转录组和蛋白质组关联分析解析巴西蕉幼苗响应低温的分子机制

doi:10.3864/j.issn.0578-1752.2024.08.012

摘要/Abstract

摘要：

【目的】低温是导致香蕉减产的主要自然灾害之一。基于转录组与蛋白质组关联分析参与香蕉抗寒的相关基因、蛋白及信号、代谢通路调控网络，探讨香蕉抗寒的分子机制。【方法】以巴西蕉为试材，在7 ℃低温下处理1 d（Cold1）和3 d（Cold3），以28 ℃培养的巴西蕉为对照（CK），基于笔者课题组前期获得的蛋白质组数据，利用转录组测序技术检测巴西蕉在低温胁迫下基因调控网络的变化，同时与蛋白质组学数据进行关联分析，共同解析巴西蕉响应低温胁迫的分子机制。【结果】转录组分析结果显示，Cold1 vs CK、Cold3 vs CK和Cold1 vs Cold3三个对比组分别鉴定出11 370、15 460和9 619个差异表达基因，对这些基因的KEGG富集分析发现，差异表达基因在光合作用信号、谷胱甘肽代谢、α-亚麻酸代谢途径和苯丙素生物合成等多个低温胁迫关键信号代谢通路中富集。对部分差异表达基因进行实时荧光定量（qRT-PCR）分析，其中DREB、MAPK和MYB等低温调控关键基因的表达量在低温处理后显著上升，所选20个基因的表达变化趋势与RNA-seq基本相符，证实了RNA-seq的准确性。转录组与蛋白质组关联分析结果显示，共鉴定到6 211个与转录本相对应的蛋白，转录组与蛋白质呈现正相关关系，共有105个转录本及其对应的蛋白表达量共同上调，有218个转录本及其对应的蛋白表达量共同下调。GO富集分析显示，差异表达基因和蛋白在光响应、叶绿体、氧化还原酶活性等功能大量富集。此外，对差异表达基因和蛋白KEGG通路的关联分析发现，低温处理抑制了苯丙素生物合成和光合作用信号途径相关基因及蛋白的表达，促进了α-亚麻酸代谢和谷胱甘肽途径的表达。【结论】运用转录组结合蛋白质组学技术绘制了基因和蛋白质水平上的香蕉抗寒调控网络，并发现香蕉响应低温的信号通路主要涉及光合作用信号、谷胱甘肽代谢、α-亚麻酸代谢途径和苯丙素生物合成等途径。

关键词: 香蕉, 低温胁迫, 转录组, 蛋白质组, 关联分析

Abstract:

【Objective】 Low temperature is a significant natural disaster that affects banana production. In this study, based on transcriptome and proteome association analysis, the regulatory network of genes, proteins, signals and metabolic pathways involved in banana cold resistance was investigated. The aim was to explore the molecular mechanism of banana cold resistance. 【Methods】 ‘Baxi’ banana (Musa nana Lour) was treated at 7 ℃ for 1 and 3 d, and a control group was treated at 28 ℃. Based on the proteome data obtained in the previous study, the transcriptome sequencing technology was used to detect changes in the gene regulatory network of banana under cold stress. Simultaneously, the correlation analysis was conducted with proteomics to analyze the molecular mechanism of banana response to cold stress. 【Result】 Transcriptome analysis revealed that 11 370, 15 460 and 9 619 differentially expressed genes were identified in the three comparison groups of Cold1 vs CK, Cold3 vs CK and Cold1 vs Cold3, respectively. KEGG enrichment analysis of these genes revealed that the differentially expressed genes were enriched in several key signaling metabolic pathways, such as photosynthesis signal, glutathione metabolism, α-linolenic acid metabolic pathway and phenylpropanoid biosynthesis under low temperature stress. Moreover, there were significant differences in the enrichment degree of glutathione metabolic pathway between Cold1 d vs CK and Cold3 d vs CK. Real-time quantitative PCR analysis (qRT-PCR) was performed on several differentially expressed genes. Among them, the expression levels of key low-temperature regulatory genes, such as DREB, MAPK and MYB, were significantly increased after the low-temperature treatment. The expression trend of the selected 20 genes was essentially consistent with that of RNA-seq, confirming the accuracy of RNA-seq. The results of the transcriptome and proteome association analysis showed a positive correlation between the transcriptome and proteomics. A total of 6 211 proteins corresponding to transcripts were identified. Among these, 105 transcripts and their proteins were up-regulated, while 218 transcripts and their proteins were down-regulated. GO enrichment analysis showed that the differentially expressed genes and proteins were enriched in functions, such as photoresponse, chloroplast and oxidoreductase activity. Furthermore, the correlation analysis of differentially expressed genes and protein KEGG pathway revealed that the low temperature treatment suppressed the expression of genes and proteins related to phenylpropanoid biosynthesis and photosynthesis signaling pathway, while promoting the expression of proteins associated with α-linolenic acid metabolism and the glutathione pathway. 【Conclusion】 The transcriptome and proteomics were used to map the regulatory network of banana cold resistance at the gene and protein levels. It was found that the signal pathway of banana response to low temperature mainly involved photosynthesis signal, glutathione metabolism, α-linolenic acid metabolism and phenylpropanol biosynthesis.

