Screening of Antagonist Against Tomato Fruit Rot and Their Preservation Qualities on Tomato

doi:10.13560/j.cnki.biotech.bull.1985.2021-0521

Abstract

Abstract:

A strain of fruit rot pathogen was isolated from storing tomatoes. In order to clarify the taxonomic status and investigate the suitable biocontrol strain,its morphology and phylogenetic analysis were conducted. The antagonistic Streptomyces was isolated and identified,and its effect on the preservation of tomato was studied. The results showed that the pathogen Qyg16-2 that caused the fruit rot of tomato was Botryosphaeria dothidea. The actinomecetes X13 with good antagonistic effects from healthy tomato rhizospheric soil on Qyg16-2 was identified as Streptomyces sampsonii by morphology,physiological and biochemical characteristics and 16S rDNA sequence. X13 significantly inhibited the colony extension and spore germination of pathogen B. dothidea,its cell-free fermentation filtrate significantly increased the activity of PPO,POD and PAL,decreased the decay index of stored tomato,and delayed the decline of post-harvest tomato fruit firmness,soluble solid content,titration acid content and vitamin C content. Conclusively,S. sampsonii X13 has significant antagonistic effects on B. dothidea,and has promising preservation effect in storing tomato fruit.

Key words: post-harvest tomato, Botryosphaeria dothidea, Streptomyces sampsonii, screening, preservation

ZHANG Hong-yan, LIN Guo-li, LI Ru-lian, JI Xiao-qi. Screening of Antagonist Against Tomato Fruit Rot and Their Preservation Qualities on Tomato[J]. Biotechnology Bulletin, 2022, 38(3): 69-78.

