Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,ex...Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,exhibiting a synergistic effect.However,it is unknown whether knock out of ZmDMP orthologs can stimulate HI in rice.In this study,a ZmPLA1 ortholog(OsPLA1)and two ZmDMP orthologs(OsDMP3 and OsDMP6)were identified in rice.All three genes encode plasma membrane-localized proteins and were highly expressed in mature anthers.Knockout of OsPLA1 in both Minghui 63 and Nipponbare resulted in reduced seed setting rate(SSR)and caused HI.The osdmp3,osdmp6 and the double mutant failed to trigger HI independently,nor increased the haploid induction rate(HIR)when combined with ospla1.Repeated pollinations operations of QX654A with the ospla1 mutant significantly improve SSR,while reducing HIR.RNA-seq profiling of mature ospla1 mutant anthers indicated that a large number of differentially expressed genes(DEGs)were enriched in redox homeostasis and lipid metabolic GO terms,plant hormone signal transduction,and MAPK signaling pathways.These findings provide important insights towards construction of an efficient DH breeding technology and study of the molecular mechanism of HI in rice.展开更多
This article summarizes the research progress inthe breeding of maize haploids, including the production and identification of maize haploids, especially the methods to produce doubled haploids and the selection of ba...This article summarizes the research progress inthe breeding of maize haploids, including the production and identification of maize haploids, especially the methods to produce doubled haploids and the selection of basic materials, as well as the applications of the maize haploids in maize breeding. Finally, this article pro- poses several issues researchers should pay attention to, and the prospects of hap- Ioids in maize breeding.展开更多
Haploid seedlings were inducted from different maize materials.At 2-3-leaf stage,maize haploids were treated with 0.06% colchicine and 2.0% dimethyl sulfoxide（DMSO） by dipping root,dripping heart leaf and acupunctur...Haploid seedlings were inducted from different maize materials.At 2-3-leaf stage,maize haploids were treated with 0.06% colchicine and 2.0% dimethyl sulfoxide（DMSO） by dipping root,dripping heart leaf and acupuncturing growing point,respectively.The doubling rate and mortality rate in different treatments were analyzed by variance analysis and multiple comparisons.The result showed that growing point acupuncturing method exhibited the highest doubling efficiency with an average doubling rate of 23%,seed-setting rate of 21.4%,and mortality rate of 16.3%.Composed with other two chemical doubling methods,growing point acupuncturing method significantly improved the doubling rate of maize haploids with a lower application dose of colchicine.This study laid the foundation for industrial application of haploid breeding techniques.展开更多
To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were n...To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were no significant differences in anther development between Ningqi No.1 and its haploids at the stage of pollen mother cell, and tetrads were formed successfully after the meiosis stage. The tetrads of Ningqi No.1 could release microspores. At the same time, the tapetal cells can provide nutrition for the development of the microspores, which eventually developed into mature pollen grains. Although the haploids could also release microspores at the tetrad stage, the tapetal cells degraded in advance, which made the released microspores unable to develop into mature pollen grains, resulting in pollen abortion of haploids.展开更多
Using seeds of inbred progenies of Cucurbita pepo L. DH12-11409 double haplolds as experimental materials, the effects of different hormone combinations, explant types and seedling ages on adventitious bud regeneratio...Using seeds of inbred progenies of Cucurbita pepo L. DH12-11409 double haplolds as experimental materials, the effects of different hormone combinations, explant types and seedling ages on adventitious bud regeneration and rooting of C. pepo L. were investigated. According to the results, inoculating cotyledonary nodes of yellow-green cotyledons from 5-d-old C. pepo L. double haploids to MS ＋ 30 g/L Suc ＋ 8 g/L Agar ＋ 2.0 mg/L 6-BA ＋ 0.1 mg/L NAA led to the best results with the induction frequency of 90.0% and differentiation coefficient of 8.5. MS medium with addition of 0.05 or 0.1 mg/L NAA led to the highest rooting rate. Regenerated seedlings with 5 - 6 true leaves exhibited the highest survival rate of 90.0%, which was the optimal period for domestication and transplanting of regenerated seedlings. This study laid a solid foundation for high-efficiency utilization of heterosis of C. pepo L.展开更多