Key words: Musa nana Lour, low temperature stress, transcriptome, proteome, comparative analysis

林蔚, 吴水金, 李跃森. 转录组和蛋白质组关联分析解析巴西蕉幼苗响应低温的分子机制[J]. 中国农业科学, 2024, 57(8): 1575-1591.

LIN Wei, WU ShuiJin, LI YueSen. Transcriptome and Proteome Association Analysis to Revealthe Molecular Mechanism of Baxi Banana Seedlings in Response to Low Temperature[J]. Scientia Agricultura Sinica, 2024, 57(8): 1575-1591.

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参考文献 39

[1]	吴烁凡, 何维弟, 李春雨, 董涛, 毕方铖, 盛鸥, 邓贵明, 胡春华, 窦同心, 高慧君, 刘思文, 易干军, 姚振, 杨乔松. 香蕉抗寒分子机制研究进展. 果树学报, 2022, 39(3): 483-494.
	WU S F, HE W D, LI C Y, DONG T, BI F C, SHENG O, DENG G M, HU C H, DOU T X, GAO H J, LIU S W, YI G J, YAO Z, YANG Q S. Research progress in molecular mechanism of cold resistance in banana. Journal of Fruit Science, 2022, 39(3): 483-494. (in Chinese)
[2]	谢江辉. 新中国果树科学研究70年: 香蕉. 果树学报, 2019, 36(10): 1429-1440.
	XIE J H. Fruit scientific research in New China in the past 70 years: banana. Journal of Fruit Science, 2019, 36(10): 1429-1440. (in Chinese)
[3]	王伟英, 邹晖, 林江波, 戴艺民. 香蕉抗寒技术研究进展. 东南园艺, 2018, 6(6): 61-66.
	WANG W Y, ZOU H, LIN J B, DAI Y M. Research progress on techniques involved in cold resistance of banana. Southeast Horticulture, 2018, 6(6): 61-66. (in Chinese)
[4]	王安邦, 金志强, 刘菊华, 贾彩红, 张建斌, 苗红霞, 徐碧玉. 香蕉寒害研究现状及展望. 生物技术通报, 2014(8): 28-33.
	WANG A B, JIN Z Q, LIU J H, JIA C H, ZHANG J B, MIAO H X, XU B Y. The current situation and prospects of banana chilling stress. Biotechnology Bulletin, 2014(8): 28-33. (in Chinese)
[5]	PLOHOVSKA S G, YEMETS A I, BLUME Y B. Influence of cold on organization of actin filaments of different types of root cells in Arabidopsis thaliana. Cytology and Genetics, 2016, 50(5): 318-323. doi: 10.3103/S0095452716050108
[6]	SONAWANE B V, KOTEYEVA N K, JOHNSON D M, COUSINS A B. Differences in leaf anatomy determines temperature response of leaf hydraulic and mesophyll CO₂ conductance in phylogenetically related C4 and C3 grass species. The New Phytologist, 2021, 230(5): 1802-1814. doi: 10.1111/nph.v230.5
[7]	SACK L, STREETER C M, HOLBROOK N M. Hydraulic analysis of water flow through leaves of sugar maple and red oak. Plant Physiology, 2004, 134(4): 1824-1833. pmid: 15064368
[8]	ZHENG G H, PAN D M, NIU X Q, WU H W, ZHANG J B. Changes in cell Ca²⁺ distribution in loquat leaves and its effects on cold tolerance. Horticultural Science and Technology, 2014, 32(5): 607-613. doi: 10.7235/hort.2014.13009
[9]	MA Y, DAI X Y, XU Y Y, LUO W, ZHENG X M, ZENG D L, PAN Y J, LIN X L, LIU H H, ZHANG D J, XIAO J, GUO X Y, XU S J, NIU Y D, JIN J B, ZHANG H, XU X, LI L G, WANG W, QIAN Q, GE S, CHONG K. COLD1 confers chilling tolerance in rice. Cell, 2015, 160(6): 1209-1221. doi: 10.1016/j.cell.2015.01.046 pmid: 25728666
[10]	LI M, DUAN X Y, GAO G, LIU T, QI H Y. CmABF1 and CmCBF4 cooperatively regulate putrescine synthesis to improve cold tolerance of melon seedlings. Horticulture Research, 2022, 9: uhac002. doi: 10.1093/hr/uhac002
[11]	耿小惠. 钙离子通道促进剂及抑制剂对香蕉抗寒性的影响及相关基因MaCNGC家族成员鉴定与表达分析[D]. 福州: 福建农林大学, 2022.
	GENG X H. Effect of calcium channel promoters and inhibitors on cold resistance in banana and identification and expression analysis of MaCNGC family members[D]. Fuzhou: Fujian Agriculture and Forestry University, 2022. (in Chinese)
[12]	刘嘉鹏. 褪黑素对香蕉抗寒性的影响及其机制研究[D]. 福州: 福建农林大学, 2022.
	LIU J P. Effects of melatonin on cold resistance of banana and study of its mechanism[D]. Fuzhou: Fujian Agriculture and Forestry University, 2022. (in Chinese)
