Germplasm Enhancement of Maize - Strategies & Synergy with Maize

Curation

Michael Blanco and Candice Gardner, USDA-ARS and Iowa State University, Ames, IA

The importance of access to and utilization of genetic plant resources for improvement of modern cultivars is widely recognized. Industry recognized the challenges of incorporating diversity into their elite breeding pools because of lack of adaptation and undesirable agronomic traits associated with exotic maize germplasm, the time required to extract valuable material, and the rapid nature of breeding cycles in commercial programs. At the conclusion of the Latin American Maize Project (LAMP) project, US participants pursued discussions of how materials identified with good potential could actually be utilized to broaden the diversity of U.S. maize. The Germplasm Enhancement of Maize (GEM) Project resulted from discussions on the need and urgency of broadening and enhancing the maize germplasm. Nearly 60 entities representing the private, public, and non-governmental organizations (NGO) in the US and nine other countries now collaborate to achieve GEM objectives.

More than 265 varieties have been released from programs based in Ames, IA, Raleigh, NC, and from university researchers, from ~ 30 maize races or tropical hybrids. Also, about 200 doubled haploid lines from nearly 60 exotic landraces are ready for release and will serve as rich resources for exploration of allelic diversity and novel traits.

Collaborative efforts are essential to provide useful germplasm for introgression, to perform breeding and testing activities, to evaluate germplasm for abiotic and biotic stress resistance, especially for pests not yet endemic to the U.S., for yield, agronomic and grain quality traits, and to conduct research to identify useful segments from exotic donor genomes.

Germplasm Enhancement of Maize - Strategies & Synergy

with Maize Curation

Michael Blanco and Candice Gardner

USDA-ARS Plant Introduction Research Unit Ames, IA

GEM Mission The mission of the GEM (Germplasm Enhancement of Maize) Project is to effectively increase the diversity of U.S. maize germplasm utilized by producers, global end-users and consumers.

The mission is achieved through collaborative efforts of USDA-ARS, public, and private research scientists by utilizing exotic, public, and proprietary maize germplasm. The resulting germplasm and associated knowledge, derived via the identification and incorporation of favorable alleles for prioritized traits from exotic sources, is shared and utilized in private and public breeding programs and ultimately contributes to increased genetic diversity of maize grown by producers and improved product performance. In addition, the GEM project provides opportunities for training plant scientists in the enhancement and utilization of un-adapted germplasm. This contributes to the global sustainability of agricultural production, economic stability and the nutrition and well being of society.

GEM History

Latin American Maize Project (LAMP) 1987-1992 American Seed Trade Association (1993)- helped secure resources and member support of GEM GEM Technical Steering Group (TSG) formed at 1993 ASTA meeting Wilfredo Salhuana TSG Chair (1993-2007) Tom Hoegemeyer TSG Chair (2008-2013) Walter Trevisan current TSG Chair

Early GEM Cooperators (1994-1995) GEM Private Cooperators:

Bo Jac Callahans Cargill CIBA seeds DeKalb FFR Garst Golden Harvest Great Lakes Growmark Gutwein Hoegemeyer Holden's Jung Farms Limagrain Madison Research NC+ Northrup King Pau Seeds Pioneer Hi-Bred Trelay Farms United AgriSeeds Wyffels Zeneca Inc (ICI)

GEM Public Cooperators:

Cornell University Iowa State University Louisiana State University Michigan State University North Carolina State University Penn State University Purdue University South Dakota State University Texas A&M University The Ohio State University University of Delaware University of Georgia University of Illinois University of Kentucky University of Minnesota University of Tennessee University of Wisconsin USDA-ARS PGRS (MO) USDA-ARS CICGRU (IA) USDA-ARS CHPRRU (MS)

GEM Cooperators (2014) GEM Private Cooperators:

3rd Millennium Genetics AgiWise, L.L.C. AgReliant Genetics, LLC BASF Plant Science Breeding, L.L.C. Beck's Superior Hybrids, Inc. Brandy Wine Seed Farms, LLC CRD Advisors, LLC DKD Genetics, Inc. Dow AgroSciences DuPont Pioneer FFR Cooperative Forage Genetics G and S Crop Services Genetic Enterprises Int'l Hoegemeyer Enterprises Illinois Foundation Seeds, Inc. Ingredion, Inc. JFS and Associates, LTD MBS Genetics, LLC. Monsanto Company PANNAR Seed Professional Seed Research, Inc. SEEDirect Summit Genetics LLC Syngenta Seeds, Inc. Terrell Seed Research Trimble Genetics International, LLC Wyffels Hybrids

