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

2019 ◽  
Vol 132 (12) ◽  
pp. 3227-3243 ◽  
Author(s):  
Vijay Chaikam ◽  
Willem Molenaar ◽  
Albrecht E. Melchinger ◽  
Prasanna M. Boddupalli
Keyword(s):  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Production Costs ◽  
Great Promise ◽  
Success Rates ◽  
Haploid Induction ◽  
Maize Breeding ◽  
Breeding Programs ◽  
Line Production

Key Message Increased efficiencies achieved in different steps of DH line production offer greater benefits to maize breeding programs. Abstract Doubled haploid (DH) technology has become an integral part of many commercial maize breeding programs as DH lines offer several economic, logistic and genetic benefits over conventional inbred lines. Further, new advances in DH technology continue to improve the efficiency of DH line development and fuel its increased adoption in breeding programs worldwide. The established method for maize DH production covered in this review involves in vivo induction of maternal haploids by a male haploid inducer genotype, identification of haploids from diploids at the seed or seedling stage, chromosome doubling of haploid (D0) seedlings and finally, selfing of fertile D0 plants. Development of haploid inducers with high haploid induction rates and adaptation to different target environments have facilitated increased adoption of DH technology in the tropics. New marker systems for haploid identification, such as the red root marker and high oil marker, are being increasingly integrated into new haploid inducers and have the potential to make DH technology accessible in germplasm such as some Flint, landrace, or tropical material, where the standard R1-nj marker is inhibited. Automation holds great promise to further reduce the cost and time in haploid identification. Increasing success rates in chromosome doubling protocols and/or reducing environmental and human toxicity of chromosome doubling protocols, including research on genetic improvement in spontaneous chromosome doubling, have the potential to greatly reduce the production costs per DH line.

Doubled haploid production in maize under Sub-montane Himalayan conditions using R1-nj-based haploid inducer TAILP1

2020 ◽  
Vol 80 (03) ◽  
Author(s):  
R. K Khulbe ◽  
A. Pattanayak ◽  
Lakshmi Kant ◽  
G. S. Bisht ◽  
M. C. Pant ◽  
...  
Keyword(s):  
Doubled Haploid ◽  
Sweet Corn ◽  
Haploid Induction ◽  
Maize Breeding ◽  
Line Production ◽  
Haploid Inducer ◽  
Breeding Programmes ◽  
Source Populations ◽  
Developmental Phases

The use of in vivo haploid induction system makes the doubled haploid (DH) technology easier to adopt for the conventional maize breeders. However, despite having played an important role in the initial developmental phases of DH technology, Indian maize research has yet to harvest its benefits. Haploid Inducer Lines (HILs) developed by CIMMYT are being widely used in maize breeding programmes in many countries including India. There, however, is no published information on the efficiency of DH line production using CIMMYT HILs in Indian maize breeding programmes. In the present study, the efficiency of DH production using CIMMYT’s tropically adapted inducer line TAILP1 was investigated with eight source populations including two of sweet corn. The average haploid induction rate (HIR) of TAILP1 was 5.48% with a range of 2.01 to 10.03%. Efficiency of DH production ranged from 0.14 to 1.87% for different source populations with an average of 1.07%. The information generated will be useful for maize breeders intending to use DH technology for accelerated development of completely homozygous lines.

scholarly journals Optimization of Cucumber Doubled Haploid Line Production Using In Vitro Rescue of In Vivo Induced Parthenogenic Embryos

2005 ◽  
Vol 130 (4) ◽  
pp. 555-560 ◽  
Author(s):  
Elisabet Claveria ◽  
Jordi Garcia-Mas ◽  
Ramon Dolcet-Sanjuan
Keyword(s):  
Flow Cytometry ◽  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Doubled Haploid Line ◽  
High Rate ◽  
Limiting Factors ◽  
Line Production ◽  
Haploid Plants

