Crops ›› 2016, Vol. 32 ›› Issue (5): 61-66.doi: 10.16035/j.issn.1001-7283.2016.05.011

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The Study on Antibacterial Effects of Different Antibiotics and Concentrations in a Genetic Transformation System of Yandao8

Wang Jingdong1,Ma Hongwen1,Fan Yunfang2,Chen Xiaojun1   

  1. 1 The Key Lab of Agricultural Biotechnology of Ningxia,Yinchuan 750002,Ningxia,China
    2 National Wolfberry Engineering Research Center,Yinchuan 750002,Ningxia,China
  • Received:2016-07-06 Revised:2016-09-27 Online:2016-10-15 Published:2018-08-26
  • Contact: Xiaojun Chen

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Abstract:

In order to establish a genetic transformation system in japonica rice, and to select the anti-callus and improve the regeneration ability in transgenic plant, two bacteriostatic agent (Carbenicillin sodium and Cefotaxime sodium ) and a selected marker (Hygromycin B) were used to optimize the culture conditions. The infected co-cultured callus was treated with 4 concentrations of each bacteriostatic agent. The results showed that the bacteriostatic activity of Carbenicillin sodium was better than that of Cefotaxime sodium. The bacteriostatic rate reached 100.0% and callus differentiation rate reached 42.2%, when the concentration of Carbenicillin sodium was 500mg/L. Meanwhile, 86.1% high quality of anti-callus was obtained and callus differentiation rate reached 39.5%, when the concentration of Hygromycin B was 30mg/L. The optimum culture condition was 500mg/L Car+30mg/L HyB in the genetic transformation system of Yandao 8, a japonica cultivar.

Key words: Transformation system, Anti-callus, Transgenic plant, Bacteriostatic agent, Selective agent, Yandao 8

Table 1

Two bacteriostat’s inhibition to agrobacterium tumefaciens and their influence on callus differentiation"

抗生素浓度(mg/L)
Antibiotic concentration		 总愈伤数
Callus number		 未污染数
Unpolluted number		 抑菌率(%)
Bacteriostatic rate		 分化愈伤数
Callus differentiation number		 分化率(%)
Differentiation rate
羧苄青霉素钠 300 50 7.3Cc 14.1Cc 0Dd 0Dd
(Car) 400 50 29.2Bb 58.1Bb 8Bb 27.5Bb
500 50 50.0Aa 100.0Aa 21Aa 42.2Aa
600 50 50.0Aa 100.0Aa 6Cc 12.3Cc
头孢噻肟钠 300 50 1.0Dd 2.3Dd 0Bb 0Bb
(Cef) 400 50 8.0Cc 16.1Cc 0Bb 0Bb
500 50 17.0Bb 34.0Bb 2Aa 11.7Aa
600 50 33.0Aa 66.2Aa 0Bb 0Bb

Table 2

The influence of selective agent concentration to selection rate of resistant callus and its differentiation"

HyB浓度(mg/L)
HyB concentration		 总愈伤块数
Callus number		 抗性愈伤数
Resistant callus number		 筛选率(%)
Screening rate		 分化愈伤数
Callus differentiation number		 分化率(%)
Differentiation rate
MMD21 20 50 46.8Aa 93.6Aa 22Aa 46.8Aa
30 50 43.0Bb 86.1Bb 17Bb 39.5Bb
40 50 16.6Cc 33.9Cc 4Cc 17.6Cc
50 50 0Dd 0Dd 0Dd 0Dd
MMD34 20 50 37.8Aa 75.5Ab 14Aa 36.8Aa
30 50 38.6Aa 77.5Aa 12Ab 30.8Bb
40 50 12.0Bb 24.0Bc 2Bc 16.7Cc
50 50 0Cc 0Cd 0Cd 0Dd
MMD43 20 50 44.7Aa 89.5Aa 22Aa 44.4Aa
30 50 40.0Bb 79.9Bb 11Bb 27.5Bb
40 50 9.2Cc 18.4Cc 1Cc 11.1Cc
50 50 0Dd 0Dd 0Cc 0Dd
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