2种内生和附生橡藻属绿藻的胞外分泌物研究

doi:10.3785/j.issn.1008-9209.2020.09.221

浙江大学学报（农业与生命科学版）

2021, Vol. 47

Issue (3): 363-370 DOI: 10.3785/j.issn.1008-9209.2020.09.221

资源利用与环境保护

2种内生和附生橡藻属绿藻的胞外分泌物研究

邓茹茹^1,²(

),马帅²,谭德冠²,付莉莉²,张家明²,孙雪飘²,刘平怀¹(

)

^1.海南大学化学工程与技术学院，海口 570228
^2.中国热带农业科学院热带生物技术研究所/中国热带农业科学院海南热带农业资源研究院/海南省热带微生物资源重点实验室，海口 571101

Studies on extracellular metabolites of two endophytic and epiphytic algae of Heveochlorella spp

Ruru DENG^1,²(

),Shuai MA²,Deguan TAN²,Lili FU²,Jiaming ZHANG²,Xuepiao SUN²,Pinghuai LIU¹(

)

^1.School of Chemical Engineering and Technology, Hainan University, Haikou 570228, China
^2.Institute of Tropical Biotechnology, Chinese Academy of Tropical Agricultural Sciences/Hainan Institute of Tropical Agricultural Resources, Chinese Academy of Tropical Agricultural Sciences/Hainan Key Laboratory of Tropical Microbe Resources, Haikou 571101, China

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摘要：

为探讨2种不同生活方式的内生和附生橡藻属（Heveochlorella）绿藻的胞外分泌物之间的差异，本实验通过测定内生海南橡藻（H. hainanensis）和附生王棕橡藻（H. roystonensis）的生物量和培养液pH，采用非靶向代谢组学分析方法对其胞外分泌物进行定性检测。结果表明：2种藻共被检测到141种胞外分泌物，其中海南橡藻拥有8种特有成分，王棕橡藻拥有6种特有成分；两者胞外分泌物中氨基酸及其衍生物、酮醇类等组分的相对含量占比相近，但有机酸的差异较明显。不加有机碳源培养15 d，内生的海南橡藻生长缓慢，培养液偏酸性，有机酸占比为3.28%；附生的王棕橡藻生长正常，培养液偏碱性，有机酸占比为2.04%。添加0.5%蔗糖作为有机碳源时，海南橡藻生物量由0.09 g/L增至3.87 g/L，有机酸占比增至4.64%，胺类及生物碱含量有所降低，培养液pH下降至4.70；王棕橡藻生物量由3.20 g/L增长到5.24 g/L，有机酸占比降至1.13%，胺类及生物碱含量有所增高，培养液pH上升至8.32。总之，2种橡藻属绿藻在培养过程中培养液的酸碱度变化相反；添加有机碳源能明显促进它们的生长繁殖，并促进海南橡藻有机酸的分泌，抑制王棕橡藻有机酸的分泌，使两者培养液酸化或碱化程度增强。本实验结果可为橡藻属的进化分析及其胞外分泌物的开发利用提供一定的参考。

关键词： 海南橡藻; 王棕橡藻; 胞外分泌物; 非靶向代谢组学

Abstract:

To investigate the differences of extracellular metabolites between endophytic algae of Heveochlorella hainangensis andepiphytic algae of Heveochlorella roystonensis, the biomass and pH of culture medium were analyzed, as well as the extracellular metabolites were qualitatively detected by non-targeted metabolomics analysis. The results showed that totally 141 compounds were detected in the culture media of the two algae species, of which eight and six ingredients specifically presented in the culture media of H. hainangensis and H. roystonensis, respectively.The relative contents of amino acids and their derivatives, ketone alcohols and other components in their extracellular metabolites were similar, but those of organic acids were remarkably different. After cultured for 15 d in the culture medium without exogenous organic carbon sources, the growth of H. hainangensis was slow, and the culture medium was slightly acidic, and the proportion of organic acids was 3.28%. However, the growth of H. roystonensis was normal, and the culture medium was alkaline, and the proportion of organic acids was 2.04%. When cultured in the media containing 0.5% sucrose for 15 d, the biomass of H. hainangensis increased from 0.09 g/L to 3.87 g/L; the proportion of organic acids increased to 4.64%; the contents of amines and alkaloids decreased; and the pH of culture medium decreased to 4.70. In the meantime, the biomass of H. roystonensis increased from 3.20 g/L to 5.24 g/L; the proportion of organic acids decreased to 1.13%; the contents of amines and alkaloids increased; and the pH of culture medium increased to 8.32. In conclusion, the change of pH in the culture media of the two species is opposite. The addition of organic carbon sources could significantly promote their growth and reproduction, promote the secretion of organic acids from H. hainangensis, inhibit the secretion of organic acids from H. roystonensis, thus enhance the degree of acidification or alkalization of the two cultures. The above results can provide a basis for the research on the evolution of Heveochlorella and the development and utilization of extracellular metabolites.

