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Complex microbial inoculant for promoting breeding of potato test-tube plantlets and preparation and use methods of complex microbial inoculant

A technology of compound inoculants and test-tube seedlings, which can be applied in the fields of botanical equipment and methods, biochemical equipment and methods, and microorganism-based methods, etc., can solve the problems of inability to obtain nutrients, prolong the breeding cycle, and cannot effectively transport nutrients, and achieve The effect of improving reproduction quantity and reproduction speed, promoting survival rate and growth, and improving the survival rate of transplanted seedlings

Active Publication Date: 2020-04-28
INNER MONGOLIA AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, repeated subcultures in the process of in vitro shoot apex culture or stem segment expansion often lead to poor development of test-tube plantlets such as slow growth, thin stems, and underdeveloped roots. Plants do not grow, leaves fall off or even die, which eventually leads to low seed potato yield, poor quality, and increased production costs
This is mainly because the root system of test-tube seedlings is not well developed, the stems are thin, and the seedlings are weak, which cannot compensate for the rapid loss of water from the stomata, cannot effectively obtain nutrients from the soil, and cannot effectively transport nutrients, etc.

Method used

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  • Complex microbial inoculant for promoting breeding of potato test-tube plantlets and preparation and use methods of complex microbial inoculant
  • Complex microbial inoculant for promoting breeding of potato test-tube plantlets and preparation and use methods of complex microbial inoculant
  • Complex microbial inoculant for promoting breeding of potato test-tube plantlets and preparation and use methods of complex microbial inoculant

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Embodiment 1: strain isolation and identification

[0041] Collect the roots of potato seedlings in the field, and separate the root-related (root surface and endophytic) plant growth-promoting traits such as nitrogen fixation, phosphorus solubilization, phytohormone production, polysaccharide production, siderophore production and ACC deaminase production by coating and streaking technology Strain function: use the isolated and purified genomic DNA as a template, use 1492R and 27F as primers, PCR amplify the 16S rRNA gene of the strain, sequence the amplified product, and the obtained sequence is classified by online EzTaxon (16S rRNA gene sequence similarity The 16S rRNA gene sequences of the obtained 180 strains of bacteria were determined to be the same species initially, and the similarity between the 16S rRNA gene sequences of the obtained 180 strains and the type species was 98%-100%. The results are shown in Table 1, which is the similarity analysis of the 16S r...

Embodiment 2

[0044] Embodiment 2: the influence of single bacterium on the growth of potato test-tube plantlets

[0045] Activation and cultivation of strains with strong plant growth-promoting ability: absorb 100 μL of glycerin preservation solution and spread it on the modified R2A solid medium, culture it at 25°C, pick a single colony with typical colony characteristics and grow rapidly, and place it in the modified R2A Liquid culture medium, cultivated at 25°C; centrifuge at 8000g centrifugal force when culturing to the middle and late logarithmic phase, and use K 2 HPO 4 -KH 2 PO 4 The buffer solution (pH7.2) was repeatedly washed 3 times, and then resuspended with the same buffer solution until the concentration of effective viable bacteria was 10. 12 CFU / mL.

[0046] Mix each suspended bacterial liquid with MS medium whose temperature has been lowered to about 50°C-55°C after high-temperature and high-pressure sterilization, shake gently to mix evenly, and quickly pour it into a...

Embodiment 3

[0053] Embodiment 3: preparation of composite bacterial agent

[0054] When inoculated alone, L10, FN14 and B28 had the highest increase in root length, stem diameter and node number of potato plantlets respectively, while other bacteria had no obvious improvement effect, or even inhibited them. Therefore, L10, FN14 and B28 were selected as the compound strains. The plate confrontation test results showed that there was no obvious mutual antagonism among the three strains, which could be used for compounding. The promotion effect after compounding with different ratios of effective viable bacteria (Table 3) was used to determine the optimal initial effective viable bacteria quantity of different strains. Table 3 shows the effect of different effective viable bacteria numbers on the growth of the bacterial strains in the compound microbial agent.

[0055] table 3:

[0056]

[0057]

[0058] Note: P<0.05.

[0059] When L10, FN14 and B28 were single-inoculated, the incr...

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Abstract

The invention provides a complex microbial inoculant for promoting breeding of potato test-tube plantlets and preparation and use methods of the complex microbial inoculant. Functional strains of thecomplex microbial inoculant comprise Fitibacillus halophilus, Bacillus gobiensis and Hymenobcter jeollabukensis, and the effective viable count ratio of the three strains is (1-3):(2-5):(2-5). The complex microbial inoculant is inoculated to potato test-tube plantlets, can significantly increase node number, root length and stem diameter of the test-tube plantlets by about 60%-80%, significantly improves the propagation quantity and propagation speed of the test-tube plantlets, significantly promotes the survival rate and growth of transplanted plantlets, increases the number of tubers, increases the volume and weight of a single tuber, enhances the resistance of seedlings to soil-borne diseases, and has the adversity potentials of resisting soil-borne diseases such as late blight and scaband saline-alkali stress.

Description

technical field [0001] The invention belongs to the technical field of microbial resource utilization and ecological agriculture, and relates to a composite microbial agent for promoting the breeding of potato test-tube seedlings and a preparation and use method thereof, which are suitable for the breeding of potato test-tube seedlings. Background technique [0002] Potato is a dual-purpose crop in the world, and it is the fourth largest staple crop after rice, wheat and corn. In potato production, disease is the most important factor affecting potato yield and quality, and many diseases can be carried and transmitted by seed potatoes (including cut tubers). Industrialization and large-scale breeding of seed potatoes through test-tube seedlings can prevent seed potatoes from carrying viruses, reduce disease transmission, and promote the development of the potato industry. [0003] Potato test-tube plantlets (rapid propagation seedlings) adopt in vitro shoot tip culture, fol...

Claims

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Application Information

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IPC IPC(8): C12N1/20A01H4/00C12R1/07C12R1/01
CPCC12N1/20A01H4/008A01H4/001C12R2001/07C12N1/205C12R2001/01
Inventor 唐凯李蘅路晓培杨杉杉冯福应袁博云欣悦郭慧玲
Owner INNER MONGOLIA AGRICULTURAL UNIVERSITY
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