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Is My Gene Important for Seed Development in Plants??

Is My Gene Important for Seed Development in Plants??. Gene: AT3G53370. Jonathan Milgrom Spring 2004. What Does My Gene Do?. Transcription Factor* Repressor of Spinach gene rps1 Active in roots Highly conserved in plant kingdom. Transcription factor. *Dao-Xiu Zhou, Cordelia Bisanz Seyer.

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Is My Gene Important for Seed Development in Plants??

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  1. Is My Gene Important for Seed Development in Plants?? Gene: AT3G53370 Jonathan Milgrom Spring 2004

  2. What Does My Gene Do? Transcription Factor* • Repressor of Spinach gene rps1 • Active in roots • Highly conserved in plant kingdom Transcription factor *Dao-Xiu Zhou, Cordelia Bisanz Seyer

  3. Is This Function Integral to Seed Development ? Knockout the gene and find out! • Engineer agrobacteria to insert tDNA • Grow mutant seeds • Genotype AT3G53370 tDNA Can an Arabidopsis plant develop without a functioning form of my gene? Agrobacteria naturally performs this task

  4. Madison & Salk Provide the Knockouts Madison SP#4 Salk Exon SP#1 5’ 3’ AT3G53370

  5. Madison project began with… Superpool Salk project began with… Mutant seeds Knockout Analysis Superpool tDNA line/ Mutant Seed PCR products amplified with RV (gene specific) and tdna primers 4/15/04 Plant Genotype Sp #4 (small length suggests insertion)

  6. SP #4 DNA pool #6 Madison Results Identified Madison DNA Pool tDNA SP#4 Madison PCR products amplified with RV (gene specific) & tdna primers 5/18/04 RV 3 3’ 5’ AT3G53370

  7. Plant # genotype 1 4 5 6 7 8 9 Wt C 1 4 5 6 7 8 9 Homozygous tDNA * * * Heterozygous * * Wild Type * * Wt 1456789 C SALK Results • Homozygous tDNA exist • Knockout is not embryo lethal 7 Plants Genotyped… Tubulin PCR products of various SALK lines amplified with FW & RV (gene specific) primers 6/4/04 PCR products of various SALK lines amplified with RV (gene specific) and tDNA primers 6/1/04 Wildtype

  8. Does My Gene Play Some Role in Seed Development? Gene activity/mRNA accumulation patterns Scarlet Runner Bean equivalent Arabidopsis RT-PCR of 6 organs Lec1 mutant 14 day old embryo

  9. Next…? • Study rsp1 spinach gene equivalent in Arabidopsis • Study lec1 gene • Identify Madison tDNA line and Genotype

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