甘蓝型油菜肌醇加氧酶基因家族鉴定与表达分析

doi:10.3785/j.issn.1008-9209.2022.09.281

浙江大学学报（农业与生命科学版）

2023, Vol. 49

Issue (4): 484-496 DOI: 10.3785/j.issn.1008-9209.2022.09.281

研究论文

甘蓝型油菜肌醇加氧酶基因家族鉴定与表达分析

陈书冰¹(

),许孜书¹,黄倩²,张慧³,张康妮²,段怡¹,孙月娥¹,周伟军²,许玲¹(

)

^1.浙江理工大学生命科学与医药学院, 浙江杭州 310018
^2.浙江大学农业与生物技术学院, 浙江杭州 310058
^3.浙江省农业技术推广中心, 浙江杭州 310020

Identification and expression analysis of myo-inositol oxygenase gene family in Brassica napus L.

Shubing CHEN¹(

),XU Zishu¹,Qian HUANG²,Hui ZHANG³,Kangni ZHANG²,Yi DUAN¹,Yue’e SUN¹,Weijun ZHOU²,Ling XU¹(

)

^1.College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, Zhejiang, China
^2.College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310058, Zhejiang, China
^3.Zhejiang Agricultural Technology Extension Center, Hangzhou 310020, Zhejiang, China

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摘要：

肌醇加氧酶（myo-inositol oxygenase, MIOX）催化肌醇转化为葡糖醛酸，在响应生物和非生物胁迫中发挥着重要作用。本研究对甘蓝型油菜MIOX家族基因（BnMIOX）进行了全基因组鉴定和表达模式分析。结果表明，BnMIOX家族包括12个成员，分布在9条染色体上；根据MIOX基因结构域特点，可以将甘蓝型油菜与拟南芥、白菜和甘蓝构建的系统进化树分为Ⅰ、Ⅱ、Ⅲ3个亚家族；且共线性分析中没有找到串联重复基因对，全部为大片段复制基因，表明片段重复和全基因组重复是甘蓝型油菜MIOX基因家族扩增的主要驱动力。转录组数据分析表明，BnMIOX基因在不同组织、不同生长发育过程中的时空表达模式不同。不同胁迫下的表达谱分析表明，BnMIOX1基因在干旱、盐胁迫下被显著诱导表达，且BnMIOX1、BnMIOX2和BnMIOX9基因对干旱、盐、脱落酸（abscisic acid, ABA）和冷胁迫响应较为明显。蛋白质互作网络分析结果进一步表明，BnMIOX与GLCAK、PIS1、VTC2、VTC4、PDF2.1等蛋白存在互作，推测BnMIOX基因在提高甘蓝型油菜抗性过程中发挥关键作用。本研究为进一步探讨BnMIOX基因的功能提供了依据。

关键词： 甘蓝型油菜; 肌醇加氧酶; 基因家族; 非生物胁迫; 基因表达

Abstract:

Myo-inositol oxygenase (MIOX) catalyzes the conversion of myo-inositol to glucuronic acid and plays an important role in response to biotic and abiotic stresses. In this study, the genome-wide identification and expression pattern analysis of the MIOX family gene in Brassica napus L.(BnMIOX) were conducted. The results showed that the BnMIOX family included 12 members distributed across nine chromosomes. According to the characteristics of MIOX gene domain, the phylogenetic tree of B. napus, Arabidopsisthaliana, B. rapa and B. oleracea could be divided into subfamilies Ⅰ, Ⅱ and Ⅲ. No tandem repeat gene pairs were found in the collinearity analysis, and all of them were large segment replication genes, demonstrating that segmental duplication and whole-genome duplication were the main driving forces for the MIOX gene family amplification in B. napus. The transcriptomic data indicated that BnMIOX genes showed different temporal and spatial expression patterns in different tissues and different growth and development processes. Expression profiles under different stresses demonstrated that the expression of BnMIOX1 gene was obviously induced under drought and salt stresses, while BnMIOX1, BnMIOX2, and BnMIOX9 genes had significant responses to drought, salt, abscisic acid (ABA), and cold stresses. The results of the protein interaction network analysis further showed that the BnMIOX interacted with proteins including GLCAK, PIS1, VTC2, VTC4, and PDF2.1, implying that BnMIOX genes play key roles in improving the resistance of B. napus. This study provides an important basis for further investigation of the function of BnMIOX genes.

