棉 花 学 报     Cotton Science     2008，20(5)：354-358

The Preliminary Study on Transformation of Cotton Pollen Using Agrobacterium - mediated Vacuum Infiltration
ZHANG Yan-hong^1，2，HUANG Le-ping^1*，ZHOU Xiao-yun¹，WANG Dong-mei¹
(1.Institute of Nuclear and Biological Technologies, Xinjiang Academy of Agricultural Sciences, Urumqi 830091, China; 2. College of Agronomy, xinjiang Agricultural University, Urumqi 830052, China)

Abstract：Upland cotton pollen as the receptor，Sucrose Synthase Gene(Susy) was transfered to Xinluzao19 by Agrobacterium-mediated vacuum infiltration, the suitable conditions of vacuum infiltration method were determined .The medium consisting of 45%（ρ）sucrose was suitable, could maintain a proper infiltration press and the lower exploded pollen rate . Under pressure of-80 Pa,OD₆₀₀ 0.6-1.2, in order to maintain the lower exploded pollen rate and higher transient transformation, the appropriate time of vacuum infiltration was 15-25 minutes. For the normal pollen, Agrobacterium infected and Agrobacterium-mediated vacuum infiltrated pollen,a transient GUS expression assay and the exploded pollen rate proved that the Agrobacterium-mediated vacuum infiltrated pollen became blue. Those activated pollen would be pollinated on the stigma emasculated, respectively. Result showed that pollen using Agrobacterium-mediated vacuum infiltration were activated and could germinate pollen tubes.
Key words：agrobacterium-mediated vacuum infiltration；upland cotton；pollen；transformation
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