CN104450560B - One plant of nematicide Sphingobacterium bacterial strain and its application - Google Patents
One plant of nematicide Sphingobacterium bacterial strain and its application Download PDFInfo
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- CN104450560B CN104450560B CN201410629500.1A CN201410629500A CN104450560B CN 104450560 B CN104450560 B CN 104450560B CN 201410629500 A CN201410629500 A CN 201410629500A CN 104450560 B CN104450560 B CN 104450560B
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Abstract
The present invention relates to one plant of nematicide Sphingobacterium (Sphingobacterium nematocida) M SX103 bacterial strain and its application, microorganism belonging to genus technical field of pesticide.Nematicide Sphingobacterium (Sphingobacterium nematocida) M SX103 bacterial strain, the preserving number of this bacterial strain is:CGMCC No.9689.Application in preparation preventing and treating tobacco bacterial wilt and notoginseng root rot microbial inoculum for nematicide Sphingobacterium (Sphingobacterium nematocida) the M SX103 bacterial strain.The beneficial effects of the present invention is:1st, use the biocontrol agent of M SX103 bacterial strain preparation to preventing and treating Nicotiana tabacum L. blue or green disease, the prevention effect of notoginseng root rot is more than 67% it is adaptable to seed treatment, transplanting time and Adult plant.There is the biocontrol bacterial strain of commercial application prospect, be that the preventing and treating of plant rot and notoginseng root rot provides new microbial resources.2nd, with the biocontrol agent of nematicide Sphingobacterium (Sphingobacterium nematocida) M SX103 bacterial strain preparation, there is low cost, preventive effect height, noresidue, easy to use, feature to person poultry safety, is the high-performance bio pesticide of preventing and treating plant-bacterial-wilt evil in organic agriculture plantation.
Description
Technical field:
The present invention relates to one plant of nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 bacterium
Strain and its application, microorganism belonging to genus technical field of pesticide.
Background technology:
Plant-bacterial-wilt (Bacterial Wilt) is to be drawn by blue or green withered Raul Salmonella (Ralstonia solanacearum)
The crushing soil occurring in a kind of world wide rising passes bacterial disease, is referred to as " cancer " of plant, can make the plant underproduction
30-100%, can infect 50 genus 200 various plants, wherein endanger main host plants heavier, distributed more widely have Nicotiana tabacum L., kind
Eggplant, Fructus Solani melongenae, Fructus Capsici etc. (Hayward, 1991;1994).Nicotiana tabacum L. (Nicotiana tabacum) is the important economic work of China
Thing, is also one of mainstay industry that Yunnan Province develops landlord economy.Tobacco bacterial wilt is tropical and subtropical region tobacco seed
The important soil-borne disease of growing area, harm is serious, and the economic loss causing is huge, has become at present and has limited the development of tobacco planting industry
Serious hindrance (Chen Ruitai etc., 1997).Radix Notoginseng (Panax notoginseng) is araliaceae ginseng plant, is China's tradition
Famous and precious genuine medicinal materials.Radix Notoginseng main product, in Wenshan Prefecture of Yunnan Province, is the living resources showing unique characteristics in Yunnan, is also that Yunnan Province is sent out
One of the mainstay industry of exhibition local economy (Wei et al., 1996).By the microbial notoginseng root rot of bacterial wilt
(Root-rot disease) is the destructive disease in notoginseng planting, and the loss that it causes accounts for the loss of Radix Notoginseng whole disease
70-85%, it has also become the serious hindrance (Dong not million etc., 1998) of restriction Radix Notoginseng industry sustainable development.Plant-bacterial-wilt and three
Seven root rot are soil-borne disease.So far, to the preventing and treating of both diseases mainly to apply based on chemical pesticide.Because chemistry is anti-
Control easily cause pesticide residues, pollution environment and pathogen produce drug resistance the problems such as, exploitation biological pesticide have become as environmental protection,
One of safety, the leading measure of the disease control of holding effect.
Nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 is deliver for 2012 new
Species, M-SX103 bacterial strain is Endophytic Bacteria of Tobacco, has good colonization ability in plant root.This microorganism is black for Nicotiana tabacum L.