[13]	GAO J, DOU T X, HE W D, SHENG O, BI F C, DENG G M, GAO H J, DONG T, LI C Y, ZHANG S, YI G J, HU C H, YANG Q S. MaMAPK3-MaICE1-MaPOD P7 pathway, a positive regulator of cold tolerance in banana. BMC Plant Biology, 2021, 21(1): 97. doi: 10.1186/s12870-021-02868-z pmid: 33596830
[14]	LIN W, WU S J, WEI M. Ubiquitylome analysis reveals the involvement of ubiquitination in the cold responses of banana seedling leaves. Journal of Proteomics, 2023, 288: 104994. doi: 10.1016/j.jprot.2023.104994
[15]	SCHEFE J H, LEHMANN K E, BUSCHMANN I R, UNGER T, FUNKE-KAISER H. Quantitative real-time RT-PCR data analysis: Current concepts and the novel “gene expression’s CT difference” formula. Journal of Molecular Medicine, 2006, 84(11): 901-910. doi: 10.1007/s00109-006-0097-6
[16]	JANMOHAMMADI M, ZOLLA L, RINALDUCCI S. Low temperature tolerance in plants: Changes at the protein level. Phytochemistry, 2015, 117: 76-89. doi: S0031-9422(15)30012-1 pmid: 26068669
[17]	GHAZALPOUR A, BENNETT B, PETYUK V A, OROZCO L, HAGOPIAN R, MUNGRUE I N, FARBER C R, SINSHEIMER J, KANG H M, FURLOTTE N, et al. Comparative analysis of proteome and transcriptome variation in mouse. PLoS Genetics, 2011, 7(6): e1001393. doi: 10.1371/journal.pgen.1001393
[18]	RODRIGUES R S, BOLDRINI-FRANÇA J, FONSECA F P P, DE LA TORRE P, HENRIQUE-SILVA F, SANZ L, CALVETE J J, RODRIGUES V M. Combined snake venomics and venom gland transcriptomic analysis of Bothropoides pauloensis. Journal of Proteomics, 2012, 75(9): 2707-2720. doi: 10.1016/j.jprot.2012.03.028
[19]	SAVITCH L V, ALLARD G, SEKI M, ROBERT L S, TINKER N A, HUNER N P A, SHINOZAKI K, SINGH J. The effect of overexpression of two Brassica CBF/DREB1-like transcription factors on photosynthetic capacity and freezing tolerance in Brassica napus. Plant & Cell Physiology, 2005, 46(9): 1525-1539.
[20]	HUNER N P A, ÖQUIST G, SARHAN F. Energy balance and acclimation to light and cold. Trends in Plant Science, 1998, 3(6): 224-230. doi: 10.1016/S1360-1385(98)01248-5
[21]	ASADA K. Production and Action of Active Oxygen Species in Photosynthetic Tissues. Boca Raton: CRC Press, 2020: 77-104.
[22]	VIJAYAKUMAR H, THAMILARASAN S K, SHANMUGAM A, NATARAJAN S, JUNG H J, PARK J I, KIM H, CHUNG M Y, NOU I S. Glutathione transferases superfamily: Cold-inducible expression of distinct GST genes in Brassica oleracea. International Journal of Molecular Sciences, 2016, 17(8): 1211. doi: 10.3390/ijms17081211
[23]	XU H Y, YU C. Transcriptomic analysis reveals crucial biological pathways associated with cold response in Camellia weiningensis in Guizhou Province, China. Scientia Horticulturae, 2022, 295: 110883. doi: 10.1016/j.scienta.2022.110883
[24]	SOUSA B, LOPES J, LEAL A, MARTINS M, SOARES C, AZENHA M, FIDALGO F, TEIXEIRA J. Specific glutathione-S- transferases ensure an efficient detoxification of diclofenac in Solanum lycopersicum L. plants. Plant Physiology and Biochemistry, 2021, 168: 263-271. doi: 10.1016/j.plaphy.2021.10.019
[25]	SUBRAMANIAN S, SOULEIMANOV A, SMITH D L. Thuricin17 production and proteome differences in Bacillus thuringiensis NEB17 cell-free supernatant under NaCl stress. Frontiers in Sustainable Food Systems, 2021, 5: 630628. doi: 10.3389/fsufs.2021.630628
[26]	SAPPL P G, CARROLL A J, CLIFTON R, LISTER R, WHELAN J, HARVEY MILLAR A, SINGH K B. The Arabidopsis glutathione transferase gene family displays complex stress regulation and co-silencing multiple genes results in altered metabolic sensitivity to oxidative stress. The Plant Journal, 2009, 58(1): 53-68. doi: 10.1111/tpj.2009.58.issue-1
