Scientia Agricultura Sinica ›› 2018, Vol. 51 ›› Issue (12): 2225-2234.doi: 10.3864/j.issn.0578-1752.2018.12.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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

LING BingQi1, BAI XingXuan2, ZHOU YongBin2, WANG ChunXiao2, XU ZhaoShi2, MA YouZhi2, CHEN Ming2, ZHANG XiaoHong1   

  1. 1College of Life Sciences, Northwest A & F University, Yangling 712100, Shaanxi; 2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081
  • Received:2018-01-29 Online:2018-06-16 Published:2018-06-16

Abstract: 【Objective】 In previous work, we had proven that a bZIP 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 L1 and L2 increased compared to wild type (WT) significantly under normal condition and low phosphorus stress. Under normal conditions the yield of transgenic wheat increase of 5.3%-8.6%, and under low phosphorus stress grain yield of transgenic wheat increased of 4.4%-7.7%. Thousand seed weight of AtTGA4 transgenic wheat increased significantly than WT. The results of physiological indexes assay in field showed that the Fv/Fm and CTD of transgenic lines L1 and L2 were significantly better than those of WT under the condition of low phosphorus, while SPAD had no significant difference. We found that under low phosphorus stress WT were in early grain filling than the transgenic wheat, and the ears turn yellow early in WT than that in transgenic wheat plants. The phosphorus content assay during the late mature stage showed that the phosphorus concentration in the stems of L1 and L2 of transgenic lines increased significantly compared with that of WT under the condition of low phosphorus, but there was no significant difference in other tissues. Under the condition of low phosphorus, the total phosphorus content in stems and leaves and grains of two transgenic lines was higher than that of WT, and the total phosphorus content in the aerial part increased by 6.38%-17.47%. qRT-PCR results of AtTGA4 in transgenic wheat showed that the expression of AtTGA4 in Line 2 (L1) was 0.69 times lower than that of Line 1 (L1). 【Conclusion】 The above results showed that under the condition of low phosphorus stress, AtTGA4 can improve the uptake and transportation of phosphorus in transgenic wheat plants, and increase the yield of transgenic plants, and further enhance the tolerance of transgenic wheat to low phosphorus stress.

Key words: transgenic wheat, AtTGA4, low phosphorus stress, yield, physiological index

