Progress 10/01/16 to 09/30/17

Outputs
Target Audience:The main target audiences were canola growers and canola industry such as Monsanto, DL Seeds Inc., Proseed, Star Seeds Inc. who were providing canola cultivars in this North Central region of USA. Changes/Problems:Weather is always a problem for field crop production. Planting time is often delayed because of early rain. Similarly, storm or rain also delayed the harvesting. Excessive rain, flood, drought, heat during crop growing time significantly affect the crop production. In 2017, due to drought and heat the experimental trials were abounded at Hettinger. What opportunities for training and professional development has the project provided? How Student Success Collaborative (SCC) Made My Job Easier. Plains Room, MU, March 01, 2017. Student Centered Learning: What a POGIL classroom can do for your students. With Laura Lavine, Washington State University. NDSU A. Glenn Hill Center, Friday, March 10, 2017. Innovating Education to Educate Innovators Colloquium with Eric Mazur, Century Theatre, Memorial Union, NDSU, April 5, 2017. Working with Industry, Hidatsa Room, MU, NDSU, November 15, 2016. Search committee training, Badland, MU, NDSU, December 15, 2016. How have the results been disseminated to communities of interest? A presentation on NDSU canola breeding research was given to 2017 canola field day at Langdon, USA on July 20, 2017. . Presentation was given to stakeholder's (Northern Canola Growers' Association) annual meeting on current canola activities and research on November 16, 2017. Radio interviews was given on current canola research at "Canola Minute" of the Red River Farm Network, Grand Forks, ND. What do you plan to do during the next reporting period to accomplish the goals?I will continue to talk to the canola growers to understand their concern and to report their questions. Research emphasis will be given to develop high seed yield, high oil content, biotic and abiotic stress tolerant canola germplasm adapted to North Central region of the USA.

Impacts
What was accomplished under these goals? Accomplishments: Released the 1st conventional canola (non-GMO) "NDOLA-01" in North Dakota. Developed Public-Private partnership research with Monsanto and Germany/Canadian based company DL Seeds Inc. for the development of joint-hybrids of canola. Plot-yield testing program expanded to seven locations with about 5,000 plot/year. Identified five potential Roundup Ready joint hybrids (NDSU-h119, NDSU-15WR10, NDSU-D1357) for release. These hybrids over performed on seed yield at NDSU-canola breeding plot testing program. Also, the hybrids showed a superior seed yield performance in canola variety trial conducted by NDSU agronomists where many commercial hybrids included in the trial. Identified two conventional open pollinated breeding lines (NDC-E12079, NDC-E12121) for release. These breeding lines showed a higher seed yield both in NDSU-canola breeding plot testing program and in canola variety trial conducted by NDSU agronomists. Identified several USDA-ARS plant introduction (PI) lines resistant/tolerant to blackleg, sclerotinia stem rot diseases, tolerant to frost and heat stress. The PI lines are in progress to use in breeding program to introgress the resistant/tolerant traits into elite breeding lines. 2017 canola breeding research activities: In the 2017 summer testing program, a total of 1,853 (Breeding lines + F2) germplasm were evaluated in 5,059 plots at 7 locations in North Dakota. The canola testing locations were distributed across the state, including, Fargo, Prosper, Langdon, Carrington, Minot, Williston and Hettinger of North Dakota. Wide area yield trial for Roundup Ready hybrid: Thirty-six hybrids (from both Monsanto and DL Seeds Inc.) including four commercial checks were evaluated in a randomized complete design with three replications at Prosper, Langdon, and Minot. Advanced yield trial for Roundup Ready hybrid: Thirty-six hybrids (from both Monsanto and DL Seeds Inc.) including three commercial checks were evaluated in a randomized complete design with two replications at Prosper and Langdon. Wide area yield trial for OP lines: Forty OP lines including four commercial hybrid checks were evaluated in a randomized complete design with three replications at Prosper, Langdon, Carrington, Minot, Williston and Hettinger. Due to severe heat stress, the trial at Hettinger was abounded. Advanced yield trial for OP lines: Ninety-six OP lines including four commercial hybrid checks were evaluated in a randomized complete design with two replications at Prosper, Langdon, Carrington and Minot. Advanced yield trial (Parental lines): Eighty-one OP parental lines including three commercial hybrid checks were evaluated in a randomized complete design with two replications at Prosper and Langdon. Early generation testing for OP lines: Four hundred OP lines including two replicated hybrid checks were evaluated in an augmented design at Prosper, Langdon and Carrington. Single-row breeding nursery for OP lines: A total of 1,320 Brassica napus breeding lines (F2 to F10 generation) and 395 Brassica napus breeding lines/accessions were planted in a single-row breeding nursery at Prosper. About 4,000 breeding lines were self-pollinated for generation advancement by using micro-perforated selfing bags and bamboo sticks in the nursery. Greenhouse activities: We have been using three greenhouse rooms in the NDSU new AES greenhouse complex for our breeding program. Crossing, backcrossing and selfing are conducting in the greenhouse. We are growing three crop cycles in the greenhouse per year. A total of about 6,000 plants were planted and harvested in three growing cycles of the year. Seed quality analysis: Northern Crops Institute (NCI), Fargo, ND has modern equipment and lab facilities for seed quality analysis. I have developed new cooperative research with NCI since 2015 to evaluate our breeding lines for oil content, protein content, and fatty acid composition. Initially, we have obtained a reference data of our 20 accessions from University of Missouri. A total of 2,400 canola samples will be analyzed by NIR for the seed oil, seed protein and other fatty acid contents such as, palmitic acid, stearic acid, oleic acid, linolenic acid, linolenic acid. Field data: The data on seed yield, seed moisture at harvest, early vigor, days to flowering, relative maturity, standability, lodging, breeder's impression were taken from six planting locations.

