转录组和代谢组联合分析阐释木薯叶片花青素合成机制

罗秀芹, 韦卓文, 蔡杰, 安飞飞, 陈松笔, 薛晶晶

中国农学通报. 2024, 40(24): 100-106

PDF(2014 KB)
PDF(2014 KB)
中国农学通报 ›› 2024, Vol. 40 ›› Issue (24) : 100-106. DOI: 10.11924/j.issn.1000-6850.casb2023-0432
生物科学

转录组和代谢组联合分析阐释木薯叶片花青素合成机制

作者信息 +

Explanation of Integrative Analysis of Metabolome and Transcriptome for Anthocyanins Synthesis Mechanism in Cassava Leaves

Author information +
History +

摘要

木薯是世界上第六大粮食作物,其块根富含淀粉但缺乏蛋白质、花青素、胡萝卜素等营养物质。为了探索木薯花青素生物合成机制,本研究选取了两种不同颜色木薯种质资源叶片(FL与PL)为材料,进行转录组和花青素靶向代谢组及其联合分析。转录组分析结果显示在FL和PL中6864个差异表达基因,其中包含4112个上调表达和2752个下调表达。代谢组分析结果显示26种显著差异代谢物在PL中显著高于FL,其中21种属于花青素类。联合分析结果显示,其中7个差异表达的基因与花青素生物合成相关,且花青素含量与差异基因的表达呈正相关,尤其是MeANS1的表达差异最大。本研究结果为阐明木薯花青素的生物合成机制提供了候选基因,同时也为提高木薯花青素含量奠定科学基础。

Abstract

Cassava is the sixth largest staple food crop in the world, which storage root is rich in starch while lack of proteins, anthocyanins and carotenes and so on. Two types of leaves of cassava with different color (FL and PL) were selected as materials to explore the mechanism of anthocyanin biosynthesis by combining with transcriptome and metabolome in this work. The transcriptome results showed that there were 6864 differentially expressed genes (DEGs) including 4112 up-regulating and 2752 down-regulating in PL compare with FL. The metabolome results showed that there were 26 significantly different metabolites (SDMs) in PL compare with FL, and 21 of them belonged to anthocyanins. The result of combination of transcriptome and metabolome showed that there were seven DEGs related to anthocyanin biosynthesis and all of them were positively correlated to anthocyanin biosynthesis. The expression of MeANS1 showed the most difference. It provided candidate genes for elucidating the mechanism of anthocyanin biosynthesis in cassava and laid a scientific foundation for enhancement content of anthocyanin in future.

关键词

木薯 / 类黄酮 / 花青素 / 转录组 / 代谢组 / 生物合成 / 差异表达基因 / MeANS

Key words

cassava / flavonoid / anthocyanins / transcriptome / metabolome / biosynthesis / differentially expressed genes / MeANS

引用本文

导出引用
罗秀芹 , 韦卓文 , 蔡杰 , 安飞飞 , 陈松笔 , 薛晶晶. 转录组和代谢组联合分析阐释木薯叶片花青素合成机制. 中国农学通报. 2024, 40(24): 100-106 https://doi.org/10.11924/j.issn.1000-6850.casb2023-0432
LUO Xiuqin , WEI Zhuowen , CAI Jie , AN Feifei , CHEN Songbi , XUE Jingjing. Explanation of Integrative Analysis of Metabolome and Transcriptome for Anthocyanins Synthesis Mechanism in Cassava Leaves. Chinese Agricultural Science Bulletin. 2024, 40(24): 100-106 https://doi.org/10.11924/j.issn.1000-6850.casb2023-0432

