Chin Bull Bot ›› 2017, Vol. 52 ›› Issue (4): 453-464.doi: 10.11983/CBB17044

Previous Articles     Next Articles

Arabidopsis Metalloprotease FtSH4 Regulates Leaf Senescence Through Auxin and Reactive Oxygen Species

Shengchun Zhang, Qingming Li, Chengwei Yang*   

  1. Guangdong Provincial Key Laboratory of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou 510631, China
  • Received:2017-03-09 Accepted:2017-05-06 Online:2017-05-05 Published:2017-07-01
  • Contact: Yang Chengwei E-mail:yangchw@scnu.edu.cn
  • About author:# Co-first authors

Abstract:

The plant metalloproteinases FtSH gene family has 12 members in Arabidopsis, and their functions are still unclear. In the present study, we analyzed the function of FtSH4 on leaf senescence using cell biology and genetics methods. The ftsh4-4 mutant displayed a premature leaf senescence phenotype with increased H2O2 content and cell death rate, decreased chlorophyll content, increased peroxidase gene expression and peroxidase activity. The ftsh4-4 leaf senescence phenotype could be rescued by applying the exogenous antioxidant AsA and endogenous or exogenous auxin by decreasing H2O2 content, peroxidase gene expression level and peroxidase activity. The expression of auxin response factor genes ARF2 and ARF7 was increased in the ftsh4-4 mutant and was reduced by exogenous auxin or AsA. Moreover, H2O2 content and the senescence phenotype of ftsh4-4 could be rescued by the arf2-8 mutant. These results indicate that FtSH4 gene plays an important role in the regulation of leaf senescence through auxin and reactive oxygen species.

Key words: FtSH4,, Arabidopsis,, reactive oxygen species,, auxin,, leaf senescence

Table 1

Primers used in this study"

Primer name Primer sequence (5ʹ-3ʹ)
PER33F TCTTCTCCATCACTTCTTCTTA
PER33R ATCCTCCAACACATATTCTCTA
PER37F CGCCAACACTCTTTGACAACAAG
PER37R ACTCATCCTTATCATTGCCTTCGC
ARF2F AATATAGCACCTTCATCTCCT
ARF2R ATCACACTCTACACTCTCAG
ARF7F GCTAATGCTAATAACAGTCCTT
ARF7R TCCACATTCTTCAGTCTCAA
SAG12F ATGATGAGCAAGCACTGATGAAGG
SAG12R TCCGTTAGTAGATTCGCCGTATCC
SAG13F GCGACAACATAAGGACGAACTCTG
SAG13R GAAGACAAAGAAATGCCACAAGCG
SAG101F GGGATGAGAGACGATGTGAGAGAG
SAG101R CGGGTGTTCATAAACTCGGTCAAG
SEN1F GGACATCCGACTAGAGCCATCAAC
SEN1R ATCGCCGTGAAGCCAGCAG
SEN4F AACCGCCAATTTCCACACTTACTC
SEN4R CTCTTGTTGCCCAATCGTCTGC
UBQ10F CCGACTACAACATTCAGAAG
UBQ10R TATCAATGGTGTCAGAACTCT

Figure 1

FtSH4 mutation causes leaf senescence of Arabi- dopsis^(A) The phenotype of premature senescence observed in the ftsh4-4 mutant (Bar=1 cm); (B) The chlorophyll content decreased in ftsh4-4 mutant; (C) The relative electrolytic leakage increased in ftsh4-4 mutant; (D) The peroxidase activities increased in ftsh4-4 mutant. * indicates significant difference at P<0.05; ** indicates significant difference at P<0.01 (Student’s t-test)."

Figure 2

Exogenous AsA rescued the leaf senescence phenotype of Arabidopsis ftsh4-4 mutant^(A) Exogenous AsA rescued the leaf senescence phenotype of ftsh4-4 mutant (Bar=1 cm); (B) Exogenous AsA reduced the H2O2 level of ftsh4-4 mutant (Bar=1 cm); (C) Exogenous AsA reduced the cell death of ftsh4-4 mutant (Bar=1 cm); (D) Exogenous AsA reduced the expression of peroxidase genes in ftsh4-4 mutant. * indicates significant difference at P<0.05; ** indicates significant difference at P<0.01 (Student’s t-test)."

