磷胁迫下羊草的响应及磷响应相关基因表达分析

doi:10.3864/j.issn.0578-1752.2019.23.003

中国农业科学 ›› 2019, Vol. 52 ›› Issue (23): 4215-4227.doi: 10.3864/j.issn.0578-1752.2019.23.003

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

磷胁迫下羊草的响应及磷响应相关基因表达分析

万东莉¹,侯向阳¹,丁勇¹,任卫波¹,王凯¹,李西良¹,万永青²()

¹ 中国农业科学院草原研究所/农业部草地生态与修复治理重点实验室,呼和浩特 010010
² 内蒙古农业大学生命科学学院/内蒙古自治区植物逆境生理与分子生物学重点实验室,呼和浩特 010011

收稿日期:2019-06-04 接受日期:2019-09-19 出版日期:2019-12-01 发布日期:2019-12-01
通讯作者: 万永青
作者简介:万东莉,E-mail：wandongli@caas.cn
基金资助:
内蒙古自治区自然科学基金(2019MS03002);中央级公益性科研院所基本科研业务费项目(1610332016018);中央级公益性科研院所基本科研业务费项目(1610332018001);内蒙古农业大学高层次人才引进科研启动项目(NDYB2018-61)

Response and the Expression of Pi-Responsive Genes in Leymus chinensis Under Inorganic Phosphate Treatment

WAN DongLi¹,HOU XiangYang¹,DING Yong¹,REN WeiBo¹,WANG Kai¹,LI XiLiang¹,WAN YongQing²()

¹ Institute of Grassland Research, Chinese Academy of Agricultural Sciences/Key Laboratory of Grassland Ecology and Restoration, Ministry of Agriculture, Hohhot 010010
² College of Life Sciences, Inner Mongolia Agricultural University/Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology, Hohhot 010011

Received:2019-06-04 Accepted:2019-09-19 Online:2019-12-01 Published:2019-12-01
Contact: YongQing WAN

摘要/Abstract

摘要：

【目的】磷是植物生长发育所必需的大量营养元素,研究无机磷酸盐（inorganic phosphate,Pi）胁迫下羊草的响应,筛选不同浓度Pi胁迫下的内参基因,并分析Pi响应相关基因的表达,为羊草Pi胁迫响应的分子机制研究提供基础数据。【方法】以羊草幼苗为材料,进行不同浓度Pi处理培养,对羊草的株高和根长进行检测,利用钒钼黄比色法检测羊草植株中Pi的含量。根据羊草转录组数据选取7个候选内参基因,从NCBI核酸序列数据库选取1个候选内参基因,对羊草不同处理材料进行总RNA提取和cDNA合成,利用qRT-PCR对候选内参基因的表达进行检测,并通过geNorm、NormFinder和Bestkeeper软件对其稳定性进行评估。根据筛选获得的较稳定内参基因,对Pi响应基因的qRT-PCR检测数据进行相对定量分析。【结果】表型观测结果显示,无论是低Pi还是高Pi胁迫都使得羊草的生长减缓;株高对低Pi或缺Pi胁迫较为敏感,根长对高Pi胁迫较为敏感;并随着处理浓度的增加羊草中Pi的含量也增加。qRT-PCR溶解曲线分析显示8个候选内参基因均具有单一的溶解峰,基因表达谱分析表明8个候选内参基因的CT值范围是17.16—26.61,其中,LcGAPDH的表达丰度最高为17.16—20.22,Lc18SrRNA的表达丰度最低为23.28—26.61,CT值变异系数最小的为LcARPT（2.09%）,最大的为LcTUA（6.8%）。综合geNorm、NormFinder和Bestkeeper稳定性排名,通过计算几何平均数,获得8个候选内参基因综合稳定性排名,其中排名靠前的3个基因分别为Lc18SrRNA、LcCAP和LcEF1α,排名最后的2个基因为LcTUA和LcTUB。分别以Lc18SrRNA、LcCAP和LcEF1α作为内参基因,qRT-PCR相对定量表达分析结果显示,与对照相比,LcPHO1-2的表达受低Pi或者缺Pi诱导;LcPAP2的表达受高Pi诱导;而LcPAP27的表达同时受低Pi和高Pi下调。【结论】低Pi和高Pi胁迫均使羊草生长受阻,羊草地上组织与地下组织对Pi胁迫响应的模式不同。筛选出3个表达较为稳定的内参基因Lc18SrRNA、LcCAP和LcEF1α。LcPHO1-2和LcPAP2分别参与了羊草对低Pi和高Pi的应答响应,LcPAP27同时参与了羊草对低Pi和高Pi的响应进程。