Publications

• Type: Journal Articles Status: Published Year Published: 2017 Citation: Rahaman M, Mamidi S, and Rahman M (2017) Genome-wide association study of heat stress-tolerance traits in spring-type Brassica napus L. under controlled conditions. The Crop Journal. https://doi.org/10.1016/j.cj.2017.08.003 (ELSEVIER).
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Rahaman M, Mamidi S, and Rahman M (2017) Association mapping of agronomic traits of canola (Brassica napus L.) subject to heat stress under field conditions. Aust J Crop Sci. 11(09): 1094-1105 doi: 10.21475/ajcs.17.11.09.pne512.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Arifuzzaman M, Mamidi S, McClean P, and Rahman M (2017) QTL mapping for root vigor and days to flowering in Brassica napus L. Can J Pl Sci. 97: 99109. dx.doi.org/10.1139/cjps-2016-0048.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Danielle F, and Rahman M (2017) Studies of freezing tolerance of rapeseed/canola (Brassica napus L.). An abstract for 2017 International Annual Meetings organized by ASA-CSSA-SSSA, October 22-26, at Tampa, FL, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Roy J, and Rahman M (2017) Association mapping for sclerotinia stem rot disease in rapeseed/canola (Brassica napus L.). An abstract for 109th Annual Meeting of the North Dakota Academy of science, April 28-29, 2017, Grand Forks, ND, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Arifuzzaman M, and Rahman M (2017) Root trait variation in spring and winter canola (Brassica napu L.) under controlled and water stressed conditions. An abstract for 109th Annual Meeting of the North Dakota Academy of science, April 28-29, 2017, Grand Forks, ND, USA.
• Type: Conference Papers and Presentations Status: Published Year Published: 2017 Citation: Danielle F, and Rahman M (2017) Genome-wide association study for electrolyte leakage in rapeseed/canola (Brassica napus L.). An abstract for 109th Annual Meeting of the North Dakota Academy of science, April 28-29, 2017, Grand Forks, ND, USA.
• Type: Journal Articles Status: Published Year Published: 2017 Citation: Arifuzzaman M, and Rahman M (2017) A Comparative Study on Root Traits of Spring and Winter Canola (Brassica napus L.) under Controlled and Water Stressed Conditions. J Agril. Sci. 9(7): 58-67.

Progress 10/01/15 to 09/30/16

Outputs
Target Audience:The main target audiences were canola growers and canola industry such as Monsanto, DL Seeds Inc., Proseed, Star Seeds Inc. who were providing canola varieties in this region. Changes/Problems:Weather is always a problem for field crop production. Planting time is often delayed because of early rain. Similarly, storm or rain also delayed the harvesting. Excessive rain and flood during crop growing time significantly affect the crop production. In 2016 and 2015, due to flood/standing water the experimental trials were abounded at Langdon and Prosper, respectively. What opportunities for training and professional development has the project provided? NDSU FORWARD event "Professional development for new faculty: Securing external funding for educational initiatives" by Bevlee Watford, Program Manager for Broadening Participation, National Science Foundation, NDSU, April 29, 2015. Faculty Professional Development Workshop (Performance Task Design Workshop) presented by the Collegiate Learning Assessment (CLA) Academy, on the University of North Dakota Campus, Grand Forks, February 4 & 5, 2016. Assessment Workshops - Connecting evidence of student learning across courses, Prairie Room, Memorial Union, NDSU, Fargo, March 07, 2016. National Alliance for Broader Impacts, Plains Ballroom, MU, NDSU, March 31, 2016 Professional Development Conference, NDSU, Fargo, August 17, 2016 How have the results been disseminated to communities of interest?Presented information at canola field days and Northern Canola Growers' Association Canola Research Conference. Also given Radio interviews on current canola research at "Canola Minute" of the Red River Farm Network, Grand Forks, ND. What do you plan to do during the next reporting period to accomplish the goals?I will continue to talk to the canola growers to understand their concern and to report their questions. Research emphasis will be given to develop high seed yield, high oil content, biotic and abiotic stress tolerant canola germplasm adapted to northern Great Plains of the USA.