Figure 3

Exogenous IAA restored the leaf senescence phenotype of Arabidopsis ftsh4-4 mutant^(A) Exogenous IAA restored the leaf senescence phenotype of ftsh4-4 mutant (Bar=1 cm); (B) Exogenous IAA reduced the cell death of ftsh4-4 mutant"

Figure 4

Increasing endogenous IAA restored the leaf senescence phenotype of Arabidopsis ftsh4-4 mutant^(A) Increasing endogenous IAA restored the leaf senescence phenotype of ftsh4-4 mutant (Bar=1 cm); (B) Cell death decreased in iaaM-ftsh4-4 (Bar=1 cm); (C) The chlorophyll content of iaaM-ftsh4-4 transgeneic line restored to the wild type level; (D) The relative electrolytic leakage of iaaM- ftsh4-4 transgeneic line restored to the wild type level; (E) Endogenous IAA decreased the expression of senescence-associated genes SAG12, SAG13, SAG101, SEN1 and SEN4 in ftsh4-4 mutant. * indicates significant difference at P<0.05; ** indicates significant difference at P<0.01 (Student’s t-test)."

Figure 5

ARF2 is involved in FtSH4-mediated leaf senescence of Arabidopsis^(A) The expression levels of ARF2 and ARF7 increased in the ftsh4-4 and were inhibited by the exogenous IAA and AsA; (B) ARF2 mutation rescued the leaf senescence of ftsh4-4 mutant (Bar=1 cm); (C) ARF2 mutation reduced the cell death of ftsh4-4 mutant (Bar=1 cm); (D) ARF2 mutation increased the chlorophyll content of ftsh4-4 mutant; (E) ARF2 mutation reduced the relative electrolytic leakage of ftsh4-4 mutant; (F) ARF2 mutation reduced the expression of senescence-associated genes SAG12, SAG13, SAG101, SEN1 and SEN4. * indicates significant difference at P<0.05; ** indicates significant difference at P<0.01 (Student’s t-test)."

Figure 6

IAA treatment reduces the H2O2 level and peroxidase activities of Arabidopsis ftsh4-4 mutant^(A) ARF2 mutation reduced the H2O2 level of ftsh4-4 mutant (Bar=1 cm); (B) ARF2 mutation reduced the peroxidase activities of ftsh4-4 mutant; (C) Exogenous IAA rescued the H2O2 level of ftsh4-4 mutant (Bar=1 cm); (D) Increasing endogenous IAA reduced the H2O2 level of ftsh4-4 mutant (Bar=1 cm); (E) Increasing endogenous IAA reduced the peroxidase activities of ftsh4-4 mutant; (F) Exogenous IAA reduced the peroxidase genes expression of ftsh4-4 mutant. * indicates significant difference at P<0.05; ** indicates significant difference at P<0.01 (Student’s t-test)."

