Knowledge Resource Center for Ecological Environment in Arid Area
| 项目编号 | 1019972 |
| Identification, characterization and utilization of microbes from the rhizosphere microbiome of Native plants in Utah to develop more resilient crops for salt and drought stress. | |
| Kaundal, Amita | |
| 主持机构 | SAES - UTAH STATE UNIVERSITY |
| 开始日期 | 2019 |
| 结束日期 | 2024 |
| 资助机构 | US-NIFA（美国食品与农业研究所） |
| 语种 | 英语 |
| 国家 | 美国 |
| 中文简介 | 1103 - Other microbiology |
| 英文简介 | Goals / Objectives Plants are sessile organisms bound to their place of origin. They cannot move to protect themselves from harsh environmental conditions. Climate change increases the incidences of extreme and harsh environments (Allen et al. 2010). The climate change refers to the rising of global temperature, elevation of carbon dioxide in the environment, change in precipitation etc. All these changes increases the incidence of drought, soil salinity, flooding, weeds and pest and disease susceptibility, which directly impacts the growth and development of crops and agriculture as a whole (FAO 2009). By 2100, the global mean surface temperature is expected to rise between 1.8 C - 4 C. These projected changes are expected to impact the growth and production of crops (de Gorter, Drabik, and Just 2013). It is a challenge to mitigate the effects of climate change and at the same time need to adapt to its consequences (FAO 2009). The other factor impacting negatively the agriculture and food production is increase in world population. The world population increased from 4.4 billion to 6.1 billion from 1980-2000. After 2000, the world population is increasing by the rate of 2% each year and reached to 7.5 billion in 2014. By 2050, it is estimated to reach 9 billion and the food demand is expected to increase 70-85% (FAO 2017, 2015; Ray et al. 2013; The World Bank, Population 2016). Various studies have shown that each degree Celsius increase in temperature could lead to 5% reduction of yield (Lobell, Schlenker, and Costa-Roberts 2011). Thus, feeding the increase in population under climate change is a challenge. This challenge demands to create more resilient crops to adverse conditions and feed the growing population. Plants have developed various mechanisms to defend themselves to the extreme conditions in which they are native, or they are exposed. One way to develop more resilient crops is the dissection of these mechanisms and exploit them to develop genetically modified crops. Besides the genetic mechanism, plants have a rhizosphere microbiome that helps them to establish and survive in their habitat. The soil directly under the influence of plant's root is known as rhizosphere. This rhizosphere is rich habitat of various microbes which directly or indirectly influence the growth and development of plants above ground. Many plant growth promoting bacteria (PGPB) have been characterized and few of them are used as successful biofertilizers. The Intermountain West (IW) region in US is rich in drought tolerant native plants that are recommended to use in low water landscaping. These plants have not been explored for their rhizosphere microbiome. Investigation of the rhizosphere microbiome of these plants can lead to open new avenues such as their use in the development of more resilient agriculture under various adverse conditions and enhance the knowledge about the potential of these plants and their microbiome to mitigate the effects of climate change.Objectives:1. Comparative study of microbiome from bulk soil, rhizosphere and endophytes in native plants Cercocarpus ledifolius (curl leaf mountain mahogany), Sheferdia rotundifolia (roundleaf buffaloberry) Sherferdia argentea (silver buffaloberry), Sphaeralcea globemallow, Ceanothus velutinus (snowbrush) from two different locations • Native location Goals / landscape for buffaloberry • drought Goals / non-drought for mahogany and snowbrush • salt Goals / non-salt for globe mallow2. Identification of novel or enriched microbes from above comparative studies.3. Isolation of identified microbes if possible, otherwise use the soil samples enriched with microbiome4. Testing and identification of microbes on native plants at landscape for Buffaloberry and on Arabidopsis and Medicago truncatula in green house for drought tolerance and salt stress.5. Transfer and testing of identified microbes on spinach, rice, maize, alfalfa and strawberry in green house and eventually in the field.Target dates:Objective 1 Collect soil and root samples of mountain mahogany and snowbrush ceanothus plants in wild populations (2019) Collect soil and root samples of round Buffaloberry and silver buffaloberry in wild populations and landscape (2020) Collect soil and root samples of Globemallow plants from wild and salt affected locations. (2021) DNA extraction, Library generation, Sequencing, and Bioinformatics analysis. (2019, 2020, 2021, 2022, 2023)Objective 2 Comparative study to identify the microbes and publication of research and file for patents if applicable (2020,2021,2022,2023) Objective 3 Isolation of identified microbes from respective rhizosphere or roots if possible, otherwise use the soil samples enriched with microbiome (2020,2021,2022,2023)Objective 4 Testing and identification of microbes on native plants at landscape for Buffaloberry and on Arabidopsis thaliana and Medicago truncatula in green house for biomass, drought tolerance and salt stress. (2020,2021,2022,2023) Objective 5 Testing of identified microbes on spinach, rice, maize, alfalfa and strawberry in green house and eventually in field. (2021,2022,2023) Project Methods Methods1. Sampling of native plants in Utah Samples will be collected for microbiome study from native plants Cercocarpus ledifolius (curl leaf mountain mahogany), Sheferdia rotundifolia (roundleaf buffaloberry) Sherferdia argentea (silver buffaloberry), Sphaeralcea globemallow, Ceanothus velutinus (snowbrush) from different locations and as mentioned elsewhere (Sriladda et al. 2012, 2014). Samples for one native plant will be collected each year. From each