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0 10 25 50 100 + Suc + ABA - Suc + ABA ABA (  M) OsAGPL3 rRNA Control 0 10 25 50 100 Fig1. Accumulation of OsAGPL3 mRNA in response to ABA Pre-cultured.

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Presentation on theme: "0 10 25 50 100 + Suc + ABA - Suc + ABA ABA (  M) OsAGPL3 rRNA Control 0 10 25 50 100 Fig1. Accumulation of OsAGPL3 mRNA in response to ABA Pre-cultured."— Presentation transcript:

1 0 10 25 50 100 + Suc + ABA - Suc + ABA ABA (  M) OsAGPL3 rRNA Control 0 10 25 50 100 Fig1. Accumulation of OsAGPL3 mRNA in response to ABA Pre-cultured cells were transfer to the medium supplemented with various concentrations of ABA as indicated. After 24 h of cultured in the dark, suspension cells were collected and total RNAs were isolated. OsAGPL3 (accession no. AK100910) mRNA levels were measured by northern blot analysis.

2 0 1 3 6 12 24 ( h ) OsAGPL3 Fig2. Time course of OsAGPL3 mRNA accumulation in response to sucrose and ABA. Pre-cultured cells were transfer to medium supplemented with 3%(w/v) sucrose + 50  M ABA. Expression of OsAGPL3 at different time points was assessed by Northern blot analysis. Time after transfer rRNA

3 Fig3. Synergistic effect of sucrose and ABA on OsAGPL3 mRNA accumulation. Pre-cultured suspension cell was used as control (control). Pre-cultured cells were transfer to medium supplemented with either 50 -  M ABA (+ABA) or 3% (w/v) sucrose (+Suc) alone, or 3%(w/v) sucrose plus 50  M ABA (+Suc+ABA). After 6 h of culture, suspension cells were collected and total RNAs were isolated. OsAGPL3 mRNA levels were measured by Northern blot analysis. control + ABA + Suc + Suc+ ABA OsAGPL3 rRNA Relative transcripts level (%) 120 100 80 60 40 20 0

4 Starch contents mg/100mg DW +Suc+Suc+ABA +ABA Control Fig4. Synergistic Effect of sugars and ABA on starch content in rice suspension cell. Pre-cultured cell was used as control (control). Pre-cultured cells were transfer to medium supplemented with either 50  M ABA (+ABA) or 3%(w/v) sucrose (+Suc) alone, or 3% (w/v) sucrose plus 50  M ABA (+Suc+ABA). After 6 h of culture, cultured cell were collected and starch contents were determined. 6 h 12h

5 Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA Control +ABA +Suc +Suc+ABA 9799 63 321 189 66 213 94 The number of bands Control +ABA +S +S+ABA (A) (B) Fig5 cDNA-AFLP display of gene expression in rice cultured cell. (A) A typical autoradiogram showing the TDFs pattern of rice cultured cells. RNAs were isolated from pre-cultured cell (control), 50  M ABA (+ABA) or 3% (w/v) sucrose (+S) alone, or 3% (w/v) sucrose plus 50  M ABA (+S+ABA) treated cells. Primer combinations for restriction endonucleases AvaII and TaqI are shown at the top of the cDNA-AFLP autoradiogram. (B) Classification of TDFs by their induction patterns. Total 11099 TDFs were identified. In total, 9799 TDFs were not different between each samples. 63, 321, 66 and 94 TDFs were increased in the cultured cells grown in the +S+ABA, + ABA, +S and +S+ABA medium, respectively. AvaII TC TC TC TC TC TC TC TC TaqI AA AT AC AG GA GT GC GG

6 TDF38 TDF41 TDF54 TDF59 TDF64 TDF65 TDF69 TDF1 TDF5 TDF9 TDF10 TDF21 TDF56 TDF67 TDF6 TDF22 TDF24 TDF28 TDF29 TDF32 TDF80 TDF79 TDF23 TDF15 TDF47 TDF75 +ABA +Suc+ABA +Suc Control Fig6 Northern blot analysis of 26 genes which is identified by cDNA-AFLP as synergistically induced by sucrose and ABA. Rice cultured cell were cultured in the medium supplement with 50  M ABA (+ABA), 3% (w/v) Sucrose ( +Suc) and 3% (w/v) Sucrose and 50  M ABA (+Suc+ABA) for 6h. Pre- cultured cell was used as control (Control). Ten  g of total RNAs from cultured cells were loaded in each lane, The blot were hybridized with 32P-labeled DNA probes. 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 (A) (B) +ABA +Suc+ABA +Suc Control

7 INV Suc Hexose HXT Hexose Hexose-P ADPG Hexose-P ADPG Starch OsAGPL3 GPT AGPase Pi ? Plastid Cytosol

8 Chloroplast Acetate Acetate-CoA C16:0 ACP C18:0 ACP C18:1 ACP fab2 C18:1 COA Endoplasmic reticulum PA 18:1 DAG PC P-EAP-MEA P-DEA P-choline PEAMT 18:1 PC 18:2 18:3 PC fad2

9 chloroplast Acetate -CoA C16:0 ACP C18:0 ACP C18:1 ACP C18:1 GLC18:2 GL fad6 fab2 C18:1 COA Endplasmic reticulum PA 18:1 DAG 18:1 PC 18:1 PC 18:2 PC 18:3 fad3 fad2 P-AI 18:1 P-EA 18:1 P-EA P-MEA P-DEA P-choline CDP-choline phosphoethanolamine N- methyltransferase

10 *1; Analyzable cDNAs, which containing both AvaII and TaqI site. *2; Unanalyzable cDNAs, which dose not containing either AvaII or TaqI or Both site. *1 *2 32127 24081 8046 70824 32127 18377 13751 36590 256 128 Total number of full length cDNAs Analyzable cDNAs Unanalyzable cDNAs Total number of TDF s Table1 In silico cDNA-AFLP analysis of Oryza sativa primers combinations

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