Key words: Brassica napus L. myo-inositol oxygenase (MIOX) gene family abiotic stress gene expression

收稿日期: 2022-09-28 出版日期: 2023-08-29

CLC:

S565.4

基金资助: 浙江省科技计划项目(2022C02034);浙江省农业重大技术协同推广计划项目(2021XTTGLY02);现代作物生产省部共建协同创新中心项目（CIC-MCP）

通讯作者: 许玲 E-mail: 2864928337@qq.com;lxu@zstu.edu.cn

作者简介: 陈书冰（https://orcid.org/0000-0001-6587-8322），E-mail：2864928337@qq.com

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引用本文:

陈书冰,许孜书,黄倩,张慧,张康妮,段怡,孙月娥,周伟军,许玲. 甘蓝型油菜肌醇加氧酶基因家族鉴定与表达分析[J]. 浙江大学学报（农业与生命科学版）, 2023, 49(4): 484-496.

Shubing CHEN,XU Zishu,Qian HUANG,Hui ZHANG,Kangni ZHANG,Yi DUAN,Yue’e SUN,Weijun ZHOU,Ling XU. Identification and expression analysis of myo-inositol oxygenase gene family in Brassica napus L.. Journal of Zhejiang University (Agriculture and Life Sciences), 2023, 49(4): 484-496.

链接本文:

https://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2022.09.281 或 https://www.zjujournals.com/agr/CN/Y2023/V49/I4/484