Impacts
What was accomplished under these goals? Developed Public-Private partnership research with Monsanto and the Germany/Canada based company DL Seeds Inc. for the development of joint-hybrids of canola. Plot-yield testing program expanded to seven locations with about 5,000 plots/year. Identified five potential Roundup Ready joint hybrids (NDSU-h119, NDSU-1227, NDSU-15WR10, NDSU-D1357, NDSU-D1355) for release. These hybrids over-performed on seed yield at NDSU-canola breeding plot testing program. Also, the hybrids showed a superior seed yield performance in canola variety trial conducted by NDSU agronomists where many commercial hybrids included in the trial. Identified two conventional open pollinated breeding lines (NDSU-14WC17 and NDSU-15WC04) for release. These breeding lines showed a higher seed yield both in NDSU-canola breeding plot testing program and in canola variety trial conducted by NDSU agronomists. Identified several USDA-ARS plant introduction (PI) lines resistant/tolerant to blackleg, sclerotinia stem rot diseases and tolerant to frost and heat stress. The PI lines will be utilized by the breeding program to introgress the resistant/tolerant traits into elite breeding lines.

Publications

• Type: Journal Articles Status: Published Year Published: 2016 Citation: Rahman M, Mamidi S, DelRio L, Ross A, Kadir MM, Rahaman MM, and Arifuzzaman M (2016) Association Mapping in Brassica napus (L.) accessions identifies a major QTL for Blackleg disease resistance on chromosome A01. Mol Breeding 36:90 (DOI 10.1007/s11032-016-0513-8). Rahman M, and de Jim�nez MM (2016) Behind the scenes of microspore-based double haploid development in Brassica napus - a review. Journal of Plant Science and Molecular Breeding 5(1): 1-9.
• Type: Journal Articles Status: Published Year Published: 2016 Citation: Fiebelkorn D, and Rahman M (2016) Development of a protocol for frost-tolerance evaluation in rapeseed/canola (Brassica napus L.). The Crop Journal 4:147-152. [doi: 10.1016/j.cj.2015.11.004 (ELSEVIER)]
• Type: Conference Papers and Presentations Status: Published Year Published: 2016 Citation: Rahaman M, Mamidi S, and Rahman M (2016) Association Mapping of Heat Stress Tolerant Traits of Canola (Brassica napus L.). An abstract (#282-7) for 2016 International Annual Meetings organized by ASA-CSSA-SSSA, November 6-9, 2016, at Phoenix, AZ, USA Rahaman M, Mamidi S, and Rahman M (2016) Genome-wide association study of high temperature stress induced phenotypes in rapeseed/canola (Brassica napus L.). An abstract for Brassica 2016 Conference, which combines the 20th Crucifer Genetics Conference, and the 19th Australian Research Assembly on Brassicas, from October 3-6th 2016, Melbourne, Australia.
• Type: Book Chapters Status: Published Year Published: 2015 Citation: Rahman M, and de Jim�nez MM (2015) Designer oil crops. In: Gupta S. (Eds.), Breeding Oilseed Crops for Sustainable Production - Opportunities and Constraints. ACADEMIC PRESS (Elsevier Lid.), Oxford, UK, pp. 359-374. (ISBN: 978-0-12-801309-0)
• Type: Conference Papers and Presentations Status: Published Year Published: 2015 Citation: Rahman M, Mamidi S, del Rio L, Ross A, Kadir MM, Rahaman MM, and Arif-Uz-Zaman M (2015) Association mapping in Brassica napus (L.) accessions identifies a major QTL for blackleg disease on chromosome A01 at 9.66 Mbp. An abstract for 2015 International Annual Meetings organized by ASA-CSSA-SSSA, November 15-19, 2015, at Minneapolis, MN, USA. de Jim�nez MM, Mamidi S, McClean P, and Rahman M (2015) Genetic diversity study of a core collection of Brassica napus accessions based on genotyping by sequencing. An abstract for Plant Biology 2015, organized by the American Society of Plant Biologists, July 26-30, 2015, at Minneapolis, MN, USA. de Jim�nez MM, Mamidi S, McClean P, and Rahman M (2015) Genome-wide characterization of genetic diversity, population structure and linkage disequilibrium in Brassica napus L. Germplasm. An abstract for 14th International Rapeseed Congress, July 5-9, 2015, at Saskatoon, Canada. Rahman M (2015) Nine years of canola breeding from scratch at North Dakota State University. An abstract for 14th International Rapeseed Congress, July 5-9, 2015, at Saskatoon, Canada.

