
水稻幼穗响应稻曲病菌毒素胁迫早期的转录组分析
收稿日期: 2021-09-06
修回日期: 2021-11-10
网络出版日期: 2022-09-09
基金资助
四川省科技计划资助项目(2021YFYZ0021);四川省科技计划资助项目(2022YFYZ0002);四川省财政农作物“卡脖子”技术攻关计划资助项目(2021ZYGG-002);四川省财政自主创新专项(2022ZZCX020)
Transcriptome Analysis of Young Rice Panicles in Early Response to Exposure to Mycotoxin of Ustilaginoidea virens
Received date: 2021-09-06
Revised date: 2021-11-10
Online published: 2022-09-09
【目的】由稻曲病菌引起的稻曲病不仅造成水稻减产,而且还会产生对动物和植物有毒的真菌毒素。探明水稻幼穗对稻曲病菌毒素胁迫响应的分子机制,可为发掘水稻抗稻曲病基因以及抗病分子育种开辟新的思路。【方法】用稻曲病菌毒素处理水稻幼穗,采用转录组测序技术对水稻幼穗进行转录组测序,以水稻9311基因组作为参考基因组进行对比,利用TPM法计算基因表达量,设定参数(差异倍数的绝对值不小于2,且q值不大于0.05)筛选差异表达基因。结合基因差异表达分析、富集功能分析,鉴定水稻响应胁迫的关键基因,并利用实时荧光定量PCR技术对差异表达基因进行验证。【结果】在稻曲病菌毒素胁迫12 h后,水稻幼穗出现2526个差异表达基因(DEG);通过GO富集、KEGG代谢途经和KOG功能分析,将差异基因划分为GO功能下的64个条目、32个代谢途径和KOG功能下23个类别,包括淀粉和蔗糖代谢、苯丙类生物合成、碳代谢、糖酵解/糖异生、氨基糖和核苷酸糖代谢等生物学过程。DEG中有66个植物转录因子,分属7种植物转录因子家族,包括WRKY和Myb两大转录因子。分析二萜类生物合成与淀粉和蔗糖代谢途径相关基因发现,OsCPS2、OsKSL4和细胞色素P450等基因表达量上调,而淀粉酶、β-呋喃果糖苷酶和UDP-焦磷酸化酶等基因表达量下调,推测这些基因在水稻响应稻曲病菌毒素胁迫时发挥重要的作用。【结论】稻曲病菌毒素作为非生物胁迫因素对水稻幼穗具有毒性;通过干扰淀粉和蔗糖代谢等途径而影响种子营养物质的合成,降低水稻抵抗病原菌侵染水稻的能力。
伏荣桃, 王剑, 陈诚, 赵黎宇, 陈雪娟, 卢代华 . 水稻幼穗响应稻曲病菌毒素胁迫早期的转录组分析[J]. 中国水稻科学, 2022 , 36(5) : 447 -458 . DOI: 10.16819/j.1001-7216.2022.210714
【Objective】 Rice false smut caused by Ustilaginoidea virens not only leads to rice yield loss, but also produces mycotoxins which is toxic to animals and plants. In order to elucidate the molecular mechanism behind the response to exposure to mycotoxin the young panicles of rice were mycotoxin-treated with U. virens mycotoxin, which may provide a new idea for exploring rice false smut resistance genes and molecular breeding for disease resistance. 【Method】Transcriptome sequencing was performed with mycotoxin-treated young panicles as materials. The genome of 9311 was used as the reference genome to align the sequences. The expression level of genes was calculated by the term of Transcripts Per Million (TPM). According to the database, the differentially expressed genes were screened by parameters (|LOG2 fold change|≥1 and q-value≤0.05). Combined with differential expression analysis and function annotation, the key genes in response to mycotoxin were identified in rice panicles. The differentially expressed genes were verified by qRT-PCR. 【Result】2526 differentially expressed genes (DEG) were identified in young panicles of rice after 12 h treated with mycotoxin. Through GO enrichment, KEGG metabolic pathway and KOG function analysis, the DEGs were divided into 64 terms under GO function, 32 metabolic pathways and 23 categories under KOG function, including starch and sucrose metabolism, phenylpropyl biosynthesis, carbon metabolism, glycolysis/gluconeogenesis, amino sugar and nucleoside sugar metabolism and other biological processes. There were 66 plant transcription factors in DEGs, which belonged to 7 family of plant transcription factors, including WRKY and Myb. The expression levels of OsCPS2, OSKSL4 and cytochrome P450 were up-regulated, while the expression levels of amylase, beta-fructofuranosidase and UGPase were down-regulated when we analyzed genes involved in diterpenoid biosynthesis and starch and sucrose metabolism pathways. Therefore, it is speculated that these genes play an important role in rice response to mycotoxin of U. virens stress. 【Conclusion】As an abiotic stress factor, mycotoxin of U. virens had a toxic effect on young panicles of rice. It affects the synthesis of nutrients in seeds by interfering with metabolism of starch and sucrose, and thus reduces the resistance of rice to pathogen infection.
