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miRNA396 AND GROWTH REGULATING FACTORS FOR CYST NEMATODE TOLERANCE IN PLANTS

a growth regulation factor and nematode technology, applied in the field ofmirna396, can solve the problems of poor understanding of the regulatory networks governing gene expression patterns in nematode-infected roots and particularly in the developing syncytium, difficult application, and high cost, so as to improve the performance and increase the root mass. , the effect of increasing the survival rate of the plan

Inactive Publication Date: 2012-11-01
IOWA STATE UNIV RES FOUND
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Benefits of technology

[0014]The present invention includes methods to alter the genetic composition of crop plants, particularly those that are susceptible to nematode infection, thereby improving tolerance to nematode infection and reducing the effects thereof in plants. This invention provides methods and compositions for modulating key pathways involved in the syncytial event of nematode infection and for preventing the cascade of differential gene expression caused by the same. Applicants have found that the microRNA miR396 acts as a master switch of syncytial gene expression changes in plants after infection, and further that miR396 and growth regulating transcription factors (GRF) with miRNA396 binding sites are connected through a negative feedback loop to establish an irreversible plant gene regulatory switch from syncytium initiation and maintenance.
[0027]In yet another embodiment, the present invention is directed to a transgenic plant or plant cells with improved performance under nematode infecting conditions, containing the nucleic acids described herein. Preferred plants containing the polynucleotides of the present invention include but are not limited to soybean, sunflower, maize, sorghum, canola, wheat, alfalfa, cotton, oat, rice, barley, tomato, cacao and millet. In another embodiment, the transgenic plant is a soybean plant or plant cells. Plants produced according to the invention can have at least one of the following phenotypes in nematode infecting conditions as compared to a non-modified control plant, including but not limited to: increased root mass, increased plant survival, increased root length, increased leaf size, increased ear size, increased seed size, absence of syncytia, smaller or decreased syncytia, or increased plant size when compared to a non-modified plant under conditions of nematode infection.

Problems solved by technology

Regulatory networks governing gene expression patterns in nematode-infected roots and particularly in the developing syncytium are very poorly understood.
However, the majority of the fumigant-type nematicides is no longer available and is also costly and difficult to apply properly under the prevailing conditions.
Unfortunately, these non-host crops are often less valuable.
Using cover crops, however, can be costly because the cover crops occupy space that could be used to grow more valuable crops.
Commercially available preparations of biological control organisms are limited in their use to regions that can support the growth of the control organism.
Moreover, the outcome of using one organism to control another is unpredictable and subject to a variety of factors such as weather and climate.

Method used

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  • miRNA396 AND GROWTH REGULATING FACTORS FOR CYST NEMATODE TOLERANCE IN PLANTS
  • miRNA396 AND GROWTH REGULATING FACTORS FOR CYST NEMATODE TOLERANCE IN PLANTS
  • miRNA396 AND GROWTH REGULATING FACTORS FOR CYST NEMATODE TOLERANCE IN PLANTS

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example 1

[0221]Pathogens alter their hosts' biology to ensure successful infection. Such modifications range from moderate to extensive, and in the case of plant pathogens, few infections result in more dramatic changes than those of sedentary endoparasitic nematodes, which include the cyst nematodes (Heterodera spp.). Maybe rivaled in complexity only by plant interactions with Agrobacterium and Rhizobia, cyst nematodes are obligate parasitic roundworms that induce the formation of novel plant cell types that are associated in a unique feeding organ, the syncytium.

[0222]Cyst nematodes infect as second-stage juveniles (J2), which initiate the induction / formation of the syncytium. During this phase, J2s begin feeding on the growing syncytium and then develop into third-stage (J3) and fourth-stage juveniles (J4) followed by the adult stage. Syncytium development can be separated into an induction / formation phase followed by a maintenance phase. Induction / formation involves effector-mediated com...

example 2

miR396 in Soybean During Cyst Nematode Infection

[0302]In order to understand the role that miR396 plays during soybean infection by the soybean cyst nematode (Heterodera glycines; SCN), expression analyses were performed on primary and mature sequences for all miR396 paralogs (miR396a, miR396b, miR396c and miR396e) and seven of its predicted target Growth Regulating Transcription Factors (GRF8, GRF9, GRF12, GRF13, GRF15, GRF16 and GRF19) using quantitative real-time PCR (qRT-PCR). Soybean seedlings were infected with SCN three days after germination and RNA was extracted 2, 4, 8 and 14 days post inoculation (dpi). RNA from both SCN infected and mock inoculated soybean seedlings was reverse-transcribed into cDNA for qRT-PCR.

[0303]Data were analyzed using the comparative Ct method with U6 snRNA as the reference gene for microRNA quantification and ubiquitin for GRFs. Significance tests were performed using the Student's t-test (p-value<0.05) and significant values are indicated on the...

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Abstract

The present invention presents methods to alter the genetic composition of crop plants susceptible to nematode infection to improve tolerance to the same. Methods and compositions for modulating key pathways involved in the syncytial event of nematode infection and for preventing the cascade of differential gene expression caused by the same as disclosed. Applicants have found that the microRNA miR396 acts as a master switch of syncytial gene expression changes in plants after infection, and further that miR396 and certain growth regulating transcription factors (GRF) are connected through feedback interaction in syncytium initiation and maintenance.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS[0001]This application claims priority under 35 U.S.C. §119 to provisional application Ser. No. 61 / 480,093 filed Apr. 28, 2011, herein incorporated by reference in its entirety.GRANT REFERENCE[0002]This invention was made with government support under Contract No. 2008-35302-18824 awarded by USDA. The government has certain rights in the invention.FIELD OF THE INVENTION[0003]The invention relates generally to the field of plant molecular biology.BACKGROUND OF THE INVENTION[0004]Nematodes are a very large group of invertebrate animals generally referred to as roundworms, threadworms, eelworms, or nemas. Some nematodes are plant parasites and can feed on stems, buds, leaves, and in particular on roots. Cyst nematodes (principally Heterodera and Globodera spp.) are key pests of major crops. Cyst nematodes are known to infect tobacco, cereals, sugar beets, potato, rice, corn, soybeans and many other crops. Heterodera schachtii principally attacks s...

Claims

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Application Information

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IPC IPC(8): A01H1/02C12N15/82A01H5/00A01H5/10C12Q1/68A01H1/00
CPCC12N15/8285C12N15/8218Y02A40/146
Inventor BAUM, THOMAS J.HEWEZI, TAREK ABDEL FATTAH
Owner IOWA STATE UNIV RES FOUND
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