糙皮侧耳乳酸脱氢酶对低温光照下菌丝生长的调控作用

doi:10.16420/j.issn.0513-353x.2023-0955

园艺学报 ›› 2024, Vol. 51 ›› Issue (12): 2886-2894.doi: 10.16420/j.issn.0513-353x.2023-0955

糙皮侧耳乳酸脱氢酶对低温光照下菌丝生长的调控作用

吴柏增¹^,², 降玉清¹, 何琪¹^,³, 黄晨阳¹, 张利姣¹, 赵梦然¹^,^*()

¹中国农业科学院农业资源与农业区划研究所，农业农村部农业微生物资源收集与保藏重点实验室，北方干旱半干旱耕地高效利用全国重点实验室，北京 100081
²吉林农业大学园艺学院，长春 130118
³吉林农业大学食药用菌教育部工程研究中心，长春 130118

收稿日期:2024-09-10 修回日期:2024-11-04 出版日期:2024-12-25 发布日期:2024-12-13
通讯作者:
^* E-mail：zhaomengran@caas.cn
基金资助:
国家自然科学基金青年基金项目(32002110); 国家自然科学基金青年基金项目(32302638); 现代农业产业技术体系建设专项资助(CARS-20)

Study on the Regulation of Lactate Dehydrogenase on Mycelia Growth of Pleurotus ostreatus Under Low Temperature and Light

WU Baizeng¹^,², JIANG Yuqing¹, HE Qi¹^,³, HUANG Chenyang¹, ZHANG Lijiao¹, ZHAO Mengran¹()

¹National Key Laboratory for Efficient Utilization of Arid and Semi-Arid Cultivated Land in North China，Key Laboratory of Microbial Resources Collection and Preservation，Ministry of Agriculture and Rural Affairs，Institute of Agricultural Resources and Regional Planning，Chinese Academy of Agricultural Sciences，Beijing 100081，China
²College of Horticulture，Jilin Agriculture University，Changchun 130118，China
³Engineering Research Center of Chinese Ministry of Education for Edible and Medicinal Fungi，Jilin Agricultural University，Changchun 130118，China

Received:2024-09-10 Revised:2024-11-04 Published:2024-12-25 Online:2024-12-13
Contact:
^* E-mail：zhaomengran@caas.cn

摘要/Abstract

摘要：

为探索乳酸脱氢酶（lactate dehydrogenase，LDH）在低温光照刺激下对糙皮侧耳（Pleurotus ostreatus）菌丝生长的影响，检测了低温光照下乳酸脱氢酶的表达特征，观测了外源添加乳酸脱氢酶抑制剂、乳酸、丙酮酸对菌丝生长的影响。结果表明，乳酸脱氢酶对15 ℃、光照12 h/黑暗12 h条件下的菌丝生长具有促进作用。进一步研究发现，抑制乳酸脱氢酶活性会导致丙酮酸脱氢酶（pyruvate dehydrogenase，PDH）活性下降15.02%，进入三羧酸循环的丙酮酸通量减少，ATP产生量降低14.78%，胞内乳酸和丙酮酸含量分别增加18.97%和12.57%；乳酸和丙酮酸的积累能够诱导乙醇脱氢酶（alcohol dehydrogenase，ADH）活性升高，胞质NAD⁺含量增加，NADH/NAD⁺比例失衡，从而抑制菌丝生长。

关键词: 糙皮侧耳, 乳酸脱氢酶, 乳酸, 糖代谢, 低温, 光照

Abstract:

In order to study the effect of lactate dehydrogenase on mycelial growth of Pleurotus ostreatus under the environmental stimulation of low temperature and light，the expression profiles of lactate dehydrogenase were analyzed，and the influences of exogenous lactate dehydrogenase inhibitor，lactic acid，and pyruvic acid on the mycelial growth were observed. The results showed that lactate dehydrogenase could accelerate mycelial growth in 12 h of light alternated with 12 h of darkness at 15 ℃. Inhibition of lactate dehydrogenase enzyme activity would cause the activity of pyruvate dehydrogenase （PDH）to be decreased by 15.02%，which giving rise to the decreasing flux of pyruvic acid into the tricarboxylic acid cycle，the production of ATP reduced by 14.78%，and the intracellular contents of lactic acid and pyruvic acid increased by 18.97% and 12.57%，respectively. The accumulation of lactic acid and pyruvic acid could induce increased activities of alcohol dehydrogenase，which leading to the increase of NAD⁺ content and the imbalance of NADH/ NAD⁺ ratio，and therefore the inhibition of mycelial growth.

