高产油绿球藻GIEC-38转录本和基因表达谱分析

doi:10.3969/j.issn.1673-5854.2019.02.004

摘要/Abstract

摘要：

通过对天然条件下生长好且含油高的绿球藻(Chlorococcum sp.) GIEC-38细胞内转录本的测定,发现:在获得的74 605条转录本、65 984个基因中共得到344条代谢途径，其中核糖体、蛋白质、核苷酸、核糖核酸、碳固定、光合作用以及脂代谢等通路都非常活跃。通过缺N培养发现GIEC-38含油脂可达50%以上，通过对比细胞表达谱发现相比于原始藻株，缺N条件下培养的GIEC-38细胞中有868个基因显著上调、1 157个基因显著下调，分布在41个生物学功能、71条代谢途径中。其中，为脂肪酸的合成提供大量原料的中间产物合成酶，以及脂类代谢相关的几个通路中关键酶的基因，表达都有明显上调，促使细胞脂类的合成，提高了细胞的油脂产量。

关键词: 绿球藻, GIEC-38, 代谢通路, 基因表达, 油脂

Abstract:

The transcript of Chlorococcum sp. GIEC-38 which exhibited high biomass and lipid yield was sequenced. 74 605 transcripts and 65 984 genes were obtained related to 344 metabolic pathways which were active in ribosome, protein, nucleotide, RNA, carbon fixation, photosynthesis, and lipid metabolism. The lipid content of Chlorococcum sp. GIEC-38 cultured without nitrogen reached up to over 50% and compared with the original strain, 868 genes of strain cultured without nitrogen significantly up-regulated and 1 157 genes significantly down-regulated which were distributed in 41 biological functions and 71 metabolic pathways. It was found that the expressions of synthetase catalyzed intermediate products synthesis which provided for lipid acid synthesis and the key enzymes in metabolic pathways related to lipid metabolism were up-regulated significantly. All those changes promoted lipid synthesis and increased the lipid content of GIEC-38.

Key words: Chlorococcum sp., GIEC-38, metabolic pathways, gene expression, lipid

中图分类号:

TQ35
TK6

冯佳,朱顺妮,许瑾,王忠铭,袁振宏. 高产油绿球藻GIEC-38转录本和基因表达谱分析[J]. 生物质化学工程, 2019, 53(2): 19-25.

Jia FENG,Shunni ZHU,Jin XU,Zhongming WANG,Zhenhong YUAN. Transcript and Gene Expression Analysis of Chlorococcum sp. GIEC-38 with High Lipid Productivity[J]. Biomass Chemical Engineering, 2019, 53(2): 19-25.

图/表 4

表1

表2

GIEC-38转录本GO功能分类统计"

	功能分类 functional classification	序列条数 sequence number	GO
分子功能 molecular function	电子载体活性electron carrier activity	78	0009055
	蛋白结合转录因子活性protein binding transcription factor activity	12	0000988
	绑定binding	3405	0005488
	转运活性transporter activity	377	0005215
	催化活性catalytic activity	4225	0003824
	金属伴侣蛋白活性metallochaperone activity	1	0016530
	鸟嘌呤核苷酸交换因子活性guanyl-nucleotide exchange factor activity	4	0005085
	营养库活性nutrient reservoir activity	6	0045735
	酶调节活性enzyme regulator activity	50	0030234
	分子转导活性molecular transducer activity	56	0060089
	结构分子活性structural molecule activity	315	0005198
	核酸结合转录因子活性nucleic acid binding transcription factor activity	60	0001071
	受体活性receptor activity	27	0004872
	抗氧化活性antioxidant activity	58	0016209
细胞成分 cellular component	细胞外基质extracellular matrix	5	0031012
	胞外区部分extracellular region part	19	0044421
	胶原三聚体collagen trimer	1	0005581
	膜部分membrane part	687	0044425
	细胞连接cell junction	346	0030054
	类核nucleoid	6	0009295
	细胞cell	3347	0005623
	突触部分synapse part	2	0044456
	共质体symplast	345	0055044
	病毒粒子部分virion part	15	0044423
	细胞器organelle	2483	0043226
	细胞部分cell part	3347	0044464
	病毒粒子virion	15	0019012
	膜蛋白腔membrane-enclosed lumen	322	0031974
	突触synapse	2	0045202
	胞外区extracellular region	228	0005576
	细胞外基质部分extracellular matrix part	3	0044420
	膜membrane	1626	0016020
	大分子复合物macromolecular complex	1289	0032991
	细胞器部分organelle part	1408	0044422
生物过程 biologicalprocess	繁殖reproduction	40	0000003
	多生物过程multi-organism process	231	0051704
	发展过程developmental process	404	0032502
	生物过程的正调控positive regulation of biological process	108	0048518
	多细胞生物过程multicellular organismal process	372	0032501
	细胞过程cellular process	4044	0009987
	激素分泌hormone secretion	1	0046879
	单组织过程single-organism process	3153	0044699
	生长growth	84	0040007
	信号signaling	230	0023052
	生物过程的负调控negative regulation of biological process	115	0048519
	代谢过程metabolic process	4686	0008152
	生物调节biological regulation	870	0065007
	免疫系统过程immune system process	65	0002376
	生殖过程reproductive process	128	0022414
	生物过程调节regulation of biological process	816	0050789
	建立定位establishment of localization	817	0051234
	细胞成分组织或生物合成cellular component organization or biogenesis	794	0071840
	节律过程rhythmic process	19	0048511
	细胞死亡cell killing	1	0001906
	刺激反应response to stimulus	1186	0050896
	生物粘附biological adhesion	7	0022610
	运动locomotion	26	0040011
	定位localization	843	0051179

