Page Banner

United States Department of Agriculture

Agricultural Research Service

Research Project: PHYSIOLOGICAL, BIOCHEMICAL AND GENETIC REGULATION OF CARBOHYDRATE METABOLISM IN CEREAL TISSUES

Location: Cereal Crops Research

2006 Annual Report


1.What major problem or issue is being resolved and how are you resolving it (summarize project aims and objectives)? How serious is the problem? Why does it matter?
This project conducts research under National Action Plan 302 Plant Biological and Molecular Processes. The successful production of cereals with value-added traits depends upon our ability to consistently obtain optimal expression of the desired traits. We cannot yet consistently produce cereal grains that meet the stringent specifications required by the processing industries that use the grain. In part, this is due to an incomplete knowledge of the physiology and biochemistry of cereals. Additional limitations result from the ill-defined and changing needs of specific industries as well as imprecise measurements of these qualities. We are working to complete our understanding of the biochemical and physiological processes that result in the conversion of starch to sugars, which is a crucial value-added trait for malting barleys. Research from this program will potentially lead to development of new or improved crop products, and it will define the properties of value-added traits in such a way that better measurements of quality result. New, more accurate measurements of malt quality will provide the basis for improved pricing of barleys shown to have high malting quality for customers. Growers who lose an anticipated premium (typically $0.25-1.00/ bushel) for malting quality barley consider crop rejection by malting/brewing industries to be a significant loss. The problem of inconsistent quality is quite serious to industrial users, as unexpected variation in the raw product (malt) requires additional expenses in downstream processing.


2.List by year the currently approved milestones (indicators of research progress)
Year 1 (FY 2006) 1. Isolate RNA, synthesize cDNA, sequence appropriate PCR products and analyze sequences from ‘MaltGenes”, which is a collection of important breeding lines used to develop malting barleys used in North America.

2. Re-establish use of the Kakefuda and Duke (1984) gels in lab and confirm use of glycogen as substrate, conduct mashing experiment in triplicate and analyze via native gels. Analyze mashing samples via western blotting.

3. Grow all cultivars at a single location.

Year 2 (FY 2007) 1. Clone, express, and evaluate consequences of SNPS in wild type rEnzymes encoded by Agl1.

2. Analyze mashing samples via western blotting.

3. Prepare manuscript.

4. Isolate RNA, synthesize cDNA, sequence appropriate PCR products and analyze sequences from ‘MaltGenes”.

5. Malt and conduct standard malt quality analyses; initiate mashing and HPLC analysis to determine RDF values; initiate TMS derivatization and GC-MS metabolite analysis.

Year 3 (FY 2008) 1. If SNPs are found that encode proteins with different biochemical characteristics, prepare manuscript. If no SNPs are found in ‘MaltGenes’, isolate RNA, synthesize cDNA, sequence PCR products from 7-10 H. spontaneum accessions.

2. Clone, express, evaluate consequences of SNPs in wild type rEnzymes encoded by Iso1.

3. Complete mashing, HPLC analysis to determine RDF values; TMS derivatization and GC-MS metabolite assays.

Year 4 (FY 2009) 1. Clone, express, evaluate consequences of SNPs in wild type rEnzymes encoded by Agl1 from H. spontaneum accessions.

2. If SNPs are found that encode proteins with different thermostabilities, prepare manuscript. If no SNPs are found in ‘MaltGenes’ , isolate RNA, synthesize cDNA, sequence appropriate PCR products and analyze sequences from 7-10 H. spontaneum accessions.

3. Analyze data.

Year 5 (FY 2010) 1. If SNPs are found that encode proteins with different biochemical characteristics in the H. spontaneum accessions, prepare manuscript.

2. Clone, express, evaluate consequences of SNPs in Iso1 from H. spontaneum. Prepare manuscript.

3. Prepare manuscript.


4a.List the single most significant research accomplishment during FY 2006.
This research addresses NP 302 Component II Biological Processes that Improve Crop Productivity and Quality, problem statement C) Developing High-Value Products. Evaluation of tools of predicting the value-added trait known as the production of fermentable sugars in malting barleys: The specific accomplishment in this reporting period was the demonstration that primary tool breeders are being encouraged to use to screen their germplasm to increase yield of fermentable sugars during processing is not reliable. The specific problem this accomplishment addresses is the development of tools that increase yield (fermentable sugars) from raw products (cereal grains) and methods that allow accurate prediction of these changes in crop quality and value-added traits. To accomplish this we studied a key enzyme in sugar production in a collection of barley that is used to develop malting cultivars. The impact is that the tool currently being promoted has been rendered less valuable than was previously thought and has made researchers aware of the need to include more detailed genetic information in addition to short gene sequences of key enzymes to their predictive models.


