2014 Annual Report of the GEM Project, Ames, IA and Raleigh, NC

Transcription

1 2014 Annual Report of the GEM Project, Ames, IA and Raleigh, NC PERSONNEL: Ames: USDA-ARS Plant Introduction Research GEM Coordinator and Maize Geneticist, Vacant Coord. Michael Peters, GEM Technician, New, Feb 2014 Technician Fred Engstrom, GEM Technician Dr. Mack Shen, IT Specialist Adam Vanous, Iowa State University, Ph.D. student Dr. Candice Gardner, Research Leader Raleigh: USDA-ARS Plant Science Res. Dr. Matt Krakowsky, Southeast GEM Dale Dowden, Agricultural Research Dr. Peter Balint-Kurti, Research Geneticist Dr. Jim Holland, Maize Research Geneticist Dr. David Marshall, Research Leader Raleigh: North Carolina State University Dr. Major Goodman, William Neal Reynolds Distinguished Professor GEM- Ames 2014 Highlights (Dr. Candice Gardner) Midwest Germplasm releases and development: Two hundred sixty-nine (269) GEM releases are now available to GEM Cooperators. Ten GEM lines from the Ames program are proposed for release to GEM Cooperators for the 201 planting year (Table 1). Yield data, and NIR results for protein, oil, and starch can be found online and in the handout for 201 released lines. Pedigree, race, percent exotic, heterotic background, agronomic characteristics, and NIR information are presented in Tables 1-3. Six of the ten releases are second cycle GEM lines from GEMN-0097, -0140, -01, and from GEMS-0147, -017, and Their parent lines derive from races that were collected at one time from Argentina, the British Virgin Islands, Brazil, Guatemala and Thailand. The remaining new releases derive from Brazilian and Thai germplasm. All new codes have respectable scores for Fusarium ear mold and for grain quality. Table Ames-GEM Germplasm Releases (10) GEM Code Pedigree Race Type GEMN-0260 (GEMN-0140/GEMN-0097)-B-B-026-B-B Dente Amarillo/FS8BT 2% Tropical NS GEMN-0261 (GEMN-0140/GEMN-0097)-B-B-027-B-B Dente Amarillo/FS8BT 2% Tropical NS (GEMS-0147/GEMS-0180)-B B- GEMS-0262 B Suwan/Tusón 2% Tropical SS GEMS-0263 BR10:S1626(GEMS-017)-B-049-B-B Composite Suwan/Tusón 2% Tropical SS GEMS-0264 BR10:S B-B Composite Suwan 2% Tropical SS GEMS-026 DKXL212:S090-B-083-B-B Hybrid Tropical 2% Tropical SS GEMN-0266 NEI9008:N0826(GEMN-01)-B-074-B-B Suwan/Cristalino Colorado 2% Tropical NS GEMN-0267 NEI9008:N0826(GEMN-01)-B-09-B-B Suwan/Cristalino Colorado 2% Tropical NS GEMS-0268 NS1:S082-B-021-B-B Suwan 2% Tropical SS GEMS-0269 NS1:S082-B-04-B-B Suwan 2% Tropical SS Het Grp 1

2 Table 2: Summary of the 10 GEM Ames Released Lines for 201 Grain GEM Code Cob Color Grain color Texture GEMN-0260 White Pale Yellow Medium Dent GEMN-0261 White Pale Yellow Medium Dent GEMS-0262 R&W Bright Yellow Dent GEMS-0263 White Orange/Yellow Semi-Flint GEMS-0264 White Yellow Semi-Flint GEMS-026 Red Yellow Dent GEMN-0266 Red Orange Semi-Flint GEMN-0267 Red Yellow Dent GEMS-0268 White Yellow Medium Dent GEMS-0269 Red Yellow Medium Dent Table 3: Topcross Maturity of 201 Ames-GEM Germplasm Releases Top Cross RM & Ear Height Estimates Average GEM Code Pedigree Estimated RM Ear Height (cm) GEMN-0260 (GEMN-0140/GEMN-0097)-B-B GEMN-0261 (GEMN-0140/GEMN-0097)-B-B GEMS-0262 (GEMS-0147/GEMS-0180)-B GEMS-0263 BR10:S1626(GEMS-017)-B GEMS-0264 BR10:S GEMS-026 DKXL212:S090-B GEMN-0266 NEI9008:N0826(GEMN-01)-B GEMN-0267 NEI9008:N0826(GEMN-01)-B GEMS-0268 NS1:S082-B GEMS-0269 NS1:S082-B Hybrid Checks HC33xLH MBS3633xMBS LH200xLH R6 ~ F8 ~ D8 ~ Summary tables of the newly released lines performance in testcross can be found in the section of this book tabbed Released Lines. 2

3 Whole grain composition analysis was generated using an NIR Infratech 1241 with sample transport module in Dr. Paul Scott s lab (USDA-ARS CICGRU). Samples were obtained from a bulk of at least 8 ears from self pollinated rows, and are presented in Table 2. Table 4. Two Year Flowering & NIR Trait Data of 201 GEM-Ames Line Releases GEM Code Pedigree Lines per se Data NIR Data 2014 Days GDU's to to Pollinatioation Pollin- Star Protein Oil ch Densi ty GEMN-0260 (GEMN-0140/GEMN-0097)-B-B-026-B-B GEMN-0261 (GEMN-0140/GEMN-0097)-B-B-027-B-B GEMS-0262 (GEMS-0147/GEMS-0180)-B B-B GEMS-0263 BR10:S1626(GEMS-017)-B-049-B-B GEMS-0264 BR10:S B-B GEMS-026 DKXL212:S090-B-083-B-B GEMN-0266 NEI9008:N0826(GEMN-01)-B-074-B-B GEMN-0267 NEI9008:N0826(GEMN-01)-B-09-B-B GEMS-0268 NS1:S082-B-021-B-B GEMS-0269 NS1:S082-B-04-B-B B Research & Breeding Activities A second year trial of 400 top crosses of BC 1 S 2 generation families of CUBA164xB73) x B73 and CUBA164xPHB47) x PHB47 were planted in yield trials at four locations at low, medium, and high population densities, collaborative research with Dr. Jode Edwards. This research is designed to provide a better understanding of the effects of exotic alleles from CUBA164, particularly for response to high plant density, and is part of an MS thesis study. Eighty eight new breeding crosses were made in the Ames nursery. GEM Cooperators made 1 additional breeding crosses as part of their in kind support. Twelve new breeding cross populations were assigned to Cooperators for in kind support. Twelve families of single seed descent (SSD) balanced bulks were advanced to S 2 in Ames, and two were advanced by Cooperators. Twenty-two populations of BC 1 s were sibbed in Puerto Rico in January 2013 as a result of TSG member support. The BC 1 -sib 1 generation was returned to Ames and random mated a second generation in 2013 to generate the BC 1 -sib 2. Four family sets of the random mated populations (BC 1 -sib 1 and BC 1 -sib 2 and the original BC 1 s were induced in 2014 in Ames to generate haploids, as well as advanced conventionally. This study was designed to compare the frequency of exotic alleles in doubled haploid lines developed directly from the BC 1 vs. from BC 1 s random mated for one or two generations, and similarly from conventionally derived lines. Families induced in summer 2014 include Cuba 164, Amargo-ARZM , Tusón-CUB 67, and Tusón-BAI III, all with the PHB47 recurrent parent. Ph.D. student Adam Vanous' research objectives are to utilize 1) selection mapping to dissect the genetic architecture of adaptation from the tropics to the U.S. Corn Belt (mass selected populations), and 2) association mapping to dissect the genetic architecture of flowering time and photoperiod response in exotic derived DH lines. Three tropical landraces, Tusón 3