Haploid breeding can shorten the breeding process and improve the breeding efficiency. Currently, in vivo haploid induction technology has been com- monly used in maize （ Zea mays L. ）. This paper briefly introduced...Haploid breeding can shorten the breeding process and improve the breeding efficiency. Currently, in vivo haploid induction technology has been com- monly used in maize （ Zea mays L. ）. This paper briefly introduced in vivo haploid induction technologies, summarized doubling methods of maize haploids and described the significance and application of maize haploids, which provided the basis for further development of haploid breeding in maize.展开更多
A large number of adventitious buds were induced from in vitro cultured young inflorescences of haploids of rice.Having been subcultured on solidified subculture media at 26 ℃ for 7 days ,the adventitious buds we...A large number of adventitious buds were induced from in vitro cultured young inflorescences of haploids of rice.Having been subcultured on solidified subculture media at 26 ℃ for 7 days ,the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45 60 min.After cooled at a rate of 1.0℃/min down to -40℃,the samples were kept in liquid nitrogen.The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38 40℃water and then plated on solidified MS medium containing 3 % sucrose,0.5 mg/L NAA and 2.0 mg/L kinetin.After plated, 23% 32 % of adventitious buds resumed growth and 15% 22 % regenerated plantlets.The results of this work indicated that the adventitious buds derived from in vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure.The effective cryoprotectant is 10 % DMSO(dimethyl sulfoximide)+0.5 mol/L sorbitol.展开更多
Dear Editor,Doubled haploid(DH)technology is the fastest and most efficient method for producing completely homozygous lines,commonly used in maize breeding(Ren et al.,2017).The discovery of Stock 6,the first maize ha...Dear Editor,Doubled haploid(DH)technology is the fastest and most efficient method for producing completely homozygous lines,commonly used in maize breeding(Ren et al.,2017).The discovery of Stock 6,the first maize haploid inducer(HI)reported in 1959,yields 2%–3%maternal haploids during self-pollination or when used as a male in outcrossing.High-efficiency HIs derived from Stock 6,with a haploid induction rate(HIR)of 7%–15%,have significantly contributed to the commercialization of maize DH breeding.Recently,the genetic basis of haploid induction has been identified with two main molecular players in maize:MTL/ZmPLA1/NLD,which triggers haploid production(Gilles et al.,2017;Kelliher et al.,2017;Liu et al.,2017),and DMP,which enhances haploid induction(Zhong et al.,2019).Importantly,MTL is conserved across cereals,and knockout of OsMATL in rice has produced 2%–6%haploids(Yao et al.,2018;Wang et al.,2019;Liu et al.,2024a),presenting promising prospects for rice breeding.However,due to the lack of high-efficiency HIs and limitations in cross-pollination,DH technology based on Osmatl remains an unreliable and inefficient practice for rice breeding.Here,we developed a high-efficiency HI with favorable agronomic performance in rice.We also present a large-scale haploid production approach achieved by combining this HI with photo-thermo-sensitive genic male sterile(PTGMS)lines(Figure 1A).展开更多
The identification of haploid induction genes has promoted the advancement of several breeding technologies.Haploid induction genes in wheat,coupled with visual marker,have led to the establishment of a novel in vivo ...The identification of haploid induction genes has promoted the advancement of several breeding technologies.Haploid induction genes in wheat,coupled with visual marker,have led to the establishment of a novel in vivo doubled-haploid(DH)technology.When combined with dominant male sterile genes,this innovative DH method presents a promising avenue for high-throughput production of DH lines.Furthermore,the application of haploid induction genes has facilitated the establishment of other innovative breeding technologies,such as HI-Edit and cyto-swapping in creating cytoplasmic male sterility lines,as well as synthetic apomixis.This review summarizes the progress of DH technology in wheat and presents examples of application of haploid induction genes in accelerating breeding practices,aiming to promote the development of these innovative technologies in wheat and enhancing wheat breeding efficiency.展开更多
Doubled haploid(DH)technology has revolutionized crop breeding by enabling the production of homozygous lines in a single generation.In vivo haploid induction(HI)offers a more widely applicable approach that can signi...Doubled haploid(DH)technology has revolutionized crop breeding by enabling the production of homozygous lines in a single generation.In vivo haploid induction(HI)offers a more widely applicable approach that can significantly improve DH breeding efficiency.ToPAR,a parthenogenesis gene,originally identified in dandelion(Taraxacum officinale),has been characterized.Researchers have successfully induced haploid embryo-like structures and haploid offspring in lettuce and foxtail millet,respectively.展开更多
