Effects of ABA and Its Synthesis Inhibitor Sodium Tungstate on Carotenoid Associated Genes and Enzymes of Cassava Tuber Root

doi:10.13560/j.cnki.biotech.bull.1985.2017-0349

Abstract

Abstract: This work aims to analyze the effect of the exogenous ABA and its inhibitor sodium tungstate（Na₂WO₄）on carotenoid associated genes and protein of cassava tuberous root. Manihot esculenta Crantz SC9 was chosen as research subject and the β-carotene of cassava tuber root was detected by HPLC,and Western blot was used to analyze the carotenoid associated protein expression and QPCR to analyze the genes expression. The results showed that exogenous ABA and sodium tungstate increased the β-carotene content in the tuber root. Both exogenous ABA and sodium tungstate increased carotenoid biosynthesis key genes PSY2 and LCYB expressions,meanwhile they restrained the degradation genes ZEP and NCED3expressions,which explained the change of β-carotene content at the transcription level. The LCYB and ZEP transcriptions were consistent with translations. Plastid associated gene PAP1 and enzyme FBN1a were up-regulated,which increased the number of plastidglobules. Exogenous ABA and sodium tungstate accelerated the reactive oxygen（ROS）related superoxide dismutase（Fe-SOD and Cu/Zn-SOD）expressions,keeping the carotenoid content stable. In conclusion,these three factors jointly increase the β-carotene content of tuber root.

Key words: cassava tuber root, carotenoid associated genes and enzymes, abscisic acid, β-carotene

DENG Chang-zhe, AN Fei-fei, LI Kai-mian, CHEN Song-bi. Effects of ABA and Its Synthesis Inhibitor Sodium Tungstate on Carotenoid Associated Genes and Enzymes of Cassava Tuber Root[J]. Biotechnology Bulletin, 2017, 33(11): 76-83.