Homozygous doubled haploid lines (DHLs) from new cucumber (Cucumis sativus L.) accessions could be useful to accelerate breeding for resistant varieties. DHLs have been generated by in vitro rescue of in vivo induced parthenogenic embryos. The protocol developed involves the following: 1) induction of parthenogenic embryos by pollinating with pollen irradiated with a Co60 γ-ray source at 500 Gy; 2) in vitro rescue of putative parthenogenic embryos identified by their morphology and localized using a dissecting scope or X-ray radiography; 3) discrimination of undesirable zygotic individuals from the homozygous plants using cucumber and melon SSR markers; 4) determination of ploidy level from homozygous plants by flow cytometry; 5) in vitro chromosome doubling of haploids; and 6) acclimation and selfing of selected lines. Codominant markers and flow cytometry confirmed the gametophytic origin of plants regenerated by parthenogenesis, since all homozygous lines were haploids. No spontaneous doubled haploid plants were rescued. Chromosome doubling of haploid plants was accomplished by an in vitro treatment with 500 μm colchicine. Rescue of diploid or chimeric plants was shown by flow cytometry, prior to their acclimation and planting in the greenhouse. Selfing of colchicine-treated haploid plants allowed for the perpetuation by seed of homozygous lines. The high rate of seed set, 90% of the lines produced seed, facilitated the recovery of inbred lines. Despite some limiting factors, parthenogenesis is routinely used in a cucumber-breeding program to achieve complete homozygosity in one generation. Breeding for new commercial hybrid cultivars will be accelerated. DHLs are ideal resources for genomic analyses.

scholarly journals Breeding Maize Maternal Haploid Inducers

Plants ◽  
2020 ◽  
Vol 9 (5) ◽  
pp. 614
Author(s):  
Henrique Uliana Trentin ◽  
Ursula K. Frei ◽  
Thomas Lübberstedt
Keyword(s):  
Poor Performance ◽  
High Price ◽  
Small Scale ◽  
Maize Breeding ◽  
Breeding Programs ◽  
Line Production ◽  
Dh Lines ◽  
Haploid Embryos ◽  
The Cost

Maize doubled haploid (DH) lines are usually created in vivo, through crosses with maternal haploid inducers. These inducers have the inherent ability of generating seeds with haploid embryos when used to pollinate other genotypes. The resulting haploid plants are treated with a doubling agent and self-pollinated, producing completely homozygous seeds. This rapid method of inbred line production reduces the length of breeding cycles and, consequently, increases genetic gain. Such advantages explain the wide adoption of this technique by large, well-established maize breeding programs. However, a slower rate of adoption was observed in medium to small-scale breeding programs. The high price and/or lack of environmental adaptation of inducers available for licensing, or the poor performance of those free of cost, might explain why smaller operations did not take full advantage of this technique. The lack of adapted inducers is especially felt in tropical countries, where inducer breeding efforts are more recent. Therefore, defining optimal breeding approaches for inducer development could benefit many breeding programs which are in the process of adopting the DH technique. In this manuscript, we review traits important to maize maternal haploid inducers, explain their genetic basis, listing known genes and quantitative trait loci (QTL), and discuss different breeding approaches for inducer development. The performance of haploid inducers has an important impact on the cost of DH line production.

scholarly journals The development of homozygous maize lines using an in vivo haploid induction in the Croatian germplasm

Poljoprivreda ◽  
2019 ◽  
Vol 25 (1) ◽  
pp. 19-25
Author(s):  
Maja Mazur ◽  
Sonja Vila ◽  
Ivan Brkić ◽  
Antun Jambrović ◽  
Domagoj Šimić
Keyword(s):  
Chromosome Doubling ◽  
Seedling Survival ◽  
Survival Rates ◽  
Misclassification Rate ◽  
Successful Implementation ◽  
Haploid Induction ◽  
Maize Breeding ◽  
Pollination Success ◽  
Self Pollination

The in vivo haploid induction has been widely applied to the maize breeding in recent decades, but it has not been used in the breeding programs in the Republic of Croatia by now. This study's objectives were to examine the haploid induction rates in the Croatian germplasm and to evaluate the properties of the D0 haploids, which are essential for a successful implementation of this method in breeding. The in vivo haploid induction was performed on 11 single-cross hybrids using the Zarodyshevy Marker Krasnodarsky (ZMK) inducer, and colchicine was used as a chromosome doubling agent. Emergence, misclassification rate, colchicine treatment survival, chromosome doubling rate and self-pollination success were examined in the D0 generation. The haploid induction rates ranged from 6.9 to 15.8%, which is consistent with the average induction rates characteristic of the ZMK inducer and the other modern ones. Significant differences were found among the populations of D0 haploids for all tested properties, except for self-pollination success. On average, the misclassification rates were lower, and the seedling survival rates were higher than those reported in other studies, indicating a possibility of a successful application of the doubled haploid method in maize breeding.

scholarly journals Evaluation of Gynogenic Responsiveness and Pollen Viability of Selfed Doubled Haploid Onion Lines and Chromosome Doubling via Somatic Regeneration