Progress 02/01/15 to 09/30/15

Outputs
Target Audience:The global demand of canola oil is increasing rapidly, and the present production is unable to meet the required demand. Its healthy attributes with a low level of saturated fat makes it desirable to the food industry and canola growers. Therefore, the target audiences for the generated information is the growers, the appropriate commodity group, stakeholders, scientists, students, and crop consultants. Changes/Problems: Nothing Reported What opportunities for training and professional development has the project provided?The project gave an opportunity to train three graduate students (2 Ph.D., 1 M.S.) in laboratory, greenhouse and field techniques including seed preparation for field planting, field layout, planting, crossing, data taking, harvesting, greenhouse management (planting, crossing, data taking, harvesting) DNA extraction, and PCR. The students presented their research at various professional meetings such as 14th International Rapeseed Congress, American Society of Plant Biologists - Midwestern Section 2015 Meeting, Dupont Pioneer and University of Minnesota Plant Breeding Symposium, Plant & Animal Genome XXIV conference. The PI attended in two workshops on professional development for new faculty, including Faculty Involvement and Academic Citizenship, and Securing External Funding for Educational Initiatives. How have the results been disseminated to communities of interest?The research results were presented at the annual meeting organized by commodity group where the participants are growers, industry personnel, public and private researchers, and crop consultants. The PI presented the results at scientific meetings, including 14th International Rapeseed Congress (held once in every four-year), 2014 ASA-CSSA-SSSA annual meeting, 2015 Plant Biology annual meeting. What do you plan to do during the next reporting period to accomplish the goals?We plan to continue with our current plan as described in the method of Hatch project. The number of breeding lines to evaluate in full-plot in the field may change based on the availability of seeds harvested from the winter nursery and from the greenhouse. To work on freezing tolerance research a new method will be developed and will be applied to identify electrolyte leakage from artificial freezing injured leaves. Combinations of the proposed method and new method will allow us to identify the best lines resistance/tolerance to freezing conditions. We will continue the germplasm screening for blackleg and sclerotinia stem rot diseases. The different phenotyping data will be aligned to genotypic data. The genome-wide association mapping study will be conducted to find the markers associated with the traits.