GEM Public Cooperators:

Cornell University Iowa State University North Carolina State University North Dakota State University South Dakota State University Texas A&M University The University of Delaware The University of Illinois The University of Nebraska The University of Tennessee The University of Wisconsin Truman State University USDA-ARS: 8 Research Units GEM NGO Cooperator:

Michael Fields Ag Institute

GEM Int’l Private Cooperators:

MayAgro Seed Corp., Turkey Nidera, Argentina Nugenplasm, LLC, Indonesia Prosemillas, Peru Seed Asia Co., Ltd., Thailand Sementes Biomatrix Ltda, Brazil Semillas Fitó, SA, Spain Semillas Tuniche, Ltda, Chile Sursem SA, Argentina GEM Int’l Public Cooperators:

Agriculture and Agri-Food, Canada Chinese Academy of Agricultural

Science, China Empresa Brasileira Pesquisa

Agropecuaria (EMBRAPA), Brazil Instituto Nacional Tecnologia

Agropecuaria (INTA), Argentina The University of Guelph, Canada

Other significant contributions

Illinois Foundation Seeds provided testers CIMMYT provided much germplasm Too many to mention….

GEM Project – Ames, IA & Raleigh, NC

Ames, IA, USDA-ARS Candice Gardner, Research Leader Mike Blanco, Midwest Coordinator Fred Engstrom, Technician Mike Peters, Technician Mack Shen, IT Specialist

Raleigh, NC, USDA-ARS

David Marshall, Research Leader

Matt Krakowsky, Southeast Coordinator

Dale Dowden, Technician

North Carolina State Univ. Major Goodman, Professor

Choice of Germplasm

LAMP Exotic Accessions-original breeding base New sources introduced:

Suwan inbreds (NEI9004, NEI9008) Proprietary tropical inbreds x GEM breeding crosses ARS lines (Mp494, Mp717, Mp313E) New tropical hybrids African germplasm (KO679Y, TZAR inbreds) CML lines

Breeding Cross Combinations

Original source: (Exotic x private line A) x private line B Exotic/Elite (Tropical or Temperate)

Other Options or Potential Options: GEM A/GEM B GEM A/Private A GEM A/Ex-PVP GEM A/Exotic //GEM B GEM A/Exotic//Ex-PVP Exotic/Private A//Elite Exotic///Private B

Exp: 121401 (2012) Tester: HC33 Reps: 5

Pedigree Yield B/A % Moist Y/ M TWT % S.L. % R.L. NEI9008:N082699ai-B-051 186.9 22.8 8.2 55.7 4.7 2.9 NEI9008:N082699ai-B-062 182.3 24.7 7.4 55.5 3.5 2.5 NEI9008:N082699ai-B-033 179.5 23.8 7.5 57.1 1.2 20.9 NEI9008:N082699ai-B-057 178.3 24.1 7.4 56.2 2.4 10.6 Pioneer 33F85 176.3 20.7 8.5 57.0 3.6 5.9 NEI9008:N082699ai-B-047 174.3 23.3 7.5 56.2 6.6 17.9 NEI9008:N082699ai-B-045 172.7 21.9 7.9 56.3 8.2 11.9 NEI9008:N082699ai-B-007 172.7 22.4 7.7 58.0 6.1 6.6 LH200xLH262 172.3 25.9 6.7 54.9 1.5 10.1 NEI9008:N082699ai-B-020 171.3 24.2 7.1 54.8 1.3 30.8 NEI9008:N082699ai-B-041 171.0 22.4 7.6 55.5 0.3 4.4 Pioneer 31D58 171.0 24.8 6.9 57.0 2.8 3.8 NEI9008:N082699ai-B-048 170.3 24.1 7.1 57.2 2.6 9.8 NEI9008:N082699ai-B-004 168.7 24.5 6.9 55.5 3.4 4.8 NEI9008:N082699ai-B-072 168.1 24.7 6.8 55.3 3.3 3.7 NEI9008:N082699ai-B-052 168.0 23.6 7.1 56.5 1.8 18.0 NEI9008:N082699ai-B-026 166.4 24.3 6.8 56.1 6.7 7.6 NEI9008:N082699ai-B-066 166.3 24.0 6.9 56.3 5.8 12.8 HC33xLH287 165.8 19.2 8.6 55.9 2.7 9.0 NEI9008:N082699ai-B-053 165.1 22.7 7.3 56.0 11.9 13.5

