藜麦幼苗响应低氮的转录组分析

doi:10.7668/hbnxb.20194769

摘要/Abstract

摘要：

为探究藜麦响应低氮的分子机制,筛选低氮响应基因,揭示藜麦响应低氮的适应性变化,从不同氮素水平处理下藜麦的生长和叶绿素合成入手,采用RNA-Seq技术从转录组水平分析藜麦对于低氮和缺氮分别处理5 d和30 d后的响应情况。结果表明,藜麦在缺氮条件下根系优先生长;低氮和缺氮均可促使老叶优先变黄或脱落而维持幼嫩叶片的叶色,且均提高了幼苗对氮素的利用效率。GO功能富集分析结果显示,低氮和缺氮分别在处理5 d和30 d后差异表达基因均主要涉及膜的重要组成部分、细胞膜、氧化还原过程、代谢过程、ATP结合及金属离子结合等。KEGG富集分析结果表明,低氮和缺氮处理相对于高氮处理5 d后比较显著的差异代谢通路在于苯丙素的生物合成和谷胱甘肽代谢;而处理30 d后比较显著的差异代谢通路在于碳代谢通路。进一步挖掘藜麦响应低氮的关键基因,结果显示,低氮和缺氮处理5 d后过氧化物酶、谷胱甘肽S-转移酶、抗坏血酸过氧化物酶基因上调,表达量较高;低氮和缺氮处理30 d后磷酸甘油酸激酶、半胱氨酸合成酶、3-磷酸甘油醛脱氢酶(NADP)、磷酸烯醇丙酮酸羧化酶基因上调,表达量较高。qRT-PCR验证结果与RNA-Seq测序结果一致。

关键词: 藜麦, 低氮, 苯丙素生物合成, 谷胱甘肽代谢, 碳代谢

Abstract:

In order to explore the molecular mechanism of low nitrogen response in quinoa, low nitrogen response genes were screened to reveal the adaptive changes in quinoa response to low nitrogen.Based on the seedling growth observation and chlorophyll synthesis detection,we analyzed the transcriptome changes of quinoa after 5 d and 30 d under nitrogen deficiency conditions.The results showed that roots were preferentially developed under nitrogen starvation condition.Older leaves turned yellow or dropped down under both low nitrogen and nitrogen starvation conditions,therefore younger leaves could maintain green.Higher NUE was shown in both low and nitrogen starvation conditions.GO enrichment analysis indicated that significantly differential expressed genes were mainly involved in integral component of membrane,membrane,oxidation-reduction process,metabolic process,ATP binding,and metal ion binding.After 5 d of low or nitrogen starvation supply,KEGG enrichment analysis showed that phenylpropyl biosynthesis and glutathione metabolism were the most significant metabolic pathways compared with high nitrogen.After 30 d of treatment,the most significant metabolic pathway was the carbon metabolic pathway.The key genes in response to low nitrogen in quinoa were further explored.The results showed that peroxidase,glutathione S-transferase and ascorbate peroxidase genes were up-regulated and their expressions were higher after 5 days of low nitrogen and nitrogen deficiency treatment.The genes of phosphoglycerate kinase,cysteine synthetase,glyceraldehyde 3-phosphate dehydrogenase (NADP)and phosphoenolpyruvate carboxylase were up-regulated and the expression levels were higher after 30 days of low and low nitrogen treatment.The results of qRT-PCR agreed with the RNA-Seq.

Key words: Quinoa, Low nitrogen, Phenylpropanoid biosynthesis, Glutathione metabolism, Carbon metabolism

杨雅舒, 郁培义, 王建华, 陕嘉楠, 裴红宾, 杨利艳. 藜麦幼苗响应低氮的转录组分析[J]. 华北农学报, 2024, 39(4): 143-153. doi: 10.7668/hbnxb.20194769.

YANG Yashu, YU Peiyi, WANG Jianhua, SHAN Jianan, PEI Hongbin, YANG Liyan. Transcriptome Analysis of Quinoa Seedlings in Response to Low Nitrogen[J]. Acta Agriculturae Boreali-Sinica, 2024, 39(4): 143-153. doi: 10.7668/hbnxb.20194769.

http://www.hbnxb.net/CN/Y2024/V39/I4/143

图/表 11

表1 荧光定量PCR验证表达基因及其引物序列

Tab.1 Fluorescent quantitative PCR was used to verify the sequence of the expressed gene and its primers