2010 ◽  
Vol 135 (1) ◽  
pp. 67-73 ◽  
Author(s):  
Marijana Jakše ◽  
Pablo Hirschegger ◽  
Borut Bohanec ◽  
Michael J. Havey
Keyword(s):  
Doubled Haploid ◽  
Chromosome Doubling ◽  
Pollen Viability ◽  
Shoot Culture ◽  
Haploid Induction ◽  
Flower Bud ◽  
Breeding Programs ◽  
Regeneration Stage ◽  
Regeneration Procedure ◽  
Dh Lines

Although haploid induction has been used in onions (Allium cepa L.) for over 20 years, several obstacles limit its use in plant breeding programs. To address these limitations, we evaluated the responsiveness of doubled haploid (DH) lines and their selfed progenies, an alternative protocol for chromosome doubling using somatic regeneration of haploid lines, and pollen viability of DH lines. Twenty-one DH lines were self pollinated and tested for haploid induction in the second generation. Among the DH lines, 18 lines showed an average of 20% decrease in gynogenic responsiveness compared with the original lines, while three lines registered an average increase of 5.7%. Using a two-step induction/regeneration procedure, 8,589 somatic regenerants were obtained from 16,170 flower buds from haploid plants, and shoot culture was established. A more laborious procedure using extraction of ovaries in the regeneration stage was found equal to flower bud culture. Chromosome doubling via somatic regeneration was found to be 83% and 100% efficient when the source material was haploid or mixoploid, respectively. Based on the results achieved in this and previous studies, an alternative protocol for chromosome doubling of gynogenic haploids is proposed.

scholarly journals Development of late temperate in vivo haploid inducers

Genetika ◽  
2021 ◽  
Vol 53 (1) ◽  
pp. 51-64
Author(s):  
Rahime Cengiz ◽  
Mesut Esmeray
Keyword(s):  
Plant Height ◽  
Haploid Induction ◽  
Maize Breeding ◽  
Branch Number ◽  
Breeding Programs ◽  
Haploid Inducer ◽  
Breeding Populations ◽  
Ssrs Markers ◽  
Maize Research Institute

In vivo doubled haploid technique has been widely used in advanced maize breeding programs due to cost, labor and time advantages and increase in efficiency. However, the number of available inducer lines in the world is sufficient. Six BC1 breeding populations including RWS and RWK-76 haploid inducer lines and late temperate ADK-451, ADK-737 and ADK-455 lines were developed by Sakarya Maize Research Institute (MRI) in Turkey. The RWS and RWK-76 haploid inducer lines were used as donors. Pedigree method was employed to develop the inducer lines. Anthocyanin coloration of plant, tassel length, branch number of tassel, plant height, days to flowering, embryo-endosperm colorfulness and haploid induction rate (HIR) were determined. The genotypes with the best characteristics were selected. The families from BC1F3 to BC1F7 were hybridized to liguleless line to determine the HIR and families with HIR over 8% were selected from BC1 populations. The HIR, plant height and days to tassel flowering values of in-1021 and in-1076 candidate haploid inducer lines were 10.5 and 12.3%, 195 and 200 cm, and 69 and 68 days, respectively. The HIR value of RWS donor haploid inducer ranged from 8.9 to 11.3% and for RWK-76 from 7.3 to 9.8%. Simple Sequence Repeats (SSRs) markers were used to identify genetic similarity between late temperate haploid inducer lines and donors. The similarity rates of in-1021 and in-1076 inducer lines to the RWS donor were 38 and 15%, and to the RWK-76 donor were 23 and 27%. The similarity rate between the two candidate inducer lines was 30%. The results indicated that the late temperate haploid inducer lines developed will increase the efficiency of maize breeding.

Optimization of chromosome doubling treatment for efficient in vivo doubled haploid production in maize

2021 ◽  
pp. 1-10
Author(s):  
Sourbh Kumar ◽  
Uttam Chandel ◽  
Satish Kumar Guleria
Keyword(s):  
Doubled Haploid ◽  
Seed Set ◽  
Chromosome Doubling ◽  
Pollen Viability ◽  
Haploid Inducer ◽  
Haploid Production ◽  
Maize Genotypes ◽  
Different Doses

Abstract An investigation to optimize the protocol for application of colchicine for enhancing the doubled haploid production in maize was done. 106 maize genotypes were used as maternal parents, whereas, pollen source involved tropically adopted haploid inducer (TAIL P1 and TAIL hybrid). After the elimination of chromosomes of inducer lines, haploid seeds were obtained from the crosses. Haploid seedlings were treated with three different doses, such as 0.04, 0.06 and 0.08 per cent of colchicines for different durations (8, 12 and 15 hours). The response of various colchicine concentrations applied for different time durations revealed significant differences at P ≤ 0.05 for various parameters viz., per cent plants survivability, stalk colour, the fertility of tassel, silk present/absent, pollen viability, seed set and per cent doubled haploid formation. In maize, colchicine doses of 0.04 per cent for 12 hours and 0.06 per cent for 8 hours, respectively were established as optimum for enhanced doubled haploid production. But among these two, 0.04 per cent for 12 hours was observed to be best dose for doubled haploid production in maize.