[1] Apel K, Hirt H (2004). Reactive oxygen species: metabolism, oxidative stress, and signal transduction.Annu Rev Plant Biol 55, 373-399.
[2] Arnon DI (1949). Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol 24, 1-15.
[3] Bashandy T, Guilleminot J, Vernoux T, Caparros-Ruiz D, Ljung K, Meyer Y, Reichheld JP (2010). Interplay bet- ween the NADP-linked thioredoxin and glutathione systems in Arabidopsis auxin signaling.Plant Cell 22 376-391.
[4] Blomster T, Salojarvi J, Sipari N, Brosche M, Ahlfors R, Keinanen M, Overmyer K, Kangasjarvi J (2011). Apo- plastic reactive oxygen species transiently decrease auxin signaling and cause stress-induced morphogenic res- ponse in Arabidopsis.Plant Physiol 157, 1866-1883.
[5] Camilleri C, Jouanin L (1991). The TR-DNA region carrying the auxin synthesis genes of the Agrobacterium rhizo- genes agropine-type plasmid pRiA4: nucleotide sequence analysis and introduction into tobacco plants. Mol Plant Microbe Interact 4, 155-162.
[6] Chen GH, Liu CP, Chen SC, Wang LC (2012). Role of ARABIDOPSIS A-FIFTEEN in regulating leaf senescence involves response to reactive oxygen species and is dependent on ETHYLENE INSENSITIVE2. J Exp Bot 63, 275-292.
[7] Chen JP, Burke JJ, Velten J, Xin ZU (2006). FtsH11 protease plays a critical role in Arabidopsis thermotolerance.Plant J 48, 73-84.
[8] Ellis CM, Nagpal P, Young JC, Hagen G, Guilfoyle TJ, Reed JW (2005). AUXIN RESPONSE FACTOR1 and AUXIN RESPONSE FACTOR2 regulate senescence and floral organ abscission in Arabidopsis thaliana. Development 132, 4563-4574.
[9] Garcia-Lorenzo M, Sjodin A, Jansson S, Funk C (2006). Protease gene families in Populus and Arabidopsis. BMC Plant Biol 6, 30.
[10] Gazarian IG, Lagrimini LM, Mellon FA, Naldrett MJ, Ashby GA, Thorneley RN (1998). Identification of skatolyl hydroperoxide and its role in the peroxidase-catalysed oxidation of indol-3-yl acetic acid.Biochem J 333, 223-232.
[11] Gibala M, Kicia M, Sakamoto W, Gola EM, Kubrakiewicz J, Smakowska E, Janska H (2009). The lack of mitochondrial AtFtsH4 protease alters Arabidopsis leaf morphology at the late stage of rosette development under short-day photoperiod.Plant J 59, 685-699.
[12] Guo Y, Gan SS (2012). Convergence and divergence in gene expression profiles induced by leaf senescence and 27 senescence-promoting hormonal, pathological and environmental stress treatments.Plant Cell Environ 35, 644-655.
[13] He J, Duan Y, Hua D, Fan G, Wang L, Liu Y, Chen Z, Han L, Qu LJ, Gong Z (2012). DEXH box RNA helicase- mediated mitochondrial reactive oxygen species production in Arabidopsis mediates crosstalk between abscisic acid and auxin signaling.Plant Cell 24, 1815-1833.
[14] Hou K, Wu W, Gan SS (2013). SAUR36, a small auxin up RNA gene, is involved in the promotion of leaf senescence in Arabidopsis.Plant Physiol 161, 1002-1009.
[15] Huang YC, Chang YL, Hsu JJ, Chuang HW (2008). Trans- criptome analysis of auxin-regulated genes of Arabidopsis thaliana. Gene 420, 118-124.
[16] Jibran R, Hunter DA, Dijkwel PP (2013). Hormonal regulation of leaf senescence through integration of developmental and stress signals.Plant Mol Biol 82, 547-561.
[17] Joo JH, Bae YS, Lee JS (2001). Role of auxin-induced reactive oxygen species in root gravitropism.Plant Physiol 126, 1055-1060.
[18] Kant S, Bi YM, Zhu T, Rothstein SJ (2009). SAUR39, a small auxin-up RNA gene, acts as a negative regulator of auxin synthesis and transport in rice.Plant Physiol 151, 691-701.
[19] Kato Y, Miura E, Ido K, Ifuku K, Sakamoto W (2009). The variegated mutants lacking chloroplastic FtsHs are defective in D1 degradation and accumulate reactive oxygen sp- ecies.Plant Physiol 151, 1790-1801.