基因号 Gene number	基因注释 Gene annotation	引物序列(5'—3') Primer sequence(5'—3')
CqEF-1a	—	F:GTACGCATGGGTGCTTGACAAACTC
		R:ATCAGCCTGGGAGGTACCAGTAAT
AUR62004699	硝酸还原酶	F:GCAGAGGAGGACATATTG
		R:TCTCAACAACATACCATACTT
AUR62029383	亚硝酸还原酶	F:ATAAGGATGATGTTGATGTTAGA
		R:TTGCTCACTCGTTGTTAC
AUR62041208	谷氨酰胺合成酶	F:CAAGCACCAGGAGAAGAC
		R:TGTTACCACCACGGAATG
AUR62042896	磷酸烯醇丙酮酸羧化酶	F:TGGTGGTGGATATGAAGA
		R:GACAGATTGAGTAGGATGAG
AUR62038255	MYB44	F:TGTAGTAGTAGCGAGAATA
		R:CTTGTTAAGGCATCATCT
AUR62043570	尿卟啉原脱羧酶	F:TGCCCTGAAACACCACTTG
		R:CAGTCCAGCCCAATCACATT

图1 处理30 d后幼苗生长情况

Fig.1 Seedling growth after 30 d of treatment

表2 不同氮素水平条件下藜麦幼苗生物量积累(n=10)

Tab.2 Biomass accumulation of quinoa seedlings under different nitrogen levels(n=10)

处理 Treatment	总生物量/g Total biomass	地上部干质量/g Aboveground dry weight	地下部干质量/g Underground dry weight	根冠比 Root-shoot ratio	氮素利用效率 NUE
HN	1.77±0.11ab	1.47±0.12b	0.30±0.02a	0.20±0.08a	2.80±0.13c
LN	1.96±0.12a	1.72±0.13a	0.24±0.02b	0.14±0.01c	4.38±0.12a
NS	0.98±0.11c	0.83±0.07c	0.15±0.01c	0.18±0.01ab	3.08±0.14b

图2 不同氮素水平下藜麦叶片叶绿素a、叶绿素b、总叶绿素含量不同小写字母表示不同处理组处理5 d在0.05水平上差异显著(P<0.05); 不同大写字母表示不同处理组处理30 d在0.05水平上差异显著(P<0.05)。

Fig.2 Chlorophyll a,Chlorophyll b,Chlorophyll(a+b)content of quinoa leaves under different nitrogen levels Different lowercase letters indicate significant difference at the 0.05 level between different treatment groups at 5 d(P<0.05); Different capital letters indicate significant difference at the 0.05 level between different treatment groups at 30 d(P<0.05).

表3 藜麦叶片样本转录组数据统计

Tab.3 Transcriptome data statistics of quinoa leaf samples

样品 Samples	全部Reads数/条 Total Reads	质控后Reads数/条 Clean Reads	质控后Bases数/bp Clean Bases	Q30碱基数占比/% Q30	GC含量/% GC content
HN5d	43 666 970	21 833 485	6 515 802 112	94.06	44.13
LN5d	39 524 672	24 305 980	7 250 637 130	93.90	43.96
NS5d	48 611 960	19 762 336	5 897 857 974	93.90	43.99
HN30d	49 542 228	24 771 114	7 382 666 816	93.64	43.83
LN30d	55 391 736	22 149 363	6 599 239 224	94.02	44.03
NS30d	44 298 726	27 695 868	8 247 014 636	94.13	44.19

表4 转录组数据与参考基因组的比对

Tab.4 Aligning transcriptome data with reference genomes

样品 Samples	所有Reads数/条 Number of Reads	比对到参考基因组的 Reads/% Compare Reads to the reference genome	比对到参考基因组单一位置的比例/% Uniquely mapped to the reference genome	比对到参考基因组多位置的比例/% Multiple mapped to the reference genome
HN5d	43 666 970	95.54	87.59	7.95
LN5d	39 524 672	95.84	86.39	9.45
NS5d	48 611 960	95.92	86.24	9.69
HN30d	49 542 228	96.14	91.06	5.08
LN30d	55 391 736	96.66	90.33	6.34
NS30d	44 298 726	96.36	88.06	8.30

图3 不同氮素水平处理组上调差异表达基因的GO分析 A:1.膜的重要组成部分;2.细胞膜;3.等离子体膜;4.核;5.细胞质;6.氧化还原过程;7.代谢过程;8.氧化应激反应;9.跨膜运输;10.镉离子响应;11.金属离子结合;12.ATP结合;13.氧化还原酶活性;14.血红素结合;15.DNA结合。B:1.膜的重要组成部分;2.叶绿体;3.叶绿体基质;4.细胞膜;5.叶绿体类囊体膜;6.氧化还原过程;7.代谢过程;8.淀粉生物合成过程;9.二磷酸异戊烯基的生物合成过程;10.类囊体膜组织;11.ATP结合;12.金属离子结合;13.氧化还原酶活性;14.血红素结合;15.DNA结合.

