Method for improving frequency of callus induction and plant differentiation rate in japonica rice anther culture

Abstract

The invention relates to a rice anther culture improvement technology, and in particular relates to a method for improving the frequency of callus induction and plant differentiation rate in japonica rice anther culture. The method comprises the following steps that: 200g / 667m<2> of 10% of paclobutrazol wettable powder and 50-60L of water are mixed for spraying treatment before sampling, the spraying of the paclobutrazol and low-temperature treatment cause multiple pollen cells in the anther to be in the later monokaryotic stage, novel cytokinin 4-FA is introduced into a callus inducing medium, under the synergistic action of the 4-FA, KT, and 2,4-D, division is easily started in the dark culture after inoculation, and the frequency of callus induction is greatly improved. By using the anther culture method, the frequency of callus induction in the japonica rice anther culture is obviously improved to exceed 85.0%. A differential medium comprises the following ingredients: an MS (Murashige and Skoog) basic culture medium, 0.5mg / L of NAA, 0.5mg / L of ZTO, 0.8mg / L of TDZ, 0.2mg / L of FA, 0.2mg / L of MET, 30g / L of saccharose, and 8g / L of agar powder; and after the novel hormones of ZT, 4-FA and TDZ are introduced, light cultivation is carried out, and plant differentiation rate can reach more than 86%.

Description

technical field [0001] The invention relates to a technique for improving rice anther culture, in particular to a method for improving the callus induction rate and green seedling differentiation rate of japonica rice anther culture, and belongs to the field of agricultural technology. Background technique [0002] The purpose of rice flower culture is to induce pollen to form haploids, and quickly obtain pure lines after chromosome doubling. It is an effective means of breeding new crop varieties. Compared with traditional breeding methods, it can greatly shorten the breeding cycle and improve selection efficiency. Accelerate the transfer of effective traits; rice anther culture technology can breed a large number of materials with stable genetic traits in a short period of time. Engineering breeding is more and more widely used. However, due to the differences in genotypes among cultivars, the responses of isolated anthers to the culture medium are very different, resulti...

AI Technical Summary

Problems solved by technology

However, due to the differences in genotypes among cultivars, the responses of isolated anthers to the culture medium are very different, resulting in problems such as low frequency of callus induction and low green shoot differentiation rate. The induction rate is generally between 30-50%

The low efficiency of flower culture, especially the low rate of callus induction and green seedling differentiation greatly affects the efficiency of flower culture and the construction of DH line genetic population, which limits the vigorous application of anther culture technology in rice breeding and rice genetic engineering. application

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