表2

表3

表4

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物种 species	同源序列数 homologous sequence number	相似度/% similarity
团藻Volvox carteri	3556	16.97
莱茵衣藻Chlamydomonas reinhardtii	3151	15.04
胶球藻Coccomyxa subellipsoidea	1710	8.16
小球藻Chlorella variabilis	1450	6.92
海枣Phoenix dactylifera	1317	6.29
毒害艾美耳球虫Eimeria necatrix	613	2.93
水稻Oryza sativa	338	1.61
和缓艾美耳球虫Eimeria mitis	235	1.12
小立碗藓Physcomitrella patens	200	0.95
布氏艾美耳球虫Eimeria brunetti	185	0.88
旋孢藻属ATCC 50920 Helicosporidium sp. ATCC 50920	182	0.87
艾美尔球虫Eimeria praecox	177	0.84
微单胞藻RCC299 Micromonas sp. RCC299	137	0.65
江南卷柏Selaginella moellendorffii	124	0.59
孔雀鱼Poecilia reticulata	116	0.55
葡萄Vitis vinifera	106	0.51
其他 others	7355	35.10

代谢通路 metabolic pathways	序列数 sequence number
核糖体ribosome	338
内质网上蛋白质合成protein processing in endoplasmic reticulum	212
嘌呤代谢purine metabolism	205
剪接体spliceosome	201
RNA运输RNA transport	190
亨廷顿氏舞蹈症huntington′s disease	163
氧化磷酸化oxidative phosphorylation	154
EB病毒感染epstein-barr virus infection	150
嘧啶代谢pyrimidine metabolism	148
淀粉和蔗糖代谢starch and sucrose metabolism	140
RNA降解RNA degradation	127
真核生物核糖体生物合成ribosome biogenesis in eukaryotes	126
糖酵解和糖质新生glycolysis/gluconeogenesis	122
细胞周期cell cycle	122
植物-病原互作plant-pathogen interaction	122
病毒致癌作用viral carcinogenesis	118
精氨酸和脯氨酸代谢arginine and proline metabolism	114
泛素介导的蛋白降解作用ubiquitin mediated proteolysis	112
过氧物酶体peroxisome	112
光合生物固碳作用carbon fixation in photosynthetic organisms	110

代谢通路 metabolism pathway	编号 ID	样品数/背景数 sample background numbers	P值 P value
光合作用-天线蛋白photosynthesis-antenna proteins	ko00196	19/45	1.81×10^-13
核糖体生物合成ribosome biogenesis in eukaryotes	ko03008	24/127	2.62×10^-10
精氨酸和脯氨酸代谢arginine and proline metabolism	ko00330	16/114	8.34×10^-6
RNA聚合酶RNA polymerase	ko03020	8/36	0.0001
嘧啶代谢pyrimidine metabolism	ko00240	15/148	0.000441
组氨酸代谢histidine metabolism	ko00340	6/27	0.000753
辅酶q和其他萜醌生物合成ubiquinone and other terpenoid-quinone biosynthesis	ko00130	7/40	0.000964
氮代谢nitrogen metabolism	ko00910	7/43	0.001403
嘌呤代谢purine metabolism	ko00230	17/206	0.001604
2-氧代羧酸代谢2-oxocarboxylic acid metabolism	ko01210	8/61	0.002262
氨基酸的生物合成biosynthesis of amino acids	ko01230	19/256	0.002722
卟啉和叶绿素代谢porphyrin and chlorophyll metabolism	ko00860	9/83	0.003981
光合作用photosynthesis	ko00195	10/110	0.007694
胞质DNA-传感通路cytosolic DNA-sensing pathway	ko04623	4/20	0.008124
异喹啉碱基合成isoquinoline alkaloid biosynthesis	ko00950	4/20	0.010837
DNA复制DNA replication	ko03030	8/82	0.01134
溶酶体lysosome	ko04142	7/66	0.011768
丙氨酸、天冬氨酸和谷氨酸代谢alanine, aspartate and glutamate metabolism	ko00250	7/69	0.014488
β-丙氨酸代谢β-alanine metabolism	ko00410	5/41	0.019276
酪氨酸代谢tyrosine metabolism	ko00350	5/48	0.033336
托烷、哌啶和吡啶生物碱合成tropane, piperidine and pyridine alkaloid biosynthesis	ko00960	3/20	0.041604
减数分裂meiosis-yeast	ko04113	6/69	0.041782

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