Fig.3 GO analysis of up-regulated differentially expressed genes in treatment groups with different nitrogen levels A:1.Integral component of membrane;2.Membrane;3.Plasma membrane;4.Nucleus;5.Cytoplasm;6.Oxidation-reduction process;7.Metabolic process;8.Response to oxidative stress;9.Transmembrane transport;10.Response to cadmium ion;11.Metal ion binding;12.ATP binding;13.Oxidoreductase activity;14.Heme binding;15.DNA binding.B:1.Integral component of membrane;2.Chloroplast;3.Chloroplast stroma;4.Membrane;5.Chloroplast thylakoid membrane;6.Oxidation-reduction process;7.Metabolic process;8.Starch biosynthetic process;9.Isopentenyl diphosphate biosynthetic process;10.Thylakoid membrane organization;11.ATP binding;12.Metal ion binding;13.Oxidoreductase activity;14.Heme binding;15.DNA binding.

图4 不同氮素水平处理组上调差异表达基因的KEGG通路富集分析 A.低氮处理5 d;B.缺氮处理5 d;C.低氮处理30 d;D.缺氮处理30 d。

Fig.4 KEGG pathway enrichment analysis of up-regulated differentially expressed genes in treatment groups with different nitrogen levels A.Low nitrogen treatment for 5 d;B.Nitrogen deficiency treatment for 5 d;C.Low nitrogen treatment for 30 d;D.Nitrogen deficiency treatment for 30 d.

表5 低氮和缺氮处理5 d苯丙素生物合成和谷胱甘肽代谢通路上调基因表达分析

Tab.5 Analysis of up-regulated gene expression of phenylpropyl biosynthesis and glutathione metabolic pathways after 5 days of low-nitrogen and no-nitrogen treatment

基因ID Gene ID	低氮VS高氮 LN VS HN		缺氮VS高氮 NS VS HN		注释 Annotation
基因ID Gene ID	FDR	Log₂FC	FDR	Log₂FC	注释 Annotation
Chenopodium_quinoa_newGene_15607	6.78E-06	2.951	1.09E-05	2.811	过氧化物酶57
gene28362	5.24E-09	1.400	1.39E-32	2.761	过氧化物酶57
gene14234	2.33E-47	5.792	3.99E-18	4.019	谷胱甘肽s-转移酶
gene33030	1.32E-68	5.237	8.41E-42	4.074	谷胱甘肽s-转移酶U8
gene8274	2.05E-23	3.013	6.12E-21	2.827	抗坏血酸过氧化物酶

表6 低氮和缺氮处理30 d碳代谢通路上调DEGs表达分析

Tab.6 Analysis of up-regulated DEGs expression in carbon metabolism pathway after 30 days of low nitrogen and nitrogen deficiency treatment

基因ID Gene ID	低氮VS高氮 LN VS HN		缺氮VS高氮 NS VS HN		注释 Annotation
基因ID Gene ID	FDR	Log₂FC	FDR	Log₂FC	注释 Annotation
gene8744	2.29E-60	11.931	1.12E-42	11.024	磷酸甘油酸激酶
gene21246	1.34E-14	5.030	6.17E-16	5.307	半胱氨酸合成酶2
gene49294	2.34E-51	4.359	8.42E-73	8.42E-73	3-羟异丁酰辅酶A水解酶
gene49295	7.03E-22	4.354	6.92E-44	5.683	3-羟异丁酰辅酶A水解酶
gene19683	9.20E-70	4.328	4.65E-77	4.607	3-磷酸甘油醛脱氢酶(NADP)
gene4686	1.04E-65	4.184	9.60E-95	5.265	3-磷酸甘油醛脱氢酶(NADP)
gene34256	3.32E-10	1.370	1.91E-20	2.029	磷酸烯醇丙酮酸羧化酶1
gene2985	2.02E-07	1.137	7.83E-16	1.756	磷酸烯醇丙酮酸羧化酶1

图5 不同氮素水平处理下AUR62004699、AUR62029383、AUR62041208、AUR62042896、AUR62038255、AUR62043570基因的表达量

Fig.5 Expression levels of AUR62004699,AUR62029383,AUR62041208,AUR62042896, AUR62038255 and AUR62043570 under different nitrogen levels

