柑橘黄龙病菌侵染对甜橙叶片糖代谢的影响

doi:10.3864/j.issn.0578-1752.2015.01.07

Abstract

Abstract: 【Objective】The objective of this experiment is to study the effect of Huanglongbing (HLB) infection on carbohydrate metabolism in Citrus sinensis, elucidate the relationship between HLB infection and starch accumulation, and provide a theoretical basis for HLB pathogenesis. 【Method】C. sinensis were used as the experimental materials, plants were inoculated by grafting three bark pieces onto the rootstock portion of each plant. For infected plants, bark pieces were derived from trees infected with Candidatus Liberibacter asiaticus, which were confirmed positive for Ca. L. asiaticus by PCR. For non-infected plants, bark pieces were obtained from healthy trees. Three plants in good growth state were chosen for successive test, mature leaves were collected every month until October. DNA and RNA were extracted, the content of soluble sugar and starch were measured immediately, the other leaf tissue was snap-frozen with liquid nitrogen and stored at -80℃ for testing carbohydrate metabolism related key enzyme activities. At the same time, the expression levels were compared using real-time quantitative PCR analysis. 【Result】The content of soluble sugar and starch tended to increase with time extended, and their peak value were respectively reached 1.59 and 3.73 times of control in June, then slightly decreased later, the ratio of soluble sugar to starch dropped down. Various key enzymes imposed different hands on carbohydrate metabolism at different stages of infection. Sucrose phosphate synthase activity rapidly rose to the top 58.44 μg?g^-1?min^-1 and had become lower than control in late infection; acid invertase had been consistently high and the difference was significant (P＜0.05), the highest activity was 2.91 times of control at middle stage; the activity of neutral invertase was kept at a lower level and had the same change trend in experimental group and control group; soluble starch synthase activity was closely related to granule-bond starch synthase, coordinating the synthesis of acting on the starch, while amylase activity had dropped to some extend at each stage, even as low as 0.38 U?g^-1 at middle stage. Real-time quantitative PCR indicated that there was a significant relationship between the expression level of sucrose phosphate synthase and its enzyme activity (P＜0.05). Among the sucrose decomposition related genes, sucrose synthase gene was not changed obviously and had lower change range while cell wall-bound invertase gene CSCWI was up-regulated with the highest value of about 7.4 times and expressed abundantly during the whole process. Starch decomposition-related genes BAM3, MEX1 and DPE2 had a relatively reduced expression and the difference was significant (P＜0.05) in different periods.【Conclusion】HLB infection has influence on the formation and transportation of photosynthate in C. sinensis and results in the disruption of carbohydrate metabolism balance, it is in deep connection with starch accumulation and the late stage symptoms.

Key words: Citrus sinensis, citrus Huanglongbing; Candidatus Liberibacter asiaticus, carbohydrate metabolism, starch accumulation

WU Yue, SU Hua-nan, HUANG Ai-jun, ZHOU Yan, LI Zhong-an, LIU Jin-xiang, ZHOU Chang-yong. Effect of Candidatus Liberibacter asiaticus Infection on Carbohydrate Metabolism in Citrus sinensis[J].Scientia Agricultura Sinica, 2015, 48(1): 63-72.

0
/ / Recommend

Add to citation manager EndNote|Reference Manager|ProCite|BibTeX|RefWorks

URL: https://www.chinaagrisci.com/EN/10.3864/j.issn.0578-1752.2015.01.07

https://www.chinaagrisci.com/EN/Y2015/V48/I1/63

References

[1] Bové J M. Huanglongbing: a destructive, newly-emerging, century- old disease of citrus. Journal of Plant Pathology, 2006, 88(1): 7-37.

[2] Schneider H. Anatomy of greening-diseased sweet orange shoots. Phytopathology, 1968, 58(1): 1155-1160.

[3] Etxeberria E, Gonzalez P, Achor D, Albrigo G. Anatomical distribution of abnormally high levels of starch in HLB-affected Valencia orange trees. Physiological and Molecular Plant Pathology, 2009, 74(1): 76-83.

[4] 夏正俊, 顾本康, 吴蔼民, 李经仪, 陈轶林, 陈小波. 棉花品种抗黄萎病性与体内生化成分相关分析. 植物保护学报, 1994, 21(4): 305-310.