Eighteen pairs of diploid-haploid twin-seedlings were identified and screened out fromspecial rice SARII-628 population. Five pairs of themwere selected and randomly designated as A, B, C, Dand E. Simple sequence repe...Eighteen pairs of diploid-haploid twin-seedlings were identified and screened out fromspecial rice SARII-628 population. Five pairs of themwere selected and randomly designated as A, B, C, Dand E. Simple sequence repeats (SSR) analysisshowed that there was no difference among 310 siteswhich indicated that there was no base mutation onDNA primary structure. DNA methylation plays animportant role in gene expression regulation duringgrowth and development stages in eukaryotes. Amodified AFLP technique (methylation-sensitiveAFLP, MSAP) was employed to detect the DNA me-thylation patterns in the 5′-CCGG sites of the fivepairs of twin-seedlings. Although no methylationmutation was detected among the five diploids,forty-three methylation mutation sites were foundfrom the corresponding haploids. The MSAP ratios,which were the ratios of MSAP sites to the total am-plified sites, in five haploids were 18.79%, 19.35%,18.49%, 18.45% and 18.75%, respectively. And cor-responding full methylation levels (5′-CmCGG indouble strands) of those haploids were 10.58%,11.3%, 10.11%, 10.09% and 10.34%, respectively.Both MSAP and full methylation levels in the fivehaploids were higher than that of their correspondingdiploids, which suggested that hypermethylation oc-curred in some 5′-CCGG sites. Five types of MASPpatterns among the five pairs of twin-seedlings weredetected as follows: (1) no changes, methylation lev-els were the same in both haploids and diploids; (2) demethylation, diploid was methylated but no me-thylation in the same site in haploid; (3) hypermethy-lation, the methylation level in haploid was higher than those in diploid; (4) hypomethylation, methyla-tion in haploid was lower than those in diploid; (5) undecided pattern, change trend of methylation lev-els in haploids was not decided. The bands of 18 sites were reclaimed, then sequenced and searched on website to determine the sites of those sequences on rice chromosomes. The result showed that the methylation mutation involved the whole rice genome and 12 pairs of chromosomes. The mutation was site-related and there were different mutation sites for different haploids. Compared to diploids, the higher methylation level in haploids might be a readjusting reaction to the decrease in ploidy for the sake of sur-vival.展开更多
In this study, microarray technique was employed to analyze the gene expression at the RNA level between haploids and corresponding diploids derived from a rice twin-seedling line SARII-628. Differ- ent degrees of exp...In this study, microarray technique was employed to analyze the gene expression at the RNA level between haploids and corresponding diploids derived from a rice twin-seedling line SARII-628. Differ- ent degrees of expression variations were observed in the plant after haploidization. The main results are as follows: (1) after haploidization, the ratio of the sensitive loci was 2.47% of the total loci designed on chip. Those loci were randomly distributed on the 12 pairs of rice chromosomes and the activated loci were more than the silenced ones. (2) Gene clusters on chromosome were observed for 33 se- quences. (3) GoPipe function classification for 575 sensitive loci revealed an involvement in the bio- logical process, cell component and molecular function. (4) RT-PCR generally validated the result from microarray with a coincidence rate of 83.78%. And for the randomly-selected activated or silenced loci in chip analysis, the coincidence rate was up to 91.86%.展开更多
The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome edit...The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome editing(GE).Recently,these techniques were combined into a GE trait delivery system called HI-Edit(Haploid Inducer-Edit).In HI-Edit,the pollen of a haploid inducer line is reprogrammed to deliver GE traits to any variety,obviating recurrent selection.For HI-Edit to operate at scale,an efficient transformable HI line is needed,but most maize varieties are recalcitrant to transformation,and haploid inducers are especially difficult to transform given their aberrant reproductive behaviors.Leveraging marker assisted selection and a three-tiered testing scheme,we report the development of new Iodent and Stiff Stalk maize germplasm that are transformable,have high haploid induction rates,and exhibit a robust,genetically-dominant anthocyanin native trait that may be used for rapid haploid identification.We show that transformation of these elite‘‘HI-Edit”lines is enhanced using the BABYBOOM and WUSCHEL morphogenetic factors.Finally,we evaluate the HI-Edit performance of one of the lines against both Stiff Stalk and non-Stiff Stalk testers.The strategy and results of this study should facilitate the development of commercially scalable HI-Edit systems in diverse crops.展开更多