参考文献 57

[1]	朱满喜, 张玉荣, 杨雅舒, 杨小兰, 王创云, 邓妍, 赵丽, 张丽光, 秦丽霞, 杨利艳. 藜麦NLP转录因子家族的鉴定及表达分析[J]. 华北农学报, 2021, 36(4):37—46.doi:10.7668/hbnxb.20191913.
	Zhu M X, Zhang Y R, Yang Y S, Yang X L, Wang C Y, Deng Y, Zhao L, Zhang L G, Qin L X, Yang L Y. Identification and expression analysis of NLP transcription factor family of Chenopodium quinoa willd[J]. Acta Agriculturae Boreali-Sinica, 2021, 36(4):37—46.
[2]	王业建. 大豆对低氮胁迫的形态和生理学响应及介导低氮胁迫miRNA的鉴定[D]. 长沙:中南大学, 2013.doi:10.7666/d.Y2687442.
	Wang Y J. Morphological and biological responses of different soybean varieties to low nitrogen and identification of low nitrogen regulated miRNA[D]. Changsha:Central South University, 2013.
[3]	赵平, 孙谷畴, 彭少麟. 植物氮素营养的生理生态学研究[J]. 生态科学, 1998, 17(2):39—44.
	Zhao P, Sun G C, Peng S L. Study on physiological ecology of plant nitrogen nutrition[J]. Ecological Science, 1998, 17(2):39—44.
[4]	刘天奇, 高红秀, 谢威, 张雪晴, 陈娜娜, 梅雪锋, 邢佳妮, 徐振华, 张忠臣. 水稻分蘖期氮素应答的转录组动态分析[J]. 华北农学报, 2021, 36(1):44—53.doi:10.7668/hbnxb.20191406.
	Liu T Q, Gao H X, Xie W, Zhang X Q, Chen N N, Mei X F, Xing J N, Xu Z H, Zhang Z C. Dynamic transcriptome analysis of rice response to nitrogen treatment at tillering stage[J]. Acta Agriculturae Boreali-Sinica, 2021, 36(1):44—53. doi: 10.7668/hbnxb.20191406
[5]	陈伟, 崔亚茹, 杨洋, 皇甫倩华, 孙从建, 张永清. 苦荞根系分泌有机酸对低氮胁迫的响应机制[J]. 土壤通报, 2019, 50(1):149—156.doi:10.19336/j.cnki.trtb.2019.01.23.
	Chen W, Cui Y R, Yang Y, Huangfu Q H, Sun C J, Zhang Y Q. Mechanism of organic acids secreted by roots of Tartary buckwheat under low nitrogen stress[J]. Chinese Journal of Soil Science, 2019, 50(1):149—156.
[6]	李强, 罗延宏, 龙文靖, 孔凡磊, 杨世民, 袁继超. 低氮胁迫对不同耐低氮性玉米品种苗期生长和生理特性的影响[J]. 草业学报, 2014, 23(4):204—212.doi:10.11686/cyxb20140425.
	Li Q, Luo Y H, Long W J, Kong F L, Yang S M, Yuan J C. Effect of low nitrogen stress on different low nitrogen tolerance maize cultivars seedling stage growth and physiological characteristics[J]. Acta Prataculturae Sinica, 2014, 23(4):204—212. doi: 10.11686/cyxb20140425
[7]	高宏伟, 杨武德, 冯美臣, 王超, Muhammad Saleem Kubar, 岳彧. 氮运筹对冬小麦氮素积累量和氮素吸收利用效率的影响[J]. 山西农业科学, 2020, 48(3):392—395.doi:10.3969/j.issn.1002-2481.2020.03.25.
	Gao H W, Yang W D, Feng M C, Wang C, Kubar M S, Yue Y. Effects of nitrogen management on nitrogen accumulation and nitrogen uptake and utilization efficiency in winter wheat[J]. Journal of Shanxi Agricultural Sciences, 2020, 48(3):392—395.
[8]	Hermans C, Hammond J P, White P J, Verbruggen N. How do plants respond to nutrient shortage by biomass allocation?[J]. Trends in Plant Science, 2006, 11(12):610—617.doi:10.1016/j.tplants.2006.10.007. pmid: 17092760
[9]	Liu Y Q, Wang H R, Jiang Z M, Wang W, Xu R N, Wang Q H, Zhang Z H, Li A F, Liang Y, Ou S J, Liu X J, Cao S Y, Tong H N, Wang Y H, Zhou F, Liao H, Hu B, Chu C C. Genomic basis of geographical adaptation to soil nitrogen in rice[J]. Nature, 2021, 590(7847):600—605.doi:10.1038/s41586-020-03091-w.
[10]	刘晓庆, 庞钦玮, 冀佳悦, 杨春, 高玲玲, 郭艳琼. 杀虫剂处理的梨小食心虫转录组测序分析及差异表达解毒酶基因筛选[J]. 山西农业科学, 2023, 51(11):1233—1244.doi:10.3969/j.issn.1002-2481.2023.11.01.
	Liu X Q, Pang Q W, Ji J Y, Yang C, Gao L L, Guo Y Q. Sequencing of transcriptome and screening of differentially expressed detoxifying enzyme genes of Grapholita molesta treated with insecticides[J]. Journal of Shanxi Agricultural Sciences, 2023, 51(11):1233—1244.
[11]	邵彩虹, 李瑶, 钱银飞, 陈金, 陈先茂, 关贤交, 刘光荣, 彭春瑞, 邱才飞. 氮素胁迫对水稻根系影响的转录组分析[J]. 华北农学报, 2018, 33(1):168—175.doi:10.7668/hbnxb.2018.01.025.