参考文献

[1]
AN F, CHEN T, LI Q, et al. Protein cross-interactions for efficient photosynthesis in the cassava cultivar SC205 relative to its wild species[J]. Journal of agricultural and food chemistry, 2019, 67(32):8746-8755.
The underlying mechanisms of the higher photosynthetic efficiency of cultivated cassava relative to its wild species are poorly understood. In the present study, proteins in leaves and chloroplasts were analyzed to compare the differences among the cultivar SC205, its wild ancestor W14, and the related species Glaziovii. The functions of differential proteins are associated with 10 ontology groups including photosynthesis, carbohydrate and energy metabolism, as well as potential signal pathway. The protein-protein networks among 41 differential proteins showed that PGK1 is a hub protein and protein cross-interactions affected the differentiation of photosynthetic rate. Anatomy patterns and PEPC detection suggested that SC205 has more C photosynthesis characteristics than Glaziovii and W14. Finally, a mechanism model of the efficient photosynthesis was proposed based on the remarkable variations in photosynthetic parameters and protein functions in the domestic cultivars.
[2]
CAI J, ZHANG J, DING Y, et al. Different fertilizers applied alter fungal community structure in rhizospheric soil of cassava (Manihot esculenta Crantz) and increase crop yield[J]. Frontiers in microbiology, 2021, 12:663781.
[3]
KUMBA Y. Genetic characterization of exotic and landraces of cassava in Ghana. M.Sc.; in Agronomy, Kwame Nkrumah[D]. University of science and technology, 2012.
[4]
OMAR N, HASSAN S, YUSOFF U, et al. Phenolics, flavonoids, antioxidant activity and cyanogenic glycosides of organic and mineral-base fertilized cassava tubers[J]. Molecules, 2012, 17(3):2378-2387.
A field study was conducted to determine the effect of organic and mineral-based fertilizers on phytochemical contents in the tubers of two cassava varieties. Treatments were arranged in a split plot design with three replicates. The main plot was fertilizer source (vermicompost, empty fruit bunch compost and inorganic fertilizer) and sub-plot was cassava variety (Medan and Sri Pontian). The amount of fertilizer applied was based on 180 kg K(2)O ha-1. The tubers were harvested and analyzed for total flavonoids, total phenolics, antioxidant activity and cyanogenic glucoside content. Total phenolic and flavonoid compounds were determined using the Folin-Ciocalteu assay and aluminium chloride colorimetric method, respectively. Different sources of fertilizer, varieties and their interactions were found to have a significant effect on phytochemical content. The phenolic and flavonoid content were significantly higher (p < 0.01) in the vermicompost treatment compared to mineral fertilizer and EFB compost. The total flavonoids and phenolics content of vermicompost treated plants were 39% and 38% higher, respectively, than those chemically fertilized. The antioxidant activity determined using the DPPH and FRAP assays were high with application of organic fertilizer. Cyanogenic glycoside levels were decreased with the application of organic fertilizer. Among the two types of compost, vermicompost resulted in higher nutritional value of cassava tubers. Medan variety with application of vermicompost showed the most promising nutritional quality. Since the nutritional quality of cassava can be improved by organic fertilization, organic fertilizer should be used in place of chemical fertilizer for environmentally sustainable production of better quality cassava.
[5]
XIAO L, CAO S, SHANG X, et al. Metabolomic and transcriptomic profiling reveals distinct nutritional properties of cassavas with different flesh colors[J]. Food chemistry: molecular sciences, 2021, 2:100016.
[6]
王定美, 王伟, 麦力文, 等. HPLC法同时测定不同采收期木薯叶片中6种类黄酮的含量[J]. 食品研究与开发, 2017, 38(18):133-137.
[7]
FU L, DING Z, TIE W, et al. Integrated metabolomic and transcriptomic analyses reveal novel insights of anthocyanin biosynthesis on color formation in cassava tuberous roots[J]. Frontiers in nutrition, 2022, 9:842693.
[8]