Shin pathogenic bacteria (Phytophthora nicotianae) has antagonistic activity, and to Panagrellus redivivus (Panagrellus
Redivivus) show 100% nematicide fatality rate (Liu et al., 2012).This microorganism to plant-bacterial-wilt opportunistic pathogen and
So far there are no open reports for the biological and ecological methods to prevent plant disease, pests, and erosion characteristic of notoginseng root rot and prevention effect.
List of references:
Hayward AC.1991.Biology and epidemiology of bacterial wilt caused by
Ralstonia solanacearum.Annu Rev Phytopathol 29:65-87.
Hayward AC.1991.The hosts of Pseudomonas solanacearum.In:Bacterial
Wilt:The Disease and its Causative Agent,Pseudomonas solanacearum.
(Eds.Hayward AC and Hartman GL.)CAB International,Wallingford,UK.pp.9-24.
Liu J,Yang LL,Xu CK,Xi JQ,Yang FX,Zhou F,Zhou Y,Mo MH,Li
WJ.2012.Sphingobacterium nematocida sp.nov.,a nematicidal endophytic
bacterium isolated from tobacco.International Journal of Systematic and
Evolutionary Microbiology 62(8):1809-1813.
Wei JX,Du YC.1996.Modern Science Research and Application of Panax
notoginseng.Yunnan Science and Technology Press,Kunming,China.
16, the whole nation of the .1997. such as Chen Ruitai main product cigarette saves (area) Nicotiana tabacum L. infectious disease investigation report. Chinese tobacco section
Learn, (4):1-7.
Dong's not million grade .1988. Yunnan notoginseng. Yunnan Science Press, Kunming, pp.103-114.
Miao makees clear grade for .2006. notoginseng root rot cause of disease research. Scientia Agricultura Sinica, 39 (7):1371-1378.
Content of the invention:
It is an object of the invention to provide one plant of nematicide Sphingobacterium (Sphingobacterium nematocida)
M-SX103 bacterial strain and its application.
Nematicide Sphingobacterium (Sphingobacterium nematocida) the M-SX10 bacterial strain of the present invention, be from
The biocontrol bacterial strain with commercial application prospect filtering out in Nicotiana tabacum L. endophyte, is the anti-of plant rot and notoginseng root rot
Control and provide new microbial resources.
The present invention is specifically realized in:
Nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 bacterial strain, is isolatable from Yunnan and bakes
In the healthy leaves of tobacco kind NC297, it is general that this bacterial strain is stored in China Microbial Culture Preservation Commission on 18th in September in 2014
Logical microorganism center, preserving number:CGMCC No.9689.
Nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 bacterial strain, in Carnis Bovis seu Bubali cream albumen
In peptone culture medium (Carnis Bovis seu Bubali cream 3g, peptone 10g, NaCl 5g, agar 20g, moisturizing to 1,000mL, pH7.2), bacterium colony is in Huang
Color, circular, edge protuberance, cell is shaft-like, 0.4-0.8 μm of diameter, and long 1.0-1.5 μm, spore does not move about, Gram-negative.
This bacterial strain grows within the temperature range of 10-40 DEG C, 37 DEG C of optimum growth temperature;Grow in the range of pH 6.0-9.0,
Suitable growth pH is 7.0;Grow in the range of the NaCl (w/v) for 0-3% for the concentration, the most suitable growth concentration is the NaCl of 0-1%.Profit
With primer 5 '-GGT TAC CTT GTT ACG ACT T-3 ', 5 '-AGA GTT TGA TCM TGG CTC AG-3 ' are through PCR expansion
Open up the 16S rRNA gene of this bacterial strain, the sequence of acquisition is submitted to GeneBank public database, its Serial No. JF731241,
This sequence is to identify the principal character foundation of this bacterial strain.The main fatty acid of M-SX103 bacterial strain is different formula 15 carbon saturated fat
Sour (57.7%), different formula 17 carbon satisfied fatty acid (7.7%) and summed feature 3 (9.6%).It is to differentiate this bacterium
Secondary feature foundation.
Nematicide Sphingobacterium (Sphingobacterium nematocida) the M-SX103 bacterial strain of the present invention is in system
Application in standby preventing and treating tobacco bacterial wilt and notoginseng root rot microbial inoculum.
The beneficial effects of the present invention is:
1st, to preventing and treating Nicotiana tabacum L. green grass or young crops disease, the prevention effect of notoginseng root rot exists the biocontrol agent using the preparation of M-SX103 bacterial strain
More than 67% it is adaptable to seed treatment, transplanting time and Adult plant.There is the biocontrol bacterial strain of commercial application prospect, be plant
The preventing and treating of rot and notoginseng root rot provides new microbial resources.
