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Endo-bacillus subtilis TR21 of plants and application thereof

A technology of Bacillus subtilis and plant pathogenic fungus, applied in the field of Bacillus subtilis TR21, can solve the problems of failure, high similarity between sequences, and inability to effectively distinguish Bacillus subtilis groups, etc., and achieve good antagonistic ability, good control effect, and good Effect of tablet antagonistic activity

Active Publication Date: 2012-05-30
珠海鼎元生态农业有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The species of the Bacillus subtilis group include Bacillus subtilis subsp.subtilis (Smith et al., 1964; Nakamura et al., 1999), B. lichenifromis (Skerman et al., 1980), B. atrophaeus (Nakamura, 1989), B. mojavensis (Robert et al., 1994), B. vallismortis (Roberts et al., 1996), B. subtilis subsp. spizizenii (Nakamura et al., 1999), B. sonorensis (Palmisano et al., 2001), B. .subtilis subsp.inaquosorum (Rooney et al., 2009), these species are closely related and can be distinguished by fatty acid composition analysis, DNA-DNA hybridization analysis and restriction enzyme analysis, but it is difficult to distinguish them by phenotypic characteristics analysis (Fritze, 2004, Wang, et al., 2007), although 16S rDNA / RNA gene sequences are widely used to identify bacteria or construct the phylogenetic relationship of bacteria, but among taxonomic groups with close kinship, due to The similarity between the sequences is too high to be invalid (Christensen et al., 1998), and the 16S rDNA sequence cannot effectively distinguish the species of the Bacillus subtilis group

Method used

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  • Endo-bacillus subtilis TR21 of plants and application thereof
  • Endo-bacillus subtilis TR21 of plants and application thereof
  • Endo-bacillus subtilis TR21 of plants and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Example 1: Separation and purification, research and identification of physicochemical properties of TR21

[0031] (1) Isolation, purification and preservation of TR21

[0032] In the greenhouse of the orchid production base of Zhuhai Agricultural Science Research Center, healthy leaves of Dendrobium bicolor were collected, rinsed with tap water, dried and treated with 75% alcohol for 1 min, then treated with 5% NaOCl for 5 min, and rinsed with sterile water 4 times; After drying, use sterile scissors and tweezers to cut the leaves into tissue pieces of about 0.2cm×0.5cm, spread them on NA (beef extract peptone medium) plates containing 50 μg / ml cycloheximide; culture at a constant temperature of 28°C , observe every day, pick out once bacteria grow out of the incision, purify and culture them on a new NA plate, and store them in the form of bacterial liquid with 20% glycerol at -20°C and -80°C. Then take 200 μl of sterile water rinsed for the fourth time and apply it ...

Embodiment 2

[0099] Embodiment 2: Antagonism test of TR21 to different Fusarium pathogenic bacteria

[0100] (1) Isolation and purification of different Fusarium pathogenic bacteria

[0101] Source of isolation: Collect typical banana fusarium wilt, celery verticillium wilt, and dendrobium leaf spot disease tissues in Zhuhai and surrounding cities to isolate pathogenic bacteria and test the antagonistic activity of TR21 against different Fusarium pathogenic bacteria.

[0102] Separation and purification method: clean the diseased tissue with clean water, dry it, cut the diseased tissue at the junction of diseased and healthy tissue with a sterile blade and cut it into an appropriate size, disinfect the surface with 70% alcohol for 30 seconds under sterile conditions, and then use available chlorine Treat with 5% NaOCl for 5 minutes, rinse with sterile water for 4 times, cut the surface-sterilized diseased tissue into tissue pieces of about 0.2 cm × 0.5 cm with sterile scissors and tweezers...

Embodiment 3

[0110] Example 3: The antagonistic activity of TR21 to other plant pathogenic fungi, the results are shown in Table 5.

[0111] The method during bioassay antagonistic activity is with embodiment 2, and wherein Alternaria, Curvularia lunata, mango stalk rot, watermelon wilt etc. are respectively the phytopathogenic fungi that Zhuhai City True Green Technology Co., Ltd. presents, and D. solani Rhizoctonia is a plant pathogenic fungus donated by Dr. Yi Runhua of Guangdong Ocean University. The larger the inhibition zone, the stronger the antibacterial effect.

[0112] Table 5TR21 plate antagonistic activity to several plant pathogenic fungi (unit: mm)

[0113] Alternaria

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Abstract

The invention relates to a bacillus subtilis str.TR21 (hereinafter referred to as 'TR21') strain having broad-spectrum antagonism to plant pathogenic fungi, in particular to the bacillus subtilis TR21 strain and a growth inhibition function thereof on the plant pathogenic fungi. The bacillus subtilis TR21 strain provided by the invention is preserved in China Center for Type Culture Collection (CCTCC) in January 20, 2010 with a preservation number of CCTCC No: M2010019, wherein the China Center for Type Culture Collection is located in Wuhan University, Lojia Mountain, Wuchang, Hubei, China. The strain provided by the invention is proved by obtained gene segments 16S rDNA, gyrA and gyrB and BLAST search and comparison on NCBI to be a novel bacillus subtilis strain having relatively higher flat antagonistic activity to a plurality of plant pathogenic fungi, particularly having high antagonism to various soil borne vascular diseases, such as fusarium oxysporum, rhizoctonia solani, alternaria and the like, and having good prevention and control effects shown in fields.

Description

technical field [0001] The present invention relates to a strain of Bacillus subtilis TR21 (subsequently referred to as TR21) with broad-spectrum antagonistic ability to plant pathogenic fungi, specifically providing Bacillus subtilis str.TR21 strain (CCTCC No: M 2010019) and Its growth inhibitory effect on plant pathogenic fungi, especially in the biological control of banana fusarium wilt and celery verticillium wilt, belongs to the category of biological pesticides and disease-resistant microbial fertilizers. Background technique [0002] The losses of horticultural crops such as fruits and vegetables in our country due to diseases are very serious. According to the results of the 1984-1986 national census of garden plant diseases and insect pests, 5508 kinds of plant diseases were found on 1256 kinds of garden plants, with an average of 4.4 kinds on each plant. Vegetables, flowers The occurrence of diseases of horticultural crops such as , fruit trees, etc. is far more s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P3/00C12R1/125
Inventor 喻国辉陈燕红黎永坚陈远凤杨紫红周林程萍
Owner 珠海鼎元生态农业有限公司
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