Figures/Tables 9

References 32

[1]	宋姝婧, 王晓拓, 王志东, 等. 5种植物精油对樱桃番茄常温保鲜效果的影响[J]. 核农学报, 2015, 29(5):932-939.
	Song SJ, Wang XT, Wang ZD, et al. Effect of five essential oils on cherry tomatoes preservation at room temperature[J]. J Nucl Agric Sci, 2015, 29(5):932-939.
[2]	Janisiewicz WJ, Conway WS. Combining biological control with physical and chemical treatments to control fruit decay after harvest[J]. Stewart Postharvest Rev, 2010, 6(1):1-16.
[3]	桑雪, 刘阳, 王锐银, 等. 一株引起番茄果腐病病原菌的分离鉴定[J]. 大连工业大学学报, 2018, 37(3):167-170.
	Sang X, Liu Y, Wang RY, et al. Isolation and identification of a pathogen causing tomato fruit rot[J]. J Dalian Polytech Univ, 2018, 37(3):167-170.
[4]	Li QL, Ning P, Zheng L, et al. Effects of volatile substances of Streptomyces globisporus JK-1 on control of Botrytis cinerea on tomato fruit[J]. Biol Control, 2012, 61(2):113-120. doi: 10.1016/j.biocontrol.2011.10.014 URL
[5]	Miedes E, Lorences EP. The implication of xyloglucan endotransglucosylase/hydrolase(XTHs)in tomato fruit infection by Penicillium expansum link. A[J]. J Agric Food Chem, 2007, 55(22):9021-9026. doi: 10.1021/jf0718244 URL
[6]	Strashnov Y, Elad Y, Sivan A, et al. Control of Rhizoctonia solani fruit rot of tomatoes by Trichoderma harzianum Rifai[J]. Crop Prot, 1985, 4(3):359-364. doi: 10.1016/0261-2194(85)90039-0 URL
[7]	Punja ZK, Rodriguez G, Tirajoh A. Effects of Bacillus subtilis strain QST 713 and storage temperatures on post-harvest disease development on greenhouse tomatoes[J]. Crop Prot, 2016, 84:98-104. doi: 10.1016/j.cropro.2016.02.011 URL
[8]	Sun C, Fu D, Jin LF, et al. Chitin isolated from yeast cell wall induces the resistance of tomato fruit to Botrytis cinerea[J]. Carbohydr Polym, 2018, 199:341-352. doi: 10.1016/j.carbpol.2018.07.045 URL
[9]	周凯强, 李志洪, 覃书漫, 等. 余甘子采后软腐病拮抗菌的筛选、鉴定及其防效[J]. 微生物学通报, 2020, 47(2):459-468.
	Zhou KQ, Li ZH, Qin SM, et al. Identification and biocontrol effect of antagonistic Streptomyces against postharvest Penicillium fruit rot of Phyllanthus emblica L.[J]. Microbiol China, 2020, 47(2):459-468.
[10]	郭雪松, 田丽波, 商桑, 等. 芒果蒂腐病拮抗放线菌A10和A17的分离、鉴定和特征化研究[J]. 浙江农业学报, 2020, 32(3):460-468.
	Guo XS, Tian LB, Shang S, et al. Isolation, identification and characterization of antagonistic actinomycetes A10 and A17 against Botryodiplodia theobromae[J]. Acta Agric Zhejiangensis, 2020, 32(3):460-468.
[11]	Zhang QM, Yong DJ, Zhang Y, et al. Streptomyces rochei A-1 induces resistance and defense-related responses against Botryosphaeria dothidea in apple fruit during storage[J]. Postharvest Biol Technol, 2016, 115:30-37. doi: 10.1016/j.postharvbio.2015.12.013 URL
[12]	方中达. 植病研究方法[M]. 第三版. 北京: 中国农业出版社, 1998.
	Fang ZD. Research methods of plant pathology[M]. 3rd. ed. Beijing: China Agriculture Press, 1998.
[13]	魏景超. 真菌鉴定手册[M]. 上海: 上海科学技术出版社, 1979.
	Wei JC. Handbook of Fungal Identification[M]. Shanghai:Shanghai Scientific & Technical Publishers, 1979.
[14]	牛晓磊, 薛泉宏, 涂璇, 等. 6株生防放线菌对辣椒疫霉的皿内拮抗研究[J]. 西北农林科技大学学报:自然科学版, 2005, 33(1):55-58.
	Niu XL, Xue QH, Tu X, et al. Study on the antagonistic activity of 6 strains of actinomycetes against Phytophthora capsici in plate[J]. J Northwest Sci Tech Univ Agric For, 2005, 33(1):55-58.
[15]	刘大群, 杨文香, 祁碧菽, 等. 拮抗链霉菌Men-myco-93-63及其发酵液对棉花黄萎病菌生长的影响[J]. 河北农业大学学报, 1999, 22(4):79-82.
	Liu DQ, Yang WX, Qi BS, et al. Effect of men-myco-93-63 and its fermentation liquid on the growth of Verticillium dahliae strains of cotton[J]. J Agric Univ Hebei, 1999, 22(4):79-82.
[16]	程丽娟, 薛泉宏. 微生物学实验技术[M]. 西安: 世界图书出版社, 2000.
	Cheng LJ, Xue QH. Experimental technique in microbiology[M]. Xi’an: World Publishing corporation, 2000.
[17]	东秀珠, 蔡妙. 常见细菌系统鉴定手册[M]. 北京: 科学出版社, 2001.
	Dong XZ, Cai MY. Manual of Determinative Bacteriology[M]. Beijing: Science Press, 2001.
[18]	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.
