贝莱斯芽孢杆菌HY19挥发物对采后柑橘的防腐效果

doi:10.16420/j.issn.0513-353x.2023-0265

摘要/Abstract

摘要：

为评估贝莱斯芽孢杆菌（Bacillus velezensis）HY19菌株挥发性有机化合物（Volatile organic compounds）作为柑橘果实防腐剂的潜力，通过对峙和二分格平板培养实验研究其对柑橘果实主要致病菌意大利青霉（Penicillium italicum）和指状青霉（P. digitatum）的抑制作用。通过气相色谱—质谱联用技术对其产生的挥发性有机化合物中的抗真菌活性物质进行定性和定量分析，并通过人工接种及常温贮藏试验研究该挥发物对采后柑橘果实病害的防治效果。结果表明，菌株HY19及其释放的挥发性有机化合物显著抑制意大利青霉和指状青霉的菌丝生长，在该挥发性有机化合物中发现2-十三酮、邻苯二甲酸二丁酯、1-壬醇、苯甲酸等18种抗真菌物质。该挥发性有机化合物诱导柑橘果皮防御酶（SOD和POD）活性，降低果皮细胞膜透性和丙二醛含量，减轻病原菌对果皮的伤害，提高果皮抗病性，对青霉和绿霉的防治效果分别达到59.65%和64.11%。柑橘果实常温贮藏15和30 d后，该挥发性有机化合物显著降低果实的腐烂率，防腐效果分别为57.04%（15 d）和58.07%（30 d），且对果实品质无影响。综上，贝莱斯芽孢杆菌HY19具有开发为采后柑橘果实防腐剂的潜力。

关键词: 柑橘, 贝莱斯芽孢杆菌, 挥发性有机化合物, 气相色谱—质谱联用, 生物防腐剂

Abstract:

To evaluate the potential of Bacillus velezensis HY19 volatile organic compounds（VOCs）as a citrus fruit preservative，the suppression of HY19 and its VOCs on the main pathogens（Penicillium italicum and P. digitatum）of postharvest diseases of citrus fruit was studied through culture experiments in confrontation and bipartite plate. Qualitative and quantitative analysis of antifungal active substances in VOCs was conducted with gas chromatography-mass spectrometry（GC-MS）. And the control efficiency of VOCs on post-harvest diseases of citrus fruit was studied through artificial inoculation tests and storage experiments at room temperature. The results showed that strain HY19 and its VOCs significantly inhibited the mycelial growth of P. italicum and P. digitatum. Eighteen antifungal substances were found in the VOCs，including 2-tridecanone，dibutyl phthalate，1-nonanol，benzoic acid，etc. The VOCs induced the activities of defense enzymes（SOD and POD）in citrus fruit peels，decreased the cell membrane permeability and malondialdehyde content，reduced the damage of pathogens to the peels，and improved the resistance of peels to diseases. The control efficacies of the VOCs against blue mold and green mold reached 59.65% and 64.11%，respectively. After 15 and 30 d of citrus fruit storage at room temperature，the VOCs considerably reduced the decay rate of citrus fruit，with the preservative efficacies of 57.04%（15 d）and 58.07%（30 d），respectively，and had no influence on fruit quality. In summary，B. velezensis HY19 had the potential to be developed as a preservative for postharvest citrus fruit.

Key words: citrus, Bacillus velezensis, volatile organic compounds, gas chromatography-mass spectrometry, bio-preservatives

李素平, 李微, 韩冷, 黄建国. 贝莱斯芽孢杆菌HY19挥发物对采后柑橘的防腐效果[J]. 园艺学报, 2024, 51(1): 162-174.

LI Suping, LI Wei, HAN Leng, HUANG Jianguo. Bacillus velezensis HY19 Volatile Organic Compounds as a Preservative in Postharvest Citrus Fruit Management[J]. Acta Horticulturae Sinica, 2024, 51(1): 162-174.

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化合物 Substances	分子式 Formula	相对含量/% Relative content	化合物 Substances	分子式 Formula	相对含量/% Relative content
1-壬醇1-Nonanol	C₉H₂OO	6.33 ± 0.71	2-癸酮2-Decanone	C₁₀H₂OO	2.07 ± 0.32
2-壬醇2-Nonanol	C₉H₂OO	5.17 ± 0.51	二十烷Eicosane	C₂₀H₄₂	4.12 ± 0.33
1-辛醇1-Octanol	C₈H₁₈O	5.04 ± 0.40	十九烷Nonadecane	C₂₀H₄₂	2.93 ± 0.14
d-薄荷醇d-Menthol	C₁₀H₂OO	3.64 ± 0.73	十六烷Hexadecane	C₁₆H₃₄	1.08 ± 0.19
香叶醇Geraniol	C₁₀H₁₈O	1.95 ± 0.21	甲酯Methyl ester	C₁₀H₁₀O₂	2.51 ± 0.45
正十五醇n-Pentadecanol	C₁₅H₃₂O	1.78 ± 0.09	邻苯二甲酸二丁酯 Dibutyl phthalate	C₁₆H₂₂O₄	2.31 ± 0.34
2-十三酮2-Tridecanone	C₁₃H₂₆O	9.57 ± 0.73	苯甲酸Benzoic acid	C₁₅H₂₂O₂	3.48 ± 0.78
2-壬酮2-Nonanone	C₉H₁₈O	7.77 ± 0.47	甲酸Formic acid	C₁₂H₂₄O₂	2.22 ± 0.19
2-十一酮2-Undecanone	C₁₁H₂₂O	5.11 ± 0.96	2,4-二叔丁基苯酚 2,4-Di-tert-butylphenol	C₁₄H₂₂O	5.52 ± 0.52