关键词: 羊草, 磷胁迫, qRT-PCR, 内参基因, 磷响应基因

Abstract:

【Objective】Phosphorus is the nutrient elements that is essential for plant growth. It would provide the basic data for molecular mechanism investigation of Leymus chinensis responding to inorganic phosphate (Pi) treatment, via analyzing the response under Pi stress in L. chinensis, and selection of the reference genes for qRT-PCR analysis, along with relative expression analysis of Pi-responsive genes. 【Method】Using L. chinensis seedlings as materials, the length of shoots and roots were measured, and the contents of Pi were examined via vanadium molybdenum yellow colorimetric method following different concentration of Pi treatments. Based on transcriptome data of L. chinensis, as well as NCBI nucleic acid sequences database, 7 and 1 candidate reference genes were selected, respectively. Total RNA was isolated from each sample after different concentration of Pi treatment. Expression level of candidate reference genes was examined through qRT-PCR, and the stability was calculated by geNorm, NormFinder and Bestkeeper. According to the most stable reference genes selected, the relative expression of Pi-responsive genes was analyzed according to the qRT-PCR results. 【Result】The phenotype observation showed that both lower and higher concentration of Pi stresses reduced the growth of L. chinensis, and the shoot growth was more sensitive to low Pi (or Pi deficiency) stress, whereas root length was more sensitive to high Pi stress. In addition, the accumulation of Pi was enhanced along with the increased concentration of Pi treatments. The melting curve analysis showed that single peak of every candidate reference gene was observed, and gene expression profiles indicated that their CT values ranged from 17.16 to 26.61, among which LcGAPDH exhibited the highest expression abundance with CT values ranged form 17.16 to 20.22, and Lc18SrRNA showed the lowest expression level with CT values ranged form 23.28 to 26.61. LcARPT (2.09%) showed the smallest expression variation, and LcTUA (6.8%) had the largest expression variation. Finally, according to the stability ranking of geNorm, NormFinder and Bestkeeper, the comprehensive stability ranking of 8 candidate reference genes were obtained via calculation geometric mean, among which the top three stable genes were Lc18SrRNA, LcCAP and LcEF1α, and the most unstable two genes were LcTUA and LcTUB. Using Lc18SrRNA, LcCAP and LcEF1α as reference genes, qRT-PCR results showed that, compared with the control, the expression of LcPHO1-2 was induced by low Pi or complete deprive of Pi, LcPAP2 was induced by high Pi, while the expressions of LcPAP27 was induced by both low and high Pi treatments.【Conclusion】Both lower and higher concentration of Pi stresses blocked the L. chinensis growth, and different responsive patterns were observed between the shoots and roots. Three relatively stable expression genes Lc18SrRNA, LcCAP and LcEF1α were selected and could be used as reference genes for qRT-PCR analysis under Pi stress. LcPHO1-2 and LcPAP2 were involved in the response of L. chinensis to low Pi and high Pi treatment, respectively, while LcPAP27 was participated in the responsive process of both low Pi and high Pi treatments.

Key words: Leymus chinensis, Pi stress, qRT-PCR, reference gene, Pi-responsive gene

万东莉,侯向阳,丁勇,任卫波,王凯,李西良,万永青. 磷胁迫下羊草的响应及磷响应相关基因表达分析[J]. 中国农业科学, 2019, 52(23): 4215-4227.