LUO X, AN F, XUE J, ZHU W, et al. Integrative analysis of metabolome and transcriptome reveals the mechanism of color formation in cassava (Manihot esculenta Crantz) leaves[J]. Frontiers in plant science, 2023, 14:1181257.
[9]
CHEN W, GAO Y, XIE W, et al. Genome-wide association analyses provide genetic and biochemical insights into natural variation in rice metabolism[J]. Nature genetics, 2014, 46(7):714-721.
Plant metabolites are important to world food security in terms of maintaining sustainable yield and providing food with enriched phytonutrients. Here we report comprehensive profiling of 840 metabolites and a further metabolic genome-wide association study based on ∼6.4 million SNPs obtained from 529 diverse accessions of Oryza sativa. We identified hundreds of common variants influencing numerous secondary metabolites with large effects at high resolution. We observed substantial heterogeneity in the natural variation of metabolites and their underlying genetic architectures among different subspecies of rice. Data mining identified 36 candidate genes modulating levels of metabolites that are of potential physiological and nutritional importance. As a proof of concept, we functionally identified or annotated five candidate genes influencing metabolic traits. Our study provides insights into the genetic and biochemical bases of rice metabolome variation and can be used as a powerful complementary tool to classical phenotypic trait mapping for rice improvement.
[10]
BOSSE M, SILVA M, OLIVEIRA N, et al. Physiological impact of flavonoids on nodulation and ureide metabolism in legume plants[J]. Plant physiology and biochemistry, 2021, 166:512-521.
Legume plants from Fabaceae family (phylogenetic group composed by three subfamilies: Caesalpinioideae, Mimosoideae, and Papilionoideae) can fix atmospheric nitrogen (N) into ammonia (NH) by the symbiotic relationship with rhizobia bacteria. These bacteria respond chemotactically to certain compounds released by plants such as sugars, amino acids and organic acids. Root secretion of isoflavonoids acts as inducers for nod genes in rhizobia and ABC transporters and ICHG (isoflavone conjugates hydrolyzing beta-glucosidase) at apoplast are related to the exudation of genistein and daidzein in soybean roots. Biological nitrogen fixation (BNF) occurs inside the nodule by the action of nitrogenase enzyme, which fixes N into NH, which is converted into ureides (allantoin and allantoic acid). In this review, we bring together the latest findings on flavonoids biosynthesis and ureide metabolism in several legume plant species. We emphasize how flavonoids induce nod genes in rhizobia, affecting chemotaxis, nodulation, ureide production, growth and yield of legume plants. Mainly, isoflavonoids daidzein and genistein are responsible for nod genes activation in the rhizobia bacteria. Flavonoids also play an important role during nodule organogenesis by acting as auxin transporter inhibitors in root cells, especially in indeterminate nodules. The ureides are the main N transport form in tropical legumes and they are catabolized in leaves and other sink tissues to produce amino acids and proteins needed for plant growth and yield.Copyright © 2021 Elsevier Masson SAS. All rights reserved.
[11]
FALCONE FERREYRA M, RIUS S, CASATI P. Flavonoids: biosynthesis, biological functions, and biotechnological applications[J]. Frontiers in plant science, 2012, 3:222.
Flavonoids are widely distributed secondary metabolites with different metabolic functions in plants. The elucidation of the biosynthetic pathways, as well as their regulation by MYB, basic helix-loop-helix (bHLH), and WD40-type transcription factors, has allowed metabolic engineering of plants through the manipulation of the different final products with valuable applications. The present review describes the regulation of flavonoid biosynthesis, as well as the biological functions of flavonoids in plants, such as in defense against UV-B radiation and pathogen infection, nodulation, and pollen fertility. In addition, we discuss different strategies and achievements through the genetic engineering of flavonoid biosynthesis with implication in the industry and the combinatorial biosynthesis in microorganisms by the reconstruction of the pathway to obtain high amounts of specific compounds.