4 (Hallauer, 1999), Tuxpeño (Hallauer, 1994), and Suwan-1, are the sources of exotic germplasm that have undergone pre-breeding and were adapted to the Midwest Corn Belt through recurrent mass selection on the basis of early female flowering. Two hundred-fiftytwo BC 1 F 1 derived doubled haploids (DH) lines represent the backcross method of introgression. Current and novel mapping techniques will be used to indentify loci associated with early flowering time in maize. Host Plant Resistance: 2014 disease data will be posted online at our website ( in January. Pathology and entomology research collaborators from the private and public sectors screened GEM lines, top crosses, and breeding crosses for Northern Leaf Blight (NLB), Southern Leaf Blight (SLB), Goss s Wilt, Gray Leafspot (GLS), Diplodia ear rot (DIPPER), Aspergillus (aflatoxin and other mycotoxins), Corn ear worm (CEW), Fall Armyworm (FAW), and Western corn root worm (WCR). Please see the quality traits sections regarding ear mold and mycotoxin studies. A wet spring and excessive rain events followed by the dry summer impacted data collection and resulted in some missing plots, but less so than in recent years. Appreciation is extended for the in-kind support and extensive efforts by private and public GEM Cooperators: CAAS (Chinese Academy of Agricultural Science) provided agronomic evaluations and disease ratings for various leaf blights including MRDV, Fusarium ear mold, and for common smut. DuPont/Pioneer Hi-Bred International (Bill Dolezal at Johnston, IA, and Mark Mancl at Woodland, CA).NLB or DIPPER data were collected at Johnston and Fusarium ear rot and Head Smut were successfully evaluated at Woodland, CA,. Professional Seed Research (Jim Dodd) for their efforts collecting data on Goss Wilt, Eyespot, GLS, NLB, and SLB. 3 rd Millennium Genetics (Ed and Raechel Baumgartner) for tropical insect evaluations on GEM lines in Puerto Rico which was particularly helpful to identify some material for Fall Armyworm (see Table following public cooperator reports). USDA-ARS-CGBRU (Xinzhi Ni) in Tifton, GA collected data for Corn ear worm (CEW), and Fall Armyworm. USDA-ARS-PSRU in Raleigh, NC provided data for SLB (Peter Balint-Kurti) and Fusarium ear rot (Jim Holland). USDA-ARS-PGRU, Columbia, MO (Bruce Hibbard) evaluated corn rootworm (CRW) USDA-ARS-CHPRRU, Mississippi State, MS (Paul Williams and Gary Windham) evaluated aflatoxin in grain. Second year host plant resistance evaluation data (2014) will be posted to our website under GEM Germplasm Releases and Key Traits. Unfortunately, there has not been time to review 1 st and 2 nd year data together at the time of this report. Appended to the end of this document are tables from P. Williams on evaluations of germplasm for aflatoxin, and from E. and R. Baumgartner for fall armyworm evaluation trials. Ames 2014 Allelic Diversity (AD) and Double Haploids (DH): The Iowa State University Doubled Haploid Facility continues to partner with the GEM Project on development of DH lines. Approximately 204 lines (representing ~2 races) were jointly released by ISU and USDA-ARS in 2014 as part of the Allelic Diversity Project. The DH lines provide unique genetic resources for a wide variety of maize research applications. Twenty sets of the doubled haploid released lines were distributed to requestors, and a subset of lines to those who wished to narrowly targeted their requests. Another seed increase was made of the 204 DH lines this summer, and phenotypic information was collected in Ames. 4

5 Approximately 237 new DH lines (DH 0 ) were advanced to DH 1 in This was part of Andrew Smelser s thesis research to study the rate of induction and doubling with exotic germplasm from the allelic diversity project. Self pollinations were made in 80 rows ranging from BC 1 F to BC 1 F 6 allelic diversity project germplasm to generate the BC 1 F 6 to BC 1 F 7 generations. This effort is part of the GEM allelic diversity project but information will be used in various thesis research projects Ames 2014 Yield Test, Nursery and Other Highlights: Approximately 1,370 plots (2,30 entries) were managed or coordinated through Ames in 2014 (increase of more than 0% over 2013). Among these, 470 entries (18.6%) were in 2nd year and 2,060 entries were in 1st year trials. Two hundred and two (202) top crosses out of 2,30 entries exceeded the mean yield of the check hybrids in Midwest trials in Of the 202 hybrids that exceeded the mean, 142 were from first year trials, and 60 from second year trials. Approximately 7,460 nursery rows for pollination, over 300 for observation, and 1,880 isolation rows were managed by Ames. One hundred new breeding crosses were observed and evaluated for breeding potential in Ames, IA, Ft. Branch, IN (AgReliant), and Memphis, TN (G&S Crop Services). Of these 100, 44 had above average scores at all three locations; 28 scored >6.0 on a 1-9 scale and will be considered for development. Quality Traits (including grain mold/mycotoxin): The newly released GEM lines were not noteworthy for protein, oil or starch traits. An unreleased line, AR1706:S1217-B-001-B showed segregation in the next generation of harvested ears for a waxy trait. This line was not sufficiently high performing to release, but is available to interested cooperators. Aflatoxin analyses show six breeding crosses with very low, promising levels of aflatoxin (Appendix Table 1). The report by Paul Williams and Gary Windham will be available online. Three newly released lines were evaluated favorably for Fusarium ear mold. New GEM Cooperators in 2014: One new private U.S. Cooperator joined the GEM Project in 2014; Summit Genetics (in cooperation with KMR) conducted yield trials in Stanton and Takama, Nebraska. New international cooperators include Global Investors, LLLP, of Des Moines, IA with their partner companies Misr Hytech (Cairo, Egypt) and Hytech Seed India Pvt. Ltd. (Hyderabad, India). These international cooperators have already provided valuable information on GEM germplasm resistant to late wilt in their respective environments, and will continue to screen germplasm for late wilt, southern rust, and other diseases as opportunities are presented.

6 2014 Annual Report of the GEM Project GEM-Raleigh Highlights Germplasm releases: Seven GEM lines from the Raleigh program are recommended for release to GEM Cooperators for 201. Yield data can be found in this book or on the GEM website and additional agronomic data can be found in the table at the end of this report. Releases include two lines derived from Dekalb tropical hybrids, two from Florida Synthetic B, one from a Suwan synthetic, one from Tuxpeno and one from St Croix. GEM code Pedigree Race Country Type Heterotic pattern GEMS-023 DK212T:S Tropical hybrid Thailand GEMN-024 FS8B(T):N11a Mixed population USA GEMS-02 FS8B(S):S Mixed population USA GEMS-026 NS1:S Suwan Thailand GEMN-027 CHIS462:N08d Tuxpeño Mexico GEMS-028 DKB830:S Tropical hybrid Mexico GEMN-029 SCR01:N11c St. Croix St. Croix 2% Tropical 0% Subtropical 0% Subtropical 2% Tropical 0% Tropical 0% Tropical 0% Tropical SSS NSSS SSS SSS NSSS SSS NSSS Germplasm development: Overall 200 GEM nursery plots and 700 GEM isolation plots were grown in Clayton, NC, with additional GEM-related nursery managed by Major Goodman. S 1 families were developed from nine breeding crosses: 89291:(LH1), 89291:(LH9), BR1039:N1, BR1039:S1, BR167:D27S21, BVIR1:S20, GEMN-0097/GEM- 0104, NS1:(GEMN-0097), and NS1:(GEMS-011). Families from eight GEM breeding crosses were advanced from S 1 to S 2: BR106:(GEMN- 0097), BR106:N42, BR106:S42, BR101:N11a, BR101:S11a, BR1721:S20, GUAT209:N19, PASCO14:S11. Eighteen GEM x GEM F 1 combinations were produced based on yield trial data. Approximately 140 new breeding crosses were developed between tropical hybrids (ATL100, SX1078 and SX2788 from Brazil) or tropical inbreds (89291 from IITA- Cameroon; CML277, CML313, CML339, CML340, CML343, CML39, CML423, CML438, CML494 and La Posta Sequia C7 F B*3 from CIMMYT and CA3402 and CA00370 from CIMMYT-Asia; N from Zimbabwe) and GEM- Ames releases (GEMN-0048, GEMN-0097, GEMS-0113, GEMN-018, GEMS-017, GEMS-0200, GEMS-0220) and ex-pvps (AS707, DKF118, DKMM402A, LH19, LH197, LH210, NK787, NKW8, NS701 and PHN46). Both groups were selected based on performance in yield trials. Germplasm Evaluation in 2014: 6

