Screening and Identification of Bacillus velezensis XC1 and Its Influence on Apple Replant Disease

doi:10.16420/j.issn.0513-353x.2020-0476

Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (3): 409-420.doi: 10.16420/j.issn.0513-353x.2020-0476

• Research Papers • Next Articles

Screening and Identification of Bacillus velezensis XC1 and Its Influence on Apple Replant Disease

YAN Zhubing¹, WANG Mei¹, MING Changjun², JIANG Yuanmao¹, SHEN Xiang¹, CHEN Xuesen¹, YIN Chengmiao¹^,^*(), MAO Zhiquan¹^,^*()

¹College of Horticulture Science and Engineering,Shandong Agricultural University/State Key Laboratory of Crop Biology,Tai'an,Shandong 271018,China
²Agrotechnical Station in Guli Town,Xintai City,Tai'an,Shandong 271215, China

Received:2020-08-21 Online:2021-03-25 Published:2021-04-02
Contact: YIN Chengmiao,MAO Zhiquan E-mail:yinchengmiao@163.com;mzhiquan@sdau.edu.cn

Abstract

Abstract:

In this study,a strain XC1 was screened from the rhizosphere soil of healthy apple trees in the replanted apple orchard,which could antagonize the main pathogenic fungi（Fusarium solani,F. proliferatum,F. oxysporum,F. moniliforme）of apple replant disease. Among them,the inhibitory rate of F. oxysporumwas the highest,reaching 79.83%. Through morphological,physiological and biochemical characteristics,16S rDNA and gyrA sequence analysis,the bacterium was identified as Bacillus velezensis,which is a new biocontrol bacterium with development potential. At present,strain XC1 has been collected by the China General Microbiological Culture Collection Center,with the preservation number CGMCC No.20057. The pot experiment showed that compared with the control（replanted soil）,XC1 treatment could significantly promote the seedlings growth of Malus hupehensisRehd. and its plant height,ground diameter,fresh and dry mass were 54.8%,34.8%,121.1% and 96.0% higher than those of the control respectively;the activities of sucrase,urease,catalase and neutral phosphatase in the replanted soil treated with XC1 fertilizer were significantly increased,which were 1.57,2.71,1.21 and 1.79 times higher than those of the control,respectively. The number of fungi in the soil were significantly reduced;and the number of bacteria and actinomycetes was increased;and the gene copy number of F. oxysporum treated with XC1 fertilizer was 76.4% lower than that of the control. This indicated that B. velezensis has obvious bacteriostatic effect,which can improve the soil environment,promote the growth of M. hupehensisRehd. seedlings,and reduce the apple replant disease to a certain extent.

Key words: apple, replant disease, Malus hupehensis, Bacillus velezensis, Fusariumspp.

CLC Number:

S661.1

YAN Zhubing, WANG Mei, MING Changjun, JIANG Yuanmao, SHEN Xiang, CHEN Xuesen, YIN Chengmiao, MAO Zhiquan. Screening and Identification of Bacillus velezensis XC1 and Its Influence on Apple Replant Disease[J]. Acta Horticulturae Sinica, 2021, 48(3): 409-420.

Figures/Tables 9

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特征 Characteristic	结果 Result	特征 Characteristic	结果 Result
V-P试验 V-P test	+	硝酸盐还原 Nitrate reduction	+
甲基红试验 Methyl red test	-	H₂S试验 H₂S test	-
淀粉水解试验 Starch hydrolysis test	+	动力学试验 Dynamic test	+
接触酶试验 Catalase test	+	明胶液化试验 Gelatin liquefaction test	+
吲哚测定 Indole test	-	D-葡萄糖 D-Glucose	+

特征 Characteristic	结果 Result	特征 Characteristic	结果 Result
V-P试验 V-P test	+	硝酸盐还原 Nitrate reduction	+
甲基红试验 Methyl red test	-	H₂S试验 H₂S test	-
淀粉水解试验 Starch hydrolysis test	+	动力学试验 Dynamic test	+
接触酶试验 Catalase test	+	明胶液化试验 Gelatin liquefaction test	+
吲哚测定 Indole test	-	D-葡萄糖 D-Glucose	+

处理 Treatment	株高/cm Plant height	地径/mm Ground diameter	鲜质量/g Fresh mass	干质量/g Dry mass
对照Control	49.56 ± 2.21 d	6.63 ± 0.09 c	25.13 ± 1.74 c	12.37 ± 0.26 c
溴甲烷Methyl bromide	87.77 ± 1.80 a	11.44 ± 0.61 a	98.07 ± 3.71 a	40.50 ± 1.71 a
菌肥载体Biofertilizer carrier	59.87 ± 3.12 c	6.84 ± 0.42 c	32.35 ± 3.69 c	15.38 ± 0.84 c
XC1菌肥XC1 biofertilize	76.70 ± 1.61 b	8.94 ± 0.33 b	55.56 ± 2.00 b	24.25 ± 1.03 b

处理 Treatment	株高/cm Plant height	地径/mm Ground diameter	鲜质量/g Fresh mass	干质量/g Dry mass
对照Control	49.56 ± 2.21 d	6.63 ± 0.09 c	25.13 ± 1.74 c	12.37 ± 0.26 c
溴甲烷Methyl bromide	87.77 ± 1.80 a	11.44 ± 0.61 a	98.07 ± 3.71 a	40.50 ± 1.71 a
菌肥载体Biofertilizer carrier	59.87 ± 3.12 c	6.84 ± 0.42 c	32.35 ± 3.69 c	15.38 ± 0.84 c
XC1菌肥XC1 biofertilize	76.70 ± 1.61 b	8.94 ± 0.33 b	55.56 ± 2.00 b	24.25 ± 1.03 b

处理 Treatment	细菌（× 10⁵ CFU·g^-1soil） Bacteria	真菌（×10³ CFU·g^-1soil） Fungi	放线菌（×10⁴ CFU·g^-1soil） Actinomyces
对照Control	20.33 ± 1.45 c	36.33 ± 3.48 a	42.33 ± 2.03 c
溴甲烷Methyl bromide	4.67 ± 0.33 d	5.00 ± 0.58 b	15.67 ± 1.76 d
菌肥载体Biofertilizer carrier	31.33 ± 2.03 b	31.33 ± 4.06 a	55.67 ± 0.67 b
XC1菌肥XC1 biofertilize	54.67 ± 2.33 a	7.67 ± 0.67 b	73.00 ± 1.53 a

Screening and Identification of Bacillus velezensis XC1 and Its Influence on Apple Replant Disease

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处理 Treatment	蔗糖酶/（mg·g^-1·d^-1） Sucrase	脲酶/（mg·g^-1·d^-1） Urease	中性磷酸酶/（mg·g^-1·d^-1） Neutral phosphatase	过氧化氢酶/（mL·g^-1） Catalase
对照Control	8.17 ± 0.16 c	0.14 ± 0.00 c	12.53 ± 0.52 b	0.19 ± 0.00 c
溴甲烷Methyl bromide	6.33 ± 0.19 d	0.08 ± 0.00 d	9.68 ± 0.61 c	0.15 ± 0.01 d
菌肥载体Biofertilizer carrier	9.42 ± 0.12 b	0.24 ± 0.00 b	13.71 ± 0.20 b	0.26 ± 0.00 b
XC1菌肥XC1 biofertilize	12.86 ± 0.03 a	0.38 ± 0.00 a	15.18 ± 0.20 a	0.34 ± 0.00 a

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