Impacts
What was accomplished under these goals? Rapeseed/canola (Brassica napus L.) germplasm lacks sufficient genetic variability to make progress in breeding from a long-term perspective. The average seed yield of spring type canola in North Dakota is 1,632 lb/a, where the average seed yield of winter type in Western Europe is 3,174 lb/a. Winter canola is not adapted to North Dakota due to its lack of winterhardiness. From a breeding and genetics point of view, the improvement of spring canola for seed yield could be achieved by introgression of favorable yield contributing characters from the winter types. Therefore, crosses were made between winter type and spring type germplasm to create genetic variability in spring canola. Canola can be attacked by disease organisms any time from seedling through maturity. There are two major diseases of canola in North Dakota, sclerotinia stem rot (white mold) and blackleg, which significantly reducesseed yield inseverely infected fields. We have established a collaborative research project with NDSU plant pathologist Dr. Luis del Rio for screening the germplasm lines in a controlled condition in the greenhouse and in the field. Spring frost can severely damage or even kill rapeseed/canola seedlings. Identifying frost tolerance in canola would be beneficial for growers, especially in North Dakota. Therefore, we initiated a research project to identify frost tolerant germplasm to use in the breeding program. A graduate student researcher has developed a method to evaluate B. napus germplasm lines under artificial frost simulation conditions in a growth chamber. Canola is very sensitive to heat stress, especially during flowering, which causes flowers abortion, pollen grain death and pollen sterility. Therefore, the development of crop cultivars tolerant to high temperature became one of the major challenges for plant breeders and plant scientists. A graduate student is conducting research to identify heat stress tolerant germplasm from our existing diversified B. napus germplasm lines to use in the heat tolerance breeding program. Association mapping (AM) is a population-based study that utilizes genetically unrelated germplasm/varieties collections and breeding lines. It is a great tool to discover multiple alleles at the genetic loci in germplasm, and thus allows one to find elite alleles for breeding. In this program, a total of 366 publicly available diversified B. napus accessions and several advanced breeding lines have been genotyped using Illumina genotyping by sequencing (GBS) platform at the Institute for Genomic Diversity at Cornell University, and obtained 42,575 high quality SNP's for trait-linked marker development. Objective 1: Develop high seed and high oil canola germplasm utilizing both spring- and winter-type canola B. napus adapted to North Dakota. Crosses and reciprocal crosses between ten winter-type and ten spring-type parents were made in the greenhouse. One hundred and sixty four F1 were advanced to F2 progenies. The F2 progenies were planted in a single-row nursery at Prosper, North Dakota. A total of 987 F1 derived double haploid (DH) lines were generated in the greenhouse of which 578 plants were fertile. Three hundred DH lines were planted in the same breeding nursery. Plant selection was made on the basis of early vigor, early flowering, standability, breeder's impression, and better seed yield. The selected breeding lines were self-pollinated by using micro-perforated selfing bags and harvested after maturity. Three hundred F2:3 lines were selected from the field and sent to the winter nursery in Santiago, Chile to generate F3:4 populations for 2016 full-plot field testing. One hundred DH lines were selected and will be grown in the greenhouse to generate enough seeds for 2016 full-plot field testing. Objective 2: Identify germplasm with enhanced disease resistance (e.g. blackleg, sclerotinia stem rot) and abiotic stress tolerance (heat, frost) traits in canola. Blackleg disease screening: An experiment was conducted to screen B. napus germplasm lines against blackleg disease during September-December 2014 greenhouse cycle. A total of 182 accessions were grown using a randomized complete block design with three replicates. Ten individual seedlings of each accession were grown in each replicate. Both cotyledon leaves of individual seedling were inoculated with the L. maculans isolates (PG-4), and thus 20 scores per accession per replicatewere scored. The L. maculans isolates (PG-4) preparation and plant inoculation procedure was conducted by the cooperation of Dr. Luis del Rio, Plant Pathology, NDSU. Disease reaction was evaluated two weeks after inoculation using a 0-9 scale as described by Williams (1985), where 0 is no lesion visible and 9 means dead seedling. Freezing tolerance study: A protocol was developed for frost tolerance evaluation under controlled environmental conditions. In this protocol, B. napus seedlings were grown for two weeks in the greenhouse, acclimated at 4°C for seven days, exposed to frost at −4°C, allowed to recover at 4°C for 24 h, and scored in the greenhouse for frost damage three days after treatment. Using this protocol, three experiments were conducted in three different greenhouse planting times. A total of 229, 206 and 162 germplasm lines were evaluated in randomized complete block design with four replicatesagainst freezing tolerance in the three experiments conducted during April, May and July, respectively. The freezing tolerance study was also conducted in the field in March 2015. Two hundred and twenty nine germplasm lines were evaluated in randomized complete block design with four replicatesin the field under natural field freezing conditions. Heat tolerance study: A total of 92 spring type B. napus germplasm lines in randomized complete block design with four replicateswere screened in the greenhouse and in a plant growth chamber under artificial heat simulation conditions. The heat stress simulation condition in the growth chamber was designed as 18C for 8 hours, temperature ramped up from 18 C to 35 C in 6 hours, followed by a constant 35 C for 4 hours, and then the temperature ramped down from 35 C to 18 C in 6 hours. The heat treatment was given to flowering plants for 5 days, and then the treated plants were brought back to a normal greenhouse growing conditions. A controlled experiment was set with the same germplasm in the greenhouse without the heat stress treatment. Objective 3: Identify molecular markers through genome-wide association mapping of agronomic traits, seed quality, disease resistance and stress tolerance traits to be used in marker assisted selection program. A total of 139 genotypes belong to 16 countries were evaluated for blackleg disease in two different greenhouse planting times. These accessions had different growth habits (75 spring, 49 winter, 12 semi-winter, and 3 rutabaga). The phenotypic data was aligned with the genotypic data for genome-wide association mapping study. One major QTL associated with the blackleg disease was identified on chromosome A01 at 9.66 Mbp. The QTL is in progress to validate to other germplasm lines to confirm the marker-trait association. The markers in the QTL could further be used for identification of phenotype that can be used in the breeding program. ?

Publications