Highly siderophore-producing bacterial mixture, production method thereof and use of same as a phytopathogen inhibitor 机翻标题: 暂无翻译,请尝试点击翻译按钮。

源语言标题
(WO2017105209) Mezcla bacteriana altamente productora de sideroforos, su proceso de obtención y su uso como agente inhibidor de fitopatogenos
公开号/公开日
WO2017105209 A2 2017-06-22 [WO2017105209] WO2017105209 A3 2017-11-09 [WO2017105209] / 2017-06-22 2017-11-09
申请号/申请日
2016WO-MX00128 / 2016-12-07
发明人
ARTEAGA GARIBAY RAMÓN IGNACIO
申请人
INSTITUTO NAC DE INVESTIGACIONES FORESTALES AGRÍCOLAS & PECUARIAS
主分类号
IPC分类号
A01N-063/00;A01P-003/00;A01P-021/00;C12N-001/20;C12P-001/04;C12R-001/38;C12R-001/425
摘要
(WO2017105209)
The present invention relates to a mixed culture of bacterial strains which have antagonist activity for pathogenic fungi in agriculturally important crops and which produce siderophores. 
The subject matter of the invention is a mixed bacterial culture isolated from maize rhizosphere consisting of a suspension of cells of the micro-organisms. 
The method for the isolation and production of the microbial mixture forms the subject matter of the present invention; hence, it is considered to be an invention in the field of biotechnology.
机翻摘要
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地址
代理人
代理机构
优先权号
2015MX-PA17262 2015-12-14
主权利要求
(WO2017105209)
1. A process for production of compounds bacterial strains () producer of siderophores highly characterized because it includes the following interstages:
. A. SHOWS PREPARATION. 25 Soil samples are taken from the area of rhizosphere a plot for the corn crop; One mixes flexible containers inside. 10 Are weighted g of the mixture and flooring they realized at least one dilution with a connection 1: 10 (1x10 (' 1) to 1x 10(7)) in physiological salt solution NaCI (0,85%). 100 Will be transferred aliquots of Μι- box dilutions Petri dish with average Pikovskaya and CAS for straightforward selecting of producers of siderophores; Also the mediums of gelosa nutritional, arginine are used glycerol, B King, for selecting of specific microbial groups. The inoculated growth media are incubated between 25 and 47 ° C by 18 to 64 h;
. B. REACTIVATION OF STRAINS. Strains of mediums Pikovskaya and CAS are selected that generated a halo orange, indicator of the production of siderophores. The solid CAS strains are revived in the middle to corroborate the production of siderophores, adjust bacterial suspensions in 5 ml physiological salt solution tubing to 0,5 in the scale of McFarland. 1 Ml of bacterial suspension is transferred and 100 ml succinic, composed acid medium inoculated by: 6,0 K2HPO4 KH2PO4 g/l,
3.
3. G/l, MgSO
0,2 g/l, (2 NH
)
to
1,0 g/l and 4,0 g/l succinic acid pH, 7.0; They are incubated at 28 ºc in an orbital shaker, 120 rpm by 24-30 h. So to get the siderophores, 10 ml of inoculated medium are centrifuge to 4696 g per 10 minutes;
. C. ISOLATION AND SEPARATION OF STRAINS. Isolations other mediums with macroscopic morphology are selected and microscopic various strains, in the same medium purify where they were insulated. The pure isolations reseal in growth media with substrates as Gelosa jelly, Gelosa Milk Gelosa, starch, Gelosa Baird- Parker, Broth nitrate, Gelosa Acetate of Fowller Gelosa, Victoria, Blue Gelosa colloidal chitin, Gelosa with lignin, among others, and assess metabolic capacities of isolations; And
. D. CRYO-PRESERVATION OF STRAINS. The characterized strains conserve long run in cryo-preservation. It is prepared I inoculate from crop in late logarithmic growth phase and previous to the stationary phase (18 to 48 h) characterized strains, it is adjusted I inoculate in SSF to turbidity 7 nephelometer of McFarland (2.1x10・ UFC/mL). Criotubo with 0,5 ml of skimmed milk to sterile 20% in an ice bath is opened and they will transfer 0,5 ml dei I inoculate adapted to tubing 7 nephelometer, labelled and cryoprotective by 30 was equilibrated min in an ice bath, then lays the tubing in a freezer at -20 ° C for 24 h, is stored the tubing in a deep freezer -80 ° C.
2.
2. A referred strain Pseudomonas spp. CM-CNRG 186, characterised as it was achieved through preparation process of claim;
3.
3. A referred strain Serratia spp. CM-CNRG 199, characterised as it was achieved through preparation process of claim;
4.
4. A referred strain Pseudomonas spp. CM-CNRG 181, characterised as it was achieved through preparation process of claim;
5.
5. A referred strain Pseudomonas spp. CM-CNRG 200, characterised as it was achieved through preparation process of claim;
6.
6. A referred strain Pseudomonas palleronian CM-CNRG 172, characterised as it was achieved through preparation process of claim;
7.
7. A referred strain Pseudomonas extremorientalis CM-CNRG 144, characterised as it was achieved through preparation process of claim;
8.
8. A referred strain Serratia grimesii CM-CNRG 197, characterised by preparation process of claim was achieved way dei;
9.
9. The compound of bacterial biomass where microorganisms were isolated, selected, preserved and deposited according to claims two, three, four, five, six and seven characterized it gets because of a process that includes the isolation cough compatible bacterial morphotypes in selective growth media, average B Arginine King, glycerol, Czapeck gelosa medium, nutritional, 37 a temperature of 35 ° C to ° C for 18 to 48 hours in aerobic conditions and dark;
10.
10. The compound of bacterial biomass of characterized claim nine because the isolation process the compatible bacterial morphotypes in selective growth media, average B Arginine King, glycerol, Czapeck gelosa medium, nutritional, to a temperature of 35 ° C to 37 ° C for 18 to 48 hours in aerobic conditions and dark, is claim one.
11.
11. A procedure for selecting the bacterial morphotypes compatible with the genus Pseudomonas and Serratia, is based on the established in the manual of systematic taxonomy Bergey for features of colonial and microscopic morphology, in addition to the sowing of strains in selective growth media claim 1 for obtainment of crops axenic;
12.
12. The compounds of claims ten and eleven done in mediums of crops formulated in characterized laboratory because it demonstrates the production capacity of siderophores and compounds with antimicrobial activity and its affixation of atmospheric nitrogen:
13.
13. The use of characterized compounds of claim twelve because they preferably take activity as inhibiting agent of phytopathogenic in crops with agricultural importance.
法律状态
(WO2017105209)
LEGAL DETAILS FOR WO2017105209