The technique of producing doublehaploid of wheat by distant hybridization between wheat and maize has characterized with better inducing effect, shorter in- ducing period, easy operation, and so on. At present, it is...The technique of producing doublehaploid of wheat by distant hybridization between wheat and maize has characterized with better inducing effect, shorter in- ducing period, easy operation, and so on. At present, it is the most efficient and has great potential of application in breeding of wheat. This article reviewed princi- ple and production process of the technique, research situation of the three key in- dicators of the technology（embryo rate, seedling rate and success rate of doubling）in recent years, and application of the technology in breeding, genetics, germplasm improvement of wheat. At last, both the achievements and the direction of further improvement and development of the technology in our program were discussed.展开更多
This study aimed to investigate the induction effect on different genotypes of maize of haploid inducers RWS, Stock6, CAU Inducer and hybridization-derived breeding lines. The results showed that 2005 had the highest ...This study aimed to investigate the induction effect on different genotypes of maize of haploid inducers RWS, Stock6, CAU Inducer and hybridization-derived breeding lines. The results showed that 2005 had the highest induction rate （2.85%） when using inducer Stock6 as male parent material; haploid induction rates were significantly different among different genotypes of maize when using RWS, Stock6, CAU Inducer and their crossbreeding materials as the male parents; inducers 2061, 2062, 2058 and 2059 led to relatively high induction rates, which could be emphatically utilized. Furthermore, during the screening of induction materials, different female parent materials should be selected based on the breeding objective.展开更多
[Purpose] The paper is to discuss the high-efficient breeding technology platform of dwarf male-sterile wheat.[Method] Conclusion was made upon the development of the creation and application of dwarf male-sterile whe...[Purpose] The paper is to discuss the high-efficient breeding technology platform of dwarf male-sterile wheat.[Method] Conclusion was made upon the development of the creation and application of dwarf male-sterile wheat,and the conception of introducing haploid breeding technology into the technological system of high-efficient breeding of dwarf male-sterile wheat was proposed.[Result] Dwarf male-sterile wheat not only retains the properties of total male abortion of Taigu Genetic Sterile Wheat,steady sterility,and high hybrid rate after open pollination,but also has the characteristic of high dwarfing ability of Aibian No.1,becoming a comparatively ideal population modification means.At present,the application of dwarf male-sterile wheat mainly includes germplasm resources platform of population modification by using dwarf male-sterile wheat,technology platform of constructing new high-efficient breeding system by using dwarf male-sterile wheat,and production platform of using dwarf male-sterile wheat to breed new varieties.Through the introduction of haploid breeding into the already established wheat high-efficient breeding system,a new system of high-efficient biological breeding technology of dwarf male-sterile wheat was constructed theoretically.[Conclusion] The study provides references to the further study and application of dwarf male-sterile wheat.展开更多
Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid indu...Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid induction rate(HIR), from the ~2% of the ancestor haploid inducer, stock6, to the ~10% of modern haploid inducers. Improvement of kernel oil content(KOC) would further enhance haploid identification efficiency. Using molecular marker-assisted selection, in combine with the number of haploids per ear as phenotypic criterion, we developed a new high-oil haploid inducer line, CHOI4, with a mean HIR of 15.8%and mean KOC of 11%. High KOC of CHOI4 can achieve a mean accuracy greater than 90% in identification of haploids of different backgrounds, with reduced false discovery rates and false negative rates in comparison with the previous high-oil haploid inducer line, CHOI3. Comparison of phenotypic selection strategies suggested that the number of haploids per ear can be used as a phenotyping criterion during haploid inducer line development. CHOI4 could further increase the efficiency of large-scale DH breeding programs with lower cost.展开更多
Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase the genetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson c...Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase the genetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson cytoplasmic sterility in pepper, a doubled haploid population of 115 pepper lines obtained from anther culture of the F1 hybrid between Yolo Wonder （sterility maintainer line） and Perennial （fertility restorer line） and the parental lines were test-crossed by 77013A （a strict cytoplasmic-genic male sterile line）. The fertility of the test-crossed lines was assessed in greenhouse and open field with the following three criteria： pollen index （PI, visual estimation of pollen amount per flower）, pollen number （PN, pollen counting under microscope）, and seed number （SN, the number of seeds per fruit in open pollination）. Correlations between the each couple of criteria within, as well as between the cultivation methods ranged from 0.55 to 0.84. Analysis of variance showed that the genotype （DH line） and environment were the significant sources of variation of the fertility. Narrow sense of heritance of fertility restoration ranged from 0.38 to 0.92, depending on the criteria and environment. The distribution of the progeny was continuous between the parental genotypes indicating the quantitative inheritance of fertility restoration. Inferred from segregation according to Snape et al.（1984）, the number of segregating genes was estimated to be that three to four genetic factors were involved in pollen traits （PI and PN） and five to eight genetic factors in seed production （SN）. The heredity analysis of the CMS will be helpful for understanding of the genetic mechanism of the fertility restoration and the exploitation of the CMS in hybrid seed production.展开更多
Field trials with a set of 108 doubled haploid lines(DHs) derived from a cross between the Chinese winter wheat cvs.CA9613 and H1488 were run at Beijing(China).Phenotypic data were recorded for major agronomic yield t...Field trials with a set of 108 doubled haploid lines(DHs) derived from a cross between the Chinese winter wheat cvs.CA9613 and H1488 were run at Beijing(China).Phenotypic data were recorded for major agronomic yield traits,i.e.grain weight per ear,grain number per ear and thousand grain weight(Tgw) in two field trials at Beijing.Based on the phenotypic data and a genetic map comprising 168 SSR markers,an analysis of quantitative trait loci(QTL) was carried out for yield and yield parameters using the composite interval mapping(CIM) approach.A total of 14 QTL were detected for these traits across two environments.Four of these QTL located on chromosomes 1A and 2B,respectively,exhibited pleiotropic effects.Loci showing pleiotropic effects will be very useful for understanding the homeologous relationships of QTL and designing an appropriate marker-assisted selection programme by multi-trait selection in order to accumulate("pyramide") favorable alleles at different loci.展开更多
A DH population derived from C49S-87/01Y1-1069 was used to study the inheritance of wheat haploid embryo production frequency（EPF） in wheat × maize cross with the mixed major gene and polygene inheritance model...A DH population derived from C49S-87/01Y1-1069 was used to study the inheritance of wheat haploid embryo production frequency（EPF） in wheat × maize cross with the mixed major gene and polygene inheritance model of quantitative traits. The results showed that the EPF of wheat × maize cross was controlled by two dominant epistatic genes and polygene with gene effects of 1.95 for the first major gene, 6.69 for the second one and 2.80 for the polygene. The inheritability of major genes was as high as 72.09%, suggesting that the differences in EPF among wheat materials were mainly influenced by genotype. However, non-genetic factors were still important, especially for wheat materials with low EPF.展开更多
基金This work was supported by the National Key Research and Development Program of China(2022YFD1200800)the China Agriculture Research System(CARS-02-05)+1 种基金Beijing Nova Program(2023067)Yunnan Province Science and Technology Department(202305AF150026).
文摘Doubled haploid(DH)technology is an important tool in crop breeding because it can significantly accelerate the breeding process.ZmPLA1/MATL/NLD and ZmDMP are two key genes controlling haploid induction(HI)in maize,exhibiting a synergistic effect.However,it is unknown whether knock out of ZmDMP orthologs can stimulate HI in rice.In this study,a ZmPLA1 ortholog(OsPLA1)and two ZmDMP orthologs(OsDMP3 and OsDMP6)were identified in rice.All three genes encode plasma membrane-localized proteins and were highly expressed in mature anthers.Knockout of OsPLA1 in both Minghui 63 and Nipponbare resulted in reduced seed setting rate(SSR)and caused HI.The osdmp3,osdmp6 and the double mutant failed to trigger HI independently,nor increased the haploid induction rate(HIR)when combined with ospla1.Repeated pollinations operations of QX654A with the ospla1 mutant significantly improve SSR,while reducing HIR.RNA-seq profiling of mature ospla1 mutant anthers indicated that a large number of differentially expressed genes(DEGs)were enriched in redox homeostasis and lipid metabolic GO terms,plant hormone signal transduction,and MAPK signaling pathways.These findings provide important insights towards construction of an efficient DH breeding technology and study of the molecular mechanism of HI in rice.
