Obtaining and analysis of flanking sequences from T-DNA transformants of Arabidopsis

Qin, Genji; Kang, Dingming; Dong, Yiyu; Shen, Yulong; Zhang, Li; Deng, Xiaohui; Yao, Zhen; Song, Li; Chen, Nan; Niu, Weidong; Chen, Cong; Liu, Peicheng; Chen, Haodong; Li, Jigang; Ren, Yanfei; Gu, Hongya; Deng, Xing‐Wang; Liu, Qu; Chen, Zhangliang

doi:10.1016/s0168-9452(03)00218-8

Cited by 56 publications

(82 citation statements)

References 24 publications

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“…The yid1 mutant is hypersensitive to iron-deficient conditions Using the T-DNA insertional mutant collection (Qin et al, 2003), we identified a chlorotic leaf mutant that displayed chlorosis between veins (Figure 1a), similar to the chlorosis phenotype caused by iron deficiency. We named this mutant yid1.…”

Section: Resultsmentioning

confidence: 87%

“…The yid1 mutant was screened from the T-DNA insertional mutant collection (Weigel et al, 2000;Qin et al, 2003). The mutants sfr6-2 and med25-2 were described previously (Knight et al, 2009;Xu and Li, 2011).…”

Section: Experimental Procedures Plant Growth Conditions and Iron-defmentioning

confidence: 99%

“…Thermal asymmetric interlaced-PCR was used to isolate the flanking sequence of the T-DNA insertion in the yid1 mutant with the specific and arbitrary degenerate primers as described previously (Qin et al, 2003). Three primers, P1, P2 and DL3, were used for co-segregation analysis.…”

Section: Thermal Asymmetric Interlaced-pcr and Plant Transformationmentioning

confidence: 99%

“…We report the isolation and characterization of an iron-deficient chlorotic mutant, yellow and sensitive to iron-deficiency 1 (yid1), from a T-DNA insertion mutant collection (Qin et al, 2003). The yid1 mutant was more sensitive to iron deficiency than wild-type plants.…”

Section: Introductionmentioning

confidence: 99%

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The Arabidopsis Mediator subunit MED16 regulates iron homeostasis by associating with EIN3/EIL1 through subunit MED25

et al. 2014

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These authors contributed equally to this work. SUMMARYIron is an essential micronutrient for plants and animals, and plants are a major source of iron for humans. Therefore, understanding the regulation of iron homeostasis in plants is critical. We identified a T-DNA insertion mutant, yellow and sensitive to iron-deficiency 1 (yid1), that was hypersensitive to iron deficiency, containing a reduced amount of iron. YID1 encodes the Arabidopsis Mediator complex subunit MED16. We demonstrated that YID1/MED16 interacted with another subunit, MED25. MED25 played an important role in regulation of iron homeostasis by interacting with EIN3 and EIL1, two transcription factors in ethylene signaling associated with regulation of iron homeostasis. We found that the transcriptome in yid1 and med25 mutants was significantly affected by iron deficiency. In particular, the transcription levels of FIT, IRT1 and FRO2 were reduced in the yid1 and med25 mutants under iron-deficient conditions. The finding that YID1/MED16 and MED25 positively regulate iron homeostasis in Arabidopsis increases our understanding of the complex transcriptional regulation of iron homeostasis in plants.

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Section: Resultsmentioning

confidence: 87%

Section: Experimental Procedures Plant Growth Conditions and Iron-defmentioning

confidence: 99%

Section: Thermal Asymmetric Interlaced-pcr and Plant Transformationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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The Arabidopsis Mediator subunit MED16 regulates iron homeostasis by associating with EIN3/EIL1 through subunit MED25

et al. 2014

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“…The DNA fragments amplified by TAIL-PCR were recovered from agarose gels and purified before sequencing using an ABI 377 automated DNA sequencer (Applied Biosystems, USA). The output sequences were then searched against the Arabidopsis genome sequence database (GenBank) using BLAST to localize the position of the T-DNA insertion as described previously [25]. …”

Section: Identifying the Flanking Sequences Of The Insertionsmentioning

confidence: 99%

Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis

et al. 2007

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Transformation by T-DNA integration causes highly sterile phenotype independent of transgenes in Arabidopsis thaliana

Ge¹,

Hai²,

Chen³

2008

Plant Cell Rep

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Agrobacterium tumefaciens-mediated gene transformation caused highly sterile phenotype in T1 transgenic populations of Arabidopsis thaliana. The phenomenon occurred independent of the genes and construct types used for transformation. The occurring frequency is less than 10% and the phenotype is inheritable. Intensive examination elucidated that the sterility is due to indehiscence or delayed dehiscence of the anthers during the flowering stage, resulting from the reduced or disordered endothecial secondary wall thickening of the anthers in the sterile flowers. Exogenous jasmonic acid application cannot rescue the sterile phenotype. Additionally, by using the Escherichia coli uidA gene encoding the beta-glucuronidase as a reporter gene, we indicated that the Cauliflower mosaic virus 35S promoter was not constitutively active as expected previously in the reproductive organs of Arabidopsis. These results contribute significantly to the plant community by suggesting that more careful examination and statistical analysis are needed while studying gain-of-function phenotypes of genes, especially for genes that might be involved in reproductive development.

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Obtaining and analysis of flanking sequences from T-DNA transformants of Arabidopsis

Cited by 56 publications

References 24 publications

The Arabidopsis Mediator subunit MED16 regulates iron homeostasis by associating with EIN3/EIL1 through subunit MED25

The Arabidopsis Mediator subunit MED16 regulates iron homeostasis by associating with EIN3/EIL1 through subunit MED25

Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis

Transformation by T-DNA integration causes highly sterile phenotype independent of transgenes in Arabidopsis thaliana

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