Isolation and identification of endophytic fungi from Cyclocarya paliurus (Batal.) Iljinskaja

doi:10.3969/j.issn.1000-2006.201812001

JOURNAL OF NANJING FORESTRY UNIVERSITY ›› 2020, Vol. 44 ›› Issue (2): 26-34.doi: 10.3969/j.issn.1000-2006.201812001

Special Issue: 青钱柳叶有效成分专题

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Isolation and identification of endophytic fungi from Cyclocarya paliurus (Batal.) Iljinskaja

KOU Xiaolin¹(), XIE Nan¹, WU Caie¹^,²^,^*(), FAN Gongjian¹, FU Xiangxiang²

1. College of Light Industry and Food Engineering, Nanjing Forestry University, Nanjing 210037, China
2. Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing 210037, China

Received:2018-12-03 Revised:2019-11-19 Online:2020-03-30 Published:2020-04-01
Contact: WU Caie E-mail:391153013@qq.com;wucaie@njfu.edu.cn

Abstract

Abstract:

【Objective】This study aimed to identify the endophytic fungi produced flavonoids in Cyclocarya paliurus (Batal.) Iljinskaja and explore a new method to extract flavonoids. 【Method】The endophytic fungi in the branches, leaves, roots, and bark of C. paliurus (Batal.) Iljinskaja were isolated using the tissue separation method. By observing the morphological characteristics of the endophytes, the fermentation products of the endophytic fungi from C. paliurus (Batal.) Iljinskaja were identified using the chromogenic reaction method, thin-layer chromatography (TLC) and high performance liquid chromatography (HPLC). The endophytic fungi produced flavonoids were screened, and the total flavonoid production was determined. Moreover, the strains were identified based on the morphological characteristics and the phylogenetic analysis of the intervening sequences of the fungal rDNA. 【Result】In total, 67 strains of endophytic fungi were obtained; through the identification of morphological characteristics, they were identified to belong to three classes, four orders, six families and ten genera. Four endophytic fungi producing flavonoids were finally identified: PZ06, CY12, CP06 and PP03. The most abundant flavonoids, with a yield of 3.61 mg/L, were produced by strain PZ06. Molecular identification combined with morphological identification indicated that PZ06, CP06 and PP03 belong to the genusAlternaria, whereas CY12 belongs to the genus Eupenicillium. The dominant flavonoid-producing endophytic fungi of C. paliurus (Batal.) Iljinskaja belong to the genus Alternaria. 【Conclusion】 The study of endophytic fungi in C. paliurus (Batal.) Iljinskaja is a key significance for the development and utilization of endophytes. In addition to identifying the endophytic fungi, the present study also provides a novel method for the extraction of active substances fromC. paliurus (Batal.) Iljinskaja plants.

Key words: Cyclocarya paliurus(Batal.) Iljinskaja, flavonoids, endophytic fungi, ITS analysis, scanning electron microscope, isolation, identification

CLC Number:

S718.81

KOU Xiaolin, XIE Nan, WU Caie, FAN Gongjian, FU Xiangxiang. Isolation and identification of endophytic fungi from Cyclocarya paliurus (Batal.) Iljinskaja[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY, 2020, 44(2): 26-34.

Figures/Tables 7

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纲classis	目order	科family	属genus	总量/株 total strain	分离频率/% separation frequency
			木霉属Trichoderma	13	19.4
			青霉属Penicillium	9	1.9
		丛梗孢科Moniliacea	拟青霉属Paecilomyces	4	6.0
			地霉属Geotrichum	2	4.5
半知菌纲Deuteromycetes	丛梗孢目Moniliales		链格孢属Alternaria	16	23.9
		束梗孢科Stilbellaceae	棒束梗霉属Stilbaceae	1	1.5
		瘤座孢科Tuberculariaceae	镰刀菌属Fusarium	10	14.9
	黑盘孢目Melanconiales	黑盘孢科Melanconiaceae	刺盘孢属Colletotrichum	6	8.9
丝孢菌纲Hyphomycetes	无孢菌目Agonomycetales	无孢菌科Agaricaceae	丝核菌属Rhizoctonia	1	1.5
结合菌纲Class Zygomycetes	毛霉目Mucorale	毛霉科Mucoraceae	根霉属Rhizoupus sp.	5	7.5

菌株 bacterial strain	异槲皮苷含量/ (μg·μL^-1) soquercitroside	槲皮素含量/ (μg·μL^-1) quercetin	山柰酚含量/ (μg·μL^-1) kaempferol	总黄酮含量/ (mg·L^-1) flavonoids of sample
CY12	0.016±0.003	0.021±0.006	0.076±0.054	1.42
PZ06	0.038±0.009	0.060±0.011	0.190±0.081	3.61
PP03	0.220±0.029	0	0	2.77
CP06	0.014±0.009	0.031±0.003	0	0.57

Isolation and identification of endophytic fungi from Cyclocarya paliurus (Batal.) Iljinskaja

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