7 Overall, 11,700 yield plots were coordinated from Raleigh, with 7300 planted in NC and the rest planted by seven cooperators at various locations throughout the Southeast and Midwest. Thirty-three entries were evaluated at eighteen locations in third-year trials (EXR1 and EXS8; see Yield Trials section) Fifty entries were evaluated in second-year trials, approximately twenty of which will be evaluated in third-year trials in entries were evaluated in first-year trials, fifty-eight of which will be evaluated in second-year trials in 201. Evaluations of GEM releases from Raleigh and Ames were conducted in two experiments (EXP8 and EXP9) for the third year to identify lines with the best potential for use in GEM x GEM crosses. EXP8 (releases from Raleigh) was planted at locations in the southern Corn Belt and southeast, while EXP9 (releases from Ames) was planted in North Carolina and the Corn Belt. A subset of the best performing GEM lines from the 2010 and 2011 trials (EX26- EX29) were planted in the 2014 trials along with more recent releases. Ex-PVPs were included in both trials for comparisons with GEM releases (see Yield Trials section). Breeding crosses were evaluated per se in the Raleigh nursery (see Breeding Crosses section). Ex-PVP inbreds were evaluated by Major Goodman at five locations in North Carolina and results for three trials conducted over three years (EXA2), two years (EXF1) or one year (EX60) will be available on the GEM website and in this booklet in Yield Trials section. Allelic Diversity: There were 300 summer nursery rows dedicated to the development of F 1 s and BC 1 s for the Allelic Diversity project, and 310 rows were planted in Homestead, FL for the 2014 winter nursery. Much of the effort in the winter nursery is focused on late maturing accessions. Twenty-two BC 1 families and two F 1 s were completed this summer in Raleigh; two BC 1 families and fifty-two F 1 s were also completed in the 2013 winter nursery. Other GEM-Raleigh recommendations: Based on 2013 and 2014 nursery observations, some of the most promising new breeding crosses include GEMN-0097 x GEM-0104, GEMN-0097 x CML449, GEMN-0097 x CML37, GEMS-0027 x GEMS-0113, GEMN-013 x GEMN-0104, and GEMN-0124 x GEMN Some of the best entries in EXP8 were GEMN-0043, GEMN-0119, GEMN-0207, GEMN-0208, and GEMN Efforts are already underway to recycle most of these GEM releases and efforts in recycling will continue in 201 with an emphasis on non- Stiff stalk GEM releases (recycling of Stiff Stalk releases was emphasized in the 2014 nursery). Some of the best entries from EXP9 were GEMS-0091, GEMN-0097, GEMS-0113, GEMN-018, and GEMS-017. GEMN-0097 had the highest value for Y/M, while GEMS-000 had the highest yield of any GEM entry but also had the highest moisture of any entry in the trial. Efforts are underway to recycle these and other GEM releases from Ames. Some of the most promising ex-pvps based on yield trial data are DKF118, DK6F629 and DKMM402A from EXA2 and DKNL001, LH197, LH198, LH204 and PHKE6 from EXF1; however, it should be noted that, as a line per se, LH198 performed very poorly in both the summer and winter 2014 nurseries and LH197 was marginally better. 7

8 Summary of the seven GEM-Raleigh Recommended Lines for 201 GEM code Pedigree Days to ANT GDU to ANT Days to Silk GDU to Silk Plant Height (cm) Ear Height (cm) GEMS-023 DK212T:S Y GEMN-024 FS8B(T):N11a Y GEMS-02 FS8B(S):S Y GEMS-026 NS1:S Y Grain color GEMN-027 CHIS462:N08d Y/W GEMS-028 DKB830:S Y/W GEMN-029 SCR01:N11c Y Check B Y 8

9 GEM TSG Meeting, September 1-16, 2014, Ames IA - Highlights: The GEM Technical Steering Group (TSG) meeting was held at the North Central Regional Plant Introduction Station in Ames, IA on September 1-16, A tour of the nurseries was hosted by Candice Gardner, Fred Engstrom, Michael Peters and Nuo (Mack) Shen. Graduate student Adam Vanous participated and discussed his research. The tour included inbreds, all generations of breeding material in the nursery, observations of Raleigh and Ames materials and the released doubled haploid lines, new potential breeding crosses, Ex-PVP lines, and some topcross trials at the nearby Uthe Farm. Several important topics included: program focus and balance between new research initiatives on breeding methodology vs. traditional germplasm development; relative focus on four program areas requesting new tropical germplasm sources, especially from company cooperators strategies for sampling of adventitious presence (AP) of transgenes, appropriate points to test, and how to deal with it if found. alternative strategies for supporting GEM programs improving maize doubled haploid systems, inducibility and spontaneous doubling potential collaborations with international organizations and entities for evaluating important diseases and their races, and the challenges involved methods to measure GEM Project success and/or impact proposal by D. Butruille to expand GEM s YT program in Ames improving GEM s technical capacity at Raleigh and Ames. 9

10 IN KIND NURSERY AND TRIAL SUPPORT MIDWEST GEM PROJECT IN 2013: Table. Private In-Kind Support Summer 2014 Table : Summer 2014 Private In-Kind Support Made Topcrosses Yield Trials Breeding Cross obs Collaborator Breeding & Nursery Support 3rd Millenium Genetics Armyworm Screening AgReliant Genetics LLC Made breeding crosses to SS and NSS lines. S 2 's BASF (Ames, IA) Disease Screening & Quality Traits NIR analyses oil, protein, starch Beck's Made S 1 's from SX1078:N(LH61)(ANTIG03:N1242-B-007-B) CAAS Cornhusker Hybrids LLC Made S 1 's from TZAR104:(LH82/GEMN-0192) CRD Advisors LLC Disease screening for MRDV, stalk rot, head smut, drought, low nitrogen tolerance etc. in multiple ecological areas. DAS DKD Genetics Made S 1 s from CML373:(PHJ40/GEMS-0162) Dow Agrosciences Advanced S 1 's to S 2 of Ki14:S21z42-B DuPont Pioneer Made breeding crosses to SS and NSS lines. Made S 1 s of SX1078:(LH123/LH61) and Forage Genetics (DJ7/CML373)/GEMS-0162 G&S Crop Services Genetic Enterprises Made breeding crosses to NSS line. Head Smut & Fusarium in Woodland, CA; Diplodia and NLB in Johnston, IA 10

11 International Illinois Foundation Seeds, Inc. Made S 1 's with PUER:S4676A(GEMS-0147) S 2 's JFS & Associates S 2 's MayAgro Made S 1 's of BR1721:S20(GEMS-0219) NLB screening Made breeding crosses to SS and NSS lines. Made S 1 's with TZAR104/LH123//ANTIG03:N1242-B-007-B and Monsanto BR1039/PHJ40//GEMS Professional Seed Research Screening for Goss' wilt, NLB, Eyespot, GLS Screen breeding crosses and lines for downy mildew, Seed Asia NLB,SCLB, and rust SEEDDirect Semillas Fito Made breeding crosses to SS and NSS lines Summit Genetics Syngenta Made breeding crosses to SS and NSS lines Make S 1 's with CML373//794/GEMS-0147 and PUER:LH61/GEMN Screen for late wilt, MRDV, and head smut Terrell Seed Research Trimble Genetics Advance S 1 's to S 2 's of CML373:S(DJ7)(GEMS-0188)-B Wyffel's Hybrids Make S 1 's of ATL100:LH82/GEMN-0097 Disease and agronomic evaluation 11

