Application of Microorganisms to Cure Plant Diseases

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8th Feb 2020 Biology Reference this

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Project Summary:

Overview: Plant disease is widespread in nature and can cause great losses to the agriculture, but the mechanism of plant diseases and what causes a disease in plants are not fully explored yet. it is known that plants can produce antibiotics to defend pathogens. Those antibiotics are controlled by gene sequences. It started in the early 20th century, the study for plants disease-resistance is more than one century. In the previous study, biologist through Cross-breeding successfully cultivated new disease-resistant varieties and greatly increased grain yield, but this method took decades to cultivate disease-resistant varieties. This project is trying to find a method to cultivate disease-resistant plants quickly and conveniently. It could shorten the time greatly and cultivate disease-resistant plant varieties efficiently. In this project, plants disease-resistant gene are extracted and inserted into plant quickly and conveniently. This project can be increasingly supplemented with plant diseases-resistance.

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Statement on Intellectual Merit:If the product from this experiment is stable and efficient, biologist don’t need to take decades to cultivate new disease-resistant varieties, this project not only can be applied in cultivating plants disease-resistant varieties but also available in other scientific fields. Such as Bio-pharmaceutical engineering, environmental engineering, Biological engineering and so on.

Statement on Broader Impacts: This project will influence human life widely. Every year, almost 40% crop reduction of output due to different kinds of plant diseases. If the losses can reduce to 10%, which means the rest food can support 2 billion people to survival. In addition to, the production organic matters and medicines are also produced from plants. More than that this project will benefit other researchers and scientists who are study on transgenic engineering, it can help them find required genes sequences quickly and provide a method to understand how protein can through various signaling pathways to defense pathogenic in the organism.

Project Description:

Section I. Personnel

PI: Hang Lin, Universal of Maryland, Graduate Student

Section II. Project

  1. Objectives:

The first objective is to compare various type of plants which are exposed to the same pathogenic factor or to infection in order to see their effects on those selected plants.

The second goal is to determine defense mechanism, find the gene sequences which can produce antibiotic from uninfected plants.

 

  1. Rationale and Significance:

 

Plant diseases through pests and mass infections (bacterial and virus) are the main causes of reduction and loss of agricultural production. Guest (2015) pointed out that “plant disease reduces the production and quality of food, fiber and biofuel crops. Losses may be catastrophic or chronic, but on average account for 42% of the production of the six most important food crops”. Besides, plant diseases will lower the quality of agricultural products, even produce some material that is harmful to human health. Currently, the chemical pesticides are used widely to control plant diseases. When skin is exposed to chemical pesticides, irritation and burns are the most common situation, the worse problem is that making people sick and some unpredictable healthy problems. Chemical pesticides can kill pests or recover plants efficiently, the disadvantage of chemical pesticides is obviously. For example, chemical pesticide residues not only harmful to human health but also damage the environment by causing soil salinization and alkalinization. It is hard for plants growing in suck kind soil. In addition to, part of the pesticides will remain on the surface of the plants, resulting in contamination in the food, vegetables, fruits and so on. The rest of pesticide will be scattered on the soil or evaporated and dispersed into the air. Through rain, those pesticides will land back to farmland drainage into rivers and lakes, polluting water resources and aquatic life. the livestock might possibly be fed with the plants that have agricultural residues on. Pesticide residues through meat can enter the human body. As a result, chemical pesticides through the air, water, soil, and food can cause various chronic or acute diseases. Furthermore, if the pesticides are used for a long-term that will cause pathogens to produce antibodies. Chemical pesticides could be a problem for human’s health and environment. For the significance of planting disease-resistant crops is very important, especial for farmer, they will contact chemical pesticides often. If there are diseases- resistant  plant, it will protect their health and save money. To increase plant diseases-resistant ability quickly is no time to delay, and some recent study have found a way to increase plant diseases-resistant ability. It was mentioned by (Hyakumachi et al 2014) “From research on utilizing specific antagonistic microorganisms, many effective biological control agents (BCAs) have been found and are increasingly implemented in integrated pest management strategies to control plant diseases”. To find the BCAs and genomes of special plants needs to be sequenced to reveal which genes are involved in defense against to diseases or have resistance against the pesticide in order to control pollution, hence, protect environment indirectly.