Test Entry Means 156.7 23.0 6.8 56.6 4.4 7.5 Check Means 163.8 21.9 7.5 56.5 2.4 6.7 CV 15.4 6.4 LSD (p=0.05) 30.3 1.8

GEM breeding cross with 4 parents

2013 GEM Photoperiod Shade House

Photo credit: Denise Costich, CIMMYT

GEM SSD Breeding protocol

How In-Kind Support Works Make Breeding Crosses Top Cross S2’s (cross to proprietary lines) Conduct Yield Trials (NS=non-stiff; SS=stiff stalk) Breeding Cross Obs Make S1’s Disease Screening Advance S1 to S2 Lab Services (Describe) Regenerations Other (Describe)

From the agreement: “It is understood that any seed or data generated as defined in this appendix is considered in-kind support and shall be returned to the GEM Coordinator. Such seed and/or data shall be shared with other GEM Cooperators, who shall have the right to freely use such seed and/or data.”

Yield Testing Protocol

Cooperators (and GEM) make top crosses in ICB Cooperators (and GEM) conduct yield trials at their managed locations First year entries tested in 5 locations on 1 tester, 2-row plots Second year entries tested in 8 locations on 2 or more testers, 2-row plots Five standard commercial checks are included in all trials Cooperator sends raw data to GEM IT Specialist for analysis Results discussed at Annual GEM Cooperator Meeting and approximately 10 GEM releases per year

Entomology Evaluation

Army Worm/CEW (USDA-ARS-CGBRU Tifton, GA) Army Worm/CEW (3rd Millennium Genetics, Puerto Rico) Corn Rootworm (USDA-ARS-PGRU, Columbia, MO; U. Illinois)

Diseases and Mycotoxin: Potential Global Threats

Late Wilt, Cephalosporium maydis Europe Maize Rough Dwarf virus (MRDV) Europe & Asia New Maize Lethal Necrosis (Africa) Gray Leaf Spot (different strain from US) and NLB (Africa) Southern Rust host differential development Aspergillus spp. and Fusarium spp. grain mycotoxins Asian Downy Mildew

Pathology Evaluation GLS, SLB, and Fusarium Ear Rot (USDA-ARS-PSRU Raleigh, NC)

GLS, SLB, Common Rust and Goss’s Wilt (Professional Seed

Research)

NLB and Diplodia Ear Rot (DuPont Pioneer Hi-Bred, Johnston, IA)

Aflatoxin (USDA-ARS CHPRRU, MS State)

Fusarium Ear Rot and Head Smut (DuPont Pioneer Hi-Bred,

Woodland, CA)

NLB and foliar diseases (Agriculture and Agri-Food, Canada)

NLB (MayAgro, Turkey)

Enhancement Accomplishments

GEMS-0002 and GEMS-0003 identified for Fusarium Ear Rot and Reduced Fumonisin (DuPont Pioneer; USDA-ARS PIRU Ames, IA; USDA-ARS PSRU Raleigh, NC) KO679Y, GEMS-0030, GEMN-0130 identified for reduced aflatoxin levels (USDA-ARS CHPRRU Mississippi State, MS; USDA-ARS PIRU Ames, IA; USDA-ARS PSRU Raleigh, NC) Antigua03:N1242 identified as potential source for Fall Armyworm resistance (3 MG, Puerto Rico; USDA-ARS PIRU Ames, IA) Multiple insect resistance identified in GEM sources GEMN-0130, GEMN-0133, and CUBA117:S15 (USDA-ARS-CGBRU Tifton, GA; USDA-ARS PIRU Ames, IA; USDA-ARS PSRU Raleigh, NC; Texas A&M University)

