三株小球藻抗旱性能评价

doi:10.13560/j.cnki.biotech.bull.1985.2024-0101

摘要/Abstract

摘要：

【目的】研究沙漠小球藻（desert Chlorella）应对干旱胁迫的生理响应，分离培养抗旱能力强的小球藻，为沙漠微藻资源的开发和生产奠定基础。【方法】以2株分离自新疆塔克拉玛干沙漠和古尔班通古特沙漠中的小球藻和1株购买自中国科学院淡水藻种库普通淡水小球藻（Chlorella vulgaris）为材料，使用Bold’s Basal Medium（BBM）作为培养基，通过聚乙二醇（PEG6000）模拟不同程度干旱胁迫，测定3株小球藻在干旱胁迫下的生长速度和生理生化指标，并通过主成分分析、隶属函数、抗旱性综合度量值等方法评价3株小球藻抗旱能力。【结果】干旱胁迫能使小球藻细胞密度（OD₆₈₀）和生物量（干重）下降，叶绿素 a、叶绿素 b 含量降低，丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性上升，但是可溶性糖、蛋白质、脯氨酸（Pro）含量和过氧化氢酶（CAT）活性则因胁迫程度和时间的变化而不同，主成分分析显示糖、Pro、SOD、CAT、叶绿素b在小球藻抗干旱过程中发挥重要作用；根据抗旱系数分析，重度干旱时沙漠小球藻可溶性糖、蛋白质、Pro、SOD、CAT比普通时作用更强。【结论】综合评价显示3株小球藻的抗旱性强弱依次为：沙漠小球藻TLD 6B>沙漠小球藻GTD 8A1>普通小球藻FACHB-2338，且沙漠小球藻在生长速度、糖含量、蛋白质含量方面相比普通小球藻更具有优势。

关键词: 沙漠小球藻, 干旱胁迫, 生理指标, 隶属函数分析, 主成分分析, 抗旱性评价

Abstract:

【Objective】In order to study the physiological response of desert Chlorella to drought stress, we isolated and cultured Chlorella with strong drought resistance, laying a foundation for the development and production of desert microalgae resources.【Method】Two strains of Chlorella vulgaris isolated from Taklimakan Desert and Gurbantunggut Desert in Xinjiang and one strain of C. vulgaris purchased from Chinese Academy of Sciences Freshwater Algae Seed Bank were used as materials, and Bold's Basal Medium（BBM）as the culture medium. The growth rate and physiological and biochemical indicators of three strains of C. vulgaris under drought stress were measured by polyethylene glycol（PEG6000）to simulate different degrees of drought stress, and the drought resistance capacity of three strains of C. vulgaris was evaluated by principal component analysis, membership function, comprehensive drought resistance measurement value and other methods.【Result】 Under drought stress, the cell density（OD₆₈₀）and biomass（dry weight）decreased, chlorophyll a and chlorophyll b content decreased, while the malondialdehyde（MDA）content and superoxide dismutase（SOD）activity increased. However, soluble sugar, protein, proline（Pro）content, and catalase（CAT）activity varied with the degree and time of stress. The principal component analysis table showed that the sugars, Pro, SOD, CAT, and chlorophyll b played important roles in the drought resistance process of C. vulgaris. According to the analysis of drought resistance coefficient, the soluble sugars, proteins, Pro, SOD, and CAT of desert Chlorella had stronger effects than ordinary ones during severe drought.【Conclusion】The comprehensive evaluation shows that the drought resistance of the three strains of Chlorella is in the following order: Desert Chlorella TLD 6B > Desert Chlorella GTD 8A1 > Common Chlorella FACHB-2338, and desert Chlorella has advantages in growth rate, sugar content, and protein content compared to common Chlorella.

Key words: desert Chlorella, drought stress, physiological index, membership function analysis, principal component analysis, drought resistance evaluation

韩凯, 周永顺, 张凯月, 王路, 高剑峰, 陈福龙. 三株小球藻抗旱性能评价[J]. 生物技术通报, 2024, 40(8): 244-254.

HAN Kai, ZHOU Yong-shun, ZHANG Kai-yue, WANG Lu, GAO Jian-feng, CHEN Fu-long. Evaluation of Drought Resistance of Three Chlorella Strains[J]. Biotechnology Bulletin, 2024, 40(8): 244-254.