	Shao C H, Li Y, Qian Y F, Chen J, Chen X M, Guan X J, Liu G R, Peng C R, Qiu C F. Transcriptional analysis of rice root under nitrogen deficiency[J]. Acta Agriculturae Boreali-Sinica, 2018, 33(1):168—175. doi: 10.7668/hbnxb.2018.01.025
[12]	周海涛, 刘浩, 么杨, 杨修仕, 高文杰, 杨才, 任贵兴. 藜麦在张家口地区试种的表现与评价[J]. 植物遗传资源学报, 2014, 15(1):222—227.doi:10.13430/j.cnki.jpgr.2014.01.034.
	Zhou H T, Liu H, Yao Y, Yang X S, Gao W J, Yang C, Ren G X. Evaluation of agronomic and quality characters of quinoa cultivated in Zhangjiakou[J]. Journal of Plant Genetic Resources, 2014, 15(1):222—227. doi: 10.13430/j.cnki.jpgr.2014.01.033
[13]	Ogungbenle H N. Nutritional evaluation and functional properties of quinoa(Chenopodium quinoa)flour[J]. International Journal of Food Sciences and Nutrition, 2003, 54(2):153—158.doi:10.1080/0963748031000084106. pmid: 12701372
[14]	齐锦秋, 池冰, 谢九龙, 陈思敏, 黄兴彦. 慈竹纤维形态及组织比量的研究[J]. 中国造纸学报, 2013, 28(3):1—4.doi:10.3969/j.issn.1000-6842.2013.03.001.
	Qi J Q, Chi B, Xie J L, Chen S M, Huang X Y. Study on variations of fiber morphology and tissue proportion of Neosinocalamus affinis culm[J]. Transactions of China Pulp and Paper, 2013, 28(3):1—4.
[15]	张乐, 郭欢, 包爱科. 盐生植物的独特泌盐结构——盐囊泡[J]. 植物生理学报, 2019, 55(3):232—240.doi:10.13592/j.cnki.ppj.2019.0028.
	Zhang L, Guo H, Bao A K. The unique salt-secreting structures of halophytes:salt bladders[J]. Plant Physiology Journal, 2019, 55(3):232—240.
[16]	翟凤强, 蔡志全, 鲁建美. 施氮量对不同藜麦品种幼苗生长的影响[J]. 应用生态学报, 2020, 31(4):1139—1145.doi:10.13287/j.1001-9332.202004.010.
	Zhai F Q, Cai Z Q, Lu J M. Effects of nitrogen application rate on the growth traits in seedlings of different quinoa cultivars[J]. Chinese Journal of Applied Ecology, 2020, 31(4):1139—1145.
[17]	Jarvis D E, Ho Y S, Lightfoot D J, Schmöckel S M, Li B, Borm T J A, Ohyanagi H, Mineta K, Michell C T, Saber N, Kharbatia N M, Rupper R R, Sharp A R, Dally N, Boughton B A, Woo Y H, Gao G, Schijlen E G W M, Guo X J, Momin A A, Negrão S, Al-Babili S, Gehring C, Roessner U, Jung C, Murphy K, Arold S T, Gojobori T, van der Linden C G, van Loo E N, Jellen E N, Maughan P J, Tester M. The genome of Chenopodium quinoa[J]. Nature, 2017, 542:307—312.doi:10.1038/nature21370.
[18]	Li H, Li M C, Luo J, Cao X, Qu L, Gai Y, Jiang X N, Liu T X, Bai H, Janz D, Polle A, Peng C H, Luo Z B. N-fertilization has different effects on the growth,carbon and nitrogen physiology,and wood properties of slow-and fast-growing Populus species[J]. Journal of Experimental Botany, 2012, 63(17):6173—6185.doi:10.1093/jxb/ers271.
[19]	Ding L, Wang K J, Jiang G M, Biswas D K, Xu H, Li L F, Li Y H. Effects of nitrogen deficiency on photosynthetic traits of maize hybrids released in different years[J]. Annals of Botany, 2005, 96(5):925—930.doi:10.1093/aob/mci244. pmid: 16103036
[20]	荣楠, 韩永亮, 荣湘民, 宋海星, 彭建伟, 谢桂先, 张玉平, 张振华. 油菜$NO_{3}^{-}$的吸收、分配及氮利用效率对低氮胁迫的响应[J]. 植物营养与肥料学报, 2017, 23(4):1104—1111.doi:10.11674/zwyf.16305.
	Rong N, Han Y L, Rong X M, Song H X, Peng J W, Xie G X, Zhang Y P, Zhang Z H. Response of $NO_{3}^{-}$ uptake and distribution and nitrogen use efficiency in oilseed rape to limited nitrogen stress[J]. Journal of Plant Nutrition and Fertilizers, 2017, 23(4):1104—1111.
