Screening， Identification and Preservation of Lactic Acid Bacteria Inhibiting the Growth of Rot-causing Fungus Lonicera caerulea L.

doi:10.13560/j.cnki.biotech.bull.1985.2025-0107

Abstract

Abstract:

Objective To screen and identify a strain of lactic acid bacteria that can inhibit rot-causing fungi in post-harvest Lonicera caerulea L., and to explore its effect on post-harvest preservation of L. caerulea L, this may provide new technical means to solve the problem of post-harvest rotting of L. caerulea L. Method After the isolation and purification of lactic acid bacteria from pickles and fermented berries, the P-2, which can inhibit the rot-causing fungi (Alternaria alternata and Rhizopus microsporus) in post-harvest L. caerulea L., were screened and identified by 16S rRNA gene sequence analysis. To evaluate the antibiotic sensitivity of P-2 and the inhibitory effect of cell-free supernatant (CFS) on rot-causing bacteria under different conditions. At last, P-2 CFS was used as a preservative to keep L. caerulea L fresh. Result Among the 22 strains of isolated and purified lactic acid bacteria, P-2 strain showed the best bacteriostatic effect and was identified as Pediococcus acidilactici. P-2 is resistant to kanamycin, gentamicin and streptomycin, sensitive or neutral to other antibiotics, its CFS has strong antifungal activity under acidic conditions, and is sensitive to protease. The MIC of P-2 CFS against A. alternata and R. microsporus were 12.8 and 25.6 mg/mL, respectively. Compared with the control group, the CFS of P-2 reduced the weight loss rate and decay rate of L. caerulea L, slowed down the decline of hardness, total phenol, flavonoid and vitamin C content of L. caerulea L during storage, and prolonged the shelf life of L. caerulea L. for at least 7 d. Conclusion The CFS of P. acidilactici P-2 can inhibit the post-harvest rot-causing bacteria of L. caerulea L., and can be used as a green preservative in the post-harvest preservation of berries.

Key words: lactic acid bacteria, Lonicera caerulea L., rot-causing fungi, antifungal activity, Alternaria alternata, Rhizopus microspora, screening, keep fresh

LI De-hai, YIN Li, ZHOU Cai-xue, WANG Ze-tong, SUN Chang-yan. Screening， Identification and Preservation of Lactic Acid Bacteria Inhibiting the Growth of Rot-causing Fungus Lonicera caerulea L.[J]. Biotechnology Bulletin, 2025, 41(8): 289-299.