[20] Kim JI, Murphy AS, Baek D, Lee SW, Yun DJ, Bressan RA, Narasimhan ML (2011). YUCCA6 over-expression demonstrates auxin function in delaying leaf senescence inArabidopsis thaliana. J Exp Bot 62, 3981-3992.
[21] Kolodziejczak M, Kolaczkowska A, Szczesny B, Urantowka A, Knorpp C, Kieleczawa J, Janska H (2002). A higher plant mitochondrial homologue of the yeast m-AAA protease-molecular cloning, localization, and putative func- tion.J Biol Chem 277, 43792-43798.
[22] Kovtun Y, Chiu WL, Tena G, Sheen J (2000). Functional analysis of oxidative stress-activated mitogen-activated protein kinase cascade in plants.Proc Natl Acad Sci USA 97, 2940-2945.
[23] Li Z, Peng J, Wen X, Guo H (2012). Gene network analysis and functional studies of senescence-associated genes reveal novel regulators of Arabidopsis leaf senescence.J Integr Plant Biol 54, 526-539.
[24] Lim PO, Kim HJ, Nam HG (2007). Leaf senescence.Annu Rev Plant Biol 58, 115-136.
[25] Lim PO, Lee IC, Kim J, Kim HJ, Ryu JS, Woo HR, Nam HG (2010). Auxin response factor 2 (ARF2) plays a major role in regulating auxin-mediated leaf longevity.J Exp Bot 61, 1419-1430.
[26] Ljung K, Hull AK, Kowalczyk M, Marchant A, Celenza J, Cohen JD, Sandberg G (2002). Biosynthesis, conjugation, catabolism and homeostasis of indole-3-acetic acid in Arabidopsis thaliana. Plant Mol Biol 49, 249-272.
[27] Malnoe A, Wang F, Girard-Bascou J, Wollman FA, de Vitry C (2014). Thylakoid FtsH protease contributes to photosystem II and cytochrome b6f remodeling in Chlamydomonas reinhardtii under stress conditions.Plant Cell 26, 373-390.
[28] Meudt WJ, Gaines TP (1967). Studies on the oxidation of indole-3-acetic acid by peroxidase enzymes. I. Colorimetric determination of indole-3-acetic acid oxidation produ- cts.Plant Physiol 144, 118-128.
[29] Mittler R, Vanderauwera S, Suzuki N, Miller G, Tognetti VB, Vandepoele K, Gollery M, Shulaev V, Van Breu- segem F (2011). ROS signaling: the new wave?Trends Plant Sci 16, 300-309.
[30] Nakagami H, Soukupova H, Schikora A, Zarsky V, Hirt H (2006). A mitogen-activated protein kinase kinase kinase mediates reactive oxygen species homeostasis in Arabidopsis.J Biol Chem 281, 38697-38704.
[31] Nolden M, Ehses S, Koppen M, Bernacchia A, Rugarli EI, Langer T (2005). The m-AAA protease defective in hereditary spastic paraplegia controls ribosome assembly in mitochondria.Cell 123, 277-289.
[32] Okushima Y, Fukaki H, Onoda M, Theologis A, Tasaka M (2007). ARF7 and ARF19 regulate lateral root formation via direct activation ofLBD/ASL genes in Arabidopsis. Pla- nt Cell 19, 118-130.
[33] Piechota J, Kolodziejczak M, Juszczak I, Sakamoto W, Janska H (2010). Identification and characterization of high molecular weight complexes formed by matrix AAA proteases and prohibitins in mitochondria ofArabidopsis thaliana. J Biol Chem 285, 12512-12521.
[34] Potters G, Pasternak TP, Guisez Y, Palme KJ, Jansen MA (2007). Stress-induced morphogenic responses: growing out of trouble?Trends Plant Sci 12, 98-105.
[35] Queval G, Issakidis-Bourguet E, Hoeberichts FA, Vandorpe M, Gakiere B, Vanacker H, Miginiac-Maslow M, Van Breusegem F, Noctor G (2007). Conditional oxidative stress responses in the Arabidopsis photorespiratory mutant cat2 demonstrate that redox state is a key modulator of daylength-dependent gene expression, and define photoperiod as a crucial factor in the regulation of H2O2- induced cell death. Plant J 52, 640-657.
[36] Ren G, Zhou Q, Wu S, Zhang Y, Zhang L, Huang J, Sun Z, Kuai B (2010). Reverse genetic identification of CRN1 and its distinctive role in chlorophyll degradation in Arabidopsis.J Integr Plant Biol 52, 496-504.
