It’s More Than Just Corn Nathan Etsitty 1, Joan Peterson 2, Candice Gardner 3, Lisa Burke 3, Lisa Pfiffner 3,and Carolyn Lawrence 2,3,4 1. Northern Arizona University, Flagstaff, AZ 2. Department of Agronomy, Iowa State University, Ames, IA 3. USDA-ARS, Ames, IA 4. Department of Development Genetics & Cell Biology, Iowa State University, Ames, IA Abstract Not only is maize (Zea mays ssp. mays; also called corn in the U.S.) consumed as food and feed, it also is an important commodity used in the manufacture of ethanol (for fuel), cosmetics, drugs, cleaners, and other products. The multiple uses of corn are made possible by the diversity available for maize germplasm. Preserving such diversity is of great importance. Here we describe our work to ensure that this diversity is maintained. Presence of the Abnormal chromosome 10 changes normal maize reproduction. We are working to devise methods to identify the chromosome 10 constitution for the purpose of devising methods to control its transmission. We also are testing and utilizing software to evaluate the phenotypic expression of diversity. Acknowledgements I would like to thank everyone who contributed to my project this summer and that includes GWC, USDA-ARS, National Science Foundation, and the Agronomy Department. Most important, I'd like to thank my family and girlfriend who have been there for me through thick and thin. With late planning and good decision making, this research project became a success. About Me Hello, my name is Nathan Etsitty and I attend Northern Arizona University where I am majoring in Athletic Training. As the youngest of my siblings, I’ve been supported and driven to do my best and that’s what keeps me motivated. My ambitions in life include having a successful career and a happy lifestyle. My mother, who is a single parent after the loss of a great father/husband, has been there for me and taken the role of both parents in my life and has taught me to drive for success and carry on that dream my late father had in store for his children. As a Native American from the Southwest, I am happy to be a part of this research because “MAIZE aka corn” is an important part of our culture and to be able to study it and find out how it functions are a plus on my end. With the information I've been able to gather from this summer’s research, I will educate those back home on what maize has to offer and why it’s marked as the number one crop on the world. Fig. 1 The Punnett Square, Normal and Abnormal 10 chromosome Methods & Materials I. Abnormal chromosome Materials: ~DNA extracts used were prepared Summer ’06 ~A new set of SSR markers was selected to provide expanded coverage of chromosome 10, bins 5-7 (Table 1). 2. Methods: ~Standard PCR protocols for testing three SSR markers and agarose gel electrophoresis were carried out. ~Photos of SSR images were captured with a FOTO/Analyst Investigator camera and filter system (Fotodyne, Inc., Hartland, WI). ~Images were processed by ImageJ software (NIH, USA). II. Phenotype Data 1. Materials: ~WinFLORA and WinSEEDLE (Regents Instruments, Inc., CN) software packages were selected for testing different maize races. ~A scanner was used to capture the images. 2. Methods: ~Five ears each from a group of six maize races were measured manually and were also scanned to capture images. ~Adobe Photoshop was used for editing a 17b 17c18a18b18c 18d19a19b 19c Fig. 3 Agarose gel of PCR products. Lanes are loaded in same order as genetic lines listed in Table 1. LaneDescription 1A1 C1 r1 N10 homozygous (Dawe lab; JM 9 sib. W23) 2R1 Ab10-I/r1 N10 (Dawe lab; JM 2-3 x 1-2 backcrossed 9x to W23) 3R1-nj N10/r1 N10 (Dawe lab; CLO4AI 9-2 W23) 4R1-st N10/r1 N10 (Dawe lab; CLO4AI 13-1 W23) 5C1 sh1 wx1 R1 K9S-I Ab10-I homozygous (stock ID 905D, mongrel background) 6r1 Ab10-I (stock ID X16B, mongrel background) 7R1 Ab10-II / r1 N10 (stock ID X16F, mongrel background) 8r1 Ab10-II / r1 N10 (stock ID X16E, mongrel background) 9R1 N10 homozygous in W22 (Weber lab; CLO4AI 11-1A) 10R1 N10 /r1-x1 in W22 (Weber lab; CLO4AI 19-2) 11H99 12B73 13Mo17 14Tx303 15CO159 16GT119 17aR1-ch Ab10-I Pl1 / r1-g N10 in W22 (stock ID X233F) 17bPI ncpo01 (original stock) 17cPI ncab01 (second generation) 18aR1-ch K10-I Pl1 homozygous in W22 (stock ID X233E) 18bPI ncpo01 (original stock) 18cPI ncai01 (second generation) 18dPI ncab01 (third generation) 19aR1-nj N10 homozygous in W22 19bPI ncei01 (original stock) 19cPI ncab01 (second generation) Table 1: Genetic background of lines presented in Fig. 3 Conclusions After conducting research on this maize project both last year and this year, I feel that I have gained even more valuable knowledge on the crop itself. I’ve learned more about the Abnormal chromosome 10 characteristics and how technology can make a huge difference in the preservation of maize. Results Abnormal chromosome 10 The SSR markers that we selected failed to distinguish between genetic lines that contain the N10 and the genetic lines that contained the Ab10. This was determined by the band patterns on gel (One of the sample gels is shown in Fig. 3). For the future, the next step would be to select different SSR markers or simply get new lines of DNA and hopefully catch the Ab10 chromosome then. Phenotype Data After careful observation and a few test runs, the software programs, WinFLORA and WinSEEDLE, seem to have shown no real benefit. Although a few areas were tweaked and revised, the data provided were not good enough compared to manually collected data to justify making these two programs a part of the germplasm curation process. Background Abnormal chromosome 10 Maize (Zea mays, spp. mays, corn) is believed to be a descendent of a tall grass named teosinte. The number one producing crop in the world, maize has many uses other than just food. Maize has been studied and grown to provide things like ethanol gas, and other valuable resources it offers to the world. In past times, corn was used by the Native American people for ceremonial purposes and for the purpose of healing the sick. Maize has ten chromosomes, the tenth chromosome has at least two variations, the normal chromosome 10 (N10) and the abnormal chromosome 10 (Ab10). Ab10 is the focus of this project because its characteristics have been shown to cause preferential segregation of traits, which could result in a change in genetic profile in seed accessions over time (Fig 1). This is a problem because maize is an important crop and if it cannot be preserved, and then other useful resources would have to be produced. We are developing SSR (simple sequence repeat) molecular markers to identify chromosome 10 constitution in maize as a tool to diagnose chromosome constitution and to serve as a tool to address this problem. Phenotype data For a second project, two software programs were tested for the purpose of capturing morphological information from a scanned image of the plant, seed, or leaf and recovering the equivalent information as if it were done by hand. Currently, information from the ears of corn are recorded by actually taking a ruler and going the extra mile to physically analyze the data to record. My job was to determine if these software programs would be beneficial enough to justify that huge transition from old to new school data entry. The process basically involves scanning the ears of corn, taking that image to the software and then comparing the software’s calculated measurements to manually captured data. The determination can then be made whether the newly processed data are accurate enough to warrant replacing the old method of phenotypic data acquisition. Fig. 2. Picture of WinFLORA which is a software I worked with.