图/表 11

图1 干旱胁迫下小球藻细胞浓度、生物量 A和E，B和F，C、G，D和H分别为0%、5%、15%和25% PEG6000胁迫时小球藻生理指标；不同小写字母表示同一处理、同一时间不同小球藻间差异的显著性（P<0.05）；CV：FACHB-2338；下同

Fig. 1 Cell concentration and biomass of Chlorella vulgaris under drought stress A and E, B and F, C and G, D and H refers to physiological indexes of Chlorella vulgaris under 0%, 5%, 15% and 25% PEG6000 stress, respectively. Different lowercase letters indicate significant difference among different chlorella at the same treatment and at the same time（P < 0.05）;CV：FACHB-2338. The same below

表1 干旱胁迫下小球藻生物量体积产率

Table 1 Biomass volume yield of C. vulgaris under drought stress

PEG6000浓度 PEG6000 concentration/%	小球藻生物量体积产率 Biomass volume yield of C. vulgaris /（g·L^-1·d^-1）
PEG6000浓度 PEG6000 concentration/%	6B	8A1	FACHB-2338
0	0.91	0.87	0.82
5	0.82	0.78	0.73
15	0.55	0.51	0.35
25	0.22	0.16	0.08

图2 干旱胁迫下小球藻可溶性糖含量 A-D分别表示0%、5%、15%和25% PEG6000胁迫处理，下同

Fig. 2 Soluble sugar content of C. vulgaris under drought stress A-D indicate 0, 5%, 15% and 25% PEG6000 stress, respectively. The same below

图3 干旱胁迫下小球藻可溶性蛋白质含量

Fig. 3 Soluble protein content of C. vulgaris under drought stress

图4 干旱胁迫下小球藻脯氨酸含量

Fig. 4 Proline content of C. vulgaris under drought stress

图5 干旱胁迫下小球藻丙二醛含量

Fig. 5 Malondialdehyde content of C. vulgaris under drought stress

图6 干旱胁迫下小球藻过氧化氢酶、超氧化物歧化酶活性

Fig. 6 Activities of catalase and superoxide dismutase in C. vulgaris under drought stress

图7 干旱胁迫下小球藻叶绿素a、叶绿素b含量

Fig. 7 Chlorophyll a and chlorophyll b content in C. vulgaris under drought stress

表2 小球藻各生理指标的抗旱系数

Table 2 Drought resistance coefficients of each physiological index of C. vulgaris

小球藻种类 Chlorella species	OD₆₈₀	干重 Dry weight	可溶性糖 Soluble sugar	可溶性蛋白质 Soluble protein	Pro	MDA	CAT	SOD	叶绿素a Chlorophyll a	叶绿素b Chlorophyll b
6B	0.269	0.412	2.812	1.434	1.569	3.034	1.440	2.384	0.583	0.565
8A1	0.197	0.361	2.383	1.465	1.394	3.365	1.474	1.891	0.555	0.592
FACHB-2338	0.169	0.307	1.880	1.286	1.280	3.592	1.332	1.742	0.530	0.549
平均值Mean	0.212	0.36	2.358	1.359	1.414	3.330	1.415	2.005	0.558	0.569

表3 小球藻各生理性状的主成分分析

Table 3 Principal component analysis of physiological characters of C. vulgaris

指标 Index	PC1	PC2
OD₆₈₀	0.94	-0.34
干重Dry weight	0.997	-0.082
可溶性糖 Soluble sugar	0.999	-0.052
可溶性蛋白质 Soluble protein	0.832	0.555
脯氨酸Pro	0.976	-0.218
丙二醛MDA	-0.979	0.204
过氧化氢酶CAT	0.792	0.611
超氧化物歧化酶SOD	0.922	-0.388
叶绿素a Chlorophyll a	0.991	-0.13
叶绿素b Chlorophyll b	0.457	0.889
特征值 Eigenvalue	8.147	1.853
贡献率 Contribution rate/%	81.473	18.527
累计贡献率 Accumulated contribution rate/%	81.473	100

表4 小球藻抗旱性综合评价排序

Table 4 Comprehensive evaluation and ranking of drought resistance of C. vulgaris

小球藻种类 Material code	D 值 D value	排序 Rank
6B	0.866	1
8A1	0.772	2
FACHB-2338	-0.079	3

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