Reducing chemical pesticides using and find a method to increase plant diseases-resistant ability have many benefits. For the public community, they want to get healthy food, vegetables and fruits caused it is important to guarantee for their health. For farmers, they want to gain considerable income and do not want to spend extra money on purchasing chemical pesticides, at the same time, not using chemical pesticide also reduce labor fee. It increases farmers income directly. The most important meaning is for environmental protection and ecologically sustainable development. Scientists has spent more than 1 hundred years to improve plant diseases-resistant, although scientific studies are very limited in this field. As John M.McDowell and Bonnie J.Woffenden (2003) referred “Although many exciting insights have emerged from recent research on plant defense signaling, our overall understanding of the process is still fragmentary”. with the development of science and technology, especially with the transgenic technology improving, it is believed that planting disease-resistant plants will be popularized in the future, which is of great significance for solving the global food crisis and environmental protection.

 

3. Research Question and Hypotheses:

Do some plant diseases only occur on certain plants?

1. Hypothesis: Only certain plants will be affected by the pathogenic factors, while other plants will be not infected or recover from the diseases.

Whether some plant diseases can cause more damage to plants that do not have a specific substance?

2. Hypothesis: The specific substance produced from uninfected plants can kill pest and bacterial.

4. Research Design:

Soil select: The biggest risk for the experiment is transferring the diseases-resistant gene into wild weed and other plants (Zapiola et al 2008), the soil in this experiment should contain enough nutrition but without other plants. The soil in this experiment could not contain other pathogenic factors. the soil needs to be heated more than 30 minutes under 100 Celsius degree. (Baker, K.F., 1957.)

Plant select: Pathogenesis-related proteins (PR-protein) is very important when plants get some diseases to defend pathogenic factors, the function of PR-protein express in antimicrobial production and antimicrobial metabolites. There are many effects of PR-proteins, which are capable of degrading fungal cell walls, cell membranes, RNA or participating in the production of metabolites that are resistant to pathogenic bacteria and by enhancing the physical barrier of cells to against diseases. (Wally, O., & Punja, Z. K. 2010). There are a lot of research on PR-protein for different plants, such as potato(Zhu B L, Chen THH, Li PH.1995)rice ( Hou MM et al,2012),and soybean (Xu P et al,2014). In this experiment, lettuce will be the testing object. There are two reasons to choose lettuce. The first reason is lettuce only need one month to grow up until harvest. Planting lettuce is neither too long to observe the experimental results nor too short to cause errors in the experimental results. The second reason is that lettuce has become one of the most important vegetables in America, especially, using in salad and hamburgers.

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RNA extraction and cDNA analysis

 For the lettuce leave tissue tubes need to be separated by couple groups under different PH and temperature. Caused the experiment need to stimulate lettuce growing in different types of environmental situation and testing will PR-protein will be produced under various condition. Trizol kit (Invitrogen, Shanghai, China) is used in this experiment for extracting RNA from lettuce leaves, the mainly RNA extraction step applied are mentioned by the research “A simple and rapid method for RNA isolation from plant tissues with high phenolic compounds and polysaccharides”. (2007). ProtoScript® II First Strand cDNA Synthesis Kit will be used to convert RNA to cDNA according to the manufacturer’s instructions. (Jiang, L et al, 2015)

References Cited:

Project Summary:

Overview: Plant disease is widespread in nature and can cause great losses to the agriculture, but the mechanism of plant diseases and what causes a disease in plants are not fully explored yet. it is known that plants can produce antibiotics to defend pathogens. Those antibiotics are controlled by gene sequences. It started in the early 20th century, the study for plants disease-resistance is more than one century. In the previous study, biologist through Cross-breeding successfully cultivated new disease-resistant varieties and greatly increased grain yield, but this method took decades to cultivate disease-resistant varieties. This project is trying to find a method to cultivate disease-resistant plants quickly and conveniently. It could shorten the time greatly and cultivate disease-resistant plant varieties efficiently. In this project, plants disease-resistant gene are extracted and inserted into plant quickly and conveniently. This project can be increasingly supplemented with plant diseases-resistance.

Statement on Intellectual Merit:If the product from this experiment is stable and efficient, biologist don’t need to take decades to cultivate new disease-resistant varieties, this project not only can be applied in cultivating plants disease-resistant varieties but also available in other scientific fields. Such as Bio-pharmaceutical engineering, environmental engineering, Biological engineering and so on.

Statement on Broader Impacts: This project will influence human life widely. Every year, almost 40% crop reduction of output due to different kinds of plant diseases. If the losses can reduce to 10%, which means the rest food can support 2 billion people to survival. In addition to, the production organic matters and medicines are also produced from plants. More than that this project will benefit other researchers and scientists who are study on transgenic engineering, it can help them find required genes sequences quickly and provide a method to understand how protein can through various signaling pathways to defense pathogenic in the organism.

Project Description:

Section I. Personnel

PI: Hang Lin, Universal of Maryland, Graduate Student

Section II. Project

  1. Objectives:

The first objective is to compare various type of plants which are exposed to the same pathogenic factor or to infection in order to see their effects on those selected plants.