Fusarium Ear Rot Inoculated GEMS-0002

Bill Dolezal, Pioneer Hi-Bred Int’l

Susceptible Line Resistant Line GEMS-0002

Diplodia Ear Rot Inoculated GEMS-0003

Bill Dolezal, Pioneer Hi-Bred Int’l

Public Cooperator Lines and Synthetics TX A&M (W. Xu)

Drought and stress resistance Tx 204, Tx 205 (drought), 5 BMR lines U of WI (N. de Leon) 7 WI releases for silage; CUBA164 + CUBA117 GEM Quality Synthetic (GQS) NDSU (M. Carena) 7 EarlyGEM inbred releases. Sources identified, lines derived from BR52051, CUBA117, CH05015, FS8B, and DKB844. (NC2000 source of earliness) U. of IL (M. Bohn) Illinois WCR synthetic- long term recurrent selection; Identification of breeding sources with reduced damage from WCR included AR16026:N1210, AR17056:N2025, CUBA117:S1520, FS8A(S):S0907, and UR13085:N0204. QTL mapping pop (AR17056 and LH51) Truman State University (M. Campbell) GEMS-0067 first public release of amylomaize VII germplasm

Racial Background of 265 GEM Releases Race Total Race TotalAlazan 1 Dente Amarelo 11Argentino 6 Dente Branco 4Camelia 8 Early Caribbean 1Cateto 11 Hybrid-tropical 94Cateto Sulino 4 Mixed 16Composite (Suwan) 6 GEM Derived* 3Composite (Tuxpeño) 1 Multiple Borer Composite 1Corn Belt Dent 1 Semidentado Riograndense 2Criollo 1 St. Croix 2Criollo/Argentino (CUBA164) 17 Suwan 8Criollo/Tuson (CUBA173) 1 Tropical Line 7Cristalino Colorado 17 Tusón 13Cuban yellow 7 Tuxpeño 9Dent. Blanco Rugoso 6 Tuxpeño/Olotillo 1Dentado Blanco 5 Unclassified 1

Totals (~30 Races)

GEM 20th Anniversary Field Day - (Sept.20, 2013)

GEM 20th Anniversary Field Day - (Sept.20, 2013)

Doubled Haploids for Release (~200) Race Country Race Country Race Country Altiplano Bolivia Cubano dentado Bolivia Olotón Guatemala

Ancashino Peru Curagua Grande Chile Onaveño Mexico

Andaqui Colombia Cuzco Peru Patillo Ecuador Aragüito Venezuela Dulcillo del Noroeste Mexico Patillo Grande Bolivia Arequipeño Peru Early Caribbean Martinique Perla Peru

Arizona Peru Elotes Occidentales Mexico Pira Colombia

Avati Moroti Guapi Paraguay Elotes Occidentales Mexico Pisankalla Bolivia

Blanco Blandito Ecuador Gordo Mexico Pojoso Chico Bolivia

Bofo Mexico Jora Peru San Geronimo Huancavelicano Peru

Cabuya Colombia Karapampa Bolivia Semi dentado paulista Paraguay

Camelia Chile Kcello Ecuatoriano Bolivia Suwan Brazil

Candela Ecuador Mishca Ecuador Tehua Mexico

Capio rosado Argentina Mixed Creole Cuba Tuxpeño Brazil

Caraja Brazil Montaña Colombia Tuxpeño Guatemala

Chandelle Cuba Morado Bolivia Vandeño Mexico

Confite Puntiagudo Peru Morado Canteño Peru Yucatan Colombia Cónico Norteño Mexico Morocho Peru Yungueño Bolivia

Cristalino amarallo Argentina Morocho Peru Yunquillano forma andaqui Ecuador

Allelic Diversity Project Update Doubled Haploid Lines

# Lines % of Total

To Be Released 204 77

Need Seed Increase

16 5

Need Verification

23 8

Discard Lines 26 10

Total (~ 60 Races)

269 100

DH lines from a Yungueno accession, NSL 286578, or Bov 362, collected in Bolivia, donated originally by the Programa Maiz Y Sorgo, Instituto Colombiano Agropecurario, and later donated to the NPGS by Dr. Major Goodman, North Carolina State University. GRIN accession status: Unavailable

