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

A callus induction and anther technology, which is applied in the agricultural field, can solve the problems of low flower cultivation efficiency, great differences in the reaction of in vitro anther medium, low callus induction frequency and green seedling differentiation rate, etc., and achieves good repeatability. , Improve work efficiency, strong operability effect

Inactive Publication Date: 2013-07-24
JIANGSU XUHUAI DISTRICT XUZHOU AGRI SCI INST
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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 f...

Method used

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

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Embodiment 1

[0015] 1. Test materials and methods

[0016] 1.1 Test material: Xudao No. 5

[0017] 1.2 Test method:

[0018] The selected material is the japonica rice variety Xudao No. 5, and 5-7 days before Yezhenping, use 10% paclobutrazol wettable powder 200g / 667m 2 Spray with 50-60L of water, and use the material without paclobutrazol as a control;

[0019] When the leaf-to-pillow distance is 3-5cm, take the young ears, treat them at 5-8°C for 5-8 days, take the young ears whose stamens have extended to 1 / 2-2 / 3 of the glume as experimental materials, and sterilize them with 70% alcohol for 45s , add 0.1%HgCl 2 Disinfect for 10-15min to remove Hgcl 2 After solution, rinse with sterile water for 3-4 times, shake while washing, pick out anthers with tweezers, inoculate to improved callus induction medium, N6 medium + KT 0.5mg / L + 2,4-D2.0 mg / L+4-FA 0-2.0mg / L+sucrose 60g / L+agar powder 8.0g / L, improved N6 medium+KT 2.0mg / L+2,4-D 2.0mg / L+sucrose 60g / L+agar powder 8.0g / L as the control...

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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...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 胡婷婷王健康丁成伟郭荣良吴玉玲徐家安王友霜赵轶鹏
Owner JIANGSU XUHUAI DISTRICT XUZHOU AGRI SCI INST
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