Barnase Barstar Gene of male sterility

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Presentation Transcript

Slide 1: 

Barnase gene…produces RNAse Barnase gene…produces RNAse TA29 Barnase gene Tissue specific (tapetum layer) promoter Herbicide Resistance (Phosphinothricin) (Gene construct) Endothecium Middle layer Tapetum Bacillus amyloliquefaciens Tapetum degenerates PMCs/pollen become sterile BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY PRINCIPLE © A.K. Chhabra 2007

Slide 2: 

Tapetum persistant PMCs/pollen are fertile Barnase gene…produces RNAse Barstar gene TA29 Barnase gene Tissue specific (tapetum layer) promoter Herbicide Resistance (Phosphinothricin) (Gene construct) Inhibits RNAse Endothecium Middle layer Tapetum Bacillus amyloliquefaciens BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY PRINCIPLE © A.K. Chhabra 2007

Slide 3: 

Barnase gene…produces RNAse Barstar gene (fertility restorer) TA29 Barnase gene Tissue specific (tapetum layer) promoter Herbicide Resistance (Phosphinothricin) (Gene construct) Inhibits RNAse Endothecium Middle layer Tapetum Bacillus amyloliquefaciens Tapetum degenerates PMCs/pollen become sterile Tapetum persistant PMCs/pollen are fertile BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY PRINCIPLE Dominant male sterility Barstar-Barnase interaction (binding) © A.K. Chhabra 2007

Slide 4: 

Barnase gene Tissue specific (tapetum layer) promoter Herbicide Resistance (Phosphinothricin) (Gene construct) BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY Male sterile Herbicide resistant Male Fertile Herbicide sensitive Herbicide spray PRINCIPLE © A.K. Chhabra 2007

Slide 5: 

BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY Male Fertile KILLED PRINCIPLE © A.K. Chhabra 2007

Slide 6: 

BARNASE-BARSTAR SYSTEM TRANSGENIC GENETIC MALE STERILITY Barnase-Bar/- - /- x Sterile Herbicide Resistant Fertile Herbicide Sensitive 50% Barnase-Bar/- (male sterile, rest.) 50% -/- (male fertile, susceptible) Phosphinothricin Spray Killed , Killed , Killed , Killed X Barstar/Barstar Fertile Restorer 50% Barnase-Bar/Barstar (male fertile) 50% -/Barstar (male fertile) 100% male fertile F1 Hybrid STEP 1 Maintenance of male sterility STEP 2 Production of Hybrid © A.K. Chhabra 2007

CHEMICALLY INDUCED MALE STERILITY : 

CHEMICALLY INDUCED MALE STERILITY MALE STERILE MALE STERILE MALE STERILE MALE FERTILE

CHEMICALLY INDUCED MALE STERILITY : 

CHEMICALLY INDUCED MALE STERILITY Induced by spray of certain chemicals GA (gibberellic acid) NAA (napthalene acetic acid) MH (Maleic Hydrazide) Etheral MALE GAMETOCIDES CHAs Chemical Hybridizing Agents SUCCESSFUL EXAMPLES CHEMICAL CROP Etherel Rice, Sugarbeet, Wheat FW 450 Cotton, Groundnut, Sugarbeet, Tomato GA (Gibberellic Acid) Lettuce, Maize, Onion, Rice, Sunflower MH (Maliec Hydrazide) Cucurbits, Onion, Tomato, Wheat NAA (Napthelene Acetic Acid) Cucurbits Sodium Methyl Arsenate Rice Zinc Methyl Arsenate Rice

CHEMICALLY INDUCED MALE STERILITY : 

CHEMICALLY INDUCED MALE STERILITY Characteristics of an Ideal CHA Female fertility should be unaffected Be non-mutagenic Consistent complete male sterility Easy and economical to apply Minimum side effects Safe to user and envt. NO SUCH CHA AVAILABLE Advantages of using CHAs Effective for the generation applied No transfer of CMS required Can be applied to the variety required only B and R are not required Saves time, labour and money to develop A, B, and R lines

CHEMICALLY INDUCED MALE STERILITY : 

CHEMICALLY INDUCED MALE STERILITY Chief limitations Pollen abortion------incomplete and variable Specific developmental stages are sensitive to CHAs Effect of CHAs are short lived Female fertility may be reduced Often undesirable side effects are produced High cost of CHAs The need for repeated applications increases the cost of hybrid seed

Transfer of cytoplasm from male sterile to a new strain : 

S Transfer of cytoplasm from male sterile to a new strain 50 % rr F rr X rr

Slide 12: 

75 % rr F X rr Transfer of cytoplasm from male sterile to a new strain S

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 87.5 % rr F X rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 93.75 % rr F X rr rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 96.87 % rr F X rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 98.94 % rr F X rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 99.47 % rr F X rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 99.73 % rr F X rr

Transfer of cytoplasm from male sterile to a new strain : 

Transfer of cytoplasm from male sterile to a new strain 99.87 % rr F X rr

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