WAN DongLi,HOU XiangYang,DING Yong,REN WeiBo,WANG Kai,LI XiLiang,WAN YongQing. Response and the Expression of Pi-Responsive Genes in Leymus chinensis Under Inorganic Phosphate Treatment[J]. Scientia Agricultura Sinica, 2019, 52(23): 4215-4227.

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参考文献 39

[1]	刘公社, 李晓霞, 齐冬梅, 陈双燕, 程丽琴 . 羊草种质资源的评价与利用. 科学通报, 2016,61(2):271-281.
	LIU G S, LI X X, QI D M, CHEN S Y, CHENG L Q . Evaluation and utilization of Leymus chinensis germplasm resources. Chinese Science Bulletin, 2016,61(2):271-281. (in Chinese)
[2]	王炜, 梁存柱, 刘钟龄, 郝敦元 . 羊草+大针茅草原群落退化演替机理的研究. 植物生态学报, 2000(4):468-472.
	WANG W, LIANG C Z, LIU Z L, HAO D Y . Mechanism of degradation succession in Leymus chinensis + Stipa grandis steppe community. Acta Phytoecologica Sinica, 2000(4):468-472. (in Chinese)
[3]	乌恩, 夏庆梅, 高娃, 邢旗 . 内蒙古天然草地磷素营养问题及其解决途径. 内蒙古草业, 2006(3):4-7.
	WU E, XIA Q M, GAO W, XING Q . Phosphorus nutrition problems and solutions in natural grassland of Inner Mongolia. Inner Mongolia Prataculture, 2006(3):4-7. (in Chinese)
[4]	杜青峰 . 内蒙古退化典型草原植被和土壤对氮磷肥配施的响应[D]. 重庆: 西南大学, 2017.
	DU Q F . The responses of degraded typical steppe vegetation and soil to nitrogen and phosphorus fertilizations in Inner Mongolia[D]. Chongqing: Southwest University, 2017. (in Chinese)
[5]	董晓兵, 郝明德, 郭胜安, 石学军, 马甜, 刘公社 . 氮磷肥配施对羊草干草产量、养分吸收及品质影响. 草地学报, 2014(6):1232-1238. doi: 10.11733/j.issn.1007-0435.2014.06.013
	DONG X B, HAO M D, GUO S A, SHI X J, MA T, LIU G S . The effects of nitrogen fertilizer and phosphate fertilizer rates on the yield, nutrient uptake and quality of Leymus chinensis. Acta Agrestia Sinica, 2014(6):1232-1238. (in Chinese) doi: 10.11733/j.issn.1007-0435.2014.06.013
[6]	WANG F, DENG M J, XU J M, ZHU X L, MAO C Z . Molecular mechanisms of phosphate transport and signaling in higher plants. Seminars in Cell & Developmental Biology, 2018,74:114-122. doi: 10.1177/0145561319879245 pmid: 31619067
[7]	WU P, MA L G, HOU X L, WANG M Y, WU Y R, LIU F Y, DENG X W . Phosphate starvation triggers distinct alterations of genome expression in Arabidopsis roots and leaves. Plant Physiology, 2003,132(3):1260-1271. doi: 10.1104/pp.103.021022 pmid: 12857808
[8]	POIRIER Y, BUCHER M . Phosphate transport and homeostasis in Arabidopsis. Arabidopsis Book, 2002,1:e0024. doi: 10.1199/tab.0024 pmid: 22303200
[9]	XU L, ZHAO H, WAN R, LIU Y, XU Z, TIAN W, RUAN W, WANG F, DENG M, WANG J, DOLAN L, LUAN S, XUE S, YI K . Identification of vacuolar phosphate efflux transporters in land plants. Nature Plants, 2019,5(1):84-94. doi: 10.1038/s41477-018-0334-3 pmid: 30626920
[10]	耿燕, 吴漪, 贺金生 . 内蒙古草地叶片磷含量与土壤有效磷的关系. 植物生态学报, 2011,35(1):1-8. doi: 10.3724/SP.J.1258.2011.00001
	GENG Y, WU Y, HE J S . Relationship between leaf phosphorus concentration and soil phosphorus availability across Inner Mongolia grassland. Chinese Journal of Plant Ecology, 2011,35(1):1-8. (in Chinese) doi: 10.3724/SP.J.1258.2011.00001
[11]	HE J S, WANG L, FLYNN D F B, WANG X P, MA W H, FANG J Y . Leaf nitrogen: Phosphorus stoichiometry across Chinese grassland biomes. Oecologia, 2008,155(2):301-310. doi: 10.1007/s00442-007-0912-y pmid: 18278518
[12]	THOMPSON K, PARKINSON J A, BAND S R, SPENCER R E . A comparative study of leaf nutrient concentrations in a regional herbaceous flora. New Phytologist, 1997,136(4):679-689. doi: 10.1046/j.1469-8137.1997.00787.x
[13]	MA W, LI J, JIMOH S O, ZHANG Y, GUO F, DING Y, LI X, HOU X . Stoichiometric ratios support plant adaption to grazing moderated by soil nutrients and root enzymes. PeerJ, 2019,7:e7047. doi: 10.7717/peerj.7047 pmid: 31218124
[14]	甄莉娜, 王润梅, 周凤, 李侠, 魏玉荣, 张英俊 . 不同施磷水平下AM真菌对羊草生长的影响. 中国草地学报, 2015(6):56-61.
