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

Previous Articles     Next Articles

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

RichHTML

3

PDF (PC)

127

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
[1] Hiei Y, Ohta S, Kumashiro T , et al. Efficient transformation of rice mediated by agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant Journal, 1994,6(2):271-282.
doi: 10.1046/j.1365-313X.1994.6020271.x
[2] 惠楠 . 单倍体小黑杨抗生素敏感试验. 林业科技, 2004,29(4):7-9.
[3] 邓君萍, 颜双春, 候佩 , 等. 不同抗生素种类及浓度对麻疯树培养的影响. 应用与环境生物学报, 2005,11(2):156-159.
doi: 10.3321/j.issn:1006-687X.2005.02.007
[4] 昊关庭, 胡张华, 郎春秀 , 等. 抗生素对高羊茅胚性愈伤组织生长与分化的影响. 核农学报, 2005,19(2):88-91.
[5] 张致平 . 中国新抗生素研究的进展. 中国抗生素杂志, 1998,23(2):81-91.
[6] 张松, 达克东, 曹辰兴 , 等. 抗生素对韭菜根尖培养植株再生的影响. 核农学报, 2003,17(2):101-l04.
[7] 洪丽兰 . MIL1介导水稻花粉母细胞形成的分子机理. 北京:中国科学院, 2012.
[8] 李倩, 张聪, 吴洁 , 等. 甘薯对潮霉素及除草剂的敏感性研究. 西南农业学报, 2013,26(5):1870-1872.
[9] 周小梅, 李君剑, 赵军良 , 等. 抗生素对农杆菌的抑制和对油菜外植体分化的影响. 西北植物学报, 2005,25(1):52-56.
doi: 10.3321/j.issn:1000-4025.2005.01.009
[10] 范正琪, 李纪元, 田敏 . 抗生素对山茶愈伤组织诱导和生长的影响. 林业科学研究, 2005,18(2):183-186.
doi: 10.3321/j.issn:1001-1498.2005.02.015
[11] Mamidala P, Nanna R S . Influence of antibiotics on regeneration efficiency in tomato. Plant Omics Journal, 2009,2(4):135-140.
doi: 10.1007/s11103-009-9494-x
[12] 张明洲, 崔海瑞, 舒庆尧 , 等. 抗生素对高粱离体营养反应的影响. 核农学报, 2004,18(2):113-117.
doi: 10.3969/j.issn.1000-8551.2004.02.009
[13] Jeyaramraja P R, Meenakshi S N . Agrobacterium tumefacients-mediated transformation of embryogenic tissues of tea [Camellia sinensis (L.) O.Kuntze]. Plant Molecular Biology Reporter, 2005,23(3):299a-299i.
doi: 10.1007/BF02772761
[14] 张明洲, 崔海瑞, 舒庆尧 , 等. 抗生素对高梁愈伤组织诱导和生长的影响. 浙江大学学报(农业与生命科学版), 2004,30(2):221-225.
doi: 10.3321/j.issn:1008-9209.2004.02.021
[15] 刘国宝, 刘昀, 郑易之 . 水稻高效再生体系的建立及其对大豆Em基因的转化. 西北植物学报, 2008,28(4):686-691.
[16] 邹莉 . 大豆子叶节对潮霉素敏感性研究. 大豆科学, 2008,27(2):94-96.
[17] 奚亚军 . 小麦遗传转化中潮霉素适宜筛选浓度的研究. 西北农林科技大学学报(自然科学版), 2003,31(1):42-45.
[18] 马丙田, 朱祯, 李平 , 等. 水稻遗传转化选择系统优化初探. 西南农业大学学报, 2003,16(1):28-31.
doi: 10.3969/j.issn.1001-4829.2003.01.007
[19] 周玲艳, 姜大刚, 吴豪 , 等. 潮霉素和PPT对水稻愈伤组织筛选效果的比较. 仲恺农业技术学院学报, 2003,16(2):10-15.
[20] 杨桥, 蔺自敏, 侯详文 , 等. 小麦遗传转化中潮霉素筛选体系的建立及应用. 麦类作物学报, 2013,34(3):304-310.
doi: 10.7606/j.issn.1009-1041.2014.03.004
[21] 康海崎, 申国安 . 水稻愈伤组织对不同培养基的适应性、潮霉素的敏感性及其在遗传转化中的利用. 中国生物工程杂志, 2009,29(10):50-54.
[22] 王关林, 方宏笃 . 植物基因工程.北京: 科学出版社, 2002.
No related articles found!
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
[1] Guangcai Zhao,Xuhong Chang,Demei Wang,Zhiqiang Tao,Yanjie Wang,Yushuang Yang,Yingjie Zhu. General Situation and Development of Wheat Production[J]. Crops, 2018, 34(4): 1 -7 .
[2] Baoquan Quan,Dongmei Bai,Yuexia Tian,Yunyun Xue. Effects of Different Leaf-Peg Ratio on Photosynthesis and Yield of Peanut[J]. Crops, 2018, 34(4): 102 -105 .
[3] Xuefang Huang,Mingjing Huang,Huatao Liu,Cong Zhao,Juanling Wang. Effects of Annual Precipitation and Population Density on Tiller-Earing and Yield of Zhangzagu 5 under Film Mulching and Hole Sowing[J]. Crops, 2018, 34(4): 106 -113 .
[4] Wenhui Huang, Hui Wang, Desheng Mei. Research Progress on Lodging Resistance of Crops[J]. Crops, 2018, 34(4): 13 -19 .
[5] Yun Zhao,Cailong Xu,Xu Yang,Suzhen Li,Jing Zhou,Jicun Li,Tianfu Han,Cunxiang Wu. Effects of Sowing Methods on Seedling Stand and Production Profit of Summer Soybean under Wheat-Soybean System[J]. Crops, 2018, 34(4): 114 -120 .
[6] Mei Lu,Min Sun,Aixia Ren,Miaomiao Lei,Lingzhu Xue,Zhiqiang Gao. Effects of Spraying Foliar Fertilizers on Dryland Wheat Growth and the Correlation with Yield Formation[J]. Crops, 2018, 34(4): 121 -125 .
[7] Xiaofei Wang,Haijun Xu,Mengqiao Guo,Yu Xiao,Xinyu Cheng,Shuxia Liu,Xiangjun Guan,Yaokun Wu,Weihua Zhao,Guojiang Wei. Effects of Sowing Date, Density and Fertilizer Utilization Rate on the Yield of Oilseed Perilla frutescens in Cold Area[J]. Crops, 2018, 34(4): 126 -130 .
[8] Pengjin Zhu,Xinhua Pang,Chun Liang,Qinliang Tan,Lin Yan,Quanguang Zhou,Kewei Ou. Effects of Cold Stress on Reactive Oxygen Metabolism and Antioxidant Enzyme Activities of Sugarcane Seedlings[J]. Crops, 2018, 34(4): 131 -137 .
[9] Jie Gao,Qingfeng Li,Qiu Peng,Xiaoyan Jiao,Jinsong Wang. Effects of Different Nutrient Combinations on Plant Production and Nitrogen, Phosphorus and Potassium Utilization Characteristics in Waxy Sorghum[J]. Crops, 2018, 34(4): 138 -142 .
[10] Na Shang,Zhongxu Yang,Qiuzhi Li,Huihui Yin,Shihong Wang,Haitao Li,Tong Li,Han Zhang. Response of Cotton with Vegetative Branches to Plant Density in the Western of Shandong Province[J]. Crops, 2018, 34(4): 143 -148 .