Xia Z J, Gu B K, Wu A M, Li J Y, Chen Y L, Chen X B. Studies on the biochemical components in cotton related to the resistance to Verticillium dahliae. Acta Phytophylacica Sinica, 1994, 21(4): 305-310. (in Chinese)

[5] 章元寿. 植物病理生理学. 南京: 江苏科学技术出版社, 1996: 34-38.

Zhang Y S. Plant Pathophysiology. Nanjing: Jiangsu Scientific and Technical Press, 1996: 34-38. (in Chinese)

[6] Duan Y P, Zhou L J, Hall D G, Li W B, Doddapanenl H, Lin H, Lin H, Liu L, Vahling C M, Gabriel D W, Williams K P, Dickerman A, Sun Y J, Gottwald T. Complete genome sequence of citrus Huanglongbing bacterium, ‘Candidatus Liberibacter asiaticus’ obtained through metagenomics. Molecular Plant-Microbe Interactions, 2009, 22(8): 1011-1020.

[7] Albrecht U, Bowman K D. Gene expression in Citrus sinensis (L.) Osbeck following infection with the bacterial pathogen Candidatus Liberibacter asiaticus causing Huanglongbing in Florida. Plant Science, 2008, 175(3): 291-306.

[8] Kim J S, Sagaram U S, Burns J K, Li J L, Wang N. Response of sweet orange (Citrus sinensis) to ‘Candidatus Liberibacter asiaticus’ infection: microscopy and microarray analyses. Phytopathology, 2009, 99(1): 50-57.

[9] Martinelli F, Uratsu S L, Albrecht U, Reagan R L, Phu M L, Britton M, Buffalo V, Fass J, Leicht E, Zhao W X, Lin D W, Souza R D, Davis C E, Bowman K D, Dandekar A M. Transcriptome profiling of citrus fruit response to Huanglongbing disease. PLos One, 2012, 7(5): e38039.

[10] Fan J, Chen C, Brlansky R H, Gmitter Jr F G, Li Z G. Changes in carbohydrate metabolism in Citrus sinensis infected with ‘Candidatus Liberibacter asiaticus’. Plant Pathology, 2010, 59: 1037-1043.

[11] Subandiyah S, Iwanami T, Tsuyumu S, Ieki H. Comparison of 16S rDNA and 16S/23S intergenic region sequences among citrus greening organisms in Asia. Plant Disease, 2000, 84(1): 15-18.

[12] 李合生. 植物生理生化实验原理和技术. 北京: 高等教育出版社, 2006.

Li H S. The Experiment Principle and Technique on Plant Physiology and Biochemistry. Beijing: Higher Education Press, 2006. (in Chinese)

[13] Nielsen T H, Skiarbak H C, Karlsen P. Carbohydrate metabolism during fruit development in sweet pepper (Capsicum annuum) plants. Physio1ogia Plantarum, 1991, 82: 311-319.

[14] Nakamura Y, Yuki K, Park S Y, Ohya T. Carbohydrate metabolism in the developing endosperm of rice grains. Plant and Cell Physiology, 1989, 30(6): 833-839.

[15] Brunner A M, Yakovlev I A, Strauss S S. Validating internal controls for quantitative plant gene expression studies. BMC Plant Biology, 2004, 4(1): 14.

[16] Livak K J, Schmittgen T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods, 2001, 25: 402-408.

[17] 李佐同, 靳学慧, 张亚玲, 吴成龙. 水稻幼苗可溶性糖及可溶性蛋白含量与抗瘟性的关系. 北方水稻, 2009, 39(4): 6-9.

Li Z T, Jin X H, Zhang Y L, Wu C L. The relationship between soluble protein, soluble sugar content and rice blast resistance of rice seedlings. North Rice, 2009, 39(4): 6-9. (in Chinese)

[18] Smeekens S. Sugar-induced signal transduction in plants. Annual Review of Plant Physiology, 2000, 51(1): 49-81.

[19] Jang J C, Sheen J. Sugar sensing in higher plants. The Plant Cell, 1994, 6(11): 1665-1679.

[20] Achor D S, Etxeberria E, Wang N, Folimonova S Y, Chung K R, Albrigo L G. Sequence of anatomical symptom observations in citrus affected with Huanglongbing disease. Plant Pathology Journal, 2010, 9(2): 56-64.

