农杆菌真空渗透法转化棉花花粉的初步研究

 以陆地棉花粉为受体，利用农杆菌真空渗透法将外源Susy基因转入新陆早19，对转化因素作了初步研究。结果表明，花粉在45%蔗糖浓度的花粉悬浮液中能维持适当渗透压，破损率最低；在-80 Pa压力下，菌液OD₆₀₀值在0.6～1.2范围内，真空渗入时间在15～25 min内，能维持较低花粉破损率及较高瞬间转化率；对自然花粉、农杆菌浸染的花粉及农杆菌真空渗透的花粉，分别进行GUS瞬时检测，未抽真空的花粉未被染色，而辅助真空处理的花粉部分变成蓝色，后将其授粉于已去雄的柱头上，观察到花粉管的伸长，说明抽真空后部分花粉仍有活力。

The Preliminary Study on Transformation of Cotton Pollen Using Agrobacterium-mediated Vacuum Infiltration

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.