Cut culture of indica rice
A technology of in vitro culture and indica rice, applied in the field of new plants, can solve the problems of test failure, continuous subculture, slow growth of callus, etc., and achieve the effect of wide pertinence and applicability
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Embodiment 1
[0019] Embodiment 1: Screening of indica rice callus subculture medium
[0020] The experimental design is divided into two parts: 1. Using MS, N 6 , B 5 and N 6 B (the medium is N 6 macronutrients and B 5 Trace and organic nutrients) based on the four most commonly used basic media (shown in Table 5), set 1 times (indicated by X, the same below), 2X, and combine with two carbon sources of sucrose and maltose to form 16 2. Based on MS macronutrients and maltose as carbon source, set MS, N 6 , B 5 trace elements, each trace element set four levels of 1X, 2X, 5X, 10X, Fe 2+ Four gradients of 1X, 1.5X, 3X, and 5X were set up to form 48 culture media. A total of 64 designed media were inoculated with indica rice callus for subculture, and each medium was inoculated with 5 bottles of callus. Incubate in the dark at 26±1°C for 20 days. By observing and comparing the growth conditions (growth speed, texture and color) of the callus on various media, the subculture medium (co...
Embodiment 2
[0021] Embodiment 2: Comparative test of indica rice callus subculture effect
[0022] The test uses four representative indica rice varieties that are widely planted in China and have large genotype differences: Minghui 63 (a three-line indica restorer line), Zhenshan 97B (a three-line indica maintainer line) ), Zhong 419 (a three-line or two-line restorer line of indica rice) and W9864S (a two-line sterile line of indica rice). J of the present invention 3 The medium and the other three are commonly used basic medium MS (coded as J 1 ), N 6 (No. J 2 ) and B 5 (No. J 4 ) composed of subculture medium, the additional components added in these medium are the same as the amount of plant hormones such as 2,4-D, proline, glutamine, maltose and plant gum, the difference is only in the composition of the basic medium and Dosage, pH are 5.9. Do a comparison test of medium performance.
[0023] Experimental design of different subculture media for indica rice:
[0024] No. J ...
Embodiment 3
[0032] Embodiment 3: indica rice callus subculture medium J 3 Effect Test on Continuous Subculture of Indica Rice Callus
[0033] The callus of above-mentioned four kinds of indica rice varieties for testing provided by the embodiment is inserted into the described J 3 Culture medium, carry out continuous subculture test, the time of each subculture is 20 days. After each subculture, weigh and inoculate each 5 bottles of each test variety callus and do the differentiation activity measurement, to evaluate the J of the present invention 3 The ability of the culture medium to maintain calligenicity in continuous subculture. The test results are shown in Table 2.
[0034] Indica Varieties
Number of subcultures
Inoculation callus a
number of differentiated plants b
Plants differentiated per gram of callus
Number of plants (strains)
Minghui 63
1
2
3
0.921
1.012
0.882
...
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