[21]	Safi A, Medici A, Szponarski W, Martin F, Clément-Vidal A, Marshall-Colon A, Ruffel S, Gaymard F, Rouached H, Leclercq J, Coruzzi G, Lacombe B, Krouk G. GARP transcription factors repress Arabidopsis nitrogen starvation response via ROS-dependent and-independent pathways[J]. Journal of Experimental Botany, 2021, 72(10):3881—3901.doi:10.1093/jxb/erab114.
[22]	顾雯雯, 胡亚婷, 韩英, 卞涵佳, 罗兵, 孙海燕, 万能, 杨志刚, 沈宗根. 植物过氧化物酶同工酶的研究进展[J]. 安徽农业科学, 2014, 42(34):12011—12013.doi:10.13989/j.cnki.0517-6611.2014.34.003.
	Gu W W, Hu Y T, Han Y, Bian H J, Luo B, Sun H Y, Wan N, Yang Z G, Shen Z G. Research progress of peroxidase isozymes in plants[J]. Journal of Anhui Agricultural Sciences, 2014, 42(34):12011—12013.
[23]	Foyer C H, Descourvières P, Kunert K J. Protection against oxygen radicals:an important defence mechanism studied in transgenic plants[J]. Plant,Cell & Environment, 1994, 17(5):507—523.doi:10.1111/j.1365-3040.1994.tb00146.x.
[24]	郭晓红. 柽柳过氧化物酶基因的序列分析及功能验证[D]. 哈尔滨: 东北林业大学, 2009.
	Guo X H. Sequence and function analysis of peroxidase in Tamarix hispida[D]. Harbin: Northeast Forestry University, 2009.
[25]	穆春. RNAi技术抑制萝卜过氧化物酶RsPrxl基因表达的初步研究[D]. 新乡:河南师范大学, 2012.doi:10.7666/d.Y2223370.
	Mu C. Primary research on RNAi-induced silencing of RsPrxl gene in radish[D]. Xinxiang:Henan Normal University, 2012.
[26]	Mittler R, Vanderauwera S, Gollery M, Van Breusegem F.Reactive oxygen gene network of plants[J]. Trends in Plant Science, 2004, 9(10):490—498.doi:10.1016/j.tplants.2004.08.009. pmid: 15465684
[27]	Wang M Q, Wang Y F, Zhang Y F, Li C X, Gong S C, Yan S Q, Li G L, Hu G H, Ren H L, Yang J F, Yu T, Yang K J. Comparative transcriptome analysis of salt-sensitive and salt-tolerant maize reveals potential mechanisms to enhance salt resistance[J]. Genes & Genomics, 2019, 41(7):781—801.doi:10.1007/s13258-019-00793-y.
[28]	Sandermann H Jr. Plant metabolism of xenobiotics[J]. Trends in Biochemical Sciences, 1992, 17(2): 82—84.doi:10.1016/0968-0004(92)90507-6. pmid: 1566333
[29]	Xu J, Tian Y S, Xing X J, Peng R H, Zhu B, Gao J J, Yao Q H. Over-expression of AtGSTU19 provides tolerance to salt,drought and methyl viologen stresses in Arabidopsis[J]. Physiologia Plantarum, 2016, 156(2):164—175.doi:10.1111/ppl.12347.
[30]	Asada K. Ascorbate peroxidase-a hydrogen peroxide-scavenging enzyme in plants[J]. Physiologia Plantarum, 1992, 85(2):235—241.doi:10.1034/j.1399-3054.1992.850216.x.
[31]	Kornyeyev D, Logan B A, Payton P, Allen R D, Holaday A S. Enhanced photochemical light utilization and decreased chilling-induced photoinhibition of photosystem Ⅱ in cotton overexpressing genes encoding chloroplast-targeted antioxidant enzymes[J]. Physiologia Plantarum, 2001, 113(3):323—331.doi:10.1034/j.1399-3054.2001.1130304.x.
[32]	Caldwell C R, Turano F J, McMahon M B. Identification of two cytosolic ascorbate peroxidase cDNAs from soybean leaves and characterization of their products by functional expression in E.coli[J]. Planta, 1998, 204(1):120—126.doi:10.1007/s004250050237. pmid: 9443387
[33]	郭楠, 赵敬会, 高天姝, 白瑞英, 黄兴琳, 陆俊杏, 张涛. 甘蓝型油菜PGK基因的克隆与表达分析[J]. 西北植物学报, 2014, 34(11):2188—2193.doi:10.7606/j.issn.1000-4025.2014.11.2188.
	Guo N, Zhao J H, Gao T S, Bai R Y, Huang X L, Lu J X, Zhang T. Cloning and expression analysis of PGK gene in Brassica napus[J]. Acta Botanica Boreali-Occidentalia Sinica, 2014, 34(11):2188—2193.
[34]	Watson H C, Walker N P, Shaw P J, Bryant T N, Wendell P L, Fothergill L A, Perkins R E, Conroy S C, Dobson M J, Tuite M F. Sequence and structure of yeast phosphoglycerate kinase[J]. The EMBO Journal, 1982, 1(12):1635—1640.doi:10.1002/j.1460-2075.1982.tb01366.x.