[27]	SEPPÄNEN M M, CARDI T, BORG HYÖKKI M, PEHU E. Characterization and expression of cold-induced glutathione S- transferase in freezing tolerant Solanum commersonii, sensitive S. tuberosum and their interspecific somatic hybrids. Plant Science, 2000, 153(2): 125-133. doi: 10.1016/S0168-9452(99)00252-6
[28]	TSUCHIYA T, TAKESAWA T, KANZAKI H, NAKAMURA I. Genomic structure and differential expression of two tandem-arranged GSTZ genes in rice. Gene, 2004, 335: 141-149. doi: 10.1016/j.gene.2004.03.020
[29]	HONG M J, JANG Y E, KIM D G, KIM J M, LEE M K, KIM J B, EOM S H, HA B K, LYU J I, KWON S J. Selection of mutants with high linolenic acid contents and characterization of fatty acid desaturase 2 and 3 genes during seed development in soybean (Glycine max). Journal of the Science of Food and Agriculture, 2019, 99(12): 5384-5391. doi: 10.1002/jsfa.v99.12
[30]	李国斌, 党林学, 郑国强, 董小云, 魏家萍, 崔俊美, 李辉, 王莹, 田海燕, 刘自刚. 甘蓝型冬油菜亚麻酸代谢参与应答低温胁迫的转录组学分析. 分子植物育种, 2023. https://kns.cnki.net/kcms2/detail/46.1068.s.20230615.1429.008.html.
	LI G B, DANG L X, ZHENG G Q, DONG X Y, WEI J P, CUI J M, LI H, WANG Y, TIAN H Y, LIU Z G. Analysis of the linolenic acid metabolism involving low temperature stress-response in leaves of brassica napus based on RNA-seq. Molecular Plant Breeding, 2023. https://kns.cnki.net/kcms2/detail/46.1068.s.20230615.1429.008.html. (in Chinese)
[31]	KOH I. Acclimative response to temperature stress in higher plants: approaches of gene engineering for temperature tolerance. Annual Review of Plant Biology, 2002, 53: 225-245. pmid: 12221974
[32]	OLSSON M, NILSSON K, LILJENBERG C, HENDRY G A F. Drought stress in seedlings: Lipid metabolism and Lipid peroxidation during recovery from drought in Lotus comiculatrus and Cerastium fontanum. Physiologia Plantarum, 1996, 96(4): 577-584. doi: 10.1111/ppl.1996.96.issue-4
[33]	DING F, WANG C, XU N, WANG M L, ZHANG S X. Jasmonic acid-regulated putrescine biosynthesis attenuates cold-induced oxidative stress in tomato plants. Scientia Horticulturae, 2021, 288: 110373. doi: 10.1016/j.scienta.2021.110373
[34]	JANNATIZADEH A. Exogenous melatonin applying confers chilling tolerance in pomegranate fruit during cold storage. Scientia Horticulturae, 2019, 246: 544-549. doi: 10.1016/j.scienta.2018.11.027
[35]	CHENG Y D, LIU L Q, ZHAO G Q, SHEN C G, YAN H B, GUAN J F, YANG K. The effects of modified atmosphere packaging on core browning and the expression patterns of PPO and PAL genes in ‘Yali’ pears during cold storage. LWT-Food Science and Technology, 2015.
[36]	SANCHEZ-BALLESTA M T, LAFUENTE M T, ZACARIAS L, GRANELL A. Involvement of phenylalanine ammonia-lyase in the response of Fortune mandarin fruits to cold temperature. Physiologia Plantarum, 2000, 108(4): 382-389. doi: 10.1034/j.1399-3054.2000.108004382.x
[37]	VOGT T. Phenylpropanoid biosynthesis. Molecular Plant, 2010, 3(1): 2-20. doi: 10.1093/mp/ssp106 pmid: 20035037
[38]	CHRISTIE P J, ALFENITO M R, WALBOT V. Impact of low- temperature stress on general phenylpropanoid and anthocyanin pathways: Enhancement of transcript abundance and anthocyanin pigmentation in maize seedlings. Planta, 1994, 194(4): 541-549. doi: 10.1007/BF00714468
[39]	JANAS K M, CVIKROVÁ M, PAŁĄGIEWICZ A, EDER J. Alterations in phenylpropanoid content in soybean roots during low temperature acclimation. Plant Physiology and Biochemistry, 2000, 38: 587-593. doi: 10.1016/S0981-9428(00)00778-6