	Cao JK, Jiang WB, Zhao YM. Experiment guidance of postharvest physiology and biochemistry of fruits and vegetables[M]. Beijing: China Light Industry Press, 2007.
[19]	罗春香, 赵玉良, 王海燕, 等. 12种杀菌剂对樱桃流胶病菌的敏感性及其防治效果研究[J]. 中国果树, 2020(4):27-30.
	Luo CX, Zhao YL, Wang HY, et al. Bactericidal activities of twelve bactericides and their field applications against pathogen of the cherry gummosis[J]. China Fruits, 2020(4):27-30.
[20]	康海婷, 凌丹燕, 吴志娟, 等. 蓝莓枝干溃疡病病原鉴定及品种抗病性研究[J]. 浙江农业学报, 2019, 31(3):436-443.
	Kang HT, Ling DY, Wu ZJ, et al. Pathogen isolation and identification of blueberry stem canker and screening of disease resistant cultivars[J]. Acta Agric Zhejiangensis, 2019, 31(3):436-443.
[21]	王亚红, 李永丽, 常乐, 等. 对三种苹果病原菌具抑制作用的新疆野苹果内生真菌的分离与鉴定[J]. 果树学报, 2020, 37(3):390-396.
	Wang YH, Li YL, Chang L, et al. Isolation and identification of endophytic fungi resistant to three apple pathogens from the branches of Malus sieversii[J]. J Fruit Sci, 2020, 37(3):390-396.
[22]	张玮, 姚晟伟, 张国军, 等. 中国葡萄主要品种对葡萄座腔菌的抗性评价[J]. 植物保护, 2017, 43(3):177-180, 212.
	Zhang W, Yao SW, Zhang GJ, et al. Resistant evaluation of main grape cultivars in China to Botryosphaeria dothidea[J]. Plant Prot, 2017, 43(3):177-180, 212.
[23]	朱琪丽, 赵会长, 谢甲涛, 等. 葡萄座腔菌侵染柑橘果实的报道[J]. 植物病理学报, 2018, 48(2):154-158.
	Zhu QL, Zhao HZ, Xie JT, et al. Report of Botryosphaeria dothidea infecting Citrus[J]. Acta Phytopathol Sin, 2018, 48(2):154-158.
[24]	Thomidis T, Michailides TJ, Exadaktylou E. Neofusicoccum parvum associated with fruit rot and shoot blight of peaches in Greece[J]. Eur J Plant Pathol, 2011, 131(4):661-668. doi: 10.1007/s10658-011-9840-0 URL
[25]	Malviya MK, Pandey A, Trivedi P, et al. Chitinolytic activity of cold tolerant antagonistic species of Streptomyces isolated from glacial sites of Indian Himalaya[J]. Curr Microbiol, 2009, 59(5):502-508. doi: 10.1007/s00284-009-9466-z pmid: 19688382
[26]	Jain PK, Jain PC. Isolation, characterization and antifungal activity of Streptomyces sampsonii GS 1322[J]. Indian J Exp Biol, 2007, 45(2):203-206.
[27]	Savi DC, Haminiuk CWI, Sora G, et al. Antitumor, antioxidant and antibacterial activities of secondary metabolites extracted by endophytic actinomycetes isolated from Vochysia divergens[J]. Int J Pharm Chem Biol Sci, 2015, 5(1):347-356.
[28]	李姝江, 朱天辉, 余琴, 等. 桑氏链霉菌突变株的抑菌活性及对杨树紫纹羽病的盆栽防效[J]. 植物保护, 2015, 41(5):61-68.
	Li SJ, Zhu TH, Yu Q, et al. Antifungal activity of mutant strains of Streptomyces sampsonii and their control effects on poplar purple root disease in potted experiment[J]. Plant Prot, 2015, 41(5):61-68.
[29]	郭虹娜, 刘佳, 奚裕婷, 等. 拮抗酵母挥发性单体桂皮醛对草莓采后灰霉病的防治[J]. 食品科学, 2020, 41(13):212-220.
	Guo HN, Liu J, Xi YT, et al. Biocontrol effect of volatile monomer trans-cinnamaldehyde with antagonistic capacity to yeast on gray mold disease of postharvest strawberry fruit[J]. Food Sci, 2020, 41(13):212-220.
[30]	孙平平, 贾晓辉, 崔建潮, 等. 梨灰霉病拮抗放线菌L-30的筛选、鉴定及作用机制研究[J]. 园艺学报, 2016, 43(12):2335-2346.
	Sun PP, Jia XH, Cui JC, et al. Selection, identification and characterization of Actinomyces L-30 for the biocontrol of pear gray mold[J]. Acta Hortic Sin, 2016, 43(12):2335-2346.
[31]	季小诗, 郭红莲, 王笑笑, 等. 拮抗菌发酵液对冬枣贮藏品质的影响[J/OL]. 食品与发酵工业, 2021. DOI: 10.13995/j.cnki.11-1802/ts.026170. doi: 10.13995/j.cnki.11-1802/ts.026170
	Ji XS, Guo HL, Wang XX, et al. Effects of an antagonistic yeast fermentation broth on the storage quality of winter jujube[J/OL]. Food and Fermentation Industry, 2021. DOI: 10.13995/j.cnki.11-1802/ts.026170. doi: 10.13995/j.cnki.11-1802/ts.026170
[32]	施俊凤, 孙常青, 张婧婷. 采前喷施洋葱伯克霍尔德菌Burkholderia contaminans对草莓采后腐烂和品质的影响[J]. 植物保护学报, 2018, 45(2):382-388.
	Shi JF, Sun CQ, Zhang JT. Effects of preharvest spraying of Burkholderia contaminans on postharvest decay and quality of strawberry[J]. J Plant Prot, 2018, 45(2):382-388.