Figures/Tables 10

References 38

[1]	Auzanneau N, Weber P, Kosińska-Cagnazzo A, et al. Bioactive compounds and antioxidant capacity of Lonicera caerulea berries: Comparison of seven cultivars over three harvesting years [J]. J Food Compos Anal, 2018, 66: 81-89.
[2]	Dong HY, Xu Y, Zhang QQ, et al. Activity and safety evaluation of natural preservatives [J]. Food Res Int, 2024, 190: 114548.
[3]	Fan LL, Lin L, Zhang Y, et al. Component characteristics and reactive oxygen species scavenging activity of anthocyanins from fruits of Lonicera caerulea L [J]. Food Chem, 2023, 403: 134391.
[4]	李山, 张彦龙, 曾伟民, 等. 蓝靛果果渣花色苷胶束的制备及其稳定性研究 [J]. 食品工业科技, 2024, 45(2): 21-29.
	Li S, Zhang YL, Zeng WM, et al. Study on preparation and stability of anthocyanin micelles from Lonicera edulis pomace [J]. Sci Technol Food Ind, 2024, 45(2): 21-29.
[5]	周静. 寒地浆果香飘四季 [N]. 黑龙江日报, 2024-10-15(5).
	Zhou J. The sweet flavor of berries in cold land [N]. Heilongjiang Daily, 2024-10-15(5).
[6]	张星, 毕金峰, 陈芹芹, 等. 蓝莓-蓝靛果复合冻干粉贮藏期品质及加工特性 [J]. 食品科学, 2022, 43(17): 240-247.
	Zhang X, Bi JF, Chen QQ, et al. Quality and processing characteristics of a mixed freeze-dried powder of blueberry and blue honeysuckle berries during storage [J]. Food Sci, 2022, 43(17): 240-247.
[7]	Chen J, Fu CL, Wang HY, et al. Combination transcriptomic and metabolomic reveal deterioration of the blue honeysuckle (Lonicera caerulea L.) fruit and candidate genes regulating metabolism in the post-harvest stage [J]. Int J Biol Macromol, 2025, 284: 138074.
[8]	李次力. 蓝靛果的壳聚糖涂膜保鲜研究 [J]. 食品科学, 2008, 29(7): 457-461.
	Li CL. Study on chitosan coating preservation of Lonicera edulis [J]. Food Sci, 2008, 29(7): 457-461.
[9]	赵倩, 张鹏, 贾晓昱, 等. 不同采收期对1-MCP处理后蓝靛果贮藏品质的影响 [J]. 包装工程, 2023, 44(11): 78-86.
	Zhao Q, Zhang P, Jia XY, et al. Effects of different harvest periods on the storage quality of Lonicera caerulea L. after 1-MCP treatment [J]. Packag Eng, 2023, 44(11): 78-86.
[10]	Adithi G, Divyashree S, Shruthi B, et al. Evaluation of Limosilactobacillus fermentum MYSAGAM1 isolated from herbal Amla juice as a probiotic candidate with antifungal characteristics against Fusarium equiseti [J]. Food Biosci, 2024, 58: 103843.
[11]	陈思宇, 金建, 赵世琳. 物理技术在果蔬保鲜中的应用研究进展 [J]. 食品研究与开发, 2023, 44(21): 167-172.
	Chen SY, Jin J, Zhao SL. Progress in application of physical technology in preservation of fruits and vegetables [J]. Food Res Dev, 2023, 44(21): 167-172.
[12]	李江阔, 高静, 张鹏, 等. 微环境气调对蓝果忍冬贮藏品质和抗氧化酶的影响 [J]. 食品与发酵工业, 2021, 47(6): 152-159.
	Li JK, Gao J, Zhang P, et al. Micro-environmental modified atmosphere on storage quality and antioxidant enzymes of blue honeysuckle fruits [J]. Food Ferment Ind, 2021, 47(6): 152-159.
[13]	Deng XZ, Wei YY, Jiang S, et al. Recent advances in the application of tea tree oil in the storage of fruit and vegetables [J]. Postharvest Biol Technol, 2025, 219: 113260.
[14]	Abouloifa H, Gaamouche S, Ghabbour N, et al. Lactic acid bacteria from Moroccan traditional foods: Techno-functional, health-promoting, nutraceutical value and application as a starter and bio-preservative agent in the food products [J]. Bioresour Technol Rep, 2024, 27: 101941.
[15]	Hernández Figueroa RH, López-Malo A, Mani-López E. Antimicrobial activity and applications of fermentates from lactic acid bacteria-a review [J]. Sustainable Food Technol, 2024, 2(2): 292-306.
[16]	Wang Q, Zhang F, Zhang YH, et al. A novel strain Lactiplantibacillus plantarum LPP95 isolated from Chinese pickles: Antifungal effect, mechanism, and potential application in yogurt [J]. Food Biosci, 2024, 58: 103640.
[17]	Wang L, Jiang SS, Zhou CX, et al. Exploring novel preservation strategies for blue honeysuckle through high-throughput sequencing and bioinformatics analysis [J]. Postharvest Biol Technol, 2025, 219: 113251.
[18]	Jiang SS, Wang L, Li DH, et al. The purification of dominant spoilage fungi on Lonicera Caeruleum and the inhibitory effects of composite essential oils against these fungi [J]. Food Biosci, 2023, 53: 102839.
[19]	Souza LV, Rodrigues da Silva R, Falqueto A, et al. Evaluation of antifungal activity of lactic acid bacteria against fungi in simulated cheese matrix [J]. LWT, 2023, 182: 114773.
[20]	Riolo M, Luz C, Santilli E, et al. Antifungal activity of selected lactic acid bacteria from olive drupes [J]. Food Biosci, 2023, 52: 102422.