[35]	Bringloe D H, Rao S K, Dyer T A, Raines C A, Bradbeer J W. Differential gene expression of chloroplast and cytosolic phosphoglycerate kinase in tobacco[J]. Plant Molecular Biology, 1996, 30(3):637—640.doi:10.1007/BF00049337. pmid: 8605311
[36]	许奕, 金志强, 宋顺, 刘菊华, 贾彩红, 张建斌, 徐碧玉. 香蕉MaCSase基因的克隆和表达分析[J]. 中国农学通报, 2012, 28(34):202—210.doi:10.3969/j.issn.1000-6850.2012.34.035.
	Xu Y, Jin Z Q, Song S, Liu J H, Jia C H, Zhang J B, Xu B Y. Molecular cloning and expression analysis of MaCSase gene in banana[J]. Chinese Agricultural Science Bulletin, 2012, 28(34):202—210.
[37]	Bogdanova N, Hell R. Cysteine synthesis in plants:protein-protein interactions of serine acetyltransferase from Arabidopsis thaliana[J]. The Plant Journal:for Cell and Molecular Biology, 1997, 11(2):251—262.doi:10.1046/j.1365-313x.1997.11020251.x.
[38]	Saito K, Miura N, Yamazaki M, Hirano H, Murakoshi I. Molecular cloning and bacterial expression of cDNA encoding a plant cysteine synthase[J]. Proceedings of the National Academy of Sciences of the United States of America, 1992, 89(17):8078—8082.doi:10.1073/pnas.89.17.8078. pmid: 1518833
[39]	Nakamura T, Yamaguchi Y, Sano H. Four rice genes encoding cysteine synthase:isolation and differential responses to sulfur,nitrogen and light[J]. Gene, 1999, 229(1/2): 155—161. doi:10.1016/s0378-1119(99)00019-0.
[40]	卢倩, 弭晓菊, 崔继哲. 植物甘油醛-3-磷酸脱氢酶作用机制的研究进展[J]. 生物技术通报, 2013(8):1—6.doi:10.13560/j.cnki.biotech.bull.1985.2013.08.028.
	Lu Q, Mi X J, Cui J Z. Research advances on the mechanism of glyceraldehydes-3-phosphate dehydrogenase in plant[J]. Biotechnology Bulletin, 2013(8):1—6.
[41]	Singh R, Green M R. Sequence-specific binding of transfer RNA by glyceraldehyde-3-phosphate dehydrogenase[J]. Science, 1993, 259(5093):365—368.doi:10.1126/science.8420004. pmid: 8420004
[42]	Hara M R, Agrawal N, Kim S F, Cascio M B, Fujimuro M, Ozeki Y, Takahashi M, Cheah J H, Tankou S K, Hester L D, Ferris C D, Hayward S D, Snyder S H, Sawa A. S-nitrosylated GAPDH initiates apoptotic cell death by nuclear translocation following Siah1 binding[J]. Nature Cell Biology, 2005, 7(7):665—674.doi:10.1038/ncb1268. pmid: 15951807
[43]	Sirover M A. Minireview. Emerging new functions of the glycolytic protein,glyceraldehyde-3-phosphate dehydrogenase,in mammalian cells[J]. Life Sciences, 1996, 58(25):2271—2277.doi:10.1016/0024-3205(96)00123-3. pmid: 8649216
[44]	Zhang X H, Rao X L, Shi H T, Li R J, Lu Y T. Overexpression of a cytosolic glyceraldehyde-3-phosphate dehydrogenase gene OsGAPC3 confers salt tolerance in rice[J]. Plant Cell,Tissue and Organ Culture(PCTOC), 2011, 107(1):1—11.doi:10.1007/s11240-011-9950-6.
[45]	齐晓花, 许学文, 罗晶晶, 高海洁, 徐强, 林肖剑, 朱碧云, 陈学好. 黄瓜3-磷酸甘油醛脱氢酶基因CsGAPDH的克隆及其涝胁迫响应分析[J]. 园艺学报, 2011, 38(9):1693—1698.doi:10.16420/j.issn.0513-353x.2011.09.009.
	Qi X H, Xu X W, Luo J J, Gao H J, Xu Q, Lin X J, Zhu B Y, Chen X H. Cloning,characterization and expression of CsGAPDH,a waterlogging responsive gene in cucumber[J]. Acta Horticulturae Sinica, 2011, 38(9):1693—1698.
[46]	李晗, 李治龙, 李晓屿, 李玉花, 蓝兴国. 羽衣甘蓝不同组织及柱头发育实时荧光定量PCR内参基因的筛选[J]. 植物研究, 2016, 36(4):565—572.doi:10.7525/j.issn.1673-5102.2016.04.012.
	Li H, Li Z L, Li X Y, Li Y H, Lan X G. Selection of reference genes for real-time fluorescence quantitative PCR in different tissues and stigma development from ornamental kale[J]. Bulletin of Botanical Research, 2016, 36(4):565—572. doi: 10.7525/j.issn.1673-5102.2016.04.012