Key words: rice; Ustilaginoidea virens; mycotoxin; transcriptome analysis
| [1] | Tanaka E, Ashizawa T, Sonoda R, Sonoda R, Tanka C. Villosiclava virens gen. nov., comb. nov., teleomorph of Ustilaginoidea virens,the causal agent of rice false smut[J]. Mycotaxon, 2008, 106(1): 491-501. |
| [2] | Fan J, Yang J, Wang Y Q, Li G B, Li Y, Huang F, Wang W M. Current understanding on Villosiclava virens, a unique flower-infecting fungus causing rice false smut disease[J]. Molecular Plant Pathology, 2016, 17(9): 1321-1330. |
| [3] | 伏荣桃, 王剑, 卢代华, 张鸿, 龚学书, 陈雪娟, 任鸿志, 毛建辉. 水稻稻曲病抗性鉴定技术及影响因子研究[J]. 中国农学通报, 2015, 31(18): 266-272. |
| [3] | Fu R T, Wang J, Lu D H, Zhang H, Gong X S, Chen X J, Ren H Z, Mao J H. Resistance identification and influence factor of rice false smut[J]. Chinese Agricultural Science Bulletin, 2015, 31(18): 266-272. (in Chinese with English abstract) |
| [4] | Hu M L, Luo L X, Wang S, Liu Y F, Li J Q. Infection processes of Ustilaginoidea virens during artificial inoculation of rice panicles[J]. European Journal of Plant Pathology, 2014, 139: 67-77. |
| [5] | Meng J J, Sun W B, Mao Z L, Dan X, Wang X, Lu S, Yang L, Zhou L, Zhang G. Main ustilaginoidins and their distribution in rice false smut balls[J]. Toxins, 2015, 7(10): 4023-4034. |
| [6] | Lai D W, Meng J J, Zhang X P, Xu D, Dai J G, Zhou L G. Ustilobisorbicillinol A, a cytotoxic sorbyl-containing aromatic polyketide from Ustilaginoidea virens[J]. Organic Letters, 2019, 21(5): 1311-1314. |
| [7] | Fu X X, Xie R S, Wang J. Development of colloidal gold-based lateral flow immunoassay for rapid qualitative and semi-quantitative analysis of ustiloxins A and B in rice samples[J]. Toxins, 2017, 9(3): 79. |
| [8] | Meng J J, Gu G, Dang P Q, Zhang X P, Wang W X, Dai J G, Liu Y, Lai D W, Zhou L G. Sorbicillinoids from the fungus Ustilaginoidea virens and their phytotoxic, cytotoxic, and antimicrobial activities[J]. Frontiers in Chemistry, 2019, 7: 435. |
| [9] | Li Y, Koiso Y, Kobayashi H, Hashimoto Y, Iwasaki S. Ustiloxins, new antimitotic cyclic peptides: Interaction with porcine brain tubulin[J]. Biochemical Pharmacology, 1995, 49(10): 1367-1372. |
| [10] | Hu Z, Dang Y, Liu C S, Zhou L, Liu H. Acute exposure to ustiloxin A affects growth and development of early life zebrafish, Danio rerio[J]. Chemosphere, 2019, 226: 851-857. |
| [11] | 陈美军, 胡东维, 徐颖. 稻曲病菌毒素的活性测定、抗体制备与细胞定位[J]. 实验生物学报, 2004, 37(4): 310-314. |
| [11] | Chen M J, Hu D W, Xu Y. Activity assay, antiserum preparation and cellular localization of ustiloxins[J]. Acta Biologiae Experimentalis Sinica, 2004, 37(4): 310-314. (in Chinese with English abstract) |
| [12] | Hamed K A, Wayne T S, Cartwright R D, Sciumbato G L. Ustilaginoidea virens infection of rice in Arkansas: Toxicity of false smut galls, their extracts and the ustiloxin fraction[J]. American Journal of Plant Sciences, 2014, 5(21): 3166-3176. |
| [13] | Wang X H, Wang J, Lai D W, Wang W X, Liu Y. Ustiloxin G, a new cyclopeptide mycotoxin from rice false smut balls[J]. Toxins, 2017, 9(2): 54-63. |
| [14] | Fu R T, Wang J, Chen C, Gong X S, Lu D H. Effect of crude toxins of Ustilaginoidea virens on rice seed germination[J]. African Journal of Microbiology Research, 2017, 11(32): 1267-1273. |
| [15] | Luduena R F, Roach M C, Prasad V. Interaction of ustiloxin A with bovine brain tubulin[J]. Biochemical Pharmacology, 1994, 47(9): 1593-1599. |
