Please wait a minute...
Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

Front. Agric. China    2009, Vol. 3 Issue (4) : 359-365     DOI: 10.1007/s11703-009-0036-8
Research articles |
Establishment of transgenic cotton lines with high efficiency via pollen-tube pathway
Haina ZHANG1,Fanghua ZHAO1,Yuxin ZHAO1,Chengjin GUO1,Cundong LI1,Kai XIAO1, 2,
1.College of Agronomy, Agricultural University of Hebei, Baoding 071001, China; 2.2010-01-14 15:20:58;
Download: PDF(8835 KB)  
Export: BibTeX | EndNote | Reference Manager | ProCite | RefWorks
Abstract  The transformation method via pollen-tube pathway has great potential roles in crop molecular breeding. In this paper, the effects of genotype, exogenous DNA concentration injected, and the flower positions in plant on cotton genetic transformation mediated by pollen-tube pathway were evaluated. It was found that there were no obvious differences on the boll setting rate and transformation efficiency between the cultivars 33B and 99B. However, the DNA concentrations and the flower positions largely affected the transformation efficiency. The putative transgenic plants derived from boll seeds injected with binary expression vector pCAMBIA3301 DNA were all to be PCR positive. On the other hand, the expression levels of reporter gene Gus varied dramatically, including high, middle, and nontranscripts detected. Both the GUS activity of the transgenic plants and the intensity of histochemical GUS staining in the representative transgenic plants were in accordance to those of the transcripts of reporter gene Gus. The selection of high quality and suitable concentration of exogenous DNA and the injection of the DNA in basal flowers at the middle fruit branches are important factors for improving the cotton transformation efficiency. The transformation method via pollen-tube pathway established in this study has provided an efficient genetic transformation technique in cotton breeding.
Keywords cotton (Gossypium hirsutum L.)      pollen-tube pathway      genetic transformation      molecular identification      
Issue Date: 05 December 2009
URL:  
http://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0036-8     OR     http://academic.hep.com.cn/fag/EN/Y2009/V3/I4/359
Chong K, Tan K H(1995). Function analysis of ver 203 gene through antisense transgenic winterwheat plants. Plant Physiol, 108(Suppl): 475―485

doi: 10.1104/pp.108.2.475
Doyle J J, Doyle J L(1989). Isolation of plant DNA from fresh tissue. Focus, 12: 13―15
Faure J, Dumas C(2001). Fertilization in flowering plants. New approaches for an old story. Plant Physiol, 125: 102―104

doi: 10.1104/pp.125.1.102
Geng H W, Zhang Q Z, He Z H, Xia L Q, Chen X M, Wang D S, Qu Y Y(2006). Molecular characterization of transgenic wheats with ectopic puroindolinegenes. Scientia Agricultura Sinica, 39(9): 1751―1755 (in Chinese)
Giordani T, Natali L, Maserti B E, Taddei S, Cavallini A(2000). Characterization and expression of DNA sequences encodingputative type-II metallothioneins in the seagrass Posidonia oceanica. Plant Physiol, 123: 1571―1582

doi: 10.1104/pp.123.4.1571
Gong Z Z, Shen W F, Zhou G Y(1988). Technique of transformationexogenous DNA into plant after pollination-DNA fragments were transferredinto embryos via pollen tube. Science inChina (B), 6: 611―614 (in Chinese)
Lei B, Yin G, Liu C, Qian H, Zhou S, Zhang K, Li X, Wang S(1992). Studies on introducingexogenous DNA into cultivated soybean, In: Zhou G, Huang J, Chen S, eds. Advancesin Molecular Breeding Research of Agriculture. Beijing: Chinese AgriculturalScience Press, 134―140 (in Chinese)
Li F G, Cui J J, Liu C L, Wu Z X, Li F L, Zhou Y, Li X L(2000). The study of insect-resistant transgenic cotton habouring double-geneand its insect-resistance. Scientia AgriculturaSinica, 33(1): 46―52 (in Chinese)
Li F G, Guo S D, Liu C L, Li F L, Cui H Z, Zhou Y, Li X L(1999). The study on the transformation and selection of insect-resistantcotton harboring double-gene. Acta GossypiiSinica, 11(2): 106―112 (in Chinese)
Li J, Han X L, Shen F F, Liu L(2005). Study on promoting the rate of pollen-tube pathway transformationin cotton. Cotton Sci, 17(2): 67―71 (in Chinese)
Liu X J, Liu Y H, Wang Z X, Wang X J, Zhang Y Q(2007). Generationof glyphosate-tolerant transgenic tobacco and cotton by transformationwith a 5-enolpyruvyl-shikimate-3-phosphate synthase (EPSPS) gene. J Agric Biotechn, 15(6): 958―963
Lord E M, Russell S D(2002). The mechanisms of pollination and fertilization in plants. Annu Rev Cell Dev Biol, 18: 81―105