	ZHEN L N, WANG R M, ZHOU F, LI X, WEI Y R, ZHANG Y J . Influence of AM fungi on the growth of Leymus chinensis under phosphorus applying at different rate. Chinese Journal of Grassland, 2015(6):56-61. (in Chinese)
[15]	钱洁鑫 . 丛枝菌根对严重退化羊草典型草原植被修复效应的研究[D]. 呼和浩特: 内蒙古农业大学, 2018.
	QIAN J X . Research on the restoration effect of arbuscular mycorrhiza in severely degraded Leymus chinensis typical steppe[D]. Huhhot: Inner Mongolia Agricultural University, 2018. (in Chinese)
[16]	SECCO D, JABNOUNE M, WALKER H, SHOU H, WU P, POIRIER Y, WHELAN J . Spatio-temporal transcript profiling of rice roots and shoots in response to phosphate starvation and recovery. The Plant Cell, 2013,25(11):4285-4304. doi: 10.1105/tpc.113.117325 pmid: 24249833
[17]	EXPOSITO-RODRIGUEZ M, BORGES A A, BORGES-PEREZ A, PEREZ J A . Selection of internal control genes for quantitative real-time RT-PCR studies during tomato development process. BMC Plant Biology, 2008,8:131. doi: 10.1186/1471-2229-8-131 pmid: 19102748
[18]	UDVARDI M K, CZECHOWSKI T, SCHEIBLE W R . Eleven golden rules of quantitative RT-PCR. The Plant Cell, 2008,20(7):1736-1737. doi: 10.1105/tpc.108.061143 pmid: 18664613
[19]	YANG Q, YIN J, LI G, QI L, YANG F, WANG R . Reference gene selection for qRT-PCR in Caragana korshinskii Kom. under different stress conditions. Molecular Biology Reports, 2014,41(4):2325-2334. doi: 10.1007/s11033-014-3086-9
[20]	胡宁宁, 郭慧琴, 李西良, 孔令琪, 武自念, 张继泽, 常春, 万东莉, 臧辉, 任卫波 . 羊草不同组织实时定量PCR内参基因的筛选. 草业科学, 2017(7):1434-1441.
	HU N N, GUO H Q, LI X L, KONG L Q, WU Z N, ZHANG J Z, CHANG C, WAN D L, ZANG H, REN W B . Selection of reference genes for quantitative real-time PCR of Leymus Chinensis in different tissues. Pratacultural Science, 2017(7):1434-1441. (in Chinese)
[21]	MISSON J, RAGHOTHAMA K G, JAIN A, JOUHET J, BLOCK M A, BLIGNY R, ORTET P, CREFF A, SOMERVILLE S, ROLLAND N, DOUMAS P, NACRY P, HERRERRA-ESTRELLA L, NUSSAUME L, THIBAUD M C . A genome-wide transcriptional analysis using Arabidopsis thaliana Affymetrix gene chips determined plant responses to phosphate deprivation. Proceedings of the National Academy of Sciences of the United States of America, 2005,102(33):11934-11939.
[22]	SUN Y, MU C, CHEN Y, KONG X, XU Y, ZHENG H, ZHANG H, WANG Q, XUE Y, LI Z, DING Z, LIU X . Comparative transcript profiling of maize inbreds in response to long-term phosphorus deficiency stress. Plant Physiology Biochemistry, 2016,109:467-481. doi: 10.1016/j.plaphy.2016.10.017 pmid: 27825075
[23]	LI L, YANG H, PENG L, REN W, GONG J, LIU P, WU X, HUANG F . Comparative study reveals insights of sheepgrass ( Leymus chinensis) coping with phosphate-deprived stress condition. Frontiers in Plant Science, 2019,10:170. doi: 10.3389/fpls.2019.00170 pmid: 30873190
[24]	万永青, 张杰, 侯向阳, 王照兰, 马玉宝, 万其号, 万东莉 . 羊草DHN3基因的克隆及其逆境响应的表达分析. 西北植物学报, 2018,38(9):1598-1604.
	WAN Y Q, ZHANG J, HOU X Y, WANG Z L, MA Y B, WAN Q H, WAN D L . Cloning of LcDHN3 in Leymus chinensis and its expression under abiotic stresses. Acta Botanica Boreali-Qccidentalia Sinica, 2018,38(9):1598-1604. (in Chinese)
[25]	VANDESOMPELE J, DE PRETER K, PATTYN F, POPPE B, VAN ROY N, DE PAEPE A, SPELEMAN F . Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biology, 2002, 3(7): research0034.1-research0034.11. doi: 10.1186/gb-2002-3-7-research0034 pmid: 12184808
[26]	ANDERSEN C L, JENSEN J L, ORNTOFT T F . Normalization of real-time quantitative reverse transcription-PCR data: A model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Research, 2004,64(15):5245-5250. doi: 10.1158/0008-5472.CAN-04-0496 pmid: 15289330
