Method and means for determining fitness in plants
View Patent ↗The present invention provides methods and means for determining parent inbred plant lines with good combining ability, for determining good combinations of parent inbred plant lines capable of yielding hybrid lines with high heterosis, and further for determining the agronomical performance of different plant lines, which can be performed in vitro by determining the electron flow in the mitochondria under control and stress conditions.
1. A method for producing a hybrid Brassica plant line, comprising the steps of:
a) assaying a collection of Brassica inbred plant lines of interest by
i) culturing a population of explants of each of said Brassica inbred plant lines of said collection on a medium comprising sucrose or on callus inducing medium comprising sucrose and plant hormones for a period of 0 to 10 days;
ii) measuring the electron flow in the mitochondrial electron transport chain in said population of explants, relative to the electron flow in the mitochondrial electron transport chain in a population of explants from a reference plant line of the same species;
b) assaying said collection of inbred lines of interest by
i) culturing a population of explants of each of said inbred Brassica plant lines of said collection on a medium comprising sucrose or on callus inducing medium comprising sucrose and plant hormones;
ii) incubating a first part of each of the populations of said cultured explants in a buffer solution;
iii) incubating a second part of each of the populations of said cultured explants in said buffer solution, further comprising a salicylic acid derivative capable of generating oxidative stress in plant cells incubated in a solution of said salicylic acid derivative;
iv) measuring the electron flow in the mitochondrial electron transport chain in each of said first and second parts of said populations;
c) selecting one inbred plant line which has a higher relative amount of electron flow in the mitochondrial electron transport chain when compared to other inbred lines from said collection as determined in step (a) and which has a negative difference between the measured electron flow in the mitochondrial electron transport chain between said second and said first part as measured in step (b);
d) crossing said selected inbred plant line with a second inbred plant line;
e) collecting seed from said crossed selected inbred plant line.
2. The method of claim 1 , wherein said second inbred line has a higher relative amount of electron flow in the mitochondrial electron transport chain when compared to other inbred lines from said collection and has a negative difference between the measured electron flow in the mitochondrial electron transport chain between said second and said first part.
3. The method any one of claim 1 or 2 , wherein said parts of said population of cultured explants are incubated in said buffer solution for about 18 hours.
4. The method of claim 1 , wherein said buffer solution comprises about 25 mM K-phosphate and about 2 to 3% sucrose.
5. The method of claim 1 , wherein said salicylic acid derivative is acetylsalicylic acid.
6. The method of claim 5 , wherein said acetylsalicylic acid is present in said buffer solution in a concentration of about 25 mg/L.
7. The method of claim 1 , wherein said explant is a hypocotyl.
8. The method of claim 1 , wherein said reference plant line of the same species is a reference plant line of the same variety.
9. The method of claim 1 , wherein said period is 4 to 6 days.
10. The method of claim 1 , wherein said period is 5 days.