化合物 Substances	分子式 Formula	相对含量/% Relative content	化合物 Substances	分子式 Formula	相对含量/% Relative content
1-壬醇1-Nonanol	C₉H₂OO	6.33 ± 0.71	2-癸酮2-Decanone	C₁₀H₂OO	2.07 ± 0.32
2-壬醇2-Nonanol	C₉H₂OO	5.17 ± 0.51	二十烷Eicosane	C₂₀H₄₂	4.12 ± 0.33
1-辛醇1-Octanol	C₈H₁₈O	5.04 ± 0.40	十九烷Nonadecane	C₂₀H₄₂	2.93 ± 0.14
d-薄荷醇d-Menthol	C₁₀H₂OO	3.64 ± 0.73	十六烷Hexadecane	C₁₆H₃₄	1.08 ± 0.19
香叶醇Geraniol	C₁₀H₁₈O	1.95 ± 0.21	甲酯Methyl ester	C₁₀H₁₀O₂	2.51 ± 0.45
正十五醇n-Pentadecanol	C₁₅H₃₂O	1.78 ± 0.09	邻苯二甲酸二丁酯 Dibutyl phthalate	C₁₆H₂₂O₄	2.31 ± 0.34
2-十三酮2-Tridecanone	C₁₃H₂₆O	9.57 ± 0.73	苯甲酸Benzoic acid	C₁₅H₂₂O₂	3.48 ± 0.78
2-壬酮2-Nonanone	C₉H₁₈O	7.77 ± 0.47	甲酸Formic acid	C₁₂H₂₄O₂	2.22 ± 0.19
2-十一酮2-Undecanone	C₁₁H₂₂O	5.11 ± 0.96	2,4-二叔丁基苯酚 2,4-Di-tert-butylphenol	C₁₄H₂₂O	5.52 ± 0.52

化合物/（μL · 皿^-1） Substances	意大利青霉P. italicum		指状青霉P. digitatum
化合物/（μL · 皿^-1） Substances	IC₅₀/（μL · 皿^-1）	95%置信区间Confidence limits	IC₅₀/（μL · 皿^-1）	95%置信区间Confidence limits
2-壬醇2-Nonanol	0.31	0.27 ~ 0.35	0.33	0.29 ~ 0.36
2-壬酮2-Nonanone	14.10	12.09 ~ 16.28	8.70	7.23 ~ 10.19
香叶醇Geraniol	49.14	44.88 ~ 53.80	47.63	44.55 ~ 50.91
苯甲酸Benzoic acid	13.80	12.20 ~ 15.59	14.45	13.49 ~ 15.47

化合物/（μL · 皿^-1） Substances	意大利青霉P. italicum		指状青霉P. digitatum
化合物/（μL · 皿^-1） Substances	IC₅₀/（μL · 皿^-1）	95%置信区间Confidence limits	IC₅₀/（μL · 皿^-1）	95%置信区间Confidence limits
2-壬醇2-Nonanol	0.31	0.27 ~ 0.35	0.33	0.29 ~ 0.36
2-壬酮2-Nonanone	14.10	12.09 ~ 16.28	8.70	7.23 ~ 10.19
香叶醇Geraniol	49.14	44.88 ~ 53.80	47.63	44.55 ~ 50.91
苯甲酸Benzoic acid	13.80	12.20 ~ 15.59	14.45	13.49 ~ 15.47

处理 Treatment	发病率/% Disease incidence		防治效果/% Control efficacies
处理 Treatment	青霉病Blue mould	绿霉病Green mould	青霉病Blue mould	绿霉病Green mould
对照Control	61.75 ± 3.86 a	62.00 ± 5.16 a	0	0
咪鲜胺Prochloraz	10.75 ± 1.89 c	8.25 ± 1.26 c	82.66 ± 2.38	86.50 ± 3.18
HY19挥发物 Volatile organic compounds	25.00 ± 4.08 b	22.25 ± 2.87 b	59.65 ± 4.46	64.11 ± 3.52