12 2014 GEM Ames Yield Trial Summary 12 Entry Yield Avg. Check Yield Avg. Overall Yield Avg. CV LSD Expt # Material Tester Total Reps Source (%) p= Ki21:N99am HC33 30 GEM NEI9004:S2899n LH GEM DKXL212:S0943b LH GEM CUBA164:S99aa99ah LH GEM CML341:S99y99ah LH GEM SCRO1:N1318 HC33 0 GEM MDI022:N99d99z HC33 0 GEM BR10:N99d99h HC33 60 GEM ANTIG01:N1699C HC33 40 GEM SCRO1:N1318 HC33 48 GEM MDI022:N99d99z HC33 48 GEM White SS x tester WSS JFS White NSS x tester WT JFS Ki21:S99al99y SGI IFS BR1039:N112 MON-SS 92 6 MON CML287:N(GEMN-01)11a SYG-SS 46 6 SYG CML287:N(GEMN-01)11a SYG-SS 42 6 SYG BR1039:S(PHJ40)11b SYG-nSS 40 6 SYG SS4/GEMS-0026 LH GEM SS4/GEMS-0031 LH GEM GEMS-0176/S49w LH GEM GEMS-011/S49w LH GEM GEMS-0176/S49x LH GEM BR201:S LH GEM (KO679Y/GEMS-011)/GEMS-0162 LH287 0 GEM Retest (Tropical) LH GEM NS2/GEMN-0133 HC33 80 GEM GEMN-0179/GEMN-020 HC33 0 GEM Retest (Tropical) HC GEM Retest MBS GEM Retest MBS GEM GEM SS x tester MBS 30 8 GEM BR101:N11a08bT4799bc GP7169GTCBLLRW 0 IFS TZAR104:N(LH82)(GEMN-020) AGR-SS 41 AGR TZAR104:N(LH82)(GEMN-020) AGR-SS 39 AGR BR101:N11a08bT47(LH82) DAS-SS 24 DAS UR13091:S99al LH287 0 GEM UR11002:S1409 LH287 0 GEM AR1603:S0209 LH GEM CHOCLERO:N99b99z HC33 70 GEM Retest (Temperate) LH GEM Retest (Temperate) HC GEM Retest MBS GEM Retest MBS GEM GEM NSS x tester MBS 40 8 GEM Early Retest NA 40 8 GEM GEMxGEM NA 40 GEM Waxy Test NA 40 8 ING

13 FIELD DAYS: Ames, IA GEM Project (C. Gardner) GEM Project 2014 Field Day: Due to severe damage to the demo field from heavy spring rains, lost population stands and lack of available fertilizer, no public field day was held in The TSG members toured the nursery plantings and one yield test location in mid-september, and numerous local and international visitors scheduled tours of the nursery as their interests dictated. Caitlin Ellingson, an undergraduate GEM employee, created a virtual tour that documents the growth and development of the GEM field demo over 11 weeks of the season. She developed a web based presentation of this as part of her journalism internship; it can be accessed at Fargo, ND North Dakota State University (M. Carena) 2014 Nursery Tours: Over 00 US and international visitors toured summer breeding nurseries with new NDSU EarlyGEM lines, hybrids, and population releases; short-season NDSU EarlyGEM releases were specifically shown. Three of the past 2014 inbred releases were from the NDSU EarlyGEM program as well as the last 10 populations releases included breeding crosses with germplasm originating in southern USA, Mexico, Cuba, St. Croix, Brazil, Chile, and Argentina Beijing, China, Chinese Academy of Agricultural Sciences (CAAS) 2014 Field Days: Aug 29 to Sep 6: The third international GEM Field Day was sponsored by CAAS and hosted by Dr. Tianyu Wang, coordinator of China s GEM Program. More than 70 attendees from the public and private section participated in this event and observed 987 elite local inbred lines and three breeding enhancement populations, as well as 196 GEM populations and selfed progenies at the Shunyi Research Station in Beijing. Breeders and scientists provided feedback on germplasm they wish to work with. PUBLIC COOPERATOR REPORTS (Specific Cooperative Agreements, or SCAs): The GEM Project funded two SCA s in 2014, Dr. Jay-lin Jane, Iowa State University, and Dr. Mark Campbell, Truman State University; their reports can be found below. A third SCA was initiated late in the fiscal year with Dr. Thomas Lübberstedt of Iowa State University, and the project is in the early stages. 13

14 Project Title: Development of GEM line starch to improve nutritional value and biofuel production Prepared by Hanyu Yangcheng and Jay-lin Jane, Department of Food Science and Human Nutrition, Iowa State University, Ames, IA 0011 Project Overview This report serves to document research conducted under a cooperative agreement between ARS and Iowa State University. Specific objectives of this research project are to (1) Develop and characterize hybrids using normal and waxy GEM lines to produce hybrids with improved yield and starch-ethanol conversion efficiency to increase ethanol yield; and (2) Develop healthy foods and healthy food ingredients using GEM lines. For Objective 1, a normal (B73) and a waxy (08GEMS0044) inbred corn and two hybrids developed by intercrossing between the normal and waxy corn were used in this study. Amylose contents ( %) of the starch negatively correlated with the dosage of the wx gene in the corn endosperm. Average amylopectin branch-chain lengths (DP ), however, showed no correlation with the wx gene dosage. The amylose content of the starch was inversely proportional to the gelatinization enthalpychange ( J/g), the gelatinization temperature-range (T c -T o ) ( C), and the raw-starch digestibility (r = -1, p<0.001), but positively correlated with the percentage retrogradation ( %) and starch pasting-temperature ( C) (r = 1, p<0.001). These results indicated that amylose molecules enhanced the integrity of starch granules, restricted granule hydration, narrowed gelatinization temperature-ranges, and reduced digestive-rates of the starch. Amylose contents, instead of the amylopectin branch-chain length distribution, showed dominant effects on starch physicochemical properties. Starch of the hybrid, 08GEMS0044 B73, displayed significantly lower amylose content and higher digestibility than the normal corn starch. The hybrid showed potential to improve the yield of ethanol production. To further investigate the dosage effects of wx gene on starch physicochemical properties, an intercross study using two pairs of isogenic lines grown in 2014 was conducted. Starch gelatinization enthalpy change was positively related with the dosage of wx gene in the endosperm. Final and setback viscosities of the starch decreased with increase of the wx gene dosage, whereas breakdown viscosities increased with increases of the wx gene dosage. Following work will be conducted to understand if the wx gene has dosage effects on starch molecular structures and digestibility. For Objective 2, the hybrid popcorn (GEM-07048) produced by crossing a popcorn (Sg133) with a high-amylose corn (GEMS-0067) was used in this study. The corn kernels of GEM were separated visually to two groups, normal popcorn with an amylose content of 36.3% and high-amylose popcorn with an amylose content of 61.4%. The high-amylose popcorn displayed the smallest expansion volume (11.8 cm/g) compared with the normal popcorn seeds (19.7 cm/g) and commercial popcorn (34.9 cm/g). The popped kernels of high-amylose popcorn showed largest resistant starch content (46.3%), indicating its potential as a healthy snack food. Publications and presentations: 1) H. Yangcheng, H. Jiang, M. Blanco and J. Jane. Characterization of starch from intercrossed normal and waxy maize. Poster presentation to Corn Utilization & Technology Conference, Louisville, KY, June 2-4, ) H. Yangcheng, H. Jiang, M. Blanco and J. Jane. Physicochemical properties of starch produced by intercrossing between normal and waxy maize. Poster presentation to American Association of Cereal Chemists International, Providence, RI, Oct.-8, Objectives Objective1: Develop and characterize hybrids using normal and waxy GEM lines to produce hybrids with improved yield and starch-ethanol conversion efficiency to increase ethanol yield. Objective 2: Develop healthy foods and healthy food ingredients using GEM lines. 14