[37] Romano CP, Robson PR, Smith H, Estelle M, Klee H (1995). Transgene-mediated auxin overproduction in Ara- bidopsis: hypocotyl elongation phenotype and interactions with the hy6-1 hypocotyl elongation and axr1 auxin-resis- tant mutants.Plant Mol Biol 27, 1071-1083.
[38] Sakamoto W, Tamura T, Hanba-Tomita Y, Murata M (2002). The VAR1 locus of Arabidopsis encodes a chloroplastic FtsH and is responsible for leaf variegation in the mutant alleles.Genes Cells 7, 769-780.
[39] Salleh FM, Evans K, Goodall B, Machin H, Mowla SB, Mur LA, Runions J, Theodoulou FL, Foyer CH, Rogers HJ (2012). A novel function for a redox-related LEA protein (SAG21/AtLEA5) in root development and biotic stress re- sponses.Plant Cell Environ 35, 418-429.
[40] Savitsky PA, Gazaryan IG, Tishkov VI, Lagrimini LM, Ruzgas T, Gorton L (1999). Oxidation of indole-3-acetic acid by dioxygen catalysed by plant peroxidases: specifi- city for the enzyme structure.Biochem J 340, 579-583.
[41] Sierla M, Rahikainen M, Salojarvi J, Kangasjarvi J, Kangasjarvi S (2013). Apoplastic and chloroplastic redox signaling networks in plant stress responses.Antioxid Re- dox Signal 18, 2220-2239.
[42] Suzuki N, Miller G, Morales J, Shulaev V, Torres MA, Mittler R (2011). Respiratory burst oxidases: the engines of ROS signaling.Curr Opin Plant Biol 14, 691-699.
[43] Wagner R, Aigner H, Pruzinska A, Jankanpaa HJ, Jansson S, Funk C (2011). Fitness analyses of Arabidopsis thaliana mutants depleted of FtsH metalloproteases and characterization of three FtsH6 deletion mutants exposed to high light stress, senescence and chilling. New Phytol 191, 449-458.
[44] Wang P, Song CP (2008). Guard-cell signaling for hydrogen peroxide and abscisic acid.New Phytol 178, 703-718.
[45] Watanabe M, Balazadeh S, Tohge T, Erban A, Giavalisco P, Kopka J, Mueller-Roeber B, Fernie AR, Hoefgen R (2013). Comprehensive dissection of spatiotemporal meta- bolic shifts in primary, secondary, and lipid metabolism during developmental senescence in Arabidopsis.Plant Physiol 162, 1290-1310.
[46] Woodward AW, Bartel B (2005). Auxin: regulation, action, and interaction.Ann Bot 95, 707-735.
[47] Xu F, Meng T, Li P, Yu Y, Cui Y, Wang Y, Gong Q, Wang NN (2011). A soybean dual-specificity kinase, GmSARK, and its Arabidopsis homolog, AtSARK, regulate leaf senescence through synergistic actions of auxin and ethyle- ne.Plant Physiol 157, 2131-2153.
[48] Yuan HM, Liu WC, Jin Y, Lu YT (2013). Role of ROS and auxin in plant response to metal-mediated stress.Plant Signal Behav 8, e24671.
[49] Zentgraf U, Laun T, Miao Y (2010). The complex regulation of WRKY53 during leaf senescence of Arabidopsis thali- ana.Eur J Cell Biol 89, 133-137.
[50] Zhang S, Wu J, Yuan D, Zhang D, Huang Z, Xiao L, Yang C (2014). Perturbation of auxin homeostasis caused by mitochondrial FtSH4 gene-mediated peroxidase accumulation regulates Arabidopsis architecture.Mol Plant 7, 856-873.
[51] Zhang S, Li C, Wang R, Chen Y, Shu S, Huang R, Zhang D, Xiao S, Yao N, Li J, Yang CW (2017). The mitochondrial protease FtSH4 regulates leaf senescence via WRKY- dependent salicylic acid signal.Plant Physiol 173, 2294-2307.
[52] Zimmermann P, Heinlein C, Orendi G, Zentgraf U (2006). Senescence-specific regulation of catalases in Arabidopsis thaliana(L.) Heynh. Plant Cell Environ 29, 1049-1060.
[1] He Zhenmei,Li Dongming,Qi Yanhua. Advances in Biofunctions of the ABCB Subfamily in Plants [J]. Chin Bull Bot, 2019, 54(6): 688-698.
[2] Zhang Shuhui,Wang Hong,Wang Wenru,Wu Xuelian,Xiao Yuansong,Peng Futian. Effects of Sucrose on Seedling Growth and Development and SnRK1 Activity in Prunus persica [J]. Chin Bull Bot, 2019, 54(6): 744-752.
[3] Xu Wanyue, Wang Yingxiang. Chromosome Behaviors of Male Meiocytes by Chromosome Spread in Arabidopsis thaliana [J]. Chin Bull Bot, 2019, 54(5): 620-624.