The second goal is to determine defense mechanism, find the gene sequences which can produce antibiotic from uninfected plants.

 

  1. Rationale and Significance:

 

Plant diseases through pests and mass infections (bacterial and virus) are the main causes of reduction and loss of agricultural production. Guest (2015) pointed out that “plant disease reduces the production and quality of food, fiber and biofuel crops. Losses may be catastrophic or chronic, but on average account for 42% of the production of the six most important food crops”. Besides, plant diseases will lower the quality of agricultural products, even produce some material that is harmful to human health. Currently, the chemical pesticides are used widely to control plant diseases. When skin is exposed to chemical pesticides, irritation and burns are the most common situation, the worse problem is that making people sick and some unpredictable healthy problems. Chemical pesticides can kill pests or recover plants efficiently, the disadvantage of chemical pesticides is obviously. For example, chemical pesticide residues not only harmful to human health but also damage the environment by causing soil salinization and alkalinization. It is hard for plants growing in suck kind soil. In addition to, part of the pesticides will remain on the surface of the plants, resulting in contamination in the food, vegetables, fruits and so on. The rest of pesticide will be scattered on the soil or evaporated and dispersed into the air. Through rain, those pesticides will land back to farmland drainage into rivers and lakes, polluting water resources and aquatic life. the livestock might possibly be fed with the plants that have agricultural residues on. Pesticide residues through meat can enter the human body. As a result, chemical pesticides through the air, water, soil, and food can cause various chronic or acute diseases. Furthermore, if the pesticides are used for a long-term that will cause pathogens to produce antibodies. Chemical pesticides could be a problem for human’s health and environment. For the significance of planting disease-resistant crops is very important, especial for farmer, they will contact chemical pesticides often. If there are diseases- resistant  plant, it will protect their health and save money. To increase plant diseases-resistant ability quickly is no time to delay, and some recent study have found a way to increase plant diseases-resistant ability. It was mentioned by (Hyakumachi et al 2014) “From research on utilizing specific antagonistic microorganisms, many effective biological control agents (BCAs) have been found and are increasingly implemented in integrated pest management strategies to control plant diseases”. To find the BCAs and genomes of special plants needs to be sequenced to reveal which genes are involved in defense against to diseases or have resistance against the pesticide in order to control pollution, hence, protect environment indirectly.

Reducing chemical pesticides using and find a method to increase plant diseases-resistant ability have many benefits. For the public community, they want to get healthy food, vegetables and fruits caused it is important to guarantee for their health. For farmers, they want to gain considerable income and do not want to spend extra money on purchasing chemical pesticides, at the same time, not using chemical pesticide also reduce labor fee. It increases farmers income directly. The most important meaning is for environmental protection and ecologically sustainable development. Scientists has spent more than 1 hundred years to improve plant diseases-resistant, although scientific studies are very limited in this field. As John M.McDowell and Bonnie J.Woffenden (2003) referred “Although many exciting insights have emerged from recent research on plant defense signaling, our overall understanding of the process is still fragmentary”. with the development of science and technology, especially with the transgenic technology improving, it is believed that planting disease-resistant plants will be popularized in the future, which is of great significance for solving the global food crisis and environmental protection.

 

3. Research Question and Hypotheses:

Do some plant diseases only occur on certain plants?

1. Hypothesis: Only certain plants will be affected by the pathogenic factors, while other plants will be not infected or recover from the diseases.

Whether some plant diseases can cause more damage to plants that do not have a specific substance?

2. Hypothesis: The specific substance produced from uninfected plants can kill pest and bacterial.

4. Research Design:

Soil select: The biggest risk for the experiment is transferring the diseases-resistant gene into wild weed and other plants (Zapiola et al 2008), the soil in this experiment should contain enough nutrition but without other plants. The soil in this experiment could not contain other pathogenic factors. the soil needs to be heated more than 30 minutes under 100 Celsius degree. (Baker, K.F., 1957.)

Plant select: Pathogenesis-related proteins (PR-protein) is very important when plants get some diseases to defend pathogenic factors, the function of PR-protein express in antimicrobial production and antimicrobial metabolites. There are many effects of PR-proteins, which are capable of degrading fungal cell walls, cell membranes, RNA or participating in the production of metabolites that are resistant to pathogenic bacteria and by enhancing the physical barrier of cells to against diseases. (Wally, O., & Punja, Z. K. 2010). There are a lot of research on PR-protein for different plants, such as potato(Zhu B L, Chen THH, Li PH.1995)rice ( Hou MM et al,2012),and soybean (Xu P et al,2014). In this experiment, lettuce will be the testing object. There are two reasons to choose lettuce. The first reason is lettuce only need one month to grow up until harvest. Planting lettuce is neither too long to observe the experimental results nor too short to cause errors in the experimental results. The second reason is that lettuce has become one of the most important vegetables in America, especially, using in salad and hamburgers.

