过表达谷子SiANT1对水稻耐盐性的影响  

Rice Overexpression of Millet SiANT1 Gene Increases Salt Tolerance

作　　者：宁蕾[1] 王曙光[1] 琚鹏举 柏星轩 葛林豪 齐欣 姜奇彦[2] 孙现军[2] 陈明[2] 孙黛珍[1] NING Lei;WANG ShuGuang;JU PengJu;BAI XingXuan;GE LinHao;QI Xin;JIANG QiYan;SUN XianJun;CHEN Ming;SUN DaiZhen(College of Agronomy, Shanxi Agricultural University, Taigu 030800, Shanxi;Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081)

机构地区：[1]山西农业大学农学院,山西太谷030800 [2]中国农业科学院作物科学研究所,北京100081

出　　处：《中国农业科学》2018年第10期1830-1841,共12页Scientia Agricultura Sinica

基　　金：国家转基因生物新品种培育科技重大专项(2016ZX08002005);谷子氮素高效利用重要基因克隆及育种价值评估(2018ZX0800924B)

摘　　要：【目的】高盐胁迫是影响作物产量的主要因素之一,谷子(Setaria italica L.)具有耐逆性强的特点,从谷子中筛选耐盐基因对于利用基因工程的手段培育耐盐作物新品种具有重要意义。【方法】分析谷子AP2/ERF类转录因子类基因SiANT1在豫谷一号幼苗期在高盐、低氮、PEG模拟干旱处理条件下的表达模式;进而利用农杆菌转化法将SiANT1转入模式作物水稻品种Kitaake;分别用0.9%盐水和自来水浸泡过表达SiANT1水稻和野生型Kitaake的种子,观察其萌芽期的表型并统计发芽率;用含0.9%Na Cl的Hogland营养液室内处理水稻幼苗10 d,65℃烘48 h之后称量干重;同时将其种植在大田,在水稻孕穗前用0.9%盐水灌溉一次,统计成熟后过表达SiANT1水稻各株系和野生型的单株籽粒重、株高、单株分蘖数,对其耐盐性进行评价;利用定量Real time-PCR方法,验证分析转基因水稻株系中SiANT1及其可能的下游基因的相对表达情况。【结果】谷子SiANT1蛋白(XP004985124.1,SiANT1)属于AP2亚族,与高粱(XP021318293.1,Sb ANT1)和玉米(XP008658933.1,Zm AP2)亲缘关系较近;豫谷一号中SiANT1的表达受高盐胁迫(100 mmol·L-1 Na Cl)诱导;将SiANT1和LP0471118-Bar-ubi-EDLL载体连接,利用农杆菌转化法将SiANT1转入到水稻基因组中,筛选得阳性T0植株,繁殖两代得T3代种子;自来水浸泡下,过表达SiANT1水稻种子萌发率和野生型相差不大,萌芽率为90%以上;0.9%盐水浸泡下,过表达SiANT1水稻种子萌发明显受到抑制,与野生型相比,过表达SiANT1水稻种子露白推迟,但最终(处理120 h)萌芽率达到80%以上;室内水稻苗期用0.9%Na Cl处理后,过表达SiANT1水稻的单株干重较受体Kitaake增重14.11%—37.42%,在1%水平上呈显著差异;大田水稻成熟后过表达SiANT1株系单株籽粒重较受体Kitaake增产56.12%—76.58%,在5%水平上呈显著差异,单株分蘖数增多、株高增加,但与野生型无显著差异;半定量RT-PCR分析表明,过�【Objective】High salt stress is one of the main factors affecting crop yield. Setaria italica L. is characterized by its strong tolerance to stress. Therefore, screening salt tolerant related genes from millet is important for salt stress tolerant improvement of crops through genetic transformation.【Method】The expression profile of AP2/ERF like gene SiANT1 were analyzed in Yugu 1 at seedling stage under high salt, low nitrogen and PEG simulated drought conditions. Millet gene SiANT1 was transformed into rice Kitaake by agrobacterium-mediated transformation method. overexpressing SiANT1 rice seeds and wild type Kitaake were digested with 0.9% saline and tap water, respectively. The phenotypes of germinating stage and germination rate were observed. Rice seedlings were treated with 0.9% Na Cl in Hogland nutrient solution for 10 days and then dried at 65℃ for 48 hours dry weights of all the rice overexpressing SiANT1 and wild type（WT） plants were analyzed. At the same time, phenotype of all plants was analyzed in the field and all plants were irrigated once with 0.9% saline before rice pregnancy. Under the similar salt treatment condition, single seed grain weigh plant height and tiller number per plant of the overexpressing SiANT1 rice lines and wild type were also analyzed. The real time-PCR method was used to verify the expression of SiANT1 and its possible downstream genes in transgenic rice lines.【Result】Setaria italica L. SiANT1 protein（XP004985124.1, SiANT1） belongs to the AP2 subfamily and has a close genetic relationship with sorghum（XP021318293.1, Sb ANT1） and maize（XP008658933.1, Zm AP2）.The gene of SiANT1 was induced by high salt stress（100 mmol·L-1 Na Cl） in Yugu 1. The SiANT1 was ligated to LP047 1118-Bar-ubi-EDLL vector, Agrobacterium transformation method was used to transfer SiANT1 into rice genome, positive T0 plants were screened, then breeded two generations and got T3 seeds. Under water immersion, the germination rate of SiANT1 overexpression rice was similar to

关 键 词：谷子 SiANT1 AP2/ERF类转录因子 耐盐性 耐盐胁迫相关基因 

分 类 号：S511[农业科学—作物学]