yangmai 12’. 1545 immature embryos of ‘Yangmai 12’ were bombarded by the particle containing pUBI∷RAR1. From the regenerated 154 plants, 18 positive transgenic individuals were identified by PCR assay in the T0 generation with a transformation frequency of 1.17%. The results of PCR and RT-PCR assay showed that the RAR1 gene could be inheritable from T0 generation to T1 one, then from T1 generation to T2 one, and could be transcribed and expressed. The resistance test showed that the RAR1 transgenic individuals in T1 and T2 generations showed high resistance to powdery mildew in their all lifecycle, suggesting that the overexpression of RAR1 enhanced the resistance to powdery mildew. 【Conclusion】The full-length cDNA of the RAR1 gene in Thinopyrum intermedium was isolated and transformed into a wheat variety ‘Yangmai 12’. The RAR1 gene could confer high resistance to powdery mildew when overexpressed, and could be used as a potential gene in wheat breeding with broad spectrum resistance to powdery mildew.
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Claims 8
Claim · model-written summary
TiRAR1 full length cDNA was isolated from Thinopyrum intermedium.
- Taxon
- Thinopyrum intermedium
Thinopyrum intermediume0 - Gene
- TiRAR1e0
- Outcome
- full-length cDNAe0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The full-length cDNA of the RAR1 gene was isolated from Thinopyrum intermedium, named TiRAR1”
Claim · model-written summary
TiRAR1 encodes a protein with two CHORD and one CCCH domain.
- Taxon
- Thinopyrum intermediumThinopyrum intermediumt0
- Gene
- TiRAR1e0
- Outcome
- 2 CHORD domains and 1 CCCH domaine0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“TiRAR1, which encodes the RAR1 protein possessing 2 CHORD domains and 1 CCCH domain”
t0“Thinopyrum intermedium”
Claim · model-written summary
TiRAR1 protein sequence is highly homologous to barley and rice RAR1 proteins.
- Taxon
- Thinopyrum intermediumThinopyrum intermediumt0
- Gene
- TiRAR1e0
- Outcome
- high homologye0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The sequence of the TiRAR1 protein shows high homology with the sequences of RAR1 proteins in barley and rice”
t0“Thinopyrum intermedium”
Claim · model-written summary
The TiRAR1 expression vector was correctly constructed and introduced into Yangmai 12 wheat.
- Taxon
- wheat
Triticum aestivume0 - Material
- ‘Yangmai 12’e0
- Gene
- TiRAR1e0
- Outcome
- correctly constructed, and transformede0
Not reportedComparatorValue
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The expression vector of TiRAR1 gene, pUBI::RAR1, was correctly constructed, and transformed into the wheat variety ‘Yangmai 12’”
Claim · model-written summary
Eighteen positive T0 transgenic wheat individuals were identified, with 1.17% transformation frequency.
- Taxon
- wheatTriticum aestivumt0
- Organ / stage
- T0 generatione0
- Outcome
- transformation frequencye0
- Value
- 1.17%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“From the regenerated 154 plants, 18 positive transgenic individuals were identified by PCR assay in the T0 generation with a transformation frequency of 1.17%”
t0“Wheat”
Claim · model-written summary
RAR1 was inherited across T0 to T2 generations and expressed.
- Taxon
- WheatTriticum aestivumt0
- Gene
- RAR1e0
- Outcome
- inheritable from T0 generation to T1 one, then from T1 generation to T2 one, and could be transcribed and expressede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the RAR1 gene could be inheritable from T0 generation to T1 one, then from T1 generation to T2 one, and could be transcribed and expressed”
t0“Wheat”
Claim · model-written summary
RAR1 transgenic T1 and T2 wheat showed high resistance to powdery mildew throughout their life cycle.
- Taxon
- WheatTriticum aestivumt0
- Treatment
- overexpression of RAR1e0
- Interactor
- powdery mildewe0pathogen
- Gene
- RAR1e0
- Outcome
- resistance to powdery mildewe0
- Direction
- enhancede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“the RAR1 transgenic individuals in T1 and T2 generations showed high resistance to powdery mildew in their all lifecycle, suggesting that the overexpression of RAR1 enhanced the resistance to powdery mildew”
t0“Wheat”
Claim · model-written summary
The authors suggest TiRAR1 as a potential gene for breeding wheat resistance to powdery mildew.
- Taxon
- wheatTriticum aestivume0
- Interactor
- powdery mildewe0pathogen
- Gene
- RAR1t0
- Outcome
- wheat breeding with broad spectrum resistance to powdery mildewe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“could be used as a potential gene in wheat breeding with broad spectrum resistance to powdery mildew”
t0“RAR1 Gene”