[27]	PFAFFL M W, TICHOPAD A, PRGOMET C, NEUVIANS T P . Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: Best Keeper-Excel-based tool using pair-wise correlations. Biotechnology Letters, 2004,26(6):509-515. doi: 10.1023/B:BILE.0000019559.84305.47
[28]	WAN D, WAN Y, YANG Q, ZOU B, REN W, DING Y, WANG Z, WANG R, WANG K, HOU X . Selection of reference genes for qRT-PCR analysis of gene expression in Stipa grandis during environmental stresses. PLoS ONE, 2017,12(1):e0169465. doi: 10.1371/journal.pone.0169465 pmid: 28056110
[29]	LIVAK K J, SCHMITTGEN T D . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta CT) method. Methods, 2001,25(4):402-408. doi: 10.1006/meth.2001.1262 pmid: 11846609
[30]	SHANE M W, LAMBERS H . Systemic suppression of cluster-root formation and net P-uptake rates in Grevillea crithmifolia at elevated P supply: A proteacean with resistance for developing symptoms of 'P toxicity'. Journal of Experimental Botany, 2006,57(2):413-423. doi: 10.1093/jxb/erj004 pmid: 16356944
[31]	SHANE M W, MCCULLY M E, LAMBERS H . Tissue and cellular phosphorus storage during development of phosphorus toxicity in Hakea prostrata (Proteaceae). Journal of Experimental Botany, 2004,55(399):1033-1044. doi: 10.1093/jxb/erh111 pmid: 15047760
[32]	DEL-SAZ N F, ROMERO-MUNAR A, CAWTHRAY G R, PALMA F, AROCA R, BARAZA E, FLOREZ-SARASA I, LAMBERS H, RIBAS-CARBO M . Phosphorus concentration coordinates a respiratory bypass, synthesis and exudation of citrate, and the expression of high-affinity phosphorus transporters in Solanum lycopersicum. Plant Cell Environment, 2018,41(4):865-875. doi: 10.1111/pce.13155 pmid: 29380389
[33]	LI Q, FU L, WANG Y, ZHOU D, RITTMANN B E . Excessive phosphorus caused inhibition and cell damage during heterotrophic growth of Chlorella regularis. Bioresource Technology, 2018,268:266-270. doi: 10.1016/j.biortech.2018.07.148 pmid: 30081286
[34]	ZHANG C, FU J, WANG Y, BAO Z, ZHAO H . Identification of suitable reference genes for gene expression normalization in the quantitative real-time PCR analysis of sweet osmanthus ( Osmanthus fragrans Lour.). PLoS ONE, 2015,10(8):e0136355. doi: 10.1371/journal.pone.0136355 pmid: 26302211
[35]	ZHANG Y T, PENG X R, LIU Y, LI Y L, LUO Y, WANG X R, TANG H R . Evaluation of suitable reference genes for qRT-PCR normalization in strawberry ( Fragaria x ananassa) under different experimental conditions. BMC Molecular Biology, 2018,19:8. doi: 10.1186/s12867-018-0109-4 pmid: 29933763
[36]	JAIN M, NIJHAWAN A, TYAGI A K, KHURANA J P . Validation of housekeeping genes as internal control for studying gene expression in rice by quantitative real-time PCR. Biochemical and Biophysical Research Communications, 2006,345(2):646-651. doi: 10.1016/j.bbrc.2006.04.140 pmid: 16690022
[37]	YANG Z, CHEN Y, HU B, TAN Z, HUANG B . Identification and validation of reference genes for quantification of target gene expression with quantitative real-time PCR for tall fescue under four abiotic stresses. PLoS ONE, 2015,10(3):e0119569. doi: 10.1371/journal.pone.0119569 pmid: 25786207
[38]	QU R, MIAO Y, CUI Y, CAO Y, ZHOU Y, TANG X, YANG J, WANG F . Selection of reference genes for the quantitative real-time PCR normalization of gene expression in Isatis indigotica fortune. BMC Molecular Biology, 2019,20(1):9. doi: 10.1186/s12867-019-0126-y pmid: 30909859
[39]	STEFANOVIC A, RIBOT C, ROUACHED H, WANG Y, CHONG J, BELBAHRI L, DELESSERT S, POIRIER Y . Members of the PHO1 gene family show limited functional redundancy in phosphate transfer to the shoot, and are regulated by phosphate deficiency via distinct pathways. The Plant Journal, 2007,50(6):982-994. doi: 10.1111/j.1365-313X.2007.03108.x pmid: 17461783