[21] Schaffer A A, Liu K C, Goldschmidt E E. Citrus leaf chlorosis induced by sink removal: starch, nitrogen, and chloroplast ultrastructure. Journal of Plant Physiology, 1986, 124(1): 111-121.

[22] 侯金锋. 大豆鲜籽粒蔗糖含量的研究及糖代谢相关基因的克隆与功能分析[D]. 南京: 南京农业大学, 2012.

Hou J F. Studies on the sucrose contents of fresh seeds and functional analysis of key genes involved in sugar accumulation in soybean[D]. Nanjing: Nanjing Agricultural University, 2012. (in Chinese)

[23] Harbron S, Foyer C, Walker D. The purification and properties of sucrose-phosphate synthetase from spinach leaves: the involvement of this enzyme and fructose bisphosphatase in the regulation of sucrose biosynthesis. Archives of Biochemistry and Biophysics, 1981, 212(1): 237-246.

[24] Tymowska-Lalanne Z, Kreis M. The plant invertases: physiology, biochemistry and molecular biology. Advances in Botanical Research, 1998, 28(1): 71-117.

[25] 钟云. Candidatus Liberibacter asiaticus诱导的柑橘转录组学及蛋白组学研究[D]. 长沙: 湖南农业大学, 2012.

Zhong Y. Research of transcription genomics and proteomics of citrus Induced by Candidatus Liberibacter asiaticus[D]. Changsha: Hunan Agricultural University, 2012. (in Chinese)

[26] Swarbrick P J, Schulze-lefert P, Scholes J D. Metabolic consequences of susceptibility and resistance (race-specific and broad-spectrum) in barley leaves challenged with powdery mildew. Plant, Cell & Environment, 2006, 29(6): 1061-1076.

[27] Smith S M, Fulton D C, Chia T, Thorneycroft D, Chapple A, Dunstan H, Hylton C, Zeeman S C, Smith A M. Diurnal changes in the transcriptome encoding enzymes of starch metabolism provide evidence for both transcriptional and posttranscriptional regulation of starch metabolism in Arabidopsis leaves. Plant Physiology, 2004, 136: 2687-2699.

[28] 安新民. 柑橘转化酶基因家族成员的分离及表达研究[D]. 杭州: 浙江大学, 2002: 10.

An X M. Studies on isolation and expression of members of invertase gene family from citrus[D]. Hangzhou: Zhejiang University, 2002: 10. (in Chinese)

[29] 李玲, 潘瑞炽. BA对花生叶片蔗糖和淀粉代谢有关酶活性的影响. 热带亚热带植物学报, 1994, 2(2): 52-57.

Li L, Pan R Z. Effect of benzyladenine on enzymatic activities of sucrose and starch metabolism in peanutleaves. Journal of Tropical and Subtropical Botany, 1997, 2(2): 52-57. (in Chinese)

[30] Andre A, Maucourt M, Moing A, Rolin D, Renaudin J. Sugar import and phytopathogenicity of Spiroplasma citri: Glucose and fructose play distinct roles. Molecular Plant-Microbe Interactions, 2005, 18(1): 33-42.

Metrics

Viewed

Full text

Abstract

Cited

Shared

Discussed

Comments

Recommended 0

No Suggested Reading articles found!

Effect of Candidatus Liberibacter asiaticus Infection on Carbohydrate Metabolism in Citrus sinensis

RichHTML

PDF

Abstract

Cite this article

share this article

References

Related Articles 3

Metrics

Comments

Recommended 0

[1]	ZHANG JiaHua,YANG HengShan,ZHANG YuQin,LI CongFeng,ZHANG RuiFu,TAI JiCheng,ZHOU YangChen. Effects of Different Drip Irrigation Modes on Starch Accumulation and Activities of Starch Synthesis-Related Enzyme of Spring Maize Grain in Northeast China [J]. Scientia Agricultura Sinica, 2022, 55(7): 1332-1345.
[2]	SHAO XiaoLong, WANG Nan, SHI XiaoZhuan, SHEN Fei, SONG Wei, ZHANG Qiang. Analysis of Moisture State and Transverse Relaxation Characteristics of Grains During the Growth Process of Rice [J]. Scientia Agricultura Sinica, 2017, 50(2): 240-249.
[3]	. Responses of Carbohydrate Metabolism to Nitrogen in Cotton Fiber Development and Its Relationships with Fiber Strength Formation [J]. Scientia Agricultura Sinica, 2009, 42(1): 93-102 .