R1-nj expression in parental inbreds as a predictor of amenability of maize hybrids to R1-nj-based doubled haploid development

2020 ◽  
Vol 79 (04) ◽  
Author(s):  
R. K. Khulbe ◽  
A. Pattanayak ◽  
Vivek Panday
Keyword(s):  
Doubled Haploid ◽  
Current Method ◽  
Distinct Pattern ◽  
Complex Nature ◽  
Maize Breeding ◽  
Early Maturity ◽  
Haploid Inducer ◽  
Source Populations ◽  
Phenotype Expression

The current method of doubled haploid (DH) development in maize involves in vivo production of haploids using R1-njbased haploid inducer lines that upon use as male render a small fraction of seed in the pollinated female ears haploid. Identification of haploid seed relies on R1-nj marker expression in the endosperm and embryo, and the degree of its expression determines efficiency of DH development process. In the present study, R1-nj expression in the endosperm was characterized in crosses of CIMMYT’s R1-nj-based haploid inducer TAILP1 with a set comprising 18 early maturity hybrids and their 23 parental inbreds. Kernel colour inhibition was observed only in a small proportion of the hybrids and inbreds. Comparison of R1-nj expression in the hybrids and their parental inbreds revealed a distinct pattern, which may be useful in identifying source populations and/or determining parental constituents for synthesizing source populations with predicted amenability to doubled haploid development using R1-nj-based haploid inducers. However, deviation from the pattern was noted in hybrids involving inbreds with higher degree of colour inhibition, which suggests complex nature of R1-nj phenotype expression and necessitates further investigation involving larger sets of germplasm for dissecting the role of maternal and paternal genetic factors in determining R1-nj phenotype expression. The hybrids found exhibiting complete kernel anthocyanin expression in present study can be used directly as source populations for DH development using R1-nj based haploid inducers. Besides, since the inbreds used in the study have originated from and/or are accessible to CGIAR/NARS maize breeding programmes, the information on their kernel anthocyanin expression can be helpful in selection of source populations or generating new source populations amenable for DH development using R1-nj based haploid inducers.

scholarly journals MYO, a Candidate Gene for Haploid Induction in Maize Causes Male Sterility

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 773
Author(s):  
Kimberly Vanous ◽  
Thomas Lübberstedt ◽  
Rania Ibrahim ◽  
Ursula K. Frei
Keyword(s):  
Male Sterility ◽  
Gene Isolation ◽  
Haploid Induction ◽  
Maize Breeding ◽  
Breeding Programs ◽  
Positional Candidate ◽  
Haploid Induction Rate ◽  
Induction Rate ◽  
Transgenic Lines ◽  
Strong Candidate

Doubled haploid technology is highly successful in maize breeding programs and is contingent on the ability of maize inducers to efficiently produce haploids. Knowledge of the genes involved in haploid induction is important for not only developing better maize inducers, but also to create inducers in other crops. The main quantitative trait loci involved in maize haploid induction are qhir1 and qhir8. The gene underlying qhir1 has been discovered and validated by independent research groups. Prior to initiation of this study, the gene associated with qhir8 had yet to be recognized. Therefore, this research focused on characterizing positional candidate genes underlying qhir8. Pursuing this goal, a strong candidate for qhir8, GRMZM2G435294 (MYO), was silenced by RNAi. Analysis of crosses with these heterozygous RNAi-transgenic lines for haploid induction rate revealed that the silencing of MYO significantly enhanced haploid induction rate by an average of 0.6% in the presence of qhir1. Recently, GRMZM2G465053 (ZmDMP) was identified by map-based gene isolation and shown to be responsible for qhir8. While our results suggest that MYO may contribute to haploid induction rate, results were inconsistent and only showing minor increases in haploid induction rate compared to ZmDMP. Instead, reciprocal crosses clearly revealed that the silencing of MYO causes male sterility.

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