Key words: Heveochlorella hainanensis Heveochlorella roystonensis extracellular metabolites non-targeted metabolomics

收稿日期: 2020-09-22 出版日期: 2021-06-25

CLC:

Q 949.211

基金资助: 国家自然科学基金(31700195)

通讯作者: 刘平怀 E-mail: 601859770@qq.com;twlph@163.com

作者简介: 邓茹茹（https://orcid.org/0000-0003-0815-6686），E-mail：601859770@qq.com

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引用本文:

邓茹茹,马帅,谭德冠,付莉莉,张家明,孙雪飘,刘平怀. 2种内生和附生橡藻属绿藻的胞外分泌物研究[J]. 浙江大学学报（农业与生命科学版）, 2021, 47(3): 363-370.

Ruru DENG,Shuai MA,Deguan TAN,Lili FU,Jiaming ZHANG,Xuepiao SUN,Pinghuai LIU. Studies on extracellular metabolites of two endophytic and epiphytic algae of Heveochlorella spp. Journal of Zhejiang University (Agriculture and Life Sciences), 2021, 47(3): 363-370.

链接本文:

http://www.zjujournals.com/agr/CN/10.3785/j.issn.1008-9209.2020.09.221 或 http://www.zjujournals.com/agr/CN/Y2021/V47/I3/363

表1 培养15 d后海南橡藻与王棕橡藻生物量和培养液pH的比较

类型 Type	胞外分泌物种类 Species of extracellular metabolites	小计 Total
有机酸类 Organic acids	羧酸^、泛酸^、乙酰乙酸、N-羧乙基-γ-氨基丁酸、5-氨基乙酰丙酸、硅酸、3，4，5-三甲氧基肉桂酸、维生素A酸、11，14-反式二十碳二烯酸、甲基乙烯酸、2-氨基十六酸、5-[5-（甲氧羰基）-5，8a-二甲基-2-亚甲基十氢-1-萘基]-3-甲基戊酸	12种
核苷酸类 Nucleotides	5 $′$ -脱氧腺苷^*、香豆苷^#、甲基腺嘌呤、2 $′$ -O-甲基腺苷、旋毛虫苷、醋酸苷、鞘氨醇半乳糖苷、D-吡喃葡萄糖苷	8种
氨基酸及其衍生物 Amino acids and their derivatives	N-乙酰-dl-蛋氨酸^*、苯丙氨酸^#、缬氨酸-丙氨酸-甘氨酸-缬氨酸-丙氨酸^#、组氨酸-苏氨酸-谷氨酰胺^#、L-亮氨酸、苏氨酸-天冬氨酸、天冬氨酸-谷氨酸、苏氨酸-脯氨酸-甘氨酸、天冬酰胺-半胱氨酸-甘氨酸、缬氨酸-亮氨酸-丝氨酸、蛋氨酸-丙氨酸-谷氨酰胺、精氨酸-丝氨酸-丝氨酸、脯氨酸-天冬酰胺-赖氨酸-亮氨酸-谷氨酰胺-缬氨酸、丝氨酸-赖氨酸-赖氨酸、丝氨酸-半胱氨酸-精氨酸、脯氨酸-谷氨酰胺-赖氨酸、赖氨酸-谷氨酰胺-异亮氨酸、组氨酸-赖氨酸-蛋氨酸、HC毒素、精氨酸-亮氨酸-赖氨酸、丝氨酸-精氨酸-酪氨酸、丙氨酸-精氨酸-色氨酸、精氨酸-蛋氨酸-蛋氨酸等	35种
糖类 Sugars	脱氧核糖^*、α-D-葡萄糖	2种
酯类 Esters	2-二甲氨基-5，6-二甲基嘧啶-4-二甲基氨基甲酸酯、磷酸十四酯、二十二碳六烯酸乙酯、热塑性聚氨酯、13，14-二氢-15-酮基-前列腺素F2α-异丙酯、17-苯基三硝基-前列腺素F2α-异丙酯、16-苯氧基四极体-前列腺素F2α-异丙酯	7种
胺类及生物碱 Amines and alkaloids	氯苯吡胺^*、乙酰苯胺、异丁酸乙酰胺、1-脱氧野尻霉素、N-去甲基丙炔苯胺、β-D-葡萄糖胺、棕榈酰胺、N-甲基二辛胺、异丙甲草胺、硬脂胺、雪花胺、1-（4-氟苄基）哌嗪、丁哌卡因、苄丙酚胺、甲哌唑、丁胺、六氢芬宁、拉贝洛尔、邻花生四烯醇乙醇胺、花生酰乙醇酰胺、R-1-甲酰胺、二十二碳六烯醇-5-羟色胺	22种
酮醇类 Ketone alcohols	内酯二醇^#、苯乙醇、D-红-鞘氨醇、鞘氨醇、1-脱氢-12-姜酮、21-乙氧基孕烯醇酮、脱氢皮质（甾）醇、1-硬脂酰-2-花生四烯醇	8种
其他 Others	F-脒^、L-硫代瓜氨酸二盐酸^、右旋异黄烷^#、2-吡咯烷、草酸铵、1，2-二苯肼、N-N′-二环己基脲、苯海拉明、6-叔丁基-3-（4，5-二氢-1H-咪唑-2-基甲基）-2，4-二甲基苯酚、瓶型酵母A、他喷他多、羟甲唑啉、洛伐他汀等	47种

表2 海南橡藻和王棕橡藻胞外分泌物种类

表3 海南橡藻与王棕橡藻胞外分泌物各种类占比 (%)

表4 海南橡藻和王棕橡藻胞外分泌物中有机酸组分占总有机酸的比例 (%)

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