15 Characterization of starch from intercrossed normal and waxy maize Two hybrids were developed by intercrossing between a normal (B73) and a waxy corn (08GEM0044). Dosages of the wx gene in endosperms of the inbred and hybrid corn are: 0 (B73), 1(B73 08GEMS0044), 2 (08GEMS0044 B73), and 3 (08GEMS0044) (Table 1). Starches of the corn lines were isolated using a wet-milling process, and the isolated starch was used for the study to understand the starch physicochemical properties and to reveal the effects of amylose contents and amylopectin structures on starch properties and digestibility. Amylose content of the starch ranged from 3.9 to 26.%, and was negatively related with the wx gene dosage in the endosperm (Table 1). Amylopectin molecular weight ranged from 1.4 to (g/mol), and showed a trend of positive correlation with the wx gene dosage in the endosperm. Table 1. Amylose content of starch and molecular weight and gyration radius of amylopectin a Sample Waxy gene dosage Amylose Amylopectin (%) M w 10 9 (g/mol) b R z (nm) c 08GEM d± ± ± GEM0044 B c± ± ±1. B73 08GEM b± ± ±1.8 B a±0.3 1.± ±3.0 a Different letters following the mean value in the same column indicate significant differences between the mean values (p<0.0). b Weight-average molecular weight (M w ). c Z-average radius of gyration. Results of amylopectin branch-chain length distribution are shown in Table 2. Average branchchain length of amylopectin ranged from DP 18.4 to 19.9, and showed no correlation with the wx gene dosage. Starch thermal properties are shown in Table 3. Starch onset-gelatinization temperatures ( ºC) negatively correlated (r=-1, p<0.001) with the percentage of short branch-chains of amylopectin ( %) (Table 2). Gelatinization enthalpy change ( J/g) negatively correlated (r=-1, p<0.001) with the amylose content (Table 1), whereas percentage retrogradation of gelatinized starch ( %) positively correlated (r=1, p<0.001) with the amylose content. The amylose molecules complexing with lipids and interacting with amylopectin to preserve integrity of the starch granule and restrict granule swelling contributed to the smaller gelatinization enthalpy change and higher percentage retrogradation of the starch. 1

16 Table 2. Molar-based branch-chain-length distribution of maize amylopectin a Sample Average CL DP 12 DP13-24 DP2-36 DP 37 (DP) b (%) (%) (%) (%) 08GEM a± b± a± a± a±1. 08GEM0044 B a± ab± a± a± a±1. B73 08GEM a± a± a± a± a±0.6 B a± ab± a± a± a±0.4 a Different letters following the mean value in the same column indicate significant differences between the mean values (p<0.0). b Average branch-chain length of amylopectin. DP = degree of polymerization. Table 3. Thermal properties of maize starches a Native starch Sample T o ( C) T p ( C) T c ( C) H (J/g) b Retrogradation (%) c 08GEM a± ± ±0. 14.a± d±0.7 08GEM0044 B b± ± ± b± c±0.2 B73 08GEM c± ± ± b± b±1.1 B73 63.d± ± ± c± a±1.4 a Samples (~3.0 mg, dsb) were mixed with deionized water (~9.0 l) before the DSC analysis. Different letters following the mean value in the same column indicate significant differences between the mean values (p<0.0). b T o, T p, T c and H are onset, peak, conclusion gelatinization temperatures and enthalpy change, respectively. c Percentage retrogradation (%) = ( H of retrograded starch / H of native starch) 100%. 16

17 Starch pasting profiles are shown in Figure 1, and the results are summarized in Table 4. Starch pasting temperature ( C) was positively related with the amylose content (Table 1), whereas the peak viscosity showed no correlation with the amylose content. Figure 1. Pasting profiles of maize starches. Table 4. Pasting properties of maize starch measured using a Rapid Visco-Analyzer a, b Sample Pasting Temp.( C) Peak (RVU) Hold (RVU) Final (RVU) Breakdown (RVU) Setback (RVU) 08GEM c± ± ± ± ± ±0.7 08GEM0044 B b± ± ± ± ± ±1.9 B73 08GEM b± ± ± ± ± ±8.9 B73 7.7a± ± ± ± ± ±0.7 a Different letters following the mean value in the same column indicate significant differences between the mean values (p<0.0). b Viscosity was reported in Rapid Visco-Units (RVU) where 1 RVU = 12 centipoise. Starch digestibility of uncooked corn starch was determined using Englyst method (1992), and the results are shown in Table. The percentages of hydrolyzed starch at 120min hydrolysis time ranged from 79.6 to 93.4%, and are inversely proportional to the amylose content (Table 1). 17

18 Table. Digestibility of maize starches a Sample Digestibility (%) 30 min 120 min 08GEM a± a±1.3 08GEM0044 B73 40.b± b±0.9 B73 08GEM c± bc±2. B c± c±0.0 a Digestibility of the uncooked maize starch was determine using Englyst s method (1992). Different letters following the mean value in the same column indicate significant differences between the mean values (p<0.0) Starch of the hybrid (08GEMS0044 B73) displayed significantly lower amylose content (21.0%) and higher digestibility (87.2% at 120min) than the normal corn starch (26.% and 79.6% at 120min, respectively), indicating its potential to improve the yield of ethanol production. To further investigate the dosage effects of wx gene on starch physicochemical properties, an intercross study using isogenic normal and waxy GEM lines was conducted. Two pairs of isogenic lines were grown in 2014, including waxy1 (wx1) and normal1 (N1) and waxy2 (wx2) and normal2 (N2). The pedigree information of these four lines and their F1 progeny obtained through reciprocal crossing is shown in Table 6. Table 6. Pedigree information of GEM corn lines Sample Pedigree wx gene dosage wx1 DKXL370:N11a B wx 3 wx1 x N1 DKXL370:N11a B wx/dkxl370:n11a b-b-b- B 2 N1 x wx1 DKXL370:N11a B-B-B-B/DKXL370:N11a B wx 1 N1 DKXL370:N11a B-B-B-B 0 wx2 AR1603:S B wx 3 wx2 x N2 AR1603:S B wx/ar1603:s b-b-b-b 2 N2 x wx2 AR1603:S B-B-B-B/AR1603:S B wx 1 N2 AR1603:S B-B-B-B 0 Starch thermal properties are shown in Table 7. Starch gelatinization enthalpy changes increased with increase of the wx gene dosage in the endosperm. Onset gelatinization temperatures showed no correlation with the wx gene dosage. The conclusion temperatures, however, showed a trend of increase with increasing wx gene dosage. 18