2nd, use the life of nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 bacterial strain preparation
Anti- microbial inoculum has low cost, preventive effect height, noresidue, easy to use, and the feature to person poultry safety, is to prevent and treat in organic agriculture plantation
The high-performance bio pesticide of plant-bacterial-wilt evil.
Specific embodiment:
Nematicide Sphingobacterium used by embodiment (Sphingobacterium nematocida) M-SX103 bacterial strain, point
In the healthy leaves from Yunnan Flue-cured Tobacco kind NC297, this bacterial strain is stored in Chinese microorganism strain on the 18th in September in 2014 and protects
Hide committee's common micro-organisms center, preserving number:CGMCC No.9689.
Embodiment 1:The culture of M-SX103 bacterial strain and microbial inoculum preparation
The culture of M-SX103 strain tube inclined-plane seed:Inoculation is oblique in beef extract-peptone solid medium
On face, culture under the conditions of 15-37 DEG C obtained M-SX103 bacterial strain inclined-plane seed after 2-3 days, and suitable condition of culture is at 32 DEG C
Culture 2 days.
The liquid culture of M-SX103 strain liquid seed:One ring M-SX103 inoculation is taken from inclined-plane seed to triangle
In beef extract-peptone fluid medium (beef extract-peptone solid medium is without agar hereinafter referred to as NB) in bottle, in
At 37 DEG C, rotating speed 200rpm, cultivates 2-3 days, prepared seed liquor.
M-SX103 strain fermentation tank mass propgation:Described seed liquor is accessed containing NB training in the ratio of 3-5% (V/V)
Cultivate in the fermentation tank of foster base, the condition of culture in 20-1,000 liter of fermentation tank controls:Temperature 32-37 DEG C, mixing speed
150-250rpm, fermentation time 48-72 hour.
The preparation of M-SX103 wettability microbial inoculum:The microbial cells of fermented tank culture acquisition and its metabolism are no as former
Medicine makes M-SX103 wettable powder biocontrol agent according to a conventional method with pesticide conventional absorbtion carrier and auxiliary agent.Microbial inoculum needs to ensure
The viable count content of M-SX103 is 1 × 1010Individual/gram more than.
Embodiment 2:The efficiency test to tobacco bacterial wilt for the M-SX103 microbial inoculum
The culture of tobacco bacterial wilt pathogen (Ralstonia solanacearum):In TTC culture medium (peptone
10.0g, casein hydrolysate 1.0g, 2,3,5- chlorinated triphenyl base four nitrogen 0.05g, glucose 10.0g, agar 20.0g, distilled water
1,000mL, pH 6.8-7.2) inoculate tobacco bacterial wilt pathogen on solid plate, after culture 3 days at 32 DEG C, picking single bacterium colony connects
Plant in NB fluid medium, at 37 DEG C, under 200rpm, shaking table culture 2 days, tobacco ralstonia solanacearum is made into 2.0 × 108cfu/
The bacteria suspension of mL concentration is standby.
Reagent agent:A. viable count content is 2.5 × 1010Individual/gram M-SX103 wettability microbial inoculum;B.25% blue or green withered net
Wettable powder (development of academy of agricultural sciences of Guangdong Province crop institute Nicotiana tabacum L. room).
Efficiency test method:During flue-cured tobacco routine floating seedlings, the M-SX103 wettability microbial inoculum of 60g is diluted with 400mL water
Educate substrate afterwards to mix with 1kg;Positive control is educated with 1kg after being processed as 25% blue or green withered net wettable powder 1g 400mL water dilution
Substrate mixes;Blank is that 400ml water educates substrate mixing with 1kg.When tobacco seedlings length is to 5-6 piece leaf, in a diameter of 20cm
, equipped with 500g soil, (mass ratio is:Sandy soil:Fertile soil=1:1) each flowerpot transplants one plant of tobacco seedlings, inoculates cigarette in cigarette root
Careless Stalk Rot 6ml, often processes three repetitions, often repeats 5 basins.Rich water quality management is carried out according to a conventional method, in room temperature greenhouse
Interior growth counts the sickness rate of the tobacco bacterial wilt of each process after 90 days, calculate sickness rate as follows, disease index and anti-
Sick effect:
Disease index=highest symptom representative value × the investigation in [∑ (at different levels symptom representative value corresponding series)] × 100/
Total strain number) × 100
Protection effect (%)=(blank disease refer to-chemicals treatment disease refer to/compare disease and refer to) × 100
Severity Scaling:0 grade is asymptomatic;L level is l-2 piece leaf withering;Wilt or cane base portion for 1/3-1/2 blade for 2 grades
Blackening;3 grades is the wilting of 2/3-3/4 blade, downright bad and cane blackening is downright bad;4 grades is that whole strain is dead.
