欢迎访问作物学报,今天是

作物学报 ›› 2009, Vol. 35 ›› Issue (3): 445-451.doi: 10.3724/SP.J.1006.2009.00445

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

蛋白和核酸合抑制剂对氮素诱导甜菜谷氨酰胺合成酶基因表达的影响

陈胜勇;侯静**;李彩凤;马凤鸣;尹春佳;黄兆峰   

  1. 东北农业大学农学院,黑龙江哈尔滨150030
  • 收稿日期:2008-06-01 修回日期:2008-10-21 出版日期:2009-03-12 网络出版日期:2009-01-16
  • 通讯作者: 李彩凤
  • 基金资助:

    本研究由国家自然科学基金项目(30471017,30771276),黑龙江省博士后科研启动基金项目(BSH-Q06103)资助

Influence of Inhibitors of Nucleic Acid Synthesis and Protein Synthesis on Glutamine Synthetase Gene Expression Induced by Nitrogen in Sugar Beet (Beta vulgaris L.)

CHEN Sheng-Yong;HOU Jing**;LI Cai-Feng;MA Feng-Ming;YIN Chun-Jie; HUANG Zhao-Feng   

  1. College of Agriculture, Northeast Agricultural University, Harbin 150030,China
  • Received:2008-06-01 Revised:2008-10-21 Published:2009-03-12 Published online:2009-01-16
  • Contact: LI Cai-Feng

摘要:

谷氨酰胺合成酶(GS)家族是甜菜等高等植物体内氨态氮同化酶, 也是氮利用与循环的核心构件。为了揭示在氮素诱导下, 放线菌素D(AMD)和放线菌酮(CHM)对甜菜GS基因调控表达的影响。采用半定量RT-PCR技术, 对甜菜的胞液型谷氨酰胺合成酶基因(GS1)和质体型谷氨酰胺合成酶基因(GS2)进行mRNA的表达检测, 同时进行GS活性的测定。结果表明, 甜菜幼苗经过低浓度AMD处理2~6 h, GS活性略有增加, 9 h后, 高和低浓度AMD处理下的GS活性都下降, 且随着浓度的增加下降幅度加大, 同时GS1mRNA和GS2mRNA的相对量随浓度的增加而下降。CHM处理甜菜幼苗9 h后, 随着浓度的增加和处理时间的延长, GS活性下降幅度增加, 但GS1mRNA和GS2mRNA的相对量在不同CHM浓度处理间变化不显著。

关键词: 甜菜, 谷氨酰胺合成酶, 氮素, 放线菌素D, 放线菌酮

Abstract:

Glutamine synthetase (GS, EC6.3.1.2) families are the key enzymes involving in nitrogen assimilation in the higher plants as well as a core elements for nitrogen use efficiency and nitrogen recycle. The objective of this study was to reveal the effect of actinomycin D (AMD) and cycloheximide (CHM) on GS gene expression and its activities induced by nitrogen in sugar beet. GS activity in sugar beet was determined under the treatment of AMD and CHM. Gene transcripts of cytosolic glutamine synthetase (GS1) and plastidic glutamine synthetase (GS2) were detected by semi-quantitative PCR. And e?ciency of GSmRNA synthesis from each sample was estimated by quantitative PCR of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The results showed that the GS activities increased slightly under the low concentration treatment of AMD for 2–6 hours, but decreased with treatment of all concentrations. The transcript level of GS1mRNA and GS2mRNA decreased with the increase of AMD concentration treated for more than 9 h. GS activities decreased fast with the increase of CHM concentrations treated for more than 9 h. There was no significant difference between GS1mRNA and GS2mRNA expressions in the treatments with all CHM concentrations for more than 9 hours.

Key words: Sugar beet(Beta vulgaris L.), Glutamine synthetase, Nitrogen, Actinonycin D, Cycloheximide