19 Table 7. Starch thermal properties of GEM corn lines T o ( C) T p ( C) T c ( C) H (J/g) wx1 6.8± ± ± ±0.2 wx1 x N1 6.6± ± ± ±0.0 N1 x wx1 6.7± ± ± ±0.1 N1 6.4± ± ± ±0.0 wx2 61.7± ± ± ±0.0 wx2 x N2 62.1± ± ± ±0.0 N2 x wx2 62.7± ± ± ±0.0 N2 61.8± ± ± ±0.1 Starch pasting properties are shown in Table 8. The final and setback viscosities decreased with increase of the wx gene dosage, whereas breakdown viscosities increased with increase of the wx gene dosage. Pasting temperatures and peak viscosities, however, showed no correlation with the wx gene dosage. Table 8. Starch pasting properties of GEM corn lines Pasting Temp.( C) Peak (RVU) Hold (RVU) Final (RVU) Breakdown (RVU) Setback (RVU) wx1 71.0± ± ± ± ± ±1.3 wx1 x N1 73.8± ± ± ± ± ±0.2 N1 x wx1 72.3± ±2. 7.1± ±1. 7.1± ±0.3 N1 71.4± ± ± ± ± ±1.4 wx2 69.0± ± ± ± ± ±1.1 wx2 x N2 72.3± ± ± ± ± ±2.2 N2 x wx2 72.± ± ± ± ± ±3.8 N2 71.3± ± ± ±2.0 2.± ±2.4 Future work will be conducted to understand if the wx gene has dosage effects on the starch content, content of amylose, molecular size of amylose and amylopectin, branchchain length of amylopectin, and starch digestibility. Relationships between the physical properties and the structures of the starches will be investigated. Kernel popping properties and starch digestibility of high-amylose popcorn GEM (Pedigree: (GEMS-0067/Sg133-B)-B) was produced by intercrossing between a popcorn line (Sg133) and a high-amylose corn (GEMS-0067). Kernels of GEM were visually separated to two groups, high-amylose popcorn (HAPC) and normal popcorn (NPC). Kernel starch content, starch amylose content, and expansion volume of popped kernels are shown in Table 9. The HAPC of GEM showed a significantly greater amylose-content (61.4%) than the NPC of GEM (36.3%). 19

20 Table 9. Kernel-starch content, starch amylose content, and expansion volume of popcorn samples Sample a Starch (%) Amylose (%) Expansion volume (cm/g) NPC 62.3 ± ± ± 2.2 HAPC 63.6 ± ± ± 1.8 CPC 60.8 ± ± ± 2.3 a NPC, GEM normal popcorn; HAPC, GEM high-amylose popcorn; CPC, Commercial popcorn. The expansion volume of GEM kernels was smaller than commercial popcorn (34.9 cm/g). The expansion volume of HAPC of GEM (11.8 cm/g) was even lower than that of the NPC kernels (19.7 cm/g). The amylose content was negatively correlated with the expansion volume (r = 0.862, p < 0.0). Starch digestibility of popcorn kernels after microwave popping is shown in Table 10. The popped kernels of HAPC showed largest resistant starch content (46.3%) and slowlydigestible starch content (19.3%) than the other two popcorn samples. The amylose content (Table 9) was positively correlated with resistant starch content and negatively correlated with rapidlydigestible starch content. Table 10. The rapidly-digestible starch (RDS), slowly-digestible starch (SDS) and resistant starch (RS) contents of popped kernel Sample a RDS (%) SDS (%) RS (%) NPC 77. ± ± ± 0. HAPC 34.4 ± ± ± 1.0 CPC 8.9 ± ± ± 1.6a a NPC, GEM normal popcorn; HAPC, GEM high-amylose popcorn; CPC, Commercial popcorn. These results indicated that high-amylose popcorn is a good source of resistant starch and slowly-digestible starch, which is potentially a healthy snack food. Further efforts, however, are needed to improve the popping characteristics of high-amylose popcorn. 20

21 2014 GEM Report: Truman State University, Annual GEM Cooperator meeting, ASTA, Dec. 10, Chicago, IL Development and Evaluation of Specialty Starch Germplasm Utilizing GEM Biodiversity to Optimize Grain Quality, Composition, and Yield. Duration Mark Campbell, Cynthia Cooper, Amanda McClure and Linsa Dean, Sushma, Shrestha and Adrianne Alexander General Objectives: Truman state has served as a public cooperator in GEM program in order for student to activity participate in demonstrating the economic value of exotic maize germplasm, educates students, the local, state and scientific community of the necessity to support the National Plant Germplasm System. Our work consist of developing parent lines and hybrids with altered starches that address current trends in industry and concerns of consumer and health specialists. Our source of used at Truman for line development relies exclusively on materials released by GEM cooperators. In addition, DH lines recently released from the allelic diversity study are being introgressed into our existing genetic materials which expands the diversity used to develop and improve novel specialty starch maize. Additionally, we have used landraces adapted to specific areas where this material could potentially serve as an affordable source of prebiotic in areas vulnerable to gastrointestinal illnesses such as cholera. Specifically, our breeding efforts fall into two categories which are believed to address the following consumer needs (1). Developing high-amylose corn which has a number of niche applications, one of which includes its use as a source of resistant starch (RS) having prebiotic properties that have well documented health benefits and functional attributes making it well suited as a nutraceutical dietary supplement (2). The second starch type involves developing sources of a slowly digested unmodified starch (SDS) from native genetic diversity. Rather than focusing on RS starch, the rate of starch hydrolysis and other components in the grain influencing retention which may determine its potential applications in controlling blood sugar. Although a number of applications exist, evaluation of our materials involved the glycemic-index (GI), glycemic load (GL), -Amylase Inhibitors and the recent observations made regarding interactions observed in human metabolomics and cereal carbohydrates. Our material may serve to assist in understanding strategies that best identify therapeutic food choices for diabetic and glycogen disorder patients suffering from nocturnal hypoglycemia. This past summer several ongoing studies and new approaches used in the development of germplasm has taken place. Highlighted are field studies that demonstrate agronomic potential of Amylomaize lines developed using only GEM parents under varied environmental conditions. Continued effort involving collaboration of Truman State with A.T. Still Medical College and ISU is being accomplished. Hybrid Evaluation Evaluation of hybrids had previously been made from crosses between SS and NS lines believed to have alleles fixed resulting in starch amylose at or exceeding 70% during the 2009/10 winter nursery with good success and the results reported in the 2012 GEM report. A series of unfortunate events have made progress in this area challenging since then. From the earlier study, an additional crossing block was prepared using two of GEM amylomaize lines exhibiting good general combining together with a number of SS and NS intended for planting during the 2011/12 winter nursery in Puerto Rico (Table 1). This seed, however, was lost in transit, eventually recovered, and planted during the 2012/13 season instead. The seed was successfully planted; however, during flowering insect damage greatly limited the amount of successful crosses. Some of this material was planted 21