[21]	de Vasconcelos Medeiros GKV, Martins ACS, Vasconcelos MG, et al. Cereus jamacaru DC. (mandacaru) fruit as a source of lactic acid bacteria with in vitro probiotic-related characteristics and its protective effects on Pediococcus pentosaceus during lyophilization and refrigeration storage [J]. Int J Food Microbiol, 2024, 417: 110695.
[22]	訾静, 王琰, 李亮亮, 等. 林麝肠道中乳酸菌的分离筛选及益生特性 [J]. 食品科学, 2024, 45(8): 79-86.
	Zi J, Wang Y, Li LL, et al. Isolation and screening of lactic acid bacteria from the gut of forest musk deer for probiotic properties [J]. Food Sci, 2024, 45(8): 79-86.
[23]	王晓宇, 吴梦娜, 于巧如, 等. 植物乳杆菌ST3.5的分离鉴定及其对霉菌的抑制作用 [J]. 食品工业科技, 2023, 44(13): 141-149.
	Wang XY, Wu MN, Yu QR, et al. Isolation and identification of lactiplantibacillus plantarum ST3.5 and its inhibitory effect on mold [J]. Sci Technol Food Ind, 2023, 44(13): 141-149.
[24]	Ma MG, Li A, Feng J, et al. Antifungal mechanism of Lactiplantibacillus plantarum P10 against Aspergillus niger and its in situ biopreservative application in Chinese steamed bread [J]. Food Chem, 2024, 449: 139181.
[25]	Shu C, Sun XX, Cao JK, et al. Antifungal efficiency and mechanisms of ethyl ferulate against postharvest pathogens [J]. Int J Food Microbiol, 2024, 417: 110710.
[26]	刘艺璇, 岳红, 司欣雨, 等. 拮抗菌复合保鲜纸的制备及其对苹果保鲜中的应用 [J]. 食品工业科技, 2025, 46(6): 342-351.
	Liu YX, Yue H, Si XY, et al. Preparation of composite preservation paper containing antagonistic fungi and its application on apple preservation [J]. Sci Technol Food Ind, 2025, 46(6): 342-351.
[27]	王成炜, 袁宇尧, 张政, 等. 基于计算流体动力学的二氧化硫压差熏蒸模型在鲜食葡萄保鲜中的应用研究 [J]. 食品与发酵工业, 2025, 51(2): 118-125.
	Wang CW, Yuan YY, Zhang Z, et al. Application research of sulfur dioxide differential pressure fumigation model based on computational fluid dynamics in table grape preservation [J]. Food Ferment Ind, 2025, 51(2): 118-125.
[28]	曹建康, 姜微波, 赵玉梅. 果蔬采后生理生化实验指导 [M]. 北京: 中国轻工业出版社, 2007: 176.
	Cao JK, Jiang WB, Zhao YM. Guidance on postharvest physiological and biochemical experiments of fruits and vegetables [M]. Beijing: China Light Industry Press, 2007: 176.
[29]	李军. 钼蓝比色法测定还原型维生素C [J]. 食品科学, 2000, 21(8): 42-45.
	Li J. Study on molybdenum blue method of L-VC test by spectrometry [J]. Food Sci, 2000, 21(8): 42-45.
[30]	史洁莹, 牟燕萍, 陆周欣, 等. 壳聚糖-植物精油纳米乳液保鲜垫的制备及其在蓝莓保鲜中的应用 [J/OL]. 食品工业科技, 2024. .
	Shi JY, Mou YP, Lu ZX, et al. Preparation and application of chitosan-essential oil nanoemulsion preservation pads for blueberry preservation [J/OL]. Sci Technol Food Ind, 2024. .
[31]	Sharma AK, Sati DM, Murti Y, et al. A comprehensive review on Chinese honeysuckle (Qusqualis indica): a Traditional Chinese plant [J]. Toxicol Rep, 2024, 13: 101768.
[32]	Islam S, Biswas S, Jabin T, et al. Probiotic potential of Lactobacillus plantarum DMR14 for preserving and extending shelf life of fruits and fruit juice [J]. Heliyon, 2023, 9(6): e17382.
[33]	Liang NY, Zhao Z, Curtis JM, et al. Antifungal cultures and metabolites of lactic acid bacteria for use in dairy fermentations [J]. Int J Food Microbiol, 2022, 383: 109938.
[34]	Liu AP, Xu RX, Zhang S, et al. Antifungal mechanisms and application of lactic acid bacteria in bakery products: a review [J]. Front Microbiol, 2022, 13: 924398.
[35]	Abouloifa H, Rokni Y, Hasnaoui I, et al. Characterization of antimicrobial compounds obtained from the potential probiotic Lactiplantibacillus plantarum S61 and their application as a biopreservative agent [J]. Braz J Microbiol, 2022, 53(3): 1501-1513.
[36]	Chen X, Wei ZR, Feng ZQ, et al. Large-scale fermentation of Lactiplantibacillus pentosus 292 for the production of lactic acid and the storage strategy based on molasses as a preservative [J]. BMC Microbiol, 2025, 25(1): 125.
[37]	Li NY, Cheng YF, Li Z, et al. An alginate-based edible coating containing lactic acid bacteria extends the shelf life of fresh strawberry (Fragaria × Ananassa Duch.) [J]. Int J Biol Macromol, 2024, 274: 133273.
[38]	王璐. 采后蓝靛果菌群分析及乳酸菌无细胞上清液对其致腐菌抑制作用 [D]. 哈尔滨: 东北林业大学, 2024.
	Wang L. Analysis of the flora of postharvest Lonicera edulis and the inhibitory effect of cell-free supernatant of lactic acid bacteria on its rot-causing bacteria [D]. Harbin: Northeast Forestry University, 2024.