References

[1] Zeevaart J, Creelman R. Metabolism and physiology of abscisic acid[J]. Annual Review of Plant Physiology and Plant Molecular Biology, 1988, 39：439-473.
[2] Taylor IB, Burbidge A, Thompson AJ. Control of abscisic acid synthesis[J]. Journal of Experiment Botony, 2000, 51：1563-1574
[3] 翁倩, 周宝利, 于洋, 等. 外源ABA、BR和ETH对番茄果实番茄红素含量的影响[J]. 沈阳农业大学学报, 2007, 38：787-784.
[4] 王贵元, 夏学仁, 周开兵. 外源ABA和GA3对红肉脐橙果皮主要色素含量变化和果皮着色的影[J]. 武汉植物学研究, 2004, 22（3）：273-276.
[5] 张振文, 李开绵, 叶剑秋, 等. 木薯光合作用特性研究[J]. 云南大学学报, 2007, 29（6）：628-632.
[6] 陈冠喜, 李开绵, 叶剑秋, 等. 6个木薯品种生长发育及产量性状的初步研究[J]. 热带农业科学, 2009, 29：6.
[7] Ortega-Flores CI, Lopes da Costa MA, Cereda P. Bioavailabilty of β-carotenein dehydrated yellow cassava（Manihot esculenta Crantz）. La biodisponibilidad de β-caroteno en yuca deshidratada hojas（Manihot esculenta Crantz）[J]. Cienc Tecnology Alim, 2003, 23：473-477.
[8] Arango J, Wust J, Welsch, et al. Characterization of phytoene synthases from cassava and their involvement in abiotic stress-mediated response[J]. Planta, 2010, 232：1251-1262.
[9] Ralf W, Jacobo A, Cornelia B, et al. Provitamin A accumulation in cassava（Manihot esculenta）roots driven by a single nucleotide polymorphism in a phytoene synthase gene[J]. Plant Cell, 2010, 22（10）：3348-3356.
[10] 王淑娟. ABA对木薯块根淀粉积累影响机制初探[D]. 海口：海南大学, 2014.
[11] Lucia M, Alcides R, Ronoel L, et al. Retention of total carotenoid and β-carotene in yellow sweet cassava（Manihot esculenta Crantz）after domestic cooking[J]. Food Nutrition Research, 2012, 56：15788.
[12] Camalho LJ, Lippolis J, Chen S, et al. Characterization of carotenoid-protein complexes and gene expression analysis associatedwith carotenoid sequestration in pigmented cassava（Manihot esculenta Crantz）storage root[J]. Open Biochemical Journal, 2012, 6：116-130.
[13] Burkhardt PK, Beyer P, Wunn J, et al. Transgenic rice（Oryza sativa）endosperm expressing daffodil（Narcissus pseudonarcissus）phytoene synthase accumulates phytoene, a key intermediate of provitamin A biosynthesis[J]. Plant Journal, 1997, 11：1071-1078.
[14] Cazzonelli C, Pogson B. Source to sink：regulation of carotenoid biosynthesis inplants[J]. Trends in Plant Science, 2010, 15：266-274.
[15] Nambara E, Marion-Poll A. Abscisic acid biosynthesis and catabolism[J]. Annual Review of Plant Biology, 2005, 56：165-185.
[16] 丁泽红, 付莉莉, 铁韦韦, 等. 木薯MeNCED3基因克隆、结构变异及其表达分析[J]. 生物技术通报, 2016, 32（10）：148-153.
[17] 刘剑锋, 张春吉, 程云清, 等. ABA及其合成抑制剂钨酸钠处理对榛胚珠发育的影响[J]. 园艺学报, 2013, 40（2）：213-220
[18] Barsan C, Zouine M, Maza E, et al. Proteomic analysis of chloroplast-to-chromoplast transition in tomatoreveals metabolic shifts coupled with disrupted thylakoid biogenesis machinery and elevatedenergy-production components[J]. Plant Physiol, 2012, 160：708-725.
[19] Marti MC, Camejo D, Olmos E, et al. Characterisation and changes in the antioxidant system of chloroplasts and chromoplast isolated from pepper fruits[J]. Plant Biology, 2009, 11：613-624.
[20] Bouvier F, Backhaus RA, Camara B. Induction and control of chromoplast-specific carotenoid genes by oxidative stress[J]. Journal of Biological Chemistry, 1998, 273：30651-30659.
[21] Egea I, Barsan C, Bian WP, et al. Chromplast differentiation：current status and perspectives[J]. Plant and Cell Physiology, 2010, 51：1601-1611.
[22] 李馨园, 杨晔, 张丽芳, 等. 外源ABA对低温胁迫下玉米幼苗内源激素含量及Asr1基因表达的调节[J]. 作物学报, 2017, 43（1）：141-148.
[23] Lu S, Van Eck J, Zhou X, et al. The cauliflower Or gene encodes a DnaJ cysteine-rich domain-containingprotein that mediates high levels of beta-carotene accumulation[J]. Plant Cell, 2006, 18：3594-3605.
[24] Li L, Paolillo DJ, Parthasarathy MV, et al. A novel gene mutation that confers abnormal patterns of β-carotene accumulation in cauliflower（Brassica oleracea var. botrytis）[J]. Plant Journal, 2001, 26（1）：59-67.
[25] Wang YQ, Yong Y, Fei ZJ, et al. Proteomic analysis of chromoplasts from six crop species reveals insights into chromoplasts function and development[J]. Journal Experiment Botony, 2013, 64：949-961
[26] Rey P, Gillet B, Romer S, et al. Over-expression of a pepper plastid lipid-associated protein in tobacco leads tochanges inplastid ultrastructure and plant development upon stress[J]. Plant Jornal, 2000, 21：483-494.
[27] Lundquist PK, Poliakov A, Bhuiyan NH, et al. The functional network of the Arabidopsis thaliana plastoglobule proteome based on quantitative proteomics and genomewide co-expression analysis[J]. Plant Physiology, 2012, 58：1172-1192.