[1]    Clarkson D T, Hanson J B. The mineral nutrition of higher plants. Plant Physiology, 1980, 31: 239-298.
[2]    Schachtman D P, Reid R J, Ayling S M. Phosphorus uptake by plants: From soil to cell. Plant Physiology, 1998, 116: 447-453.
[3]    郭再华, 贺立源, 徐才国. 不同耐低磷水稻基因型秧苗对难溶性磷的吸收利用. 作物学报, 2005, 31(10): 1322-1327.
Guo Z H, He L Y, Xu C G. Uptake and use of sparingly soluble phosphorus by rice genotypes with different P-efficiency. Acta Agronomica Sinica, 2005, 31(10): 1322-1327. (in Chinese)
[4]    李玉京, 李滨, 李继云, 李振声. 植物有效利用土壤磷特性的遗传学研究进展. 遗传, 1998, 20(3): 38-41.
Li Y J, Li B, Li J Y, Li Z S. Advances in genetic study on character of plant efficiently utilizing soil phosphorus. Hereditas, 1998, 20(3): 38-41. (in Chinese)
[5]    廖红, 严小龙. 菜豆根构型对低磷胁迫和适应性变化及其基因型差异. 植物学报, 2000, 42(2): 158-163.
Liao H, Yan X L. Adaptive changes and genotypic variation for root architecture of common bean in response to phosphorus deficiency. Acta Botanica Sinica, 2000, 42(2): 158-163. (in Chinese)
[6]    Davies T G E, Ying J, Xu Q, Li Z S, Li J, Gordon-weeks R. Expression analysis of putative high-affinaty phosphate transporters in Chinese winter wheat. Plant Cell and Environment, 2002(25): 1325-1340.
[7]    Su J Y, Zheng Q, Li H W, Li B, Jing R L, Tong Y P, Li Z S. Detection of QTLs for phosphorus use efficiency in relation to agronomic performance of wheat grown under phosphorus sufficient and limited conditions. Plant Science, 2009(176): 824-836.
[8]    Wang J, Sun J H, Miao J, Guo J K, Shi Z L, He M Q, Chen Y, Zhao X Q, Li B, Han F P, Tong Y P, Li Z S. A phosphate starvation response regulator Ta-PHR1 is involved in phosphate signalling and increases grain yield in wheat. Annals of Botany, 2013, 111: 1139-1153.
[9]    Rubio V, Linhares F, Solano R, Martin A C, Iglesias J, Leyva A, Paz-Ares J. A conserved MYB transcription fator involved in phophate starvation signaling both in vascular plants and in unicellular algae. Genes Development, 2001, 15: 2122-2133.
[10]   Devaiah B N, Karthikeyan A S, Raghotthama K G. WRKY75 transcription factor is a modulator of phosphate acquisition and root development in Arabidopsis. Plant Physiology, 2007, 143: 1789-1801.
[11]   Sano T, Nagata T. The possible involvement of a phosphate- induced transcription factor encoded by Phi-2 gene from tobacco in ABA signaling pathways. Plant Cell Physiology, 2002, 43: 12-20.
[12]   Yi K K, Wu Z C, Zhou J, Du L M, Guo L B, Wu Y R, Wu P. OsPTF1, a novel transcription factor involved in tolerance to phosphate starvation in rice. Plant Physiology, 2005, 138: 2087-2096.
[13]   Zhang J Y, Zhou X, Xu Y, Yao M L, Xie F B, Gai J Y, Li Y, Yang S P. Soybean SPX1 is an important component of the response to phosphate deficiency for phosphorus homeostasis. Plant Science, 2016, 248: 82-91.
[14]   Zhong L, Chen D D, Min D H, Li W W, Xu Z S, Zhou Y B, Li L C, Chen M, Ma Y Z. AtTGA4, a bZIP transcription factor, confers drought resistance by enhancing nitrate transport and assimilation in Arabidopsis thaliana. Biochemical and Biophysical Research Communications, 2015, 457: 433-439.
[15]   王娟, 孙爱平, 王开营, 沈永平, 张延成, 王锦贵. 土壤样品采集的原则与方法. 现代农业科技, 2011, 21: 300-301.
Wang J, Sun A P, Wang K Y, Shen Y P, Zhang Y C, Wang J G. Principles and methods to collect soil samples. Modern Agricultural Science and Technology, 2011, 21: 300-301. (in Chinese)
[16]   郭胜利, 党延辉, 刘守赞, 郝明德. 磷素吸附特性演变及其与土壤磷素形态、土壤有机碳含量的关系. 植物营养与肥料学报, 2005, 11(1): 33-39.
Guo S L, Dang Y H, Liu S Z, Hao M D. Changes in characterization of phosphorus sorption in relation to it’s forms and soil organic carbon. Plant Nutrition and Fertilizing Science, 2005, 11(1): 33-39. (in Chinese)
[17]   宇万太, 张璐, 沈善敏, 廉鸿志. 辽西褐土施肥及养分循环再利用中长期试验Ⅲ. 磷和氮在堆腐过程中的循环率及有机肥料中养分利用率. 应用生态学报, 2002, 11(13): 1407-1409.
Yu W T, Zhang L, Shen S M, Lian H Z. A long-term field trial on fertilization and on use of recycled nutrients in farming systems Ⅲ. Recycling rate of P and N through a composting cycle and recoveries of P and N in organic manure. Chinese Journal of Applied Ecology, 2002, 11(13): 1407-1409. (in Chinese)
[18]   李科江, 张素芳, 贾文竹, 宋平忠, 刘树庆, 霍习良, 王玉朵. 半干旱区长期施肥对作物产量和土壤肥力的影响. 植物营养与肥料学报, 1999, 5(1): 21-25.
Li K J, Zhang S F, Jia W Z, Song P Z, Liu S Q, Huo X L, Wang Y D. Effect of long-term fertilization on crop yield and soil fertility in semi-arid area. Plant Nutrition and Fertilizing Science, 1999, 5(1): 21-25. (in Chinese)
[19]   王旭刚, 李建民, 张春霞. 氮磷配施对旱地小麦产量和吸肥特性的影响. 西北农林科技大学学报(自然科学版), 2007, 35(2): 138-142.
Wang X G, Li J M, Zhang C X. Effects of fertilizer N, P on wheat yield and nutrients uptake characters in dryland of Loess Plateau. Journal of Northwest A & F University (Natural Science Edition), 2007, 35(2): 138-142. (in Chinese)