[1]Migge A, Carrayol E, Hirel B, Becher T W. Leaf-specific overexpression of plastidic glutamine synthetase stimulates the growth of transgenic tobacco seedlings. Planta, 2000, 210: 252–260
[2]Tobin A K, Yamaya T. Cellular compartmentation of ammonium assimilation in rice and barley. J Exp Bot, 2001, 52: 591–604
[3]Weber A, Flugge U I. Interaction of cytosolic and plastidic nitrogen metabolism in plants. J Exp Bot, 2002, 53: 865–874
[4]Li C-F(李彩凤), Ma F-M(马凤鸣), Zhao Y(赵越). Effects of nitrogen forms on key enzyme activities and related products in sugar and nitrogen metabolism of sugar beet (Beta vulgaris L.). Acta Agron Sin (作物学报), 2003, 29(1): 128–132(in Chinese with English abstract)
[5]Zhao Y(赵越), Ma F-M(马凤鸣), Zhang D-Y(张多英). Study on the absorption kinetics of different nitrogen in sugar beet. J Northeast Agric Univ(东北农业大学学报), 2006, 37(3): 294–298 (in Chinese with English abstract)
[6]Wu X-P(吴小平), Wang P-H(汪沛洪), Zhang H(张惠). Effects of osmotic stress on protease activities of two winter wheat cultivars with different drought resistance. Acta Agric Boreali-Occident Sin (西北农业学报), 1994, 3(1): 49–53(in Chinese with English abstract)
[7]Xue Y-C(薛永常), Chen Y-J(陈永军), Cao M(曹敏). Effect of actinomycin D and cycloheximide on ABA accumulation induced by water stress in winter wheat seedling. J Hebei Norm Univ(Nat Sci)(河北师范大学学报×自然科学版), 1997, 21(3): 317–319(in Chinese with English abstract)
[8]Wang X-K(王学奎), Li H-S(李合生), Liu W-D(刘武定). Preliminary study on mechanism of light regulated glutamine synthetase in wheat leaves. J Huazhong Agric Univ(华中农业大学学报), 2000, 19(2): 102–105(in Chinese with English abstract)
[9]Yin L-P(印莉萍), Chai X-Q(柴晓清), Liu X-L(刘祥林). Influence of chloroplast development and light on the gene expression of glutamine synthetase in wheat leaves. Acta Bot Sin(植物学报), 1994, 36(8): 597–602(in Chinese with English abstract)
[10]Schumidt S, Mobr H. Regulation of the appearance of glutamine synthetase in mustard (Sinap isalba L.) cotyledons by light, nitrate and ammonium. Planta, 1989, 177: 526–534
[11]Hirel B, Cadal P. Glutamine synthetase in rice: A comparative study of the enzymes from roots and leaves. Plant Physiol, 1980, 66: 619–623
[12]Peterman T K, Goodman H M. The glutamine synthetase gene family of Arabidopsis thaliana: Light-regulation and differential expression in leaves, root and seeds. Mol Gen Genet, 1991, 230: 145–154
[13]Chen Y(陈煜), Zhu B-G(朱保葛), Zhang J(张敬). Effects of different nitrogens on activities of nitrate reductase, glutamine synthetase and seed protein contents in soybean vultivars. Soybean Sci(大豆科学), 2004, 23(2): 143–146(in Chinese with English abstract)
[14]Dai T-B(戴廷波), Cao W-X(曹卫星), Sun C-F(孙传范). Effect of enhanced ammonium nutrition on photosynthesis and nitrate reductase and glutamine synthetase activities of winter wheat. Chin J Appl Ecol (应用生态学报), 2003, 14(9): 1529–1532(in Chinese with English abstract)
[15]Mack G. Organ-specific changes in the activity and subunit composition of glutamine synhetase isoforms of barley (Hordeum vulgare L.) after growth on different levels of NH4+. Planta, 1995, 196: 231–238
[16]Zhang C F, Peng S B, Peng X X. Response of glutamine synthetase isoforms to nitrogen sources in rice (Oryza sativa L.) roots. Plant Sci, 1997, 125: 163–170
[17]Lam H M, Coschigano K T, Oliveira I C. The molecular genetics of nitrogen assimilation into amino acids in higher plants. Annu Rev Plant Physiol Plant Mol Biol, 1996, 47: 569–593
[18]Li C-J(李常健), Lin Q-H(林清华), Zhang C-F(张楚富). Effect of NaCl stress on activity and isozymes of glutamine synthetase in rice plants. J Wuhan Univ(Nat Sci Edn)(武汉大学学报×自然科学版), 1999, 45(4): 497–500(in Chinese with English abstract)
