Cotton leafstalk tissue cultivation and high-differentiation cotton material selective breeding method
A tissue culture, high differentiation rate technology, applied in horticultural methods, botanical equipment and methods, plant cells, etc., can solve the problems of genetic research and high differentiation rate material selection that cannot be used for differentiation rate, and shorten the transformation period. The effect of improving transformation efficiency and simple screening method
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Embodiment 1
[0023] Example 1, the screening of high differentiation rate strains in China Cotton Institute 24
[0024] Callus induction medium: Added indole acetic acid 0.1mg / L, kinetin 0.1mg / L, 2,4-D 0.1mg / L, glucose 30g / L, Gelrite 2mg / L on the basis of MSB basic medium L, pH 5.8.
[0025] Differentiation medium: Indole acetic acid 0.005mg / L, kinetin 0.005mg / L, glucose 30g / L, Gelrite 2mg / L, pH 6.8 were added on the basis of MSB basic medium.
[0026] Embryoid body germination and seedling growth medium: on the basis of MSB basic medium, kinetin 0.01mg / L, 6-BA 0.01mg / L, sucrose 30g / L, Gelrite 1.5-2.5mg / L, pH 6.8 were added.
[0027] The hormones and other components in the above-mentioned culture medium were all added to the culture medium before sterilization. Before the culture medium was sterilized, the pH value was adjusted with 1M KOH, and then at 121°C, 1.1kg / cm 2 High temperature and high pressure sterilization for 14min.
[0028] Taking Zhongmian Institute 24 as an example, the...
Embodiment 2
[0051] The breeding of high differentiation rate material in embodiment 2, Zhong 394
[0052] Use the same method as in Example 1 to screen 394 individual plants out of 150 strains, and then continue to use the same method to screen out strains with higher differentiation rates for the selected individual plants with high differentiation rates. More than 80% of the lines (Z1-Z7). In 2003, he went to Jiadai in Hainan. In 2004, he multiplied in the experimental field of China Cotton Research Institute in Anyang, Henan, and used the petiole as the explant to screen with the same method. As a result, the lines Z3 and Z7, Z1-Z7 with a differentiation rate of more than 90% were obtained. The differentiation rates of the strains are shown in Table 5.
[0053] material code
[0054] In 2004, the high differentiation rate Z7 was used as the female parent, and the hard-to-differentiate Zhongmiansuo 12 was used as the male parent, and F 1 Dai Dong sent Hainan Jiadai to get F...
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