Method for Inducing Tobacco Embryoid Body with Lingfa
A technology of stimulant and embryoid body, which is applied in the field of plant bioengineering tissue culture, can solve the problems of complex procedures, low efficiency, and long induction period of tobacco embryoid body, and achieve fast formation speed, good induction effect, and high yield and the effect of quality improvement
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Embodiment 1
[0019] A method for inducing tobacco embryoid bodies with linguine, comprising the following steps:
[0020] Step 1. Preparation of induction medium: use MS as the basic medium, add 3.0 mg / L of lingfalin, adjust the pH of the medium to 5.8-6.0, and sterilize it for later use;
[0021] Step 2, explant preparation: take the sterile tobacco plants obtained from tobacco seeds, under aseptic conditions, cut the tobacco leaves into square slices as explants, and set aside, the square slices can be cut into about 5mm×5mm in size ;
[0022] Step 3, inoculation and induction culture: insert the explants of tobacco outer leaves obtained in Step 2 into the induction medium of Step 1, and culture for 25 days.
Embodiment 2
[0024] Step 1. Preparation of induction medium: use MS as the basic medium, add 4.5 mg / L of lingfalin, adjust the pH of the medium to 5.8-6.0, and sterilize it for later use;
[0025] Step 2, explant preparation: take the sterile tobacco plant obtained from tobacco seeds, when it grows to 5 or 6 leaves, take the third leaf from top to bottom, and cut it into square pieces under aseptic conditions As an explant, spare, the square piece can be cut into a size of about 5mm×5mm;
[0026] Step 3, inoculation and induction culture: insert the explants of tobacco outer leaves obtained in Step 2 into the induction medium of Step 1, and culture for 25 days.
[0027] When the sterile tobacco plant grows to 5 or 6 leaves, the third leaf from top to bottom is a fully mature and robust functional leaf, with vigorous cell metabolism and strong vitality. Differentiate and redifferentiate to form embryoid bodies.
Embodiment 3
[0029] Step 1. Preparation of induction medium: use MS as the basic medium, add 5.0 mg / L of Lingfalin, adjust the pH of the medium to 5.8-6.0, and sterilize it for later use;
[0030] Step 2, explant preparation: take the sterile tobacco plant obtained from tobacco seeds, when it grows to 5 or 6 leaves, take the third leaf from top to bottom, and cut off 2 mm above the petiole under aseptic conditions and the blade part of 2mm below the tip, leave the middle part of the blade, and cut it into 5mm × 5mm square slices along the two sides of the main vein as explants for future use. It should be noted that the aforementioned cutting along the main vein is Refers to discarding the leaves on both sides of the leaves near the edge, as long as the leaves on both sides of the main vein;
[0031] Step 3, inoculation and induction culture: insert the explants of tobacco outer leaves obtained in Step 2 into the induction medium of Step 1, and culture for 25 days.
[0032] When the steri...
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