表1 BnMIOX家族的序列特征

图1 甘蓝型油菜、拟南芥、白菜和甘蓝中MIOX蛋白序列的系统发育树代表拟南芥；代表甘蓝；代表白菜；代表甘蓝型油菜。

图3 BnMIOX 启动子中预测的顺式作用元件

表2 预测的部分顺式作用元件信息

图2 BnMIOX家族的基因结构和保守基序A. BnMIOX家族的基因结构；B. BnMIOX家族的保守基序。

图4 BnMIOX 基因在染色体上的分布每条染色体以0.1 Mb为单位划分，该区域的颜色代表基因密度（蓝色：低；白色：中；红色：高）

图5 BnMIOX 家族的共线性分析灰色区域表示甘蓝型油菜基因组中所有的共线性区域，黄色线表示BnMIOX基因的复制事件。

图6 不同植物间 MIOX 基因的共线性分析灰色线表示甘蓝型油菜与其他物种所有基因的共线性，红色线表示具有共线关系的MIOX基因对。

图7 BnMIOX 基因在不同组织和器官中的表达热图FPKM：每千个碱基的转录每百万映射读取的片段。

图8 不同胁迫下 BnMIOX 基因的表达热图

表3 BnMIOX蛋白预测结构之间的均方根偏差

图9 BnMIOX蛋白的三维结构结构模型为小鼠肌醇加氧酶与底物复合物的晶体结构。

图10 BnMIOX蛋白与其他蛋白质之间的互作网络预测圆圈代表各类蛋白质，连接线代表蛋白质-蛋白质相互作用。

4	杨楠.杨树肌醇代谢关键酶基因的表达及功能研究[D].山东,烟台:鲁东大学,2017. YANG N. Expression and function of inositol metabolism key enzymes gene in poplar[D]. Yantai, Shandong: Ludong University, 2017. (in Chinese with English abstract）
5	LOEWUS F A, MURTHY P P N. myo-inositol metabolism in plants[J]. Plant Science, 2000, 150: 1-19.
6	YE W X, REN W B, KONG L Q, et al. Transcriptomic proling analysis of Arabidopsis thaliana treated with exogenous myo-inositol[J]. PLoS ONE, 2016, 11(9): e161949. DOI: 10.1371/journal.pone.0161949 doi: 10.1371/journal.pone.0161949
7	ARNER R J, PRABHU K S, THOMPSON J T, et al. myo-inositol oxygenase: molecular cloning and expression of a unique enzyme that oxidizes myo-inositol and D-chiro-inositol[J]. Biochemical Journal, 2001, 360: 313-320.
8	LU Y, LIU C, MIAO X, et al. Increased expression of myo-inositol oxygenase is involved in the tubulointerstitial injury of diabetic nephropathy[J]. Experimental and Clinical Endo-crinology & Diabetes, 2009, 117(6): 257-265. DOI: 10.1055/s-2008-1081212 doi: 10.1055/s-2008-1081212
9	YANG B M, HODGKINSON A, MILLWARD B A, et al. Polymorphisms of myo-inositol oxygenase gene are associated with type 1 diabetes mellitus[J]. Journal of Diabetes and its Complications, 2010, 24(6): 404-408. DOI: 10.1016/j.jdiacomp.2009.09.005 doi: 10.1016/j.jdiacomp.2009.09.005
10	XIE P, SUN L, OATES P J, et al. Pathobiology of renal-specific oxidoreductase/myo-inositol oxygenase in diabetic nephropathy: its implications in tubulointerstitial fibrosis[J]. American Journal of Physiology-Renal Physiology, 2010, 298(6): F1393-F1404. DOI: 10.1152/ajprenal.00137.2010 doi: 10.1152/ajprenal.00137.2010
11	ARNER R J, PRABHU K S, KRISHNAN V, et al. Expression of myo-inositol oxygenase in tissues susceptible todiabetic complications[J]. Biochemical and Biophysical Research Communications, 2006, 339: 816-820. DOI: 10.1016/j.bbrc.2005.11.090 doi: 10.1016/j.bbrc.2005.11.090
12	傅小娟,陈雪梅,周钦,等. Miox基因在爪蟾胚胎发育中的时间和空间表达谱分析[J].第二军医大学学报,2014,35(8):832-836. DOI:10.3724/SP.J.1008.2014.00832 FU X J, CHEN X M, ZHOU Q, et al. Analysis of temporal and spatial expression patterns of Miox genes during development of Xenopus laevis embryos[J]. Academic Journal of Second Military Medical University, 2014, 35(8): 832-836. (in Chinese with English abstract） doi: 10.3724/SP.J.1008.2014.00832
13	DUAN J Z, ZHANG M H, ZHANG H L, et al. OsMIOX, a myo-inositol oxygenase gene, improves drought tolerance through scavenging of reactive oxygen species in rice (Oryza sativa L.)[J]. Plant Science, 2012, 196: 143-151. DOI: 10.1016/j.plantsci.2012.08.003 doi: 10.1016/j.plantsci.2012.08.003
