拟南芥钾吸收调控基因AtCIPK23、AtCBL9和AtAKT1农杆菌介导共转化甘蔗

Agrobacterium-mediated Co-transformation of Arabidopsis Functional Regulating Potassium Uptake Genes AtCIPK23,AtCBL9 and AtAKT1 into Sugarcane

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,At CIPK23,At CBL9,and At AKT1 from Arabidopsis thaliana that play key roles in regulating plant K＋uptake under low-K＋condition were co-transformed into sugarcane variety YT00-236（Saccharum spp.） mediated by Agrobacterium Tumefaciens to improve sugarcane tolerance to low K＋stress.PCR analysis showed that a total of 36 transgenic clones carrying At CBL9,At CIPK23 and At AKT1 were obtained in this study.Southern blot analysis by using specific sequence of At CBL9 as probe showed that 2 to 5 copies of fragments were integrated into sugarcane genome.The transgenic clones showed much enhanced K＋uptake ability than that of the control under low-K＋stress condition.In the 200 μmol/L K＋solution,the average K＋content in roots and shoots of transgenic clones was 37% and 23% higher than that of non-transgenic control,respectively.Moreover,the average leaf relative conductivity of transgenic lines was about 6% lower than that of non-transgenic control,which indicated that less plasma membrane injury should be found in transgenic lines.The results indicated that over-expression of At CIPK23,At CBL9,and At AKT1 might enhance sugarcane tolerance to low-K＋stress.