[47]	赵晓, 马会勤, 陈尚武, 徐海英. 葡萄果实发育后期半定量RT-PCR内参基因的优选[J]. 中国农业大学学报, 2010, 15(3):7—14.doi:10.11841/j.issn.1007-4333.2010.03.002.
	Zhao X, Ma H Q, Chen S W, Xu H Y. Internal reference gene selection for semi quantitative RT-PCR of genes in the second half of grape berry development[J]. Journal of China Agricultural University, 2010, 15(3):7—14.
[48]	赵晋锋, 杜艳伟, 王高鸿, 李颜方, 赵根有, 王振华, 王玉文, 余爱丽. 谷子PEPC基因的鉴定及其对非生物逆境的响应特性[J]. 作物学报, 2020, 46(5):700—711.doi:10.3724/SP.J.1006.2020.94107.
	Zhao J F, Du Y W, Wang G H, Li Y F, Zhao G Y, Wang Z H, Wang Y W, Yu A L. Identification of PEPC genes from foxtail millet and its response to abiotic stress[J]. Acta Agronomica Sinica, 2020, 46(5):700—711.
[49]	魏绍巍, 黎茵. 植物磷酸烯醇式丙酮酸羧化酶的功能及其在基因工程中的应用[J]. 生物工程学报, 2011, 27(12):1702—1710.doi:10.13345/j.cjb.2011.12.010.
	Wei S W, Li Y. Functions of plant phosphoenolpyruvate carboxylase and its applications for genetic engineering[J]. Chinese Journal of Biotechnology, 2011, 27(12):1702—1710.
[50]	焦进安. 植物磷酸烯醇式丙酮酸羧化酶的多生理功能[J]. 植物生理学通讯, 1987, 23(1):40—43.doi:10.13592/j.cnki.ppj.1987.01.016.
	Jiao J A. Multiple functions of phosphoenolpyruvate carboxylase in plants[J]. Plant Physiology Communications, 1987, 23(1):40—43.
[51]	丁在松, 周宝元, 孙雪芳, 赵明. 干旱胁迫下PEPC过表达增强水稻的耐强光能力[J]. 作物学报, 2012, 38(2):285—292.doi:10.3724/SP.J.1006.2012.00285.
	Ding Z S, Zhou B Y, Sun X F, Zhao M. High light tolerance is enhanced by overexpressed PEPC in rice under drought stress[J]. Acta Agronomica Sinica, 2012, 38(2):285—292.
[52]	焦德茂, 李霞, 黄雪清, 迟伟, 匡廷云, 古森本. 转PEPC基因水稻的光合CO₂同化和叶绿素荧光特性[J]. 科学通报, 2001, 46(5):414—418.doi:10.1360/csb2001-46-5-414.
	Jiao D M, Li X, Huang X Q, Chi W, Kuang T Y, Gu S B. Photosynthetic CO₂ assimilation and chlorophyll fluorescence characteristics of PEPC transgenic rice[J]. Chinese Science Bulletin, 2001, 46(5):414—418.
[53]	方立锋, 丁在松, 赵明. 转ppc基因水稻苗期抗旱特性研究[J]. 作物学报, 2008, 34(7):1220—1226.doi:10.3724/SP.J.1006.2008.01220.
	Fang L F, Ding Z S, Zhao M. Characteristics of drought tolerance in ppc overexpressed rice seedlings[J]. Acta Agronomica Sinica, 2008, 34(7):1220—1226.
[54]	Jeanneau M, Gerentes D, Foueillassar X, Zivy M, Vidal J, Toppan A, Perez P. Improvement of drought tolerance in maize:towards the functional validation of the Zm-Asr1 gene and increase of water use efficiency by over-expressing C4-PEPC[J]. Biochimie, 2002, 84(11):1127—1135.doi:10.1016/s0300-9084(02)00024-x. pmid: 12595141
[55]	González M C, Sánchez R, Cejudo F J. Abiotic stresses affecting water balance induce phosphoenolpyruvate carboxylase expression in roots of wheat seedlings[J]. Planta, 2003, 216(6):985—992.doi:10.1007/s00425-002-0951-x.
[56]	Sánchez R, Flores A, Cejudo F J. Arabidopsis phosphoenolpyruvate carboxylase genes encode immunologically unrelated polypeptides and are differentially expressed in response to drought and salt stress[J]. Planta, 2006, 223(5):901—909.doi:10.1007/s00425-005-0144-5.
[57]	García-Mauriño S, Monreal J A, Alvarez R, Vidal J, Echevarr a C. Characterization of salt stress-enhanced phosphoenolpyruvate carboxylase kinase activity in leaves of Sorghum vulgare:independence from osmotic stress,involvement of ion toxicity and significance of dark phosphorylation[J]. Planta, 2003, 216(4):648—655.doi:10.1007/s00425-002-0893-3. pmid: 12569407

选择文件类型/文献管理软件名称

选择包含的内容