[12]
SHEN J, ZOU Z, ZHANG X, et al. Metabolic analyses reveal different mechanisms of leaf color change in two purple-leaf tea plant (Camellia sinensis L.) cultivars[J]. Horticulture research, 2018, 5:7.
[13]
FORKMANN G. Flavonoids as flower pigments: the formation of the natural spectrum and its extension by genetic engineering[J]. Plant breeding, 2010, 106:1-26.
[14]
SHARMA A, SHAHZAD B, REHMAN A, et al. Response of phenylpropanoid pathway and the role of polyphenols in plants under abiotic stress[J]. Molecules, 2019, 24(13):2452.
[15]
ZAYNAB M, FATIMA M, ABBAS S, et al. Role of secondary metabolites in plant defense against pathogens[J]. Microbial pathogenesis, 2018, 124:198-202.
Pathogens get entry into host cell, reproduce there and use biological machinery of host plants which is threat to global crop production. Integrated management strategies based upon minimizing population and use of resistant cultivars can address this potential problem. In developing world farmers are less likely to adopt these approaches instead they prefer the use of chemical pesticides. Reckless use of chemical pesticides is destroying our ecosystem. That's why it is required to explore ecofriendly alternatives, like plant based metabolites to control pathogens. Studies conducted on different plant-metabolites reported that these metabolite can potentially combat plant pathogens. In this study we have also discussed some of plant secondary metabolites including alkaloids, flavonoids and phenolics. In this review we tried to highlight the new trends in utilizing secondary metabolites for controlling bacterial, viral and fungal pathogens with the hope that upcoming drugs will be human and ecosystem friendly.Copyright © 2018 Elsevier Ltd. All rights reserved.
[16]
BAI L, LI X, HE L, et al. Antidiabetic potential of flavonoids from traditional chinese medicine: a review[J]. American journal of Chinese medicine, 2019, 47(5):933-957.
[17]
PARK M, KIM Y, HA S, et al. Function and application of flavonoids in the breast cancer[J]. International journal of molecular sciences, 2022, 23(14):7732.
[18]
CIUMARNEAN L, MILACIU M, RUNCAN O, et al. The effects of flavonoids in cardiovascular diseases[J]. Molecules, 2020, 25(18):4320.
[19]
QUATTROCCHIO F, BAUDRY A, LEPINIEC L, et al. The regulation of flavonoid biosynthesis[A]. The science of flavonoids, 2006:97-122.
[20]
JIANG T, ZHANG M, WEN C, et al. Integrated metabolomic and transcriptomic analysis of the anthocyanin regulatory networks in Salvia miltiorrhiza Bge. flowers[J]. BMC plant biology, 2020, 20(1):349.
[21]
LI J, LV X, WANG L, et al. Transcriptome analysis reveals the accumulation mechanism of anthocyanins in ‘Zijuan’ tea (Camellia sinensis var. asssamica (Masters) kitamura) leaves[J]. Plant growth regulation, 2017, 81:51-61.
[22]
BEN-SIMHON Z, JUDEINSTEIN S, TRAININ T, et al. A "white" anthocyanin-less pomegranate (Punica granatum L.) caused by an insertion in the coding region of the leucoanthocyanidin dioxygenase (LDOX; ANS) gene[J]. Plos one, 2015, 10(11):e0142777.
[23]
王硕, 郑秀文, 王琪, 等. 果树中花青素合成及其分子调控机制研究进展[J/OL]. 分子植物育种, 2022(7), https://kns.cnki.net/kcms/detail/46.1068.S.20220728.1710.014.html.
[24]
LI H, LIU J, PEI T, et al. Overexpression of SmANS enhances anthocyanin accumulation and alters phenolic acids content in Salvia miltiorrhiza and Salvia miltiorrhiza Bge f. alba plantlets[J]. International journal of molecular sciences, 2019, 20(9):2225.
[25]
ZHANG H, ZHAO X, ZHANG J, et al. Functional analysis of an anthocyanin synthase gene StANS in potato[J]. Scientia horticulture, 2020, 272:109569.
[26]
YANG B, WEI Y, LIANG C, et al. VvANR silencing promotes expression of VvANS and accumulation of anthocyanin in grape berries[J]. Protoplasma, 2022, 259(3):743-753.

基金

海南省自然科学基金青年基金“MeZDS和MeCRTISO互作促进木薯类胡萝卜素生物合成的分子机制”(321QN321)
中国热带农业科学院基本科研业务费专项(1630032022007)
PDF(2014 KB)

Accesses

Citation

Detail

段落导航
相关文章

/