Doubled haploid technology for line development in maize: technical advances and prospects

Chaikam, Vijay; Molenaar, Willem S.; Melchinger, Albrecht E.; Prasanna, Boddupalli M.

doi:10.1007/s00122-019-03433-x

Cited by 162 publications

(199 citation statements)

References 119 publications

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“…In this study, the transcriptomic response of pre-selected doubled haploid (DH) lines derived from the European flint landrace "Petkuser Ferdinand Rot" towards cold treatment was evaluated towards the goal to improve cold tolerance. DH technology is an efficient method to generate homozygous DH inbred lines by chromosome doubling of haploid cells [26].…”

Section: Introductionmentioning

confidence: 99%

Transcriptomic diversity in seedling roots of European flint maize in response to cold

et al. 2020

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Background: Low temperatures decrease the capacity for biomass production and lead to growth retardation up to irreversible cellular damage in modern maize cultivars. European flint landraces are an untapped genetic resource for genes and alleles conferring cold tolerance which they acquired during their adaptation to the agroecological conditions in Europe. Results: Based on a phenotyping experiment of 276 doubled haploid lines derived from the European flint landrace "Petkuser Ferdinand Rot" diverging for cold tolerance, we selected 21 of these lines for an RNA-seq experiment. The different genotypes showed highly variable transcriptomic responses to cold. We identified 148, 3254 and 563 genes differentially expressed with respect to cold treatment, cold tolerance and growth rate at cold, respectively. Gene ontology (GO) term enrichment demonstrated that the detoxification of reactive oxygen species is associated with cold tolerance, whereas amino acids might play a crucial role as antioxidant precursors and signaling molecules. Conclusion: Doubled haploids representing a European maize flint landrace display genotype-specific transcriptome patterns associated with cold response, cold tolerance and seedling growth rate at cold. Identification of cold regulated genes in European flint germplasm, could be a starting point for introgressing such alleles in modern breeding material for maize improvement.

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Section: Introductionmentioning

confidence: 99%

Transcriptomic diversity in seedling roots of European flint maize in response to cold

et al. 2020

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“…Higher genetic variance and higher heritabilities when using DH lines compared to conventional selfed lines in per se and testcross evaluation facilitate accurate selection decisions. The major steps in maize DH line production pipelines are in vivo induction of haploids, separation of haploids from diploids, doubling the genome in haploid (D 0 ) plants, and production of D 1 seed from D 0 plants [3,4]. Development of protocols for large-scale DH line production and continuous process improvement in the last three decades resulted in higher efficiencies at different steps of DH line development [3].…”

Section: Introductionmentioning

confidence: 99%

“…Chromosome doubling protocols involves several labor-intensive steps and use of expensive chemicals. Hence, increasing chromosomal doubling efficiency is critical to reduce the cost per DH line [3].…”

Section: Introductionmentioning

confidence: 99%

“…However, colchicine is more commonly used in DH production pipelines because the colchicine is more readily available for purchase in large quantities and protocols are well-established. Several methods of colchicine application to haploids were described in literature that include (a) treating the seed; (b) immersing germinating haploid seedlings in colchicine solution; (c) immersing the roots of haploid seedlings; or (d) injecting colchicine into shoot apical meristem [3]. Among these, the seedling immersion method is widely adapted for large-scale DH line production as it showed a better success rate compared to seed treatment [23], root treatment [22], or injection into shoot apical meristem [26].…”

Section: Introductionmentioning

confidence: 99%

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Improving the Efficiency of Colchicine-Based Chromosomal Doubling of Maize Haploids