菌株编号 Strain No.	D/mm	T	抑菌率 Inhibition rate/%	孢子萌发抑制率 Inhibition of seed germination/%	菌株编号 Strain No.	D/mm	T	抑菌率 Inhibition rate/%	孢子萌发抑制率Inhibition of germination/%
X1	20.31±2.12a	+++	68.15±1.25b	73.89±4.01b	X54	10.67±0.34d	++	-	-
X13	26.53±1.03a	+++	82.53±0.93a	98.37±3.54a	X60	11.22±0.51d	+++	-	-
X19	18.11±2.00b	+++	61.25±1.20b	62.12±5.13c	X75	14.64±0.39d	+++	-	-
X20	13.45±1.09c	++	-	-	X85	18.92±1.11ab	+++	49.21±0.35c	39.12±3.25e
X43	19.06±1.32b	+++	59.00±0.65b	46.98±4.20e	X90	19.46±1.20ab	+++	47.35±1.04c	54.99±2.84cd
CK	-	-	-	0.81±0.15f

菌株编号 Strain No.	D/mm	T	抑菌率 Inhibition rate/%	孢子萌发抑制率 Inhibition of seed germination/%	菌株编号 Strain No.	D/mm	T	抑菌率 Inhibition rate/%	孢子萌发抑制率Inhibition of germination/%
X1	20.31±2.12a	+++	68.15±1.25b	73.89±4.01b	X54	10.67±0.34d	++	-	-
X13	26.53±1.03a	+++	82.53±0.93a	98.37±3.54a	X60	11.22±0.51d	+++	-	-
X19	18.11±2.00b	+++	61.25±1.20b	62.12±5.13c	X75	14.64±0.39d	+++	-	-
X20	13.45±1.09c	++	-	-	X85	18.92±1.11ab	+++	49.21±0.35c	39.12±3.25e
X43	19.06±1.32b	+++	59.00±0.65b	46.98±4.20e	X90	19.46±1.20ab	+++	47.35±1.04c	54.99±2.84cd
CK	-	-	-	0.81±0.15f

测定内容Measured item	结果Result	碳源利用Carbon utilization	结果Result
明胶液化Gelatin liquefaction	+	D-蔗糖D-sucrose	+
牛奶胨化Milk coagulation	+	L-鼠李糖L-rhamnose	-
硝酸盐还原Nitrate reduction	+	肌醇Inositol	-
黑色素产生Black pigment	-	D-葡萄糖D-glucose	+
酪氨酸酶Tyrosinase	-	D-木糖D-xylose	+
H₂S	-	D-果糖D-fructose	+

测定内容Measured item	结果Result	碳源利用Carbon utilization	结果Result
明胶液化Gelatin liquefaction	+	D-蔗糖D-sucrose	+
牛奶胨化Milk coagulation	+	L-鼠李糖L-rhamnose	-
硝酸盐还原Nitrate reduction	+	肌醇Inositol	-
黑色素产生Black pigment	-	D-葡萄糖D-glucose	+
酪氨酸酶Tyrosinase	-	D-木糖D-xylose	+
H₂S	-	D-果糖D-fructose	+

处理 Treatment
处理 Treatment	4	8	12	16	20
对照CK	0	9.81±1.05aD	25.26±3.15aC	48.03±1.21aB	68.32±1.20aA
5%	0	4.98±1.35bC	18.35±1.41bB	25.10±1.27cA	26.11±1.02bA
10%	0	4.36±1.04bC	14.26±1.50bB	22.14±1.00bA	23.32±1.84cA
20%	0	2.05±0.68cC	10.35±1.36cB	20.16±1.35dA	22.02±0.65cA