doi: 10.1146/annurev.cellbio.18.012502.083438
Luo Z X, Wu R(1988). A simple method for the transformation of rice via the pollen-tubepathway. Plant Molecular Biology Reporter, 6: 165―174

doi: 10.1007/BF02669590
Russell S D(1993). The egg cell: development and rolein fertilization and early embryogenesis. Plant Cell, 5: 1349―1359
Saeesd N A, Zafar Y, Malik K A(1997). A simple procedureof Gossypium meristem shoot tipculture. Plant Cell Tissue Organ Culture, 51(3): 201―207

doi: 10.1023/A:1005958812583
Song X M, Gu Y H, Qin G Y(2006). Application of atransformation method via the pollen-tube pathway in agriculture molecularbreeding. Life Sci J, 4(1): 77―79 (in Chinese)
Wang L H, Liu M, Su Q, Gao X R, An L J(2004). Moleculartoken of transferred soybean via Pollen-tube Pathway. Molecular Plant Breeding, 2: 193―196 (in Chinese)
Wang X F, Chi J N, Ma Z Y(2006). Advances on genetictransformation of cotton shoot apical meristem via gene-gun bombardment. Cotton Sci, 18(1): 53―57 (in Chinese)
Weterings K, Russell S D(2004). Experimental analysis of the fertilization process. Plant Cell, 16(Suppl): S107―S118

doi: 10.1105/tpc.016873
Xiao K, Zhang C, Harrison M, Wang Z Y(2005). Isolation and characterization of a novel plant promoterthat directs strong constitutive expression of transgenes in plants. Molecular Breeding, 15: 221―231

doi: 10.1007/s11032-004-5679-9
Xie D X, Fan Y L, Ni P C(1991). Obtaining transgenicrice plants by introducing Bacillus thuringiensis insecticidal gene into rice variety Zhonghua 11. Science in China (Series B), 21: 830―834 (in Chinese)
Zhou G Y, Gong Z Z, Wang Z F(1979). The molecular basisof remote hybridization: an evidence of the hypothesis that DNA segmentsof distantly related plants may be hybridized. Acta Genetica Sinica, 6(4): 405―413 (in Chinese)
[1] Guiqin LI, Jing QI, Yuxing ZHANG, Zhihua GAO, Dongqian XU, Huixuan LI, Chenmin HUO. Construction and transformation for the antisense expression vector of the polyphenol oxidase gene in Yali pear[J]. Front Agric Chin, 2011, 5(1): 40-44.
[2] Chengjin GUO, Jinfeng ZHAO, Cundong LI, Kai XIAO, Haina ZHANG, Chuanfan SUN, Juntao GU, . Identification of senescence-related genes by cDNA-AFLP in cotton ( Gossypium hirsutum L.)[J]. Front. Agric. China, 2010, 4(3): 308-316.
[3] Jun ZHANG, Jinmao WANG, Haiyong LIANG, Minsheng YANG, . A poplar chimera with stable differentiation regulation[J]. Front. Agric. China, 2009, 3(4): 486-492.
[4] Chunli SONG, Junlian MA, Xia TANG, Zide ZHANG, Zhixia HOU, . Strawberry FaEtr2 gene RNAi expression vector construction and genetic transformation[J]. Front. Agric. China, 2009, 3(4): 419-424.
[5] HAN Shengfang, GU Juntao, XIAO Kai. Improving organic phosphate utilization in transgenic white clover by overexpression of Aspergillus niger PhyA gene[J]. Front. Agric. China, 2007, 1(3): 265-270.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed