Cotton Ovules Culture and Analysis
棉花胚珠的培养和分析
发布: 2013年11月20日第3卷第22期 DOI: 10.21769/BioProtoc.972 浏览次数: 10641
评审: Tie Liu
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参见作者原研究论文
The authors used this protocol in:
May 2013
Abstract
The cotton ovules culture was innovated by Beasley and Ting (1973), and named after them. It is a convenient system to analyze the effect of chemical or environmental treatment on fiber development directly on ovules. This protocol was generated according to previous published papers and our practical experience.
Keywords: Cotton (棉)
Ovule culture (胚珠培养)
Plant hormone (植物激素)
Fiber (纤维)
Materials and Reagents
- Flowers (-1 to 1 DPA of flowers are easy to manipulate)
- 0.1% (w/v) HgCl2 (caution: HgCl2 is very dangerous. Please select an advantageous reagent to sterilize according to your lab condition. Otherwise 75% ethanol and N,N'-Dicyclohexylcarbodiimide (DCCS) could be the alternatives)
- Toluidine blue O (Sigma-Aldrich, catalog number: T3260 )
- Glacial acetic acid-ethanol-water (10:95:5, v/v)
- Vitamins:
VB1(Vitamin B1, Thiamine) (Sigma-Aldrich, catalog number: T3902 )
VB6 (Vitamin B6, Pyridoxine) (Sigma-Aldrich, catalog number: P8666 )
VB3 (Vitamin B3, Nicotinic acid) (Sigma-Aldrich, catalog number: N0765 ) - Inositol (Sigma-Aldrich, catalog number: I3011 )
- IAA (Sigma-Aldrich, catalog number: I2886 )
- GA3 (Sigma-Aldrich, catalog number: G7645 )
- KH2PO4 (Sigma-Aldrich)
- CaCl2.2H2O (Sigma-Aldrich)
- MgSO4.7H2O (Sigma-Aldrich)
- KNO3 (Sigma-Aldrich)
- H3BO3 (Sigma-Aldrich)
- Na2MoO4.2H2O (Sigma-Aldrich)
- KI (Sigma-Aldrich)
- CoCl2.6H2O (Sigma-Aldrich)
- MnSO4.H2O (Sigma-Aldrich)
- ZnSO4.7H2O (Sigma-Aldrich)
- CuSO4.5H2O (Sigma-Aldrich)
- FeSO4.7H2O (Sigma-Aldrich)
- Na2EDTA (Sigma-Aldrich)
- BT medium preparation (see Recipes)
- Hormone preparation (see Recipes)
- Working BT medium preparation (see Recipes)
Equipment
- Erlenmeyer flask (50 ml)
- 1.5 ml microcentrifuge tubes
- Spectrophotometer (Beckman Coulter, model: DU 800 ) or microplate reader (Tecan Trading AG, model: infinite® M200 )
- Clean bench laminar air-flow-hood with burner (Harbin East Electronic Technology, model: HD-1360 )
- Common plant tissue culture equipment (Measuring cylinder/volumetric flask, Beaker, Weighing machine, Autoclave, pH-meter, Scalpel, Fine forceps)
Software
- ImageJ
Procedure
文章信息
版权信息
© 2013 The Authors; exclusive licensee Bio-protocol LLC.
如何引用
Readers should cite both the Bio-protocol article and the original research article where this protocol was used:
- Tan, J., Deng, F., Tang, W., Han, J., Kai, G., Tu, L. and Zhang, X. (2013). Cotton Ovules Culture and Analysis. Bio-protocol 3(22): e972. DOI: 10.21769/BioProtoc.972.
- Tan, J., Tu, L., Deng, F., Hu, H., Nie, Y. and Zhang, X. (2013). A genetic and metabolic analysis revealed that cotton fiber cell development was retarded by flavonoid naringenin. Plant Physiol 162(1): 86-95.
Download Citation in RIS Format
分类
植物科学 > 植物发育生物学 > 形态建成
植物科学 > 植物生理学 > 植物生长
植物科学 > 植物细胞生物学 > 组织分析
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