基因名称（基因ID） Gene name (Gene ID)	引物序列 Primers sequence (5′-3′)
基因名称（基因ID） Gene name (Gene ID)	正向 Forward	反向 Reverse
MaMYB (LOC103972162)	CTGCCCTTTCGTCTGTCTTC	CAGTTCTCGGCAGGAGGTAG
MaDREB (LOC103990508)	GACAGAGAGCTCAGCACACG	AGCATCGACATCATCCCAAT
MaWRKY45 (LOC108952331)	AGTGGGTCAGAGCCTTGAGA	TGCACTCCGTCGTAAGTGAC
MaMYBP (LOC103998926)	GCTCATCATCAAGCTCCACA	CTCGAAATGGGTGGCTGTAT
MaWRKY33 (LOC103989687)	CGGGTTGTCTTCCATGAGAT	CCTCCAGTTGTATCCGTCGT
MaMYC (LOC103985317)	TCTCCATGACCCAGTCCTTC	GAAGAGGACGCTGATCTTGC
MaCPK29 (LOC103995457)	GCAATTCAGAGCGATGAACA	CCAGCCTTCTCAGTCCAGTC
MaMAPK5 (LOC103996929)	AACCCGAGCTTGGATTTCTT	AGAAAGGGTCCATGCAAGTG
MaWRKY24 (LOC103989687)	CGGGTTGTCTTCCATGAGAT	CCTCCAGTTGTATCCGTCGT
MaGST U17 (LOC103994768)	TGGTTTGACAGCCACCATTA	CACTTCCGCCATCTCTTCTC
MaGST F12 (LOC103981216)	CAACAAGGTCCTGGAGGTGT	TTGACGTGCTTCTTGTCGTC
MaGST F10 (LOC103982403)	TGGTCGAACAGAGCATGAAG	TCCCACCACCTGTACACCTT
MaGST zeta (LOC103995971)	TCGGCTCGAGTATTCAACCT	TTTGGAAGCGCACTAATCCT
MaGST T1 (LOC103973270)	TTTCGAGGAGGTGAGGATTG	GATACCAATGGTCCGGAATG
Ma14-3-3 (LOC103985456)	TGAGGATCATGTTTCCGTGA	GCCTCCTTCCTTTCATTTCC
MaMKK5 (LOC103981573)	GGATCAACACCGACCTCAAC	CAGATGGCCACCATGAGAC
MaCML49 (LOC103972901)	CCCAAAGGAAGGGAAGACTC	TGTTGCAGCTCCTTGTCATC
MaCML18 (LOC103994211)	GCTTTGAACGCGTCTTCTTC	GGTCAAGCAAGGAGACCTCA
MaCML31 (LOC104000196)	TGATTCTCGTCAATCCAACG	TTTGTCGTGCTGCTTATTGG
MaCML10 (LOC103995933)	TTCGTCGAACTCAACACCAG	GGTCATCATGGCCTTGAACT
MaACTIN (LOC103992883)	CGTAGCACCAGAAGAACA	CATAAAGGGAGAGGACAG