Key words: Pleurotus ostreatus, lactate dehydrogenase, lactate, glycometabolism, low temperature, light

吴柏增, 降玉清, 何琪, 黄晨阳, 张利姣, 赵梦然. 糙皮侧耳乳酸脱氢酶对低温光照下菌丝生长的调控作用[J]. 园艺学报, 2024, 51(12): 2886-2894.

WU Baizeng, JIANG Yuqing, HE Qi, HUANG Chenyang, ZHANG Lijiao, ZHAO Mengran. Study on the Regulation of Lactate Dehydrogenase on Mycelia Growth of Pleurotus ostreatus Under Low Temperature and Light[J]. Acta Horticulturae Sinica, 2024, 51(12): 2886-2894.

图/表 6

表1 荧光定量PCR引物

Table 1 Primers for qPCR

引物 Primer	正向序列（5′-3′） Forward sequence	反向序列（5′-3′） Reverse sequence
qPCR-LDH1	GGTCCTGCTTTGTCTCGTAG	GGGAAAGGATGGAGTACAGATG
qPCR-LDH2	AGAGGAATTGGAAAGACATGGG	ATACGGAATGACGGGCATAAG
qPCR-LDH3	CTTGGGAGATGTGATACTGGG	GTTCTCGGTTTGAATTCGCTG
qPCR-LDH4	GCGATTAGTGGCAAGGTTTAC	CATCCAACATGAAATCGGCG
qPCR-LDH5	CACCCGCTCTACCAATTCTG	GCCATTCCTTTACGCTCAATC
qPCR-LDH6	TGCTTCTACCCGTTTCTGTC	ATATGACCCAATACATCCACCG
qPCR-LDH7	TGAACTTGCGTCTACTTGGTG	CAGGCGGAGAGGTTGATATG
qPCR-LDH8	GGCGGAGGTGTCAAGATTC	ATGGCGGAACGAAGGATATG

图1 低温光照刺激下糙皮侧耳菌落表型（A）和胞内ATP含量（B）的变化 Dark：24 h黑暗；Light：光照12 h /黑暗12 h。不同小写字母表示使用邓肯氏分析方法处理间差异显著（P < 0.05）。下同。

Fig. 1 Colonial phenotype（A）and intracellular ATP content（B）of Pleurotus ostreatus under the stimulation of low temperature and light Dark：24 h in dark；Light：12 h in light /12 h in dark. Different lowercase letters indicated significant difference among treatments based on Duncan’s method（P < 0.05）. The same below.

图2 低温光照刺激下糙皮侧耳菌丝中乳酸脱氢酶（LDH）基因表达量（A）和酶活性（B）

Fig. 2 Gene expression and enzyme activity of LDH in mycelia of Pleurotus ostreatus under stimulation of low temperature and light

图3 乳酸脱氢酶抑制剂Galloflavin对28 ℃黑暗培养6 d的糙皮侧耳菌丝中乳酸脱氢酶活性、菌丝生长速率和菌落形态的影响

Fig. 3 Effects of exogenous LDH inhibitors on LDH activity，mycelial growth rate and colony morphology of Pleurotus ostreatus at 28 ℃ in dark for six days

图4 乳酸脱氢酶抑制剂Galloflavin对28 ℃黑暗培养6 d的糙皮侧耳菌丝中乳酸、丙酮酸等相关指标的影响 t 检验，*代表P < 0.05，**代表P < 0.01，***代表P < 0.001，ns代表无显著差异。

Fig. 4 Effects of exogenous LDH inhibitors on LA，PA and other related indexes in mycelia of Pleurotus ostreatus at 28 ℃ in dark for six days Student’s t test，* for P < 0.05，** for P < 0.01，*** for P < 0.001，ns for no significant difference.

图5 外源丙酮酸（PA）和乳酸（LA）对28 ℃黑暗培养6 d的糙皮侧耳菌落直径和乙醇脱氢酶活性的影响

Fig. 5 The effects of exogenous PA and LA on mycelial growth rate and ADH enzyme activity of Pleurotus ostreatus at 28 ℃ in dark for six days

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糙皮侧耳乳酸脱氢酶对低温光照下菌丝生长的调控作用

Study on the Regulation of Lactate Dehydrogenase on Mycelia Growth of Pleurotus ostreatus Under Low Temperature and Light

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糙皮侧耳乳酸脱氢酶对低温光照下菌丝生长的调控作用

Study on the Regulation of Lactate Dehydrogenase on Mycelia Growth of Pleurotus ostreatus Under Low Temperature and Light

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