4b.List other significant research accomplishment(s), if any.
This research addresses NP 302 Component II Biological Processes that Improve Crop Productivity and Quality, problem statement C) Developing High-Value Products. Evaluation of tools to increase the value-added trait known as the production of fermentable sugars in malting barleys: One specific accomplishment in this reporting period was the demonstration that North American malting barley germplasm contains extremely limited variation in the genetics of one key enzyme in the production of fermentable sugars and that potentially useful variation is present in germplasm collected in the Fertile Crescent of Middle Eastern countries. We identified opportunities to use these non-native barleys as a source of increased enzyme production that may provide increased yield of product from raw material. Evaluation of methods of predicting the value-added trait known as the production of fermentable sugars in malting barleys: A second specific accomplishment is this reporting period was the development of use of osmolyte concentrations as a measure of and a predictor of malt extract. This provides a novel method for the assessment of the single most important metric of malting barley quality that is used to determine the commercial value of all malting barleys.


4c.List significant activities that support special target populations.
None.


5.Describe the major accomplishments to date and their predicted or actual impact.
This research addresses NP 302 Component II Biological Processes that Improve Crop Productivity and Quality, problem statement C) Developing High-Value Products. Evaluation of tools of predicting the value-added trait known as the production of fermentable sugars in malting barleys: The specific accomplishment in this reporting period was the demonstration that primary tool breeders are being encouraged to use to screen their germplasm to increase yield of fermentable sugars during processing is not reliable. The specific problem this accomplishment addresses is the development of tools that increase yield (fermentable sugars) from raw products (cereal grains) and methods that allow accurate prediction of these changes in crop quality and value-added traits. To accomplish this we studied a key enzyme in sugar production in a collection of barley that is used to develop malting cultivars. The impact is that the tool currently being promoted has been rendered less valuable than was previously thought and has made researchers aware of the need to include genetic information in addition to simple gene sequences of key enzymes to their predictive models.

This research addresses NP 302 Component II Biological Processes that Improve Crop Productivity and Quality, problem statement C) Developing High-Value Products. Evaluation of tools to increase the value-added trait known as the production of fermentable sugars in malting barleys: One specific accomplishment in this reporting period was the demonstration that North American malting barley germplasm contains extremely limited variation in the genetics of key enzymes in the production of fermentable sugars and that potentially useful variation is present in germplasm collected in the Fertile Crescent of Middle Eastern countries. We identified opportunities to use these non-native barleys as a source of increased enzyme production that may provide increased yield of product from raw material.

Evaluation of methods of predicting the value-added trait known as the production of fermentable sugars in malting barleys: A second specific accomplishment is this reporting period was the development of use of osmolyte concentrations as a measure of and a predictor of malt extract. This provides a novel method for the assessment of the single most important metric of malting barley quality that is used to determine the commercial value of all malting barleys.


6.What science and/or technologies have been transferred and to whom? When is the science and/or technology likely to become available to the end-user (industry, farmer, other scientists)? What are the constraints, if known, to the adoption and durability of the technology products?
The osmolyte assay method for measure of malt extract has been presented at an international meeting and a manuscript has been accepted for publication in the Journal of the American Society of Brewing Chemists. This journal, which is of limited distribution, is targeted specifically to the audience for which the assay was developed. This is a simple, rapid and non-subjective assay that is quantitative. This assay is expected to supplement the traditional measure of extract by providing novel physical measures of malt components.


Review Publications
Tanaka, Y., Duke, S., Henson, C.A. 2006. Alpha-glucosidases from the glycoside hydrolase family 31 in germinating seeds and seedling leaves of barley. XXIIIrd International Carbohydrate Symposium, July 23-28, 2006, Whistler, Canada. 2006 CDROM.

Last Modified: 4/17/2014
Footer Content Back to Top of Page