[19]Hoshi H, Tanaka Y, Hibino T. Enhanced tolerance to salt stress in transgenic rice that overexpress chloroplast glutamine synthetase. Plant Mol Biol, 2000, 43: 103–111
[20]Miflin B J, Lea P J. Ammonia Assimilation in the Biochemistry of Plants: Amino Acids Their Derivatives. New York: Academic Press, 1980. pp 169–202
[21]Oliveira L C, Coruzzi G M. Carbon and Amino acids reciprocally modulate the expression of glutamine synthetase in Arabidopsis. Plant Physiol, 1999, 121(1): 301–309
[22]Chen S-Y(陈胜勇), Li C-F(李彩凤), Ma F-M(马凤鸣), Yang D-G(杨德光), Hou J(侯静), Sun S-C(孙世臣), Yin C-J(尹春佳), Huang Z-F(黄兆峰), Zhao L-Y(赵丽影), Chen Y-T(陈业婷), Yue P(越鹏). Expression analysis of glutamine synthetase gene under different nitrogen conditions in sugar beet (Beta vulgaris L.). Crops (作物杂志), 2008, (4): 64–67(in Chinese with English abstract)
[23]Kang S M, Titus J S. Activity pro?les of enzymes involved in glutamine and glutamate metabolism in the apple during autumnal snescence. Physiol Plant, 1980, 50: 291–297
[24]Kang S M, Titus J S. Increased proteolysis of senescing rice leaves in the presence of NaCl and KCl. Plant Physiol, 1989, 91: 1232–1237
[25]Kar M, Feierabend J. Changes in the activities of enzymes involved in amino acid metabolism during the senescence of detached wheat leaves. Physiol Plant, 1984, 62: 39–44
[26]Yuang H, Hou C A. comparative study of two forms of glutamine synthetase from rice leaves. Bot Bull Acad Sin, 1987, 28: 91–108
[27]Han N(韩娜), Ge R-C(葛荣朝), Zhao B-C(赵宝存). Research development of the glutamine synthetase in plants. J Hebei Norm Univ(Nat Sci Edn)(河北师范大学学报×自然科学版), 2004, 28(4): 407–410 (in Chinese with English abstract)
[28]Becker K, Pan D, Whitley C B. Real-time quantitative polymerase chain reaction to assess gene transfer. Human Gene Therapy, 1999, 10: 2559–2566
[1] 杨宗桃, 杨婷, 王禹童, 艾静, 李燕烨, 刘家勇, 邓军, 赵勇, 张跃彬. 甘蔗CLC基因家族鉴定与表达分析[J]. 作物学报, 2026, 52(3): 722-734.
[2] 于天一, 王春晓, 肖丽, 钟召迪, 王宣仓, 赵勇, 路亚, 吴月, 吴正锋. 不同结瘤特性花生品种氮素累积、产量及品质特性对氮肥用量的响应[J]. 作物学报, 2026, 52(3): 881-894.
[3] 刘宁, 樊平, 王成, 陈琪琪, 成庆悦, 铁夏娜, 汤菁莎, 刘彬彬, 谢鸿堃, 王嘉悦, 施园青, 马均. 减氮配施有机肥对机插稻产量形成与氮素利用的影响[J]. 作物学报, 2026, 52(3): 866-880.
[4] 王婷, 段武丽, 王蕊, 刘海岚. 植物胆碱单加氧酶基因家族的进化与表达分析[J]. 作物学报, 2026, 52(1): 44-55.
[5] 尤根基, 谢昊, 梁毓文, 李龙, 王玉茹, 蒋晨炀, 郭剑, 李广浩, 陆大雷. 氮肥减施措施对江淮春玉米产量和氮素吸收利用的影响[J]. 作物学报, 2025, 51(8): 2152-2163.
[6] 闫喆林, 任强, 樊志龙, 殷文, 孙亚丽, 范虹, 何蔚, 胡发龙, 闫丽娟, 柴强. 氮肥后移优化绿洲灌区小麦间作玉米种间关系提高氮素利用效率[J]. 作物学报, 2025, 51(8): 2190-2203.
[7] 霍建喆, 于爱忠, 王玉珑, 王鹏飞, 尹波, 刘亚龙, 张冬玲, 姜科强, 庞小能, 王凤. 有机肥替代化肥对绿洲灌区甜玉米产量、品质及氮素利用的影响[J]. 作物学报, 2025, 51(7): 1887-1900.
[8] 孟凡琦, 房孟颖, 罗艺, 卢霖, 董学瑞, 王亚菲, 郭丽娜, 闫鹏, 董志强, 张凤路. 乙烯利-甜菜碱-水杨酸合剂对夏玉米耐热性和产量的调控效应[J]. 作物学报, 2025, 51(5): 1299-1311.
[9] 闫秉春, 万雪, 钟敏, 刘宇奇, 赵艳泽, 姜红芳, 刘雅, 刘惠玲, 马沁春, 高继平, 张文忠. 氮素水平对北方优良食味粳米品质及精碾磨粉颗粒特性的影响[J]. 作物学报, 2025, 51(2): 503-515.
[10] 黄绒, 周渠晨, 陈楚铭, 罗倩, 易东, 杜常欢, 黄祥宇, 盛锋, 杜雪竹. 过表达BnNRT2.3-like对油菜氮素吸收利用及产量的影响[J]. 作物学报, 2025, 51(12): 3184-3197.
[11] 王晶卿, 陈惠哲, 兰天明, 李慧, 唐承翰, 马昕伶, 张玉屏, 王亚梁. 不同播期下氮素穗肥对大穗型籼粳杂交稻甬优538颖花退化的调控作用[J]. 作物学报, 2025, 51(10): 2759-2774.
[12] 郝琪, 陈天陆, 王富贵, 王振, 白岚方, 王永强, 王志刚. 基于无人机多光谱数据和氮素空间分异的玉米冠层氮浓度估算[J]. 作物学报, 2025, 51(1): 189-206.
[13] 王媛, 许佳茵, 董二伟, 王劲松, 刘秋霞, 黄晓磊, 焦晓燕. 有机肥替代化肥氮对谷子氮素累积、产量及品质的影响[J]. 作物学报, 2025, 51(1): 149-160.
[14] 刘陈, 王昆昆, 廖世鹏, 杨佳群, 丛日环, 任涛, 李小坤, 鲁剑巍. 氮肥用量对玉米-油菜和水稻-油菜轮作模式下油菜产量及氮素吸收利用的影响[J]. 作物学报, 2024, 50(8): 2067-2077.
[15] 宋志文, 赵蕾, 毕俊国, 唐清芸, 王国栋, 李玉祥. 滴灌条件下施氮量对不同氮效率水稻品种物质积累及养分吸收的影响[J]. 作物学报, 2024, 50(8): 2025-2038.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!