RNA extraction and cDNA analysis

 For the lettuce leave tissue tubes need to be separated by couple groups under different PH and temperature. Caused the experiment need to stimulate lettuce growing in different types of environmental situation and testing will PR-protein will be produced under various condition. Trizol kit (Invitrogen, Shanghai, China) is used in this experiment for extracting RNA from lettuce leaves, the mainly RNA extraction step applied are mentioned by the research “A simple and rapid method for RNA isolation from plant tissues with high phenolic compounds and polysaccharides”. (2007). ProtoScript® II First Strand cDNA Synthesis Kit will be used to convert RNA to cDNA according to the manufacturer’s instructions. (Jiang, L et al, 2015)

References Cited:

  • Mcdowell, J. M., & Woffenden, B. J. (2003). Plant disease resistance genes: Recent insights and potential applications. Trends in Biotechnology,21(4), 178-183. doi:10.1016/s0167-7799(03)00053-
  • Klein, T., Zihlmann, D., Derlon, N., Isaacson, C., Szivak, I., Weissbrodt, D. G., & Pronk, W. (2016). Biological control of biofilms on membranes by metazoans. Water Research,88, 20-29. doi:10.1016/j.watres.2015.09.050
  • Ortiz-Urquiza, A., Luo, Z., & Keyhani, N. O. (2014). Improving mycoinsecticides for insect biological control. Applied Microbiology and Biotechnology,99(3), 1057-1068. doi:10.1007/s00253-014-6270-x
  • Postma, J. (2009). The Status of Biological Control of Plant Diseases in Soilless Cultivation. Recent Developments in Management of Plant Diseases,133-146. doi:10.1007/978-1-4020-8804-9_11
  • Thiebaut, F., Grativol, C., Hemerly, A. S., & Ferreira, P. C. (2015). MicroRNA Networks in Plant-Microorganism Interactions. Tropical Plant Biology,8(1-2), 40-50. doi:10.1007/s12042-015-9149-9
  • Zapiola, M.L.; Campbell, C.K.; Butler, M.D.; Mallory-Smith, C.A. Escape and establishment of transgenic glyphosate-resistant creeping bentgrass Agrostis stolonifera in Oregon, USA: A 4-year study. J. Appl. Ecol.2008, 45, 486–494
  • Baker, K.F. Editor. 1957. The U.C. System for Producing Healthy Container Grown Plants, Manual 23. University of California, Division of Agricultural Sciences, Agricultural Experiment Station Extension Service.
  • Park C J, Kim K J, Shin R, Park J M, Shin Y C, Paek K H. Pathogenesis-related protein 10 isolated from hot pepper functions as a ribonuclease in an antiviral pathway. Plant J. 2004; 37: 186–198.
  • Xie Y R, Chen Z Y, Brown R L, Bhatnagar D. Expression and functional characterization of two pathogenesis-related protein 10 genes from Zea mays . J. Plant Physiol. 2010; 67: 121–130.
  • Wally, O., & Punja, Z. K. (2010). Genetic engineering for increasing fungal and bacterial disease resistance in crop plants. GM Crops,1(4), 199-206. doi:10.4161/gmcr.1.4.13225
  • Zhu B L, Chen T H H, Li P H. Expression of three osmotin-like protein genes in response to osmotic stress and fungal infection in potato. Plant Mol. Biol. 1995; 28: 17–26.
  • Hou MM, Xu WJ, Bai H, Liu Y M, Li L Y. Characteristic expression of rice pathogenesis-related proteins in rice leaves during interactions with Xanthomonas oryzae pv. Oryzae . Plant Cell Rep. 2012; 31: 895–904. doi: 10.1007/s00299-011-1210-z
  • Xu PF, Jiang LY, Wu JJ, Li WB, Fan SJ, Zhang SZ. Isolation and characterization of a pathogenesis-related protein 10 gene (GmPR10) with induced expression in soybean (Glycine max) during infection with Phytophthora sojae . Mol. Biol. Rep. 2014; 41: 4899–4909. doi: 10.1007/s11033-014-3356-6 
  • Jiang, L., Wu, J., Fan, S., Li, W., Dong, L., Cheng, Q., . . . Zhang, S. (2015). Isolation and Characterization of a Novel Pathogenesis-Related Protein Gene (GmPRP) with Induced Expression in Soybean (Glycine max) during Infection with Phytophthora sojae. Plos One,10(6). doi:10.1371/journal.pone.0129932

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