| [16] | 武斌, 温雪玮, 李衫衫, 胡东维, 梁五生. 稻曲病菌毒素对水稻幼根转录组的影响[J]. 农业生物技术报, 2018, 26(7):1093-1106. |
| [16] | Wu B, Wen X W, Li S S, Hu D W, Lang W S. Influences of mycotoxins of Villosiclava virens on the transcriptome of rice (Oryza sativa) seedling roots[J]. Journal of Agricultural Biotechnology, 2018, 26(7): 1093-1106. (in Chinese with English abstract) |
| [17] | Yuan Z, Zhang Y, Xu G, Bi D, Qu H, Zou X, Gao X, Yang H, He H, Wang X, Bao J, Zuo S, Pan X, Zhou B, Wang G, Qu S. Comparative transcriptome analysis of Rhizoctonia solani-resistant and -susceptible rice cultivars reveals the importance of pathogen recognition and active immune responses in host resistance[J]. Journal of Plant Biology, 2018, 61(3):143-158. |
| [18] | 楚乐乐, 罗成科, 李芳兰, 路旭平, 马天利, 李培富. 盐胁迫下OsDSR2 RNAi转基因水稻的生理特性及转录组学分析[J]. 植物遗传资源学报, 2020, 21(4): 954-965. |
| [18] | Chu L L, Luo C K, Li F L, Lu X P, Ma T L, Li P F. Analysis of the physiological characteristics and transcriptome profiles of OsDSR2 RNAi transgenic rice under salt stress[J]. Journal of Plant Genetic Resources, 2020, 21(4): 954-965. (in Chinese with English abstract) |
| [19] | Benjamini Y, Yekutieli D. The control of the false discovery rate in multiple testing under dependency[J]. The Annals of Statistics, 2001, 29(4): 1165-1188. |
| [20] | Chao J, Jin J, Wang D, Han R, Zhu R S, Zhu Y G, Li S Q, Sun M X. Cytological and transcriptional dynamics analysis of host plant revealed stages specific biological processes related to compatible rice Ustilaginoidea virens interaction[J]. PLoS ONE, 2014, 9(3): e91391. |
| [21] | Livak K J, Schmittgen T D. Analysis of relative gene expression data using realtime quantitative PCR and the 2−ΔΔCT method[J]. Methods, 2001, 25(4): 402-408. |
| [22] | Ismaiel A, Papenbrock J. Mycotoxins: Producing fungi and mechanisms of phytotoxicity[J]. Agriculture, 2015, 5: 492-537. |
| [23] | Janardhanan K K, Husain A. Phytotoxic activity of tenuazonic acid isolated from Alternaria alternata (Fr.) Keissler causing leaf blight of Datura innoxia Mill. and its effect on host metabolism[J]. Journal of Phytopathology, 1984, 111(3-4): 305-311. |
| [24] | McLean M. The phytotoxicity of Fusarium metabolites: An update since 1989. Mycopathologia, 1996, 133: 163-179. |
| [25] | Ismaiel A A, Tharwat N A. Antifungal activity of silver ion on ultrastructure and production aflatoxin B1 and patulin by two mycotoxigenic strains, Aspergillus flavus OC1 and Penicillium vulpinum CM1[J]. Journal de Mycologie Medical, 2014, 24(3): 193-204. |
| [26] | Fujita K, Arase S, Hiratsuka H, Honda Y, Nozu M. The role of toxin(s) produced by germinating spores of Pyricularia oryzae in pathogenesis[J]. Journal of Phytopathology, 1994, 142(3-4): 245-252. |
| [27] | Vidhyasekaran P, Ruby Ponmalar T, Samiyappan R, Velazhahan R, Muthukrishnan S. Host-specific toxin production by Rhizoctonia solani, the rice sheath blight pathogen[J]. Phytopathology, 1997, 87(12): 1258-1263. |
| [28] | Samuel A T, Valentine I T. Effect of total aflatoxin on the growth characteristics and chlorophyll level of sesame (Sesamum indicum L.)[J]. New York Science Journal, 2014, 7: 8-13. |
| [29] | Aver’yanov A A, Lapikova V P, Lebrun M H. Tenuazonic acid, toxin of rice blast fungus, induces disease resistance and reactive oxygen production in plants[J]. Russian Journal of Plant Physiology, 2007, 54: 749-754. |