[4] Hu Kongqin, Ding Zhaojun. A TIR1-independent Auxin Signaling Module [J]. Chin Bull Bot, 2019, 54(3): 293-295.
[5] Dai Yujia,Luo Xiaofeng,Zhou Wenguan,Chen Feng,Shuai Haiwei,Yang Wenyu,Shu Kai. Plant Systemic Signaling Under Biotic and Abiotic Stresses Conditions [J]. Chin Bull Bot, 2019, 54(2): 255-264.
[6] Cui Shengnan, Zhang Yihan, Xu Fan. Heterologous Overexpression of Rice OsSAPP3 Gene Promotes Leaf Senescence in Transgenic Arabidopsis [J]. Chin Bull Bot, 2019, 54(1): 46-57.
[7] Ma Danying, Ji Dongchao, Xu Yong, Chen Tong, Tian Shiping. Advances in the Regulation on Autophagy by Reactive Oxygen Species in Plant Cells [J]. Chin Bull Bot, 2019, 54(1): 81-92.
[8] Zhang Xian-sheng. Chinese Scientists Have Made a Great Breakthrough in the Mechanism of Programmed Cell Death [J]. Chin Bull Bot, 2018, 53(4): 445-446.
[9] Zhao Xijuan, Qian Lichao, Liu Yule. Chinese Scientists Made Breakthrough Progresses in Plant Programmed Cell Death [J]. Chin Bull Bot, 2018, 53(4): 447-450.
[10] He Guangming, Deng Xingwang. Death Signal Transduction: Chloroplast-to-Mitochondrion Communication Regulates Programmed Cell Death in Plants [J]. Chin Bull Bot, 2018, 53(4): 441-444.
[11] Qianqian Zhang, Tong Zheng, Qian Yu, Lei Ge. Auxin and the Maintenance of Root Stem Cell Niches in Plants [J]. Chin Bull Bot, 2018, 53(1): 126-138.
[12] Haiwei Shuai, Yongjie Meng, Feng Chen, Wenguan Zhou, Xiaofeng Luo, Wenyu Yang, Kai Shu. Phytohormone-mediated Plant Shade Responses [J]. Chin Bull Bot, 2018, 53(1): 139-148.
[13] Guangchao Liu , Zhaojun Ding. Auxin Regulates Plant Growth and Development by Mediating Various Environmental Cues [J]. Chin Bull Bot, 2018, 53(1): 17-26.
[14] Xinlu Xu, Dandan Li, Yuandan Ma, Jianyun Zhai, Jianfei Sun, Yan Gao, Rumin Zhang. Responses of the Antioxidant Defense System of Osmanthus fragrans cv. ‘Tian Xiang TaiGe’ to Drought, Heat and the Synergistic Stress [J]. Chin Bull Bot, 2018, 53(1): 72-81.
[15] Wang Yu, He Yikun. The Molecular Mechanism of Nitric Oxide-mediated S-nitrosylation Coordinating with Protein Methylation During Abiotic Stress Responses [J]. Chin Bull Bot, 2017, 52(6): 681-684.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
[1] Zhang Zhen-jue. Some Principles Governing Shedding of Flowers and Fruits in Vanilla fragrans[J]. Chin Bull Bot, 1985, 3(05): 36 -37 .
[2] Qian Gao;Yuying Liu;Yinan Fei;Dapeng Li;Xianglin Liu* . Research Advances into the Root Radial Patterning Gene SHORT-ROOT[J]. Chin Bull Bot, 2008, 25(03): 363 -372 .
[3] Wang Bao-shan;Zou Qi and Zhao Ke-fu. Advances in Mechanism of Crop Salt Tolerance and Strategies for Raising Crop Salt Tolerance[J]. Chin Bull Bot, 1997, 14(增刊): 25 -30 .
[4] HE Feng WU Zhen-Bin. Application of Aquatic Plants in Sewage Treatment and Water Quality Improvement[J]. Chin Bull Bot, 2003, 20(06): 641 -647 .
[5] ZHANG Yan FANG Li LI Tian-Fei YAO Zhao-BingJIANG Jin-Hui. Effect of Calcium on the Heat Tolerance and Active Oxygen Metabolism of Tobacco Leaves[J]. Chin Bull Bot, 2002, 19(06): 721 -726 .
[6] JIA Hu-Sen LI De-QuanHAN Ya-Qin. Cytochrome b-559 in Chloroplasts[J]. Chin Bull Bot, 2001, 18(02): 158 -162 .
[7] Wei Sun;Chonghui Li;Liangsheng Wang;Silan Dai*. Analysis of Anthocyanins and Flavones in Different-colored Flowers of Chrysanthemum[J]. Chin Bull Bot, 2010, 45(03): 327 -336 .
[8] . Phosphate_Stress Protein and Iron_Stress Protein in Plants[J]. Chin Bull Bot, 2001, 18(05): 571 -576 .
[9] ZHANG Da-Yong, JIANG Xin-Hua. An Ecological Perspective on Crop Prduction[J]. Chin J Plan Ecolo, 2000, 24(3): 383 -384 .
[10] Gui Ji-xun, Zhu Ting-cheng. Study of Energy Flow Between Litter and Decomposers in Aneurolepidium chinese Grassland[J]. Chin J Plan Ecolo, 1992, 16(2): 143 -148 .