ACQUISITION RELATED GENES INVOLVED IN EARLY STAGE IRON DEFICIENCY RESPONSE IN FOXTAIL MILLET (Setaria italica)
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Date
2017
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Indira Gandhi Krishi Vishwavidhyalaya, Raipur
Abstract
Foxtail millet (Setaria italica) is minor millet known to have high grain nutritive value especially iron compare to that of major staple cereal rice. Any increase in quality of millets might have significant role in combating micronutrient malnutrition for human health over the world. For the development of nutria rich rice, isolation of the genes involves in the metal homeostasis is the proven and successful approach for beneficial malnourished population. In our study 13 genes involved in the iron acquisition and transport were used for gene expression analysis by using semi quantitative RT-PCR. Leaf and root tissue were collected from Fe sufficient and Fe deficient grown Foxtail millet CO (Te)-7 plant at early stage. The expression level of VIT1,NAAT,FER1,FRO2,OsIRO2,YSL2 SiNAS2,and OsIDFE1genes show up regulation in Fe deficient leaves.VIT1 and Si022298 genes show up regulation in Fe deficient roots.NAAT,FER1,OsIRO2,YSL2, SiNAS2 and Si036196 genes show down regulation in Fe deficient roots.On the basis of above expression analysis we have selected SiNAS2 gene for further cloning and characterization using PCR based amplification. Amplification of full length SiNAS2 gene was performed on tissue collected from CO (Te)-7 genotype using SiNAS2 specifically designer primer. These result in generating an amplicon of 2.8 kb full length for genomic DNA. These amplified fragments were clone in Pcambia1301 followed by transformation of Agrobacterium cell. The positive clone was screen and conforms for SiNAS2 insert by colony PCR. The amplified fragments were sequence by Sanger di-deoxy chain termination method that results of generating 2262 bp sequence. This sequence was analyzed by nucleotide BLAST which showed high level of sequence similarity of SiNAS2 of with Zea mays nicotianamine synthase 2-like gene from maize (89% sequence similarity). The presence of functional domain analyzed at pfam database revealed the presence of two intact NAS domain in derived SiNAS2 protein sequence. Plant transformation and over expression studies using this gene construct will be useful for enhancing grain iron content in staple crops like rice and genetically simple solution to Fe deficiency disorders affecting billions of people throughout the world.
Description
ACQUISITION RELATED GENES INVOLVED IN EARLY STAGE IRON DEFICIENCY RESPONSE IN FOXTAIL MILLET (Setaria italica)
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Citation
ACQUISITION RELATED GENES INVOLVED IN EARLY STAGE IRON DEFICIENCY RESPONSE IN FOXTAIL MILLET (Setaria italica)