[20]   赵亚妮. 不同氮磷和种植模式下小麦的生长动态、产量及养分效率研究[D]. 雅安: 四川农业大学, 2016.
Zhao Y N. Wheat growth dynamic, yield and nutrient use efficiency associated with N and P in different planting patterns[D]. Yaan: Sichuan Agricultural University, 2016. (in Chinese)
[21]   胡梦芸, 李辉, 庞建周, 刘茜, 张颖君, 孙丽静. 过量表达蔗糖转运蛋白基因增强转基因小麦的耐寒性. 中国农业科学, 2015, 48(8): 1473-1483.
Hu M Y, Li H, Pang J Z, Liu Q, Zhang Y J, Sun L J. Overexpression of sucrose transporter (TaSUT1A) improves drought tolerance in transgenic wheat. Scientia Agricultura Sinica, 2015, 48(8): 1473-1483. (in Chinese)
[22]   Vance C P, Uhde-Stone C, Allen D L. Phosphorus acquisition and use: Critical adaptation by plants for securing a nonrenewable resource. New Phytologist, 2003, 157: 423-447.
[23]   彦君, 熊又升, 黄丽, 袁家富, 徐祥玉, 刘晔. 施氮对不同品种冬小麦氮素积累和转运的影响. 西北植物学报, 2010, 10: 2040-2046.
Chai Y J, Xiong Y S, Huang L, Yuan J F, Xu X Y, Liu Y. Effects of nitrogen application on nitrogen accumulation, distribution and translocation of different winter wheat varieties. Acta Botanica Boreali-Occidentalia Sinica, 2010, 10: 2040-2046. (in Chinese)
[24]   Jakoby M. bZIP transcription factors in Arabidopsis. Trends in Plant Science, 2002, 7(3): 106-111.
[25]   田义, 张彩霞, 康国栋, 李武兴, 张利益, 丛佩华. 植物TGA转录因子研究进展. 中国农业科学, 2016, 49(4): 632-642.
Tian Y, Zhang C X, Kang G D, Li W X, Zhang L Y, Cong P H. Progress on TGA transcription factors in plant. Scientia Agricultura Sinica, 2016, 49(4): 632-642. (in Chinese)
[26]   Katagiri F, Lam E, Chua N H. Two tobacco DNA-bind proteins with homology to the nuclear factor CREB. Nature, 1989, 340: 723-730.
[27]   喻修道, 徐兆师, 陈明, 李连成, 马有志. 小麦转基因技术研究及其应用. 中国农业科学, 2010, 43(8): 1539-1553.
Yu X D, Xu Z S, Chen M, Li L C, Ma Y Z. The progress and application of wheat transformation technology. Scientia Agricultura Sinica, 2010, 43(8): 1539-1553. (in Chinese)
[28]   Muller R, Morant M, Jarmer H, Nilsson L, Nielsen T H. Genome-wide analysis of the Arabidopsis leaf transcription reveals interaction of phosphate and sugar metabolism. Plant Physiology, 2007, 143: 156-171.
[29]   Nilsson L, Muller R, Nielsen T H. Dissecting the plant transcriptome and the regulatory responses to phosphate deprivation. Physiologia Plantarum, 2010, 139: 129-143.
[30]   Brant A N, Chen Y H Y. Patterns and mechanisms of nutrient resorption in plants. Critical Reviews in Plant Sciences, 2015, 34(5): 471-486.
[31]   Freschet G T, Cornelissen J H C, Van Logtestijn R S P, Aerts R. Substantial nutrient resorption from leaves, stems and roots in a subarctic flora: what is the link with other resource economics traits? New Phytologist, 2010, 186(4): 879-889.
[32]   Aerts R. Nutrient resorption from senescing leaves of perennials: are there general patterns? The Journal of Ecology, 1996, 84(4): 597-608.
[33]   Mao R, Song C C, Zhang X H, Wang X W, Zhang Z H. Response of leaf, sheath and stem nutrient resorption to 7 years of N addition in freshwater wetland of northeast China. Plant and Soil, 2013, 364(1): 385-394.
[34]   Nilsson L, Muller R, Nielsen T H. Increased expression of the MYB-related transcription factor, PHR1, leads to enhanced phosphate uptake in Arabidopsis thaliana. Plant, Cell and Environment, 2007, 30: 1499-1512.
[35]   Zhou J, Jiao F C, Wu Z C, Li Y Y, Wang X M, He X W, Zhong W Q, Wu P. OsPHR2 is involved in phosphate-starvation signaling and excessive phosphate accumulation in shoots of plants. Plant Physiology, 2008, 146: 1673-1686.
[36]   Jacob J, Lawlor D W. In vivo photosynthetic electron transport does not limit photosynthetic capacity in phosphate-deficient sunflower and maize leaves. Plant, Cell and Environment, 1993, 16: 785-795.
[37]   许大全. 光合速率、光合效率与作物产量. 生物学通报, 1999, 34(8): 8-10.
Xu D Q. Photosynthetic rate, photosynthetic efficiency and crop yield. Bulletin of Biology, 1999, 34(8): 8-10. (in Chinese)
[38]   王菲, 曹翠玲. 磷水平对不同磷效率小麦叶绿素荧光参数的影响. 植物营养与肥料学报, 2010, 16(3): 758-762.
Wang F, Cao C L. Effects of phosphorus levels on chlorophyll fluorescence parameters of wheat (Triticum aestivum L.) with different phosphorus efficiencies. Plant Nutrition and Fertilizer Science, 2010, 16(3): 758-762. (in Chinese)
[39]   黄晓林, 李妍, 李国强. 冠层温度与作物水分状况关系研究进展. 安徽农业科学, 2009, 4: 1511-1515.
Huang X L, Li Y, Li G Q. Research advance in relationship between canopy temperature and crop water status. Journal of Anhui Agricultural Sciences, 2009, 4: 1511-1515. (in Chinese)
[40]   梁鸡保, 张元香, 王涛. 提高作物水分利用的研究. 北京农业, 2012, 27: 149.
Liang J B, Zhang Y X, Wang T. Study on improving water used of crop. Beijing Agriculture, 2012, 27:149. (in Chinese)
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