location, three different types of samples from 10 plants will be collected; 1) bulk soil sample, 2) rhizosphere soil sample, 3) root sample for endophytes. Geological location from each sampling will be recorded for future reference.2. DNA extraction from all samplesDNA from the soil samples will be extracted using Dneasy PowerSoil Kit (Qiagen) and for the roots samples, DNA will be extorted using Dneasy PowerPlant Pro Kit (Qiagen) (McPherson et al. 2018) and quantified.3. Library preparation and shotgun metagenomics sequencingLibrary preparation and shotgun metagenomics sequencing will be done at the Genomics Core facility of the Center for Integrated BioSystems (CIB) at the Utah State University.4. Bioinformatics analysis and comparative study of rhizosphere microbiomeObtained sequencing data will be analyzed at the Bioinformatics Core Facility of the Center for Integrated BioSystems (CIB) at Utah State University.List of microbes will be developed from the comparative metagenomic study of native plants based on different parameters and potential microbes will be chosen for further isolation and evaluation.5. Isolation of identified microbesSoil contains over one million of bacteria per gram of soil. Dilution and spread plating will be used to isolate and purify selected bacteria from different samples. R2A agar media will be used to grow and isolate mixture of heterotrophic and autotrophic bacteria and Glycerol-casein agar media will be used to isolate actinomycetes according to protocol described elsewhere (Schmitz et al. 2015). Selective media and specific protocols for specific selected microorganisms will be used to isolate and culture. Endophytes will be isolated by following protocol and media described (Yaish, Antony, and Glick 2015). Frankia will be isolated from the root nodules by following the protocol described in (Benson 1982). Isolation of nodule associated bacteria will be done by protocol described (Koli and Swarnalakshmi 2017). The fungal endophytes will be cultured and isolated by following method (Wolters et al. 2006). The cultured microbes will then be identified by extracting DNA and sequencing 16SrRNA genes. If a microorganism is found to be not culturable, then the rhizosphere and bulk soil from the native plants from its original location will be used as inoculum in testing the model plants as well as mentioned crops in the proposal. All experiments will be done in project leader's lab at USU.6. Evaluation and identification of microbes on native plants at landscape for Buffaloberry and on Arabidopsis thaliana and Medicago truncatula in green house for drought tolerance and salt stressThe identified and selected microbes from above objectives will be used to test on native and model plants. The selected microbes from Buffaloberry plants will be tested on the roundleaf buffaloberry and silver buffaloberry in the greenhouse to enhance the propagation and survival of these plants. The seedlings or rooted cutting of roundleaf buffaloberry and silver buffaloberry will be developed in green house as per recommend protocols (Rupp and Wheaton 2014). The roots of the plants will be cleaned and inoculated by dipping in the microbial suspensions. The inoculated (Treatment) and uninoculated (control) plants will then be transferred into potting mix and observed for biomass and survival. On regular intervals of one-week, samples will be collected from inoculated plants and screened for the bacterial count by serial dilution. Further, the identified and selected microbes from all of the native plants will be tested on model plant Arabidopsis thaliana and Medicago truncatula with drought and salt stress. Plants will be germinated in greenhouse and seedlings be cleaned and inoculated with bacterial suspension or fungal suspension depending upon the mode of infection of selected microbe. The inoculated (Treatment) and uninoculated plants (control) will be transferred in soil and allowed to grow. The inoculum will be tested for colonization and enrichment every week. For drought and salinity experiments, the drought and salt treatment will be imposed after 2 weeks or colonization of microbes (varies for each microbe depending upon their nature). Different ECs ranging from 5 dS m−1- 20 dS m−1 will be used for salt tolerance. The plants after drought treatment will be evaluated for electrolyte leakage, biomass, relative water contents, water loss, stomatal conductance and survival, Gene Expression for drought induced components such as ABA biosynthesis pathway and stomatal regulation pathway etc. The plants after salt stress will be evaluated for regular intervals for electrolyte leakage, biomass, relative water contents, ion contents, net photosynthetic rate, stomatal conductance, survival and gene expression for salt induced components such as HKT1, SOS pathway and NHX family genes. All experiments will be conducted in the green house facilities at USU.7. Evaluation of identified microbes on spinach, rice, maize, alfalfa and strawberry in green house and eventually in fieldThe identified microbes from above studies will be tested on different crops for drought and salt stress. Collaborations will be developed with different experts in drought and salt stress in all over the US and overseas to test these microbes on different crops in field and greenhouse studies. |
| 英文关键词 | rhizosphere microbiome native plants abiotic stress drought salinity |
| 来源学科分类 | 1103 - Other microbiology |
| 资源类型 | 项目 |
| 条目标识符 | http://119.78.100.177/qdio/handle/2XILL650/356130 |
| 推荐引用方式 GB/T 7714 | Kaundal, Amita.Identification, characterization and utilization of microbes from the rhizosphere microbiome of Native plants in Utah to develop more resilient crops for salt and drought stress..2019. |
| 条目包含的文件 | 条目无相关文件。 | |||||
| 个性服务 |
| 推荐该条目 |
| 保存到收藏夹 |
| 导出为Endnote文件 |
| 谷歌学术 |
| 谷歌学术中相似的文章 |
| [Kaundal, Amita]的文章 |
| 百度学术 |
| 百度学术中相似的文章 |
| [Kaundal, Amita]的文章 |
| 必应学术 |
| 必应学术中相似的文章 |
| [Kaundal, Amita]的文章 |
| 相关权益政策 |
| 暂无数据 |
| 收藏/分享 |
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。