基因名称 Gene name	基因描述 Gene description	引物序列 Primer sequences (5′-3′)	引物扩增效率 Amplification efficiency (%)	产物大小 Amplicon length (bp)	溶解温度 Product melting temperature (℃)
LcTUA	α-微管蛋白Alpha-tubulin	F: ACGAGGCCATCTACGACATCT	95.0	135	87.6
		R: CCACGTTCAGGGCACCAT
LcTUB	β-微管蛋白Beta-tubulin	F: TCCCTCGCCTCCACTTCTT	107.2	197	88.8
		R: CTCCTTTGTGCTCATCTTCCC
LcGAPDH	甘油醛3-磷酸脱氢酶Glyceraldehyde-3-phosphate dehydrogenase	F: AAACGCAACGAAGCAACTACA	97.7	105	82.2
	甘油醛3-磷酸脱氢酶Glyceraldehyde-3-phosphate dehydrogenase	R: AGAGCACCAAGCCGCAAG
LcEF1α	延伸因子1-α Elongation factor 1-alpha	F: CCGCCATCAAGAAGAAATAGG	103.9	98	82.6
	延伸因子1-α Elongation factor 1-alpha	R: CGCCGAAGCAGGAATAAAA
Lc18SrRNA	18S核糖体RNA 18S ribosomal RNA	F: TTGAAGCTGATAGGAGGGAGG	96.7	98	82.8
	18S核糖体RNA 18S ribosomal RNA	R: CTGAAGGTTGGGCGGAATA
LcCAP	腺苷酰环化酶相关蛋白 Adenylyl cyclase-associated protein	F: AGGACGGGCAGTTCACCA	103.6	114	83.1
	腺苷酰环化酶相关蛋白 Adenylyl cyclase-associated protein	R: ACAAGGCAAAGGCAGCATC
LcAPRT	腺嘌呤磷酸核糖转化酶 Adenine phosphoribosyl transferase	F: GAATCCTCTGGCTTCAACACC	101.4	118	83.7
	腺嘌呤磷酸核糖转化酶 Adenine phosphoribosyl transferase	R: GGACATCACAACCTTGCTTCTT
LcACT	肌动蛋白Actin	F: ATTGTGCTCAGTGGTGGGTCA	106.5	137	85.5
		R: CCAATCCAAACACTGTACTTCCTC
LcPHO1-2	磷酸盐1-2 Phosphate1-2	F: CTCGCCTACTGGATTTCTCCC	-	97	82.5
	磷酸盐1-2 Phosphate1-2	R: CCAACATTGTTCAAGTGTTCGTTC
LcPAP2	紫色酸性磷酸酶2 Purple acid phosphatase2	F: CTCGCGTCCGCACACATAAT	-	83	82.9
	紫色酸性磷酸酶2 Purple acid phosphatase2	R: TCCGCTTCCAGCCACTTATA
LcPAP27	紫色酸性磷酸酶27 Purple acid phosphatase27	F: CGACTCCTTCACCATCCACA	-	172	88.3
	紫色酸性磷酸酶27 Purple acid phosphatase27	R: TTCACGCACACGAGATTCTTT