样品名称 Sample name	过滤后序列 Clean reads	过滤后总数据量 Clean bases	碱基错误率 Error rate (%)	Q20 (%)	Q30 (%)	GC (%)
CK1	55962510	8251833288	0.0266	97.39	92.72	54.51
CK2	54693020	7946130178	0.0267	97.38	92.69	53.64
CK3	46556340	6760589819	0.0273	97.16	92.15	53.30
Cold1-1	43765858	6433985244	0.0268	97.34	92.49	50.46
Cold1-2	49835534	7319445856	0.0275	97.07	91.89	50.12
Cold1-3	45665780	6692564466	0.0272	97.21	92.18	50.38
Cold3-1	43171118	6347773723	0.0275	97.07	91.9	50.69
Cold3-2	47440180	6960515302	0.0276	97.04	91.83	50.43
Cold3-3	43284622	6385904142	0.0276	97.06	91.86	50.56

样品名称 Sample	过滤序列总数 Clean reads	总比对（对比率） Totalmapped (Mapping ratio, %)	多方比对（对比率） Multiple mapped (Mapping ratio, %)	唯一比对（对比率） Uniquely mapped (Mapping ratio, %)
CK1	55962510	51136284 (91.38%)	853090 (1.52%)	50283194 (89.85%)
CK2	54693020	49654543 (90.79%)	792871 (1.45%)	48861672 (89.34%)
CK3	46556340	42271280 (90.8%)	647597 (1.39%)	41623683 (89.4%)
Cold1-1	43765858	38887517 (88.85%)	553404 (1.26%)	38334113 (87.59%)
Cold1-2	49835534	44029762 (88.35%)	605472 (1.21%)	43424290 (87.14%)
Cold1-3	45665780	40473620 (88.63%)	548796 (1.2%)	39924824 (87.43%)
Cold3-1	43171118	38239493 (88.58%)	596851 (1.38%)	37642642 (87.19%)
Cold3-2	47440180	42055866 (88.65%)	728177 (1.53%)	41327689 (87.12%)
Cold3-3	43284622	38397112 (88.71%)	702198 (1.62%)	37694914 (87.09%)

功能分类 Functional classification	Unigene数量 Unigene number
功能分类 Functional classification	Cold1/CK	Cold3/CK	Cold1/Cold3
RNA加工和修饰 RNA processing and modification	29	58	25
染色质结构与动力学 Chromatin structure and dynamics	74	98	53
能源生产和转换 Energy production and conversion	185	250	164
细胞周期控制、细胞分裂 Cell cycle control, cell division	118	154	89
氨基酸运输和代谢 Amino acid transport and metabolisS	255	385	251
核苷酸运输和代谢 Nucleotide transport and metabolism	63	87	59
碳水化合物运输和代谢 Carbohydrate transport and metabolism	505	604	413
辅酶转运与代谢 Coenzyme transport and metabolism	85	123	74
脂质运输和代谢 Lipid transport and metabolism	195	275	179
核糖体结构与生物发生 Ribosomal structure and biogenesis	240	417	242
转录 Transcription	1093	1396	898
复制、重组和修复 Replication, recombination and repair	136	228	145
细胞壁、膜、包膜生物发生 Cell wall, membrane,envelope biogenesis	168	224	148
细胞运动 Cell motility	5	5	5
翻译后修饰 Posttranslational modification	763	1176	760
无机离子运输和代谢 Inorganic ion transport and metabolism	176	250	165
次生代谢产物生物合成 Secondary metabolites biosynthesis	166	235	169
信号传导机制 Signal transduction mechanisms	784	1107	673
胞内运输 Intracellular trafficking	338	522	315
防御机制 Defense mechanisms	80	99	72
核结构 Nuclear structure	0	2	1
细胞骨架 Cytoskeleton	133	173	98