| [30] | 齐俊生, 李怀方. 一种检测棉花黄萎菌毒素致萎性的新方法: 叶片针刺涂抹法[J]. 棉花学报, 2006, 18(4): 228-232. |
| [30] | Qi J S, Li H F. A new detection method of wilting induction by phytotoxin from V. dahliae on cotton through leaf pricking and spreading[J]. Cotton Science, 2006, 18(4): 228-232. (in Chinese with English abstract) |
| [31] | 杨艳丽, 肖浪涛, 胡先奇. 马铃薯晚疫病菌与寄主品种抗性关系研究. 中国农业科学, 2009, 42(6): 2202-2210. |
| [31] | Yang Y L, Xiao L T, Hu X Q. Study on the relationship between the toxin of Phytophthora infestans and resistance of potato[J]. Scientia Agricultura Sinica, 2009, 42(6): 2202-2210. (in Chinese with English abstract) |
| [32] | Ambawat S, Sharma P, Yadav N R. MYB transcription factor genes as regulators for plant responses: An overview[J]. Physiology and Molecular Biology of Plants, 2013, 19(3): 307-321. |
| [33] | Wang Y M, Kwon S J, Wu J N, Choi J Y, Lee Y H, Agrawai G K, Tamogami S, Rakwal R, Park S R, Kim B G, Jung K H, Kang K Y, Kim S G, Kim S T. Transcriptome analysis of early responsive genes in rice during Magnaporthe oryzae infection[J]. The Plant Pathology Journal, 2014, 30(4): 343-354. |
| [34] | Han Y Q, Zhang K, Yang J, Zhang N, Zhang Y, Liu Y F, Chen Z Y, Hsiang T, Sun W X. Differential expression profiling of the early response to Ustilaginoidea virens between false smut resistant and susceptible rice varieties[J]. BMC Genomics, 2015, 16: 955. |
| [35] | 张艺丹, 曾英, 卢山. 水稻二萜合成途径中代谢流调控机制研究进展[J]. 植物生理学报, 2019, 55(12): 1762-1768. |
| [35] | Zhang Y D, Zeng Y, Lu S. Recent progress in the study of metabolic flux regulation in rice diterpene biosynthesis[J]. Plant Physiology Journal, 2019, 55(12): 1762-1768. (in Chinese with English abstract) |
| [36] | Peters R J. Uncovering the complex metabolic network underlying diterpenoid phytoalexin biosynthesis in rice and other cereal crop plants[J]. Phytochemistry, 2006, 67: 2307-2317 |
| [37] | Zi J, Mafu S, Peters R J. To gibberellins and beyond Surveying the evolution of (di)terpenoid metabolism[J]. Annual Review of Plant Biology, 2014, 65: 259-286. |
| [38] | 韩彦卿, 韩渊怀, 张春来, 孙文献. 水稻幼穗与Ustilaginoidea virens互作早期的转录组分析[J]. 植物病理学报, 2019, 49(3): 296-305. |
| [38] | Han Y Q, Han Y H, Zhang C L, Sun W X. Transcriptomic analysis of early interaction between rice young spikelets and Ustilaginoidea virens[J]. Acta Phytopathologica Sinica, 2019, 49(3): 296-305. (in Chinese with English abstract) |
| [39] | 彭波, 彭宇, 彭娟, 孔冬艳, 何璐璐, 孙艳芳, 黄雅琴, 宋世枝. 水稻种子主要营养物质合成及调控研究与展望. 热带作物学报, 2018, 39(6): 1241-1251. |
| [39] | Peng B, Peng Y, Peng J, Kong D M, He L L, Sun Y F, Huang Y Q, Song S Z. Research advancement and prospects of main nutritious substances synthesis and regulation in rice seeds[J]. Chinese Journal of Tropical Crops, 2018, 39(6): 1241-1251. (in Chinese with English abstract) |
| [40] | Thitisaksakul M, Jiménez R C, Arias M C, Beckles D M. Effects of environmental factors on cereal starch biosynthesis and composition[J]. Journal of Cereal Science, 2012, 56(1): 67-80. |
| [41] | Fujita N. Starch biosynthesis in rice endosperm[J]. Agri-bioscience Monographs, 2014, 4(1): 1-18. |
| [42] | Fan J, Yang L, Zheng A P, Wang W M, Guo X Y, Li L, Huang F, Sun W X, Yan L, Huang Y Y. Infection of Ustilaginoidea virens intercepts rice seed formation but activates grain-filling-related genes[J]. Journal of Integrative Plant Biology, 2015, 57(6): 577-590. |
/
| 〈 |
|
〉 |