22 in Ames, IA during the summer of The results of this study are provided in Table 2. Due to the unfortunate loss of a fellow GEM private cooperator and friend, no winter crossing blocks have been made since. It will soon be necessary to identify an alternative plan, perhaps through in kind support of a GEM cooperator to ensure continued hybrid field experiments as new inbreds are developed. Table 1. Crossing block grown during the 2012/13 winter nursery and numbers of successful ears recovered following crossing are shown. No Crossing block, Puerto Rico, 2012/ CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS ears harvested - tassel damage army worm 2 CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS-67 3 Male Entry 3 CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS-67 1 CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS /97_DK888N11F2S3_ b-b/////CH001:N B-B////DKXL370:N11a B-B- SIB///GEMS BARBGP2:N08a B-B-B/////CH001:N B-B////DKXL370:N11a B-B-SIB///GEMS FS8B(T):N11a b-b-sib-b-b/////CH001:N B-B////DKXL370:N11a B-B-SIB///GEMS CHO01:N b-b-b/////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS DKXL370:N11a B-B-B-Sib/////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS BR167:N /////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS MDI022:N21-B ///// DKXL370:N11a B-B-B////DKXL370:N11a B-B-SIB///GEMS CH01:N b-b-b/////UR1308:N B-B-B-B////DKXL370:N11a B-B-SIB///GEMS DK888:N11-B B-001/////DREP10:N2011d B-B////DKXL370:N11a B-B-SIB///GEMS DREP10:N2011d B-B////DKXL370:N11a B-B-SIB///GEMS CHO01:N b-b-b/////UR1308:N B///GEMS /97_DK888N11F2S3_ b-b////UR1308:N B///GEMS BARBGP2:N08a b-b-b/////UR1308:N B///GEMS BR167:N ////UR1308:N B///GEMS CL-G1607(CML420):N ////UR1308:N B///GEMS CH01:N b-b-b////UR10001:S ///GEMS CH001:N B-B-B/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS CH001:N B-B-B/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS DK212T:N11a B-B-B/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS DK212T:N11a B-B-B/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS DKXL370:N11a B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS BR1403(PE001):N16-B C-001////CH001:N1-3-1-B-B///GEMS DKXL370:N11a B-B-B-Sib////FS8A(S):S B///GEMS DKXL380:N11-B B-002/////CHRIS77:S1911b B-B-B////CUBA164:S B///GEMS-67 0 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B- B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B- B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B- B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B- B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B- B///GEMS-67 Male Entry _SE32_S17_F2S4_9148/////CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00/////CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEMS CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEMS CUBA164:S B-B////AR1603:S B-B///GEMS DKB844:S B-B-B-B-B/////CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEMS BR10:S B-B/////DKB844:S B-B-B////AR1603:S B-B///GEMS

23 41 DKB844:S B-B-B////AR1603:S B-B///GEMS _DK212T_S11_F2S4_917-Blk29/00-sib-B-B-B/////DKB844:S B-B-B////AR1603:S B- B///GEMS _SE32_S17_F2S4_9148-Blk22/00////AR1603:S B-B///GEMS GUAT209:S13 08a B-B////CHIS740:S1411a B-B////AR1603:S B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00-sib///GEMS CHIS77:S1911b B-B-B-B//// _SE32_S17_F2S4_9148-Blk22/00-sib///GEMS _SE32_S17_F2S4_9148-Blk22/00//// _SE32_S17_F2S4_9148-Blk22/00-sib///GEMS CUBA164:S B-B-Sib//// _SE32_S17_F2S4_9148-Blk22/00-sib/// GEMS CHIS740:S11411a b-b-b//// _SE32_S17_F2S4_9148-Blk22/00-sib///GEMS CUBA164:S B///GEMS CHRIS77:S1911b B-B-B////CUBA164:S B///GEMS CUBA164:S111b B-B-B-B-B-Sib/////CHRIS77:S1911b B-B-B//// CUBA164:S B///GEMS FS8A(S):S B///GEMS _SE32_S17_F2S4_9148-Blk22/00////FS8A(S):S B///GEMS-67 1 CHIS740:S11411a b-b-b/////CUBA164:S111b-32-1-B-B////FS8A(S):S B///GEMS BVIR1:S B-B////UR10001:S ///GEMS CHIS740:S11411a b-b-b////UR10001:S ///GEMS DKB844:S B-B-B-B-B////UR10001:S ///GEMS UR10001:S ///GEMS _SE32_S17_F2S4_9148-Blk22/00////UR10001:S ///GEMS UR11003:S b-b-b////UR10001:S ///GEMS BR167:N ////UR10001:S ///GEMS BR2060:S b-b-b////UR10001:S ///GEMS GUAT209:S13 08a B-B////UR10001:S ///GEMS CUBA164:S2008c B-B/////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS H99 ae///gems-67 1 Results from the yield evaluation grown in Ames, IA in 2013 are provided in Table 2. Seed obtained from the 2012/13 crossing block (Table 1) was intended to be used as a source for hybrid entries which are indicated by the purple entry numbers in Table 2. The delayed arrival and extensive insect damage required that seed from 2009/10 was used (geen entry numbers) to complete the experiment. Entry 47 (AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEM67 X CUBA164:S B///GEM67) displayed the greatest yield with a plot average of 16.0 bu/ac. Of the entries from the later crossing block, entry 16 (BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B- B////CH001:N1-3-1-B-B///GEMS-67) displayed the greatest yield at 13.2 bu/ac. In fact, a number of independent ears having similar parentage were used as independent entries due to the insect damage that resulted in a lack of successful crosses. This provides useful information regardless of the poor hybrid cross attempts since BVIR1:S B-B////UR10001:S ///GEMS-67 had not been used in any experimental hybrids in the past and suggests that lines derived from this pedigree be further prioritized for increasing yield performance. Table 3 was made to examine SS lines that had a common parent constituting 0% of their pedigree. The lines possessing _DK212T_S11_F2S4_917-Blk29/00-sib-B-B-B/////DKB844:S B- B-B////AR1603:S B-B///GEMS-67 averaged the greatest yield of bu/ac. In addition, lines with GUAT209:S1308a B-B and BVIR1:S B-B////UR10001:S ///GEMS-67 consistently performed well. During the summer of 2014, an effort was made to include remaining seed of experimental hybrids that had not yet been examined. Stand counts suffered for many as a result of as a result of seed aging. Early planting if followed by cool damp conditions also make emergence of amylomaize stands suffer. There were a few hybrids that yielded fairly well. In some cases, seeding rates routinely used 23

24 may be excessive for amylomaize VII material and, therefore, yields may have increased with decrease seed viability. CHIS77:S1911b B-B-B////AR1603:S B-B/// GEM67 X AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B/// GEM67 Yield %stand CHIS77:S1911b B-B-B////AR1603:S B-B/// GEM67 X DKB844:S B-B-B-B-B/////CUBA164:S111b-32-1-B-B////AR1603:S B-B/// GEM67 Yield %stand CH001:N B-B////(CH001:N1-3-1-B-B///GEM67 X CUBA164:S111b-32-1-B-B////AR16303:S B- B///GEM67 Yield %stand

25 Table 2. Experimental amylomaize GEMN x GEMS hybrids grown in a randomized complete block design near Ames, IA 2013 Hybrid (Amylomaize GEMN x GEMS) Yield Evaluation, Entry 09/10 12/13 WT Yield (bu/ac) MOIST Y/M TWT AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS-67 x CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS-67 x CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS-67 x CHIS77:S1911b B-B-B////AR1603:S B-B///GEMS-67 AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS-67 x UR11003:S b-b-b////UR10001:S ///GEMS _SE32_S17_F2S4_9148-Blk22/00/////CUBA164:S111b-32-1-B- B////AR1603:S B-B///GEMS-67 x AR0306:N B-B-B- Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 BR10:S B-B/////DKB844:S B-B-B////AR1603:S B- B///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B- B////CH001:N1-3-1-B-B///GEMS _DK212T_S11_F2S4_917-Blk29/00-sib-B-B-B/////DKB844:S B-B- B////AR1603:S B-B///GEMS-67 x AR0306:N B-B-B- Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00//// _SE32_S17_F2S4_9148-Blk22/00- sib///gems-67x AR0306:N B-B-B-Sib/////AR0306:N B-B- B////CH001:N1-3-1-B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00//// _SE32_S17_F2S4_9148-Blk22/00- sib///gems-67x AR0306:N B-B-B-Sib/////AR0306:N B-B- B////CH001:N1-3-1-B-B///GEMS-67 CUBA164:S2008c B-B/////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B- B////CH001:N1-3-1-B-B///GEMS-67 CUBA164:S2008c B-B/////AR0306:N B-B-B////DKXL370:N11a B-B-SIB///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B

