转AtTGA4小麦田间耐低磷胁迫的功能分析  

Functional Analysis of AtTGA4 Transgenic Wheat Tolerance to Low Phosphorus Stress in Field

作　　者：凌炳琦 柏星轩 周永斌[2] 王春霄 徐兆师[2] 马有志[2] 陈明[2] 张小红[1] LING BingQi;BAI XingXuan;ZHOU YongBin;WANG ChunXiao;XU ZhaoShi;MA YouZhi;CHEN Ming;ZHANG XiaoHong(College of Life Sciences, Northwest A＆F University, Yangling 712100, Shaanxi;Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081)

机构地区：[1]西北农林科技大学生命科学学院,陕西杨凌712100 [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12期2225-2234,共10页Scientia Agricultura Sinica

基　　金：转基因生物重大专项(2016ZX08002-005)

摘　　要：【目的】将bZIP类转录因子基因AtTGA4转化小麦创制耐低磷转基因小麦新材料,同时分析AtTGA4提高小麦抗逆性的生理机制,为小麦耐低磷胁迫分子育种奠定基础。【方法】采用最小表达框基因枪转化法将AtTGA4和筛选标记基因Bar共转化受体小麦石4056,通过PCR检测筛选出无Bar并能稳定遗传AtTGA4的转基因小麦新株系。基于试验地土壤养分含量状况施用不同水平的磷肥,形成一定程度的正常和低磷营养胁迫,对AtTGA4转基因小麦新株系进行低磷胁迫耐受性试验。在开花期进行了光系统Ⅱ原初光能转化效率(light efficiency of the light systemⅡ,Fv/Fm),叶绿素相对含量(soil and plant analyzer development readings,SPAD值)和气冠温差(canopy temperature depression,CTD)等生理指标的测定,在成熟期进行了株高、分蘖数、穗粒数等农艺性状的调查,并在小麦收获后进行了产量及不同组分(根、茎、叶、籽粒)磷浓度和磷吸收、残留总量的测定和统计。【结果】PCR分析结果证明,AtTGA4已在石4056小麦中稳定遗传至T4代,共获得4个稳定转基因株系。根据土壤养分含量测定结果,在正常条件地块施加812.39 kg·hm-2的过磷酸钙,低磷处理地块不施磷肥。产量及农艺性状统计结果显示,AtTGA4转基因株系L1和L2在正常和低磷胁迫条件下的产量相对于受体对照小麦显著增加,正常条件下产量增幅为5.3%—8.6%,低磷胁迫下产量增幅为4.4%—7.7%。在低磷胁迫条件下,过表达AtTGA4的转基因小麦种子千粒重显著比受体显著增加。开花期田间生理指标测定结果显示,转基因株系L1和L2在低磷条件下的Fv/Fm和CTD明显优于受体,而SPAD值没有明显差异。田间调查时发现,低磷条件下受体比转基因材料提早结束灌浆,表现在穗子提早变黄。成熟末期磷含量测定结果显示,转基因株系L1和L2在低磷条件下茎杆磷浓度比受体显著提高,在其他组织中则无显著差【Objective】 In previous work, we had proven that a b ZIP type transcription factor gene, AtTGA4 can improve the drought resistance and low nitrogen tolerance of transgenic Arabidopsis. In this study, AtTGA4 was transformed into wheat to identify the tolerance of transgenic wheats to low phosphorus stress in field. At the same time, the physiological mechanism of AtTGA4 gene to improve the stress resistance of transgenic wheats was analyzed, which laid the foundation for the molecular breeding of wheat tolerant to low phosphorus stress. 【Method】Used particle bombardment method of minimal expression box to co-transform AtTGA4 and marker gene Bar into wheat variety Shi4056. After transformation, we screened many transgenic wheat lines without Bar gene and with AtTGA4 gene through PCR assay. In field experiment, base on fertilizer nutrient content in soil we applied different levels of phosphate, which led to low phosphorus stress in soil. AtTGA4 transgenic wheat lines were tested under low phosphorus. During the flowering stage some physiological indexes were identified such as light efficiency of the light system Ⅱ（Fv/Fm）, the relative content of chlorophyll（SPAD） and crown the temperature difference（CTD）, and other agronomic traits were investigated, during mature stage such as plants height, tiller number, grains per spike in mature period, and some values such as the grain yield and phosphorus concentration and phosphorus uptake in different components in wheat（root, stem, leaf and grain） were measured and data statistic analysis were completed. 【Result】 PCR analysis showed that AtTGA4 gene had been stable heritability for T4 generation in Shi4056 and four stable transgenic lines were obtained. According to the results of soil nutrient content, 812.39 kg·hm^-2 of superphosphate was applied to normal plots, and no phosphorus was applied in low phosphorus treatment plots. The statistical results of yield and agronomic characters showed that the grain yield of AtTGA4 transgenic lines

关 键 词：转基因小麦 AtTGA4 低磷营养胁迫 产量 生理指标 

分 类 号：S512.1[农业科学—作物学]