Result of the test shows:The disease index of blank is 77.4;The state of an illness that 25% blue or green withered net wettability microbial inoculum is processed
Index is 25.4, and preventive effect is 67.1%;The disease index that M-SX103 wettability microbial inoculum is processed is 23.7%, and preventive effect is 69.4%,
Suitable with the prevention effect of the blue or green withered net wettable powder of chemical pesticide 25%.
Embodiment 3:The field test of notoginseng root rot prevented and treated by M-SX103 microbial inoculum
This test is experimental field carried out in the graduate booth of Wenshan Prefecture of Yunnan Province Radix Notoginseng, this experimental field continuous cropping Radix Notoginseng 20
Year, the sickness rate of notoginseng root rot is more than 80%.In Radix Notoginseng nursery, 100g thalline content is 2.5 × 1010The M- of individual/g
SX103 wettability microbial inoculum 1L water-solubleization, adds 1kg Panax notoginseng seeds to mix, so that microbial inoculum is sticked on seed, then routinely educate
Plant management by methods.During transplanting every mu of Radix Notoginseng Seedling be watered the uniform pouring root of 200kg with 10kg microbial inoculum, use same dose after transplanting 1 year
Microbial inoculum and mode apply again once.Comparison medicament is 50% vinclozolin wettable powder (BASF Aktiengesellschaft), consumption
For every mu of 2kg, application process is with M-SX103 powder.Blank does not apply any medicament, often processes three repetitions, often repeats
100 young plants.Rich water quality management after Radix Notoginseng is transplanted routinely is carried out, and counts the sickness rate of each process, by following public affairs after transplanting 2 years
Formula calculates sickness rate and preventive effect:
Sickness rate (%)=morbidity strain number/investigation strain number × 100
The sickness rate of the sickness rate-chemicals treatment of prevention effect (%)=blank
Result of the test shows:The sickness rate of blank is 100%;The sickness rate that vinclozolin is processed is 30.6%, preventive effect
For 69.4%;The sickness rate that M-SX103 wettable powder is processed is 28.2%, and preventive effect is 71.8%, can with 50% vinclozolin
WP preventive effect is suitable.
Claims (1)
1. nematicide Sphingobacterium (Sphingobacterium nematocida) M-SX103 bacterial strain is in preparation preventing and treating Nicotiana tabacum L.
Application in bacterial wilt and notoginseng root rot microbial inoculum, the preserving number of this bacterial strain is:CGMCC No.9689.
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CN108004165A (en) * | 2017-12-11 | 2018-05-08 | 云南中烟工业有限责任公司 | The application of one plant of bacterium and its tunning in preventing that cigarette feed liquid is putrid and deteriorated |
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CN108467839B (en) * | 2018-03-12 | 2021-09-28 | 山西大学 | Astragalus root growth-promoting endophyte and growth promoting method and application thereof |
CN108753659B (en) * | 2018-06-21 | 2021-06-08 | 云南大学 | Carbendazim degrading bacterium and application thereof |
CN108841764B (en) * | 2018-07-23 | 2021-07-09 | 云南大学 | Biocontrol bacterium and application thereof |
CN110964675B (en) * | 2020-01-02 | 2021-10-08 | 山东农业大学 | Application of Sphingobacterium montanense in prevention and treatment of parasitic nematode disease |
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CN101032249A (en) * | 2006-03-09 | 2007-09-12 | 文山壮族苗族自治州三七科学技术研究所 | Preparation for protecting panax notoginseng from root rot disease and using method |
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Non-Patent Citations (1)
Title |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108004165A (en) * | 2017-12-11 | 2018-05-08 | 云南中烟工业有限责任公司 | The application of one plant of bacterium and its tunning in preventing that cigarette feed liquid is putrid and deteriorated |
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