26 B////CH001:N1-3-1-B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS _SE32_S17_F2S4_9148-Blk22/00////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N B-B///GEMS-67 BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 BVIR1:S B-B////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 CHIS740:S11411a b-b-b////UR10001:S ///GEMS-67x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS _DK212T_S11_F2S4_917-Blk29/00-sib-B-B-B/////DKB844:S B-B B////AR1603:S B-B///GEMS-67 x AR0306:N B-B-B- Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 amylomaize I amylomaize II UR11003:S b-b-b////UR10001:S ///GEMS-67 x AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEM AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X UR10001:S ///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X GUAT209:S1308a B-B////UR10001:S ///GEM

27 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X DKB844:S B-B-B-B-B/////CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X UR1308:N B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X DK212T:N11a B-B-B/////CH001:N B-B////DK L370:N11a B-B- SIB///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X GUAT209:S1308a B-B/////CHIS740:S1411a B-B////AR1603:S B- B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X CH001:N B-B////DKXL370:N11a B-B-SIB/// GEM67 CHIS740:S11411a b-b-b////UR10001:S ///GEMS-67 X AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 X AR0306:N B-B-B////(CH001:N1-3-1-B-B/// GEMS-67 CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 X AR0306:N B-B-B////(CH001:N1-3-1-B-B/// GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X UR1308:N B///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X CUBA164:S B///GEM67 CUBA164:S111b B-B-B-B-B-Sib/////CHRIS77:S1911b B-B-B//// CUBA164:S B///GEMS-67 X AR0306:N B-B-B- Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEMS-67 CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 X DKXL370:N11a B-B-SIB//GEM67 CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 X FS8B(T):N11a B-B////DKXL370:N11a B-B-SIB///GEM67 CUBA164:S111b-32-1-B-B////AR1603:S B-B/// GEM67 X DREP10:N2011d B-B////DKXL370:N11a B-B-SIB/// GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X AR0306:N B-B-B////DK L370:N11a B-B-SIB///GEM

28 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X DREP10:N2011d B-B////DK L370:N11a B-B-SIB///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X _SE32_S17_F2S4_9148-Blk22/00-sib///GEM67 CH001:N B-B////(CH001:N1-3-1-B-B///GEM67 X CUBA164:S111b B-B////AR1603:S B-B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X AR0306:N B-B-B//// CH001:N1-3-1-B-B///GEM67 AR0306:N B-B-B///DKX L370:N11a B-B-SIB///GEM67 X CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X CUBA164:S B///GEM67 AR0306:N B-B-B////DKX L370:N11a B-B-SIB///GEM67 X CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X DK212T:N11a B-B-B/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEM67 CHIS77:S1911b B-B-B////AR1603:S B-B///GEM67 X AR0306:N B-B-B-Sib/////AR0306:N B-B-B////CH001:N1-3-1-B-B///GEM mean S CV% Entry ** ** ** ** ** ** Block ** ** ** ns ** ** 28

29 Table 3. Amylomaize VII SS lines grouped having in their pedigree at least half being derived from a common ancestor. In other words, these are average performance values of line having a common parent. Mean hybrid performance sharing the following SS lines making up at least 0% of it background Entry WT Yield (bu/ac) MOIST Y/M TWT _SE32_S17_F2S4_9148-Blk22/00- sib///gem67 1 mean S n = 7 CV% _DK212T_S11_F2S4_917-Blk29/00-sib-B-B- B 2 mean S n =2 CV% Amylomaize VII Checks 3 mean S n =2 CV% BR10:S B-B 4 mean S n =6 CV% CHIS740:S11411a b-b-b mean S n =2 CV% CHIS77:S1911b B-B-B 6 mean S n =14 CV% CUBA164:S B///GEM67 7 mean S n =12 CV% GUAT209:S1308a B-B 8 mean

30 n=1 GUAT209:S1308a B-B 9 mean S n =2 CV% UR10001:S ///GEM67 10 mean n=1 UR11003:S b-b-b 11 mean S n =2 CV%

31 Table 4. Hybrid performance of amylomaize VII GEMN x GEMS lines in Ames, IA 2014 from seed of lines produced in a crossing block grown in a 2009/10 P.R. where most hybrids entries have been included in previous yield trials. Ames, IA 2014 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X CHIS740:S1411a B- B////AR1603:S B-B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B////(CH001:N1-3-1-B-B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X (UR1308:N B///GEMS-67 WT Yield (bu/ac) MOIST Y/M TWT %stand %skldg %rtldg AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X (CUBA164:S B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X (CUBA164:S B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X DKB844:S B-B-B-B- B/////CUBA164:S111b-32-1-B-B////AR1603:S B- B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X GUAT209:S1308a B- B/////CHIS740:S1411a B-B////AR1603:S B- B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B- B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B- B////DKXL370:N11a B-B-SIB///GEMS

32 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X FS8B(T):N11a B- B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X DREP10:N2011d B- B////DKXL370:N11a B-B-SIB///GEMS CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X DKXL370:N11a B-B-SIB///GEMS CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X DK212T:N11a B-B- B/////CH001:N B-B////DKXL370:N11a B- B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B- Sib/////AR0306:N B-B-B////(CH001:N1-3-1-B- B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X AR1603:S B-B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X CUBA164:S111b-32-1-B B////AR1603:S B-B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X UR10001:S ///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X ( _SE32_S17_F2S4_9148-Blk22/ sib///gems-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X (UR1308:N B///GEMS CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B- B////DKXL370:N11a B-B-SIB///GEMS

33 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X DKXL370:N11a B-B B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X CH001:N B B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B B////(CH001:N1-3-1-B-B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X AR1603:S B-B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X AR1603:S B-B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X DKB844:S B-B-B-B- B/////CUBA164:S111b-32-1-B-B////AR1603:S B B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X CUBA164:S111b-32-1-B B////AR1603:S B-B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X UR10001:S ///GEMS CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X UR1308:N B///GEMS AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X _SE32_S17_F2S4_9148-Blk22/00- sib///gems

34 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X DKXL370:N11a B-B B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X CH001:N B B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B////CH001:N B-B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B////CH001:N B-B///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B////CH001:N B-B///GEMS-67 AmyVI Check AmyVI Check CUBA164:S111b-32-1-B-B////AR1603:S B- B///GEM67 X DKXL370:N11a B-B-SIB//GEM CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X FS8B(T):N11a B- B////DKXL370:N11a B-B-SIB///GEMS-67 CHIS77:S1911b B-B-B////AR1603:S B- B///GEMS-67 X AR0306:N B-B-B////(CH001:N1-3-1-B-B///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X UR10001:S ///GEMS-67 AR0306:N B-B-B////DKXL370:N11a B-B- SIB///GEMS-67 X CHIS740:S1411a B- B////AR1603:S B-B///GEMS-67 CH001:N B-B////CH001:N1-3-1-B-B///GEM- 67 X CUBA164:S111b-32-1-B-B////AR1603:S B-B///GEM

35 401/3042 DREP10:N2011d B- B////DKXL370:N11a B-B-SIB///GEMS-67 X _DK212T_S11_F2S4_917-Blk29/00-sib-B-B B/////DKB844:S B-B-B////AR1603:S B- B///GEMS /4038 GUAT209:S13 08a B- B////CHIS740:S1411a B-B////AR1603:S B- B///GEMS-6 X CUBA164:S B-B////AR1603:S B-B///GEMS-67 CUBA164:S111b-32-1-B-B////AR1603:S B- B///GEM67 X FS8B(T):N11a B-B////DKXL370:N11a B-B-SIB///GEM /4016 CHIS77:S1911b B-B-B-B//// _SE32_S17_F2S4_9148-Blk22/00-sib///GEMS-67 X CHO01:N b-b-b/////UR1308:N B///GEMS-67 Normal (Asgrow) Mean S CV% Max min entry ** ** ** ** ** ns ** ns ns ns ns ns ns ns 3

36 Accessions currently being used to introduce adaptation genes to various regions in Nepal. Map made using Excel Fusion and Google maps. Page 36 of 41