Chaikam

Gowda

Martinez

et al. 2020

Plants

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Production and use of doubled haploids (DH) is becoming an essential part of maize breeding programs worldwide as DH lines offer several advantages in line development and evaluation. One of the critical steps in maize DH line production is doubling the chromosomes of in vivo-derived haploids so that naturally sterile haploids become reproductively fertile diploids (DH) to produce seed. This step of artificially doubling the chromosomes is labor-intensive and costly; hence, optimizing protocols to improve the doubling success is critical for achieving efficiencies in the DH production pipelines. Immersion of 3–4-day old germinating haploid seedlings in colchicine solution is commonly used for chromosome doubling in large-scale maize DH line production. This manuscript presents a new method of colchicine application to haploid seedlings that showed superior doubling rates compared to other methods like standard seedling immersion, seed immersion, root immersion, and direct application of colchicine solution to the seedlings at V2 stage in the greenhouse trays. The new method involves immersing the crown region of the haploid seedlings along with all the seedling roots at V2 stage in the colchicine solution. Further experiments to optimize this method indicated that increasing colchicine concentration had a very positive effect on overall success rate in chromosomal doubling, while not drastically affecting survival rate. The optimized method showed on average 5.6 times higher overall success rate (OSR) compared to the standard haploid seedling immersion method which was the second-best method in our experiments. This improved method of colchicine application saves resources by reducing the number of haploids to be generated and handled in a maize DH production pipeline.

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“…These cloned genes and associated molecular evidence allowed researchers to re-evaluate the two competing hypotheses explaining the maternal haploid induction: (1) regular double fertilization followed by male chromosome elimination and (2) impaired double fertilization or single fertilization (Li et al 2009a;Tian et al 2018). It eventually led to a unified hypothesis that both fertilization defects and chromosome elimination could be involved in the maternal haploid induction (Chaikam et al 2019b;Jacquier et al 2020). However, more evidence is needed to elucidate the detailed molecular mechanisms for the phenomena.…”

Section: Introductionmentioning

confidence: 99%

Genomic selection to optimize doubled haploid-based hybrid breeding in maize

Cheng

Guo

et al. 2020

Preprint

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Crop improvement, as a long-term endeavor, requires continuous innovations in technique from multiple perspectives. Doubled haploid (DH) technology for pure inbred production, which shaves years off of the conventional selfing approach, has been widely used for breeding. However, the final success rate of in vivo maternal DH production is determined by four factors: haploids induction, haploids identification, chromosome doubling, and successful selfing of the fertile haploid plants to produce DH seeds. Traits in each of these steps, if they can be accurately predicted using genomic selection methods, will help adjust the DH production protocol and simplify the logistics and save costs. Here, a hybrid population (N=158) was generated based on an incomplete half diallel design using 27 elite inbred lines. These hybrids were induced to create F1-derived haploid families. The hybrid materials, as well as the 27 inbreds, the inbred-derived haploids (N=200), and the F1-derived haploids (N=5,000) were planted in the field to collect four DH-production traits, three yield-related traits, and three developmental traits. Quantitative genetics analysis suggested that in both diploids and haploid families, most of the developmental traits showed high heritability, while the DH-production and developmental traits exhibited intermediate levels of heritability. By employing different genomic selection models, our results showed that the prediction accuracy ranged from 0.52 to 0.59 for the DH-production traits, 0.50 to 0.68 for the yield-related traits, and 0.44 to 0.87 for the developmental traits. Further analysis using index selection achieved the highest prediction accuracy when considering both DH production efficiency and the agronomic trait performance. Furthermore, the long-term responses through simulation confirmed that index selection would increase the genetic gain for targeted agronomic traits while maintaining the DH production efficiency. Therefore, our study provides an optimization strategy to integrate GS technology for DH-based hybrid breeding.

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Doubled haploid technology for line development in maize: technical advances and prospects

Cited by 162 publications

References 119 publications

Transcriptomic diversity in seedling roots of European flint maize in response to cold

Transcriptomic diversity in seedling roots of European flint maize in response to cold

Improving the Efficiency of Colchicine-Based Chromosomal Doubling of Maize Haploids

Genomic selection to optimize doubled haploid-based hybrid breeding in maize

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