Actual or expected expiration date=2019-06-14   
Legal state=ALIVE   
Status=PENDING 



Event publication date=2016-12-07 
Event code=WO/APP 
Event indicator=Pos 
Event type=Examination events 
Application details

Application country=WO WOMX2016000128 
Application date=2016-12-07 
Standardized application number=2016WO-MX00128 



Event publication date=2017-06-22 
Event code=WO/A2 
Event type=Examination events 
International application published with declaration under Article 17 (2) (a)

Publication country=WO 
Publication number=WO2017105209 
Publication stage Code=A2 
Publication date=2017-06-22 
Standardized publication number=WO2017105209 



Event publication date=2017-11-09 
Event code=WO/A3 
Event indicator=Pos 
Event type=Examination events 
Later publication of ISR with revised front page

Publication country=WO 
Publication number=WO2017105209 
Publication stage Code=A3 
Publication date=2017-11-09 
Standardized publication number=WO2017105209 
LEGAL DETAILS FOR DESIGNATED STATE EP

Actual or expected expiration date=2036-12-07   
Legal state=ALIVE   
Status=PENDING 

Corresponding cc: 
Designated or member state=EP Corresponding appl: EP16876106



Event publication date=2017-08-23 
Event code=WO/121 
Event type=Designated states 
EP: The EPO has been informed by wipo that ep was designated in this application
Corresponding cc: 
Designated or member state=EP
专利类型码
A2 A3
国别省市代码
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