14	CHEN C, SUN X L, DUANMU H Z, et al. Ectopic expression of a Glycine soja myo-inositol oxygenase gene (GsMIOX1a) in Arabidopsis enhances tolerance to alkaline stress[J]. PLoS ONE, 2015, 10(6): e129998. DOI: 10.1371/journal.pone.0129998 doi: 10.1371/journal.pone.0129998
1	王博.冬、春和半冬性甘蓝型油菜的遗传特征和基因组分化[D].湖北,武汉:华中农业大学,2019. WANG B. Genetic characteristics and genomic divergence among winter, spring and semi-winter oilseed rape[D]. Wuhan, Hubei: Huazhong Agricultural University, 2019. (in Chinese with English abstract）
15	ENDRES S, TENHAKEN R. myo-inositol oxygenase controls the level of myoinositol in Arabidopsis, but does not increase ascorbic acid[J]. Plant Physiology, 2009, 149(2): 1042-1049. DOI: 10.1104/pp.108.130948 doi: 10.1104/pp.108.130948
16	GIBON Y, USADEL B, BLAESING O E, et al. Integration of metabolite with transcript and enzyme activity profiling during diurnal cycles in Arabidopsis rosettes[J]. Genome Biology, 2006, 7(8): R76. DOI: 10.1186/gb-2006-7-8-r76 doi: 10.1186/gb-2006-7-8-r76
17	OSUNA D, USADEL B, MORCUENDE R, et al. Temporal responses of transcripts, enzyme activities and metabolites after adding sucrose to carbon-deprived Arabidopsis seedlings[J]. The Plant Journal, 2007, 49: 463-491. DOI: 10.1111/j.1365-313x.2006.02979.x doi: 10.1111/j.1365-313x.2006.02979.x
18	MUNIR S, MUMTAZ M A, AHIAKPA J K, et al. Genome-wide analysis of myo-inositol oxygenase gene family in tomato reveals their involvement in ascorbic acid accumulation[J]. BMC Genomics, 2020, 21: 284. DOI: 10.1186/s12864-020-6708-8 doi: 10.1186/s12864-020-6708-8
19	YANG J, YANG J L, ZHAO L L, et al. Ectopic expression of a Malus hupehensis Rehd. myo-inositol oxygenase gene (MhMIOX2) enhances tolerance to salt stress[J]. Scientia Horticulturae, 2021, 281: 109898. DOI: 10.1016/j.scienta.2021.109898 doi: 10.1016/j.scienta.2021.109898
20	SUN F M, FAN G Y, HU Q, et al. The high-quality genome of Brassica napus cultivar ‘ZS11’ reveals the introgression history in semi-winter morphotype[J]. The Plant Journal, 2017, 92(3): 452-468. DOI: 10.1111/tpj.13669 doi: 10.1111/tpj.13669
21	XUE T T, WANG D, ZHANG S Z, et al. Genome-wide and expression analysis of protein phosphatase 2C in rice and Arabidopsis [J]. BMC Genomics, 2008, 9: 550. DOI: 10.1186/1471-2164-9-550 doi: 10.1186/1471-2164-9-550
22	ALI G M, KOMATSU S. Proteomic analysis of rice leaf sheath during drought stress[J]. Journal of Proteome Research, 2006, 5(2): 396-403. DOI: 10.1021/pr050291g doi: 10.1021/pr050291g
23	SARDA X, TOUSCH D, FERRARE K, et al. Two TIP-like genes encoding aquaporins are expressed in sunflower guard cells[J]. The Plant Journal, 1997, 12(5): 1103-1111.
24	CHUNG B Y W, SIMONS C, FIRTH A E, et al. Effect of 5 ´ UTR introns on gene expression in Arabidopsis thaliana [J]. BMC Genomics, 2006, 7: 120. DOI: 10.1186/1471-2164-7-120 doi: 10.1186/1471-2164-7-120
25	JEFFARES D C, PENKETT C J, BÄHLER J. Rapidly regulated genes are intron poor[J]. Trends in Genetics, 2008, 24(8): 375-378. DOI: 10.1016/j.tig.2008.05.006 doi: 10.1016/j.tig.2008.05.006
26	吴楠,杨君,张艳,等.过表达棉花葡萄糖醛酸激酶基因GbGlcAK促进拟南芥细胞伸长[J].中国农业科技导报,2022,24(6):36-46. DOI:10.13304/j.nykjdb.2021.0806 WU N, ZHANG J, ZHANG Y, et al. Overexpression of a cotton glucuronokinase gene GbGlcAK promotes cell elongation in Arabidopsis thaliana [J]. Journal of Agricultural Science and Technology, 2022, 24(6): 36-46. (in Chinese with English abstract） doi: 10.13304/j.nykjdb.2021.0806
