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本研究通过农杆菌介导方法将拟南芥调控钾离子吸收的三个重要基因At AKT1、At CBL9和At CIPK23共转化到甘蔗品种粤糖00-236中,以期提高甘蔗耐低钾能力。PCR分析结果表明共获得36份携带At CBL9、At CIPK23和At AKT1三个基因的转基因无性系;Southern blot分析结果表明外源At CBL9基因在甘蔗基因组中的拷贝数为2~5个;在钾离子浓度为200μmol/L的营养液中进行低钾胁迫试验,转基因品系的根系和地上部的平均钾含量分别比对照高37%和23%,叶片平均相对电导率比对照低6%,说明超表达At CIPK23、At CBL9和At AKT1显著提高了甘蔗耐低钾胁迫能力,降低了因低钾胁迫造成的质膜伤害,是创制耐低钾甘蔗种质的有效途径。
In this study, Agrobacterium-mediated transformation of Arabidopsis thaliana regulated potassium uptake of three important genes At AKT1, At CBL9 and At CIPK23 were co-transformed into sugarcane variety Yueshun 00-236 in order to improve low potassium tolerance of sugarcane. PCR analysis showed that a total of 36 transgenic clones carrying At CBL9, At CIPK23 and At AKT1 were obtained. Southern blot analysis showed that the copy number of exogenous At CBL9 gene in sugarcane genome ranged from 2 to 5; Potassium concentration of 200μmol / L nutrient solution for low potassium stress test, the average potassium content of roots and shoots of transgenic lines were 37% and 23% higher than the control, the average relative conductivity of leaves was 6% lower than the control, indicating that super The expression of At CIPK23, At CBL9 and At AKT1 significantly increased the ability of sugarcane to tolerate low potassium stress and reduced the plasma membrane damage caused by low potassium stress, which is an effective way to create low potassium tolerant sugarcane germplasm.