乳酸菌 Lactic acid bacteria	链格孢霉 A. alternata	小孢根霉 R. microspora
P-1	-	-
P-2	+++	++
P-3	-	-
P-4	++	++
P-5	++	+++
P-6	+++	-
P-7	-	+
P-8	-	-
P-9	++	+
P-10	+	-
P-11	+	-
J-12	+++	++
J-13	++	+++
J-14	-	-
J-15	-	++
J-16	++	-
J-17	+	-
J-18	-	-
J-19	++	++
J-20	+	-
J-21	++	++
J-22	-	-

乳酸菌 Lactic acid bacteria	链格孢霉 A. alternata	小孢根霉 R. microspora
P-1	-	-
P-2	+++	++
P-3	-	-
P-4	++	++
P-5	++	+++
P-6	+++	-
P-7	-	+
P-8	-	-
P-9	++	+
P-10	+	-
P-11	+	-
J-12	+++	++
J-13	++	+++
J-14	-	-
J-15	-	++
J-16	++	-
J-17	+	-
J-18	-	-
J-19	++	++
J-20	+	-
J-21	++	++
J-22	-	-

抗生素 Antibiotic	抑菌直径 Inhibitory diameter （mm）			P-2抑菌直径 P-2 inhibitory diameter （mm）	结果 Result
抗生素 Antibiotic	敏感 Susceptible （S）	中介 Intermediary （I）	耐受 Resistance （R）	P-2抑菌直径 P-2 inhibitory diameter （mm）	结果 Result
四环素 Tetracycline	≥19	15-18	≤14	24.43±1.39^e	S
氯霉素 Chloramphenicol	≥18	13-17	≤12	40.83±0.67^b	S
氨苄西林 Ampicillin	≥17	14-16	≤13	32.97±0.73^c	S
万古霉素 Vancomycin	≥17	15-16	≤14	15.50±0.12^f	I
庆大霉素 Gentamicin	≥15	13-14	≤12	0.00±0.00^g	R
卡那霉素 Kanamycin	≥18	14-17	≤13	0.00±0.00^g	R
链霉素 Streptomycin	≥15	12-14	≤11	0.00±0.00^g	R
红霉素 Erythromycin	≥23	14-22	≤13	29.50±0.70^d	S
克林霉素 Clindamycin	≥21	15-20	≤14	42.97±0.58^a	S

