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Setosphaeria turcica attenuated strain STAM-226 and application thereof

A technology of Zea mays, CGMCCNO.20269, applied in the field of phytopathology

Inactive Publication Date: 2021-01-22
TANGSHAN NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no relevant research on the use of attenuated protection of attenuated strains for the prevention and treatment of maize leaf spot

Method used

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  • Setosphaeria turcica attenuated strain STAM-226 and application thereof
  • Setosphaeria turcica attenuated strain STAM-226 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Preparation of attenuated strain of maize leaf spot fungus:

[0030] Pick a single colony of Agrobacterium (containing the pBHtl plasmid) and place it in LB liquid medium, shake it at 28°C at 120rpm for 48h, take 1mL of the bacterial liquid and centrifuge at 4°C at 10,000rpm for 1min, and discard the supernatant. Wash twice with induction medium (IM), and dilute to OD600≈0.15 with IM medium, add AS (acetophthalate syringone) with a final concentration of 200-400 μM, and activate at 28° C. for 7 hours at 180 rpm. Take 1 mL of spore suspension in a 40 mL centrifuge tube, add 3 mL of Fries liquid medium, culture at 25°C for 24 h with vibration at 100 rpm, and add 1 mL of activated Agrobacterium for co-culture for 48 h. Take 500 μL of the co-culture of Rhizoma spp. and Agrobacterium, smear it on the PDA medium containing hygromycin B (50 μg / mL), cefotaxime sodium and carbenicillin (each 200 μg / mL), and culture at 25 °C for 1 about a week. A single colony was picked and cu...

Embodiment 2

[0036] Embodiment 2 screens the attenuated bacterial strain from the ATMT mutant library of Leptosphaeria maize:

[0037] Add a small amount of sterile water to each mutant strain cultivated on the PDA medium for 10 days, scrape gently with a sterilized glass slide, then filter with 2 layers of gauze to remove the fragments of the medium, and make a spore suspension (10 5 spores / ml). When the corn seedlings grow to 4-6 leaves, the prepared spore suspensions are evenly sprayed on the leaves of the susceptible corn inbred line OH43 for friction inoculation. After inoculation, the film should be kept moist for 24 hours, and after removing the moisturizing film, it should be managed according to the conventional method, and the indoor temperature should be controlled between 20 and 30°C. The incidence investigation was carried out 2 weeks after the inoculation, and the attenuated and virulent strains of P. maize were determined. The results showed that maize leaf spot bacteria S...

Embodiment 3

[0039] Analysis of the cross-protective effect of attenuated corn leaf spot disease:

[0040] (1) Corn inoculation methods in different periods

[0041] The corn seedling stage (7-8 leaf stage) is inoculated with acupuncture: the needle point of the micro-syringe is used to make wounds. The acupuncture points are required to be compact. The 4 wounds are diamond-shaped, and the wounds cannot penetrate the leaves. Then, the 7d-cultivated plate (diameter 1.4 cm) of Leptosphaeria maize was covered on the micro-wound with the mycelium surface contacting the corn leaf, and fixed with a pin.

[0042] Inoculate with sorghum grains at the maize tassering stage: Inoculate 3 plates (diameter 1.4 cm) of S. maize rot bacteria cultured for 7 days into autoclaved sorghum grains (100 g sorghum grains + 100 g water), and store in the dark at 23°C to 25°C. After culturing for 5 days, shake the sorghum grain medium evenly to break the hyphae, and then culture in the dark at 23°C to 25°C for 5 day...

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Abstract

The invention relates to phytopathology, in particular to a setosphaeria turcica attenuated strain STAM-226 and application thereof, and particularly relates to the setosphaeria turcica attenuated strain and application thereof in prevention and treatment of northern leaf blight of corn. Through ATMT conversion, screening, identification, pot culture experiments and field experiments, a setosphaeria turcica attenuated strain with a significant control effect on the northern leaf blight of corn is obtained, and the preservation number is CGMCC NO.20269. Test results show that the setosphaeria turcica attenuated strain is firstly inoculated, the setosphaeria turcica virulent strain STAM-YC is inoculated after 1-3 days, and a remarkable mild strain cross protection (MSCP) effect is shown in each growth period. The STAM-YC is inoculated after the setosphaeria turcica attenuated strain is inoculated for 2 d, so that the prevention and control effect on the northern leaf blight of corn is most remarkable, reaching 89.6%. The setosphaeria turcica attenuated strain for preventing and treating the northern leaf blight of corn is safe to people, livestock, insects and crops, and is environmentally friendly.

Description

Technical field: [0001] The invention relates to phytopathology, in particular to an attenuated bacterium of maize leaf spot (STAM-226) and an application thereof, in particular to an attenuated bacterium of maize leaf spot and its application in the prevention and treatment of maize leaf spot. Background technique: [0002] In my country, maize spot disease is mainly distributed in the north and high-altitude mountainous areas, which directly affects the yield and quality of maize and causes serious economic losses. Bacteria maize belongs to Ascomycota subphylum Thinomycetes. Pseudomonas maize is a filamentous fungus that has been found to mutate rapidly so far. Aiming at the resistance gene in maize, it will quickly produce a virulence gene that can overcome it, thus forming a new physiological race. In recent years, due to the continuous emergence of new races, maize leaf spot is still an important disease in production, and it gradually spreads from northeast to north C...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14A01N63/30A01P3/00C12R1/645
CPCA01N63/30C12R2001/645C12N1/145
Inventor 张淑红张运峰武秋颖高凤菊石洪凌李艳梅范永山
Owner TANGSHAN NORMAL UNIV
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