基因 Gene	平均数 Mean	标准差 SD	变异系数 CV(%)
LcACT	20.10	0.97	4.82
LcTUA	23.15	1.58	6.80
LcTUB	20.75	1.37	6.60
LcAPRT	23.32	0.49	2.09
LcEF1α	21.11	0.92	4.35
Lc18SrRNA	25.27	0.93	3.68
LcCAP	22.56	0.89	3.95
LcGAPDH	18.61	0.92	4.96

排名 Rank	geNorm		NormFinder		Bestkeeper
排名 Rank	基因Gene	M值M value	基因Gene	稳定值Stability value	基因Gene	标准差Std dev (± CP)
1	LcCAP	0.42	Lc18SrRNA	0.138	LcAPRT	0.40
2	Lc18SrRNA	0.42	LcCAP	0.231	Lc18SrRNA	0.73
3	LcEF1α	0.49	LcEF1α	0.255	LcCAP	0.75
4	LcACT	0.52	LcACT	0.342	LcEF1α	0.76
5	LcGAPDH	0.54	LcGAPDH	0.390	LcGAPDH	0.76
6	LcAPRT	0.60	LcTUB	0.457	LcACT	0.81
7	LcTUB	0.68	LcAPRT	0.470	LcTUB	1.04
8	LcTUA	0.78	LcTUA	0.692	LcTUA	1.16

排名Rank	基因Gene	几何平均数Geomean
1	Lc18SrRNA	1.260
2	LcCAP	1.817
3	LcEF1α	3.302
4	LcAPRT	3.476
5	LcACT	4.579
6	LcGAPDH	4.642
7	LcTUB	6.649
8	LcTUA	8.000

磷胁迫下羊草的响应及磷响应相关基因表达分析

Response and the Expression of Pi-Responsive Genes in Leymus chinensis Under Inorganic Phosphate Treatment

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