Bov 903 references PI 485364, also collected in Bolivia, donated through the Instituto Colombiano Agropecurario, CIMMYT, and Major Goodman. Race: Altiplano GRIN status: Unavailable

DH lines derived from VEN 678, PI 485364, cultivated material of the Araguito race. Collected in Venezuela, deposited in the NPGS by Major Goodman, via CIMMYT. GRIN status: Unavailable

CUZ 13, or Cuzco 13, is an example of the ‘Confite Puntiagudo’ race collected from Peru, and part of the Goodman Maize Racial collection. GRIN ID = Ames 28653 Status: Unavailable

DH lines from the race Conico Norteno, PI 515577, from Mexico. GRIN status: Available

Derived from PI 485274, collected in Urubamba, Cuzco, Peru, Elevation: 3200 meters. Primary race: Cuzco Obtained via CIMMYT and Wilfredo Salhuana.

DH lines derived from Elotes Occidentales Nay 29, from Nayarit, Mexico ()

DH lines derived from Gordo race (PI 484406), collected in state of Chihuahua, Mexico

GEM -0235-N derived from GUA 456, Ames 28567, from El Progreso, Guatelmala; and others from PI 445514, from Colombia; donated to the NPGS by Major Goodman, NCSU.

Note: Most of the images you observed indicated the status of tropical landrace accessions is generally ‘unavailable.’ Without sufficient resources to maintain tropical accessions within the maize collection, they will continue to be unavailable .

Current GEM Efforts

57 external partners help accomplish breeding, evaluation and genetic characterization research objectives

Traditional GEM population and line development

•Introgression of unadapted germplasm • Testing, selection, and S2 line release

Allelic Diversity Project – develop unselected inbred lines representative of all of the races of maize for use as research resources.

•Conventional breeding •DH breeding in partnership with the ISU Doubled Haploid Facility

Education of plant breeders, especially in use of exotic germplasm

With the assistance of GEM TSG members, a set of germplasm has been generated to study whether random mating of the allelic diversity BC1’s prior to selection and selfing is important when working with exotic germplasm.

20 families - (exotic/PHZ51)PHZ51 or (exotic/PHB47)/PHB47 - BC1s - BC1Sib1 (generated in PR) - BC1Sib2 (generated in Ames)

Germplasm will be induced, doubled, and the genetic contributions of the exotic donors in DH and traditionally derived lines compared.

The Next Chapter of GEM

GEM (Ames and Raleigh) and the Maize Curation Project at the NCRPIS will continue to develop synergies and efficiencies to understand and capture the value of exotic germplasm. Curation and utilization are complementary.

Balance priorities with available resources to provide useful germplasm that offers unique alleles and traits. •Exploratory – including allelic diversity •Introgressing tropical inbreds and hybrids •Recycling superior GEM germplasm •Identifying sources of DR/IN resistance •Identify useful genetic segments •Database and information delivery

Partnerships will be sought to develop DH lines, to understand the nuances involved in applying DH technologies to exotic germplasm, and genetic consequences

International collaborations will increase to characterize germplasm in unique environments to provide information on performance, biotic and abiotic stress resistance and other traits

International disease nurseries will serve to monitor host- pathogen interactions, especially for diseases not yet in North America.

GEM will contribute to addressing vulnerabilities associated with climate change and global crop production challenges

Thoughts to Consider What is the dividend from the GEM Project beyond insurance? GEM germplasm and research are leveraged to improve plant breeding knowledge and improve crop production. The magnitude of the dividend will be determined by the quality of the germplasm, the science, and our collaborations.

Acknowledgements

Raleigh, NC GEM Project: Major Goodman, Matt Krakowsky, Dale Dowden Ames, IA GEM Project: Nuo (Mack) Shen, Fred Engstrom Maize Curation Project Members: Mark Millard, Brady North, Bruce Hall Iowa State University Doubled Haploid Facility: Thomas Lubberstedt, Uschi Frei, and Ph.D. student Adam Vanous GEM Cooperators and GEM Technical Steering Group Members A very special thank you to Wilfredo Salhuana,

Major Goodman, Marty Carson and Linda Pollak for their many contributions to maize

germplasm enhancement !

Mike Blanco, Germplasm Enhancement Breeder

Thanks for 14 years of Service and Leadership of GEM!