27	彭丽娟,彭正松,杨在君,等.小麦三雌蕊基因Pis1的多效性研究初报[J].西华师范大学学报(自然科学版),2011,32(1):12-15. DOI:10.16246/j.issn.1673-5072.2011.01.002 PENG L J, PENG Z S, YANG Z J, et al. Preliminary study on heritable pleiotropy of three pistils gene Pis1 in wheat[J]. Journal of China West Normal University (Natural Sciences), 2011, 32(1): 12-15. (in Chinese with English abstract） doi: 10.16246/j.issn.1673-5072.2011.01.002
28	董祥雨,陈丽华,涂泽行,等.棉花GhVTC2基因促进烟草BY2悬浮细胞和拟南芥主根伸长[J].生物学杂志,2022,39(5):32-37. DOI:10.3969/j.issn.2095-1736.2022.05.032 DONG X Y, CHEN L H, TU Z X, et al. GhVTC2 gene promotes elongation of tobacco BY2 suspension cells and Arabidopsis main roots[J]. Journal of Biology, 2022, 39(5): 32-37. (in Chinese with English abstract) doi: 10.3969/j.issn.2095-1736.2022.05.032
29	BOYCE K J, KRETSCHMER M, KRONSTAD J W. The vtc4 gene influences polyphosphate storage, morphogenesis, and virulence in the maize pathogen Ustilago maydis [J]. Eukaryotic Cell, 2006, 5(8): 1399-1409. DOI: 10.1128/EC.00131-06 doi: 10.1128/EC.00131-06
30	张曼,羊杏平,王薇薇,等.西瓜防御素基因ClPDF2.1的克隆及表达分析[J].西北植物学报,2013,33(5):872-877. DOI:10.3969/j.issn.1000-4025.2013.05.003 ZHANG M, YANG X P, WANG W W, et al. Cloning and expression analysis of plant defensin-like gene (ClPDF2.1) from watermelon[J]. Acta Botanica Boreali-Occidentalia Sinica, 2013, 33(5): 872-877. (in Chinese with English abstract） doi: 10.3969/j.issn.1000-4025.2013.05.003
31	CRAMER G R, URANO K, DELROT S, et al. Effects of abiotic stress on plants: a systems biology perspective[J]. BMC Plant Biology, 2011, 11: 163. DOI: 10.1186/1471-2229-11-163 doi: 10.1186/1471-2229-11-163
32	YANG Y J, MA C, XU Y J, et al. A zinc finger protein regulates flowering time and abiotic stress tolerance in chrysanthemum by modulating gibberellin biosynthesis[J]. The Plant Cell, 2014, 26(5): 2038-2354. DOI: 10.1105/tpc.114.124867 doi: 10.1105/tpc.114.124867
33	LIU S M, KANDOTH P K, WARREN S D, et al. A soybean cyst nematode resistance gene points to a new mechanism of plant resistance to pathogens[J]. Nature, 2012, 492(7428): 256-260. DOI: 10.1038/nature11651 doi: 10.1038/nature11651
34	MITCHUM M G. Soy bean resistance to the soybean cyst nematode Heterodera glycines: an update[J]. Phytopathology, 2016, 106(12): 1444-1450. DOI: 10.1094/PHYTO-06-16-0227-RVW doi: 10.1094/PHYTO-06-16-0227-RVW
35	王学敏,杨若巍,王超,等.大豆肌醇加氧酶基因响应线虫胁迫的表达分析[J].中国油料作物学报,2017,39(6):778-784. DOI:10.7505 /j.issn.1007-9084.2017.06.008 WANG X M, YANG R W, WANG C, et al. Expression analysis of GmMIOX gene response to SCN stress[J]. Chinese Journal of Oil Crop Sciences, 2017, 39(6): 778-784. (in Chinese with English abstract） doi: 10.7505 /j.issn.1007-9084.2017.06.008
2	CHALHOUB B, DENOEUD F, LIU S Y, et al. Early allopolyploid evolution in the post-neolithic Brassica napus oilseed genome[J]. Science, 2014, 345(6199): 950-953. DOI: 10.1126/science.1253435 doi: 10.1126/science.1253435
3	张梦,谢益民,杨海涛,等.肌醇在植物体内的代谢概述:肌醇作为细胞壁木聚糖和果胶前驱物的代谢途径[J].林产化学与工业,2013,33(5):106-114. DOI:10.3969/j.issn.0253-2417.2013.05.021 ZHANG M, XIE Y M, YANG H T, et al. Myo-inositol metamolism as the precursor of xylan and pectin in plants[J]. Chemistry and Industry of Forest Products, 2013, 33(5): 106-114. (in Chinese with English abstract） doi: 10.3969/j.issn.0253-2417.2013.05.021

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