抗生素 Antibiotic	抑菌直径 Inhibitory diameter （mm）			P-2抑菌直径 P-2 inhibitory diameter （mm）	结果 Result
抗生素 Antibiotic	敏感 Susceptible （S）	中介 Intermediary （I）	耐受 Resistance （R）	P-2抑菌直径 P-2 inhibitory diameter （mm）	结果 Result
四环素 Tetracycline	≥19	15-18	≤14	24.43±1.39^e	S
氯霉素 Chloramphenicol	≥18	13-17	≤12	40.83±0.67^b	S
氨苄西林 Ampicillin	≥17	14-16	≤13	32.97±0.73^c	S
万古霉素 Vancomycin	≥17	15-16	≤14	15.50±0.12^f	I
庆大霉素 Gentamicin	≥15	13-14	≤12	0.00±0.00^g	R
卡那霉素 Kanamycin	≥18	14-17	≤13	0.00±0.00^g	R
链霉素 Streptomycin	≥15	12-14	≤11	0.00±0.00^g	R
红霉素 Erythromycin	≥23	14-22	≤13	29.50±0.70^d	S
克林霉素 Clindamycin	≥21	15-20	≤14	42.97±0.58^a	S

处理组 Processing group	时间 Time （d）	外观 Appearance （Scores）	质地 Texture （Scores）	味道 Taste （Scores）	整体可接受性 Overall acceptability （Scores）
CK	1	4.20±0.00^a	4.80±0.45^a	4.60±0.55^a	5.00±0.00^a
	7	4.00±0.00^a	1.80±0.84^b	1.60±0.55^b	2.20±0.45^b
	14	-	1.20±0.45^b	-	-
	21	-	0.20±0.45^c	-	-
	28	-	-	-	-
1/2 MIC	1	4.60±0.55^a	5.00±0.00^a	4.80±0.45^a	4.60±0.55^a
	7	4.20±0.84^a	4.00±0.71^b	3.20±1.64^a	4.40±0.89^a
	14	3.20±1.30^ab	2.40±0.55^c	1.20±0.84^b	2.80±0.84^b
	21	1.60±1.14^bc	0.80±0.45^d	0.20±0.45^b	1.20±0.84^c
	28	0.2±0.45^c	0.20±0.45^d	-	0.40±0.89^c
MIC	1	4.60±0.55^a	4.80±0.45^a	4.60±0.55^a	4.80±0.45^a
	7	4.20±0.45^ab	3.60±0.55^a	3.20±0.45^b	3.80±0.45^a
	14	3.40±0.55^b	2.00±1.00^b	1.20±0.84^c	2.40±0.55^b
	21	2.20±0.84^c	1.40±1.14^b	0.40±0.55^cd	1.00±1.00^c
	28	0.40±0.55^d	0.60±0.55^b	-	0.20±0.45^c
2 MIC	1	4.60±0.55^a	4.8±0.45^a	4.60±0.55^a	5.00±0^a
	7	4.40±0.55^a	4.00±0.71^a	3.80±0.45^a	4.20±0.45^a
	14	3.60±0.55^a	2.00±1.00^b	1.80±0.84^b	2.80±0.45^b
	21	2.20±0.84^b	1.40±1.14^b	1.00±0.71^bc	1.40±0.89^c
	28	0.60±0.5^c	0.80±0.45^b	-	1.00±0.71^c