基金Supported by Key Science and Technology Research and Development Program of Anhui Province(1604a0702021)Seed Program of Anhui Academy of Agricultural Sciences(16D0911)+1 种基金the General Program of Special Foundation of the President of Anhui Academy of Agricultural Sciences for Young Scholars(16B0916,14B0946)Discipline Construction Program of Anhui Academy of Agricultural Sciences(14A0925)
文摘This article summarizes the research progress inthe breeding of maize haploids, including the production and identification of maize haploids, especially the methods to produce doubled haploids and the selection of basic materials, as well as the applications of the maize haploids in maize breeding. Finally, this article pro- poses several issues researchers should pay attention to, and the prospects of hap- Ioids in maize breeding.
文摘Haploid seedlings were inducted from different maize materials.At 2-3-leaf stage,maize haploids were treated with 0.06% colchicine and 2.0% dimethyl sulfoxide（DMSO） by dipping root,dripping heart leaf and acupuncturing growing point,respectively.The doubling rate and mortality rate in different treatments were analyzed by variance analysis and multiple comparisons.The result showed that growing point acupuncturing method exhibited the highest doubling efficiency with an average doubling rate of 23%,seed-setting rate of 21.4%,and mortality rate of 16.3%.Composed with other two chemical doubling methods,growing point acupuncturing method significantly improved the doubling rate of maize haploids with a lower application dose of colchicine.This study laid the foundation for industrial application of haploid breeding techniques.
基金Supported by Natural Science Foundation of Ningxia Hui Autonomous Region(NZ16097)Special Agricultural Breeding Project of Ningxia Hui Autonomous Region(2013NYYZ0105,2018NYYZ01-03)
文摘To reveal the sterility characteristics of Lycium barbarum haploids, cytological observations were made on the anthers of Ningqi No.1 and its haploids obtained from anther culture. The results showed that there were no significant differences in anther development between Ningqi No.1 and its haploids at the stage of pollen mother cell, and tetrads were formed successfully after the meiosis stage. The tetrads of Ningqi No.1 could release microspores. At the same time, the tapetal cells can provide nutrition for the development of the microspores, which eventually developed into mature pollen grains. Although the haploids could also release microspores at the tetrad stage, the tapetal cells degraded in advance, which made the released microspores unable to develop into mature pollen grains, resulting in pollen abortion of haploids.
基金Supported by National Staple Vegetable Industry Technical System(CARS-25)Special Fund for Agro-scientific Research in the Public Interest(201303112)
文摘Using seeds of inbred progenies of Cucurbita pepo L. DH12-11409 double haplolds as experimental materials, the effects of different hormone combinations, explant types and seedling ages on adventitious bud regeneration and rooting of C. pepo L. were investigated. According to the results, inoculating cotyledonary nodes of yellow-green cotyledons from 5-d-old C. pepo L. double haploids to MS ＋ 30 g/L Suc ＋ 8 g/L Agar ＋ 2.0 mg/L 6-BA ＋ 0.1 mg/L NAA led to the best results with the induction frequency of 90.0% and differentiation coefficient of 8.5. MS medium with addition of 0.05 or 0.1 mg/L NAA led to the highest rooting rate. Regenerated seedlings with 5 - 6 true leaves exhibited the highest survival rate of 90.0%, which was the optimal period for domestication and transplanting of regenerated seedlings. This study laid a solid foundation for high-efficiency utilization of heterosis of C. pepo L.
基金Supported by Project of Modern Crop Breeding[Guangdong Finance of Agriculture(2014)No.492]2012 Innovation and Entrepreneurship Training Program for University Students of Guangdong Province(1134712062)
文摘Haploid breeding can shorten the breeding process and improve the breeding efficiency. Currently, in vivo haploid induction technology has been com- monly used in maize （ Zea mays L. ）. This paper briefly introduced in vivo haploid induction technologies, summarized doubling methods of maize haploids and described the significance and application of maize haploids, which provided the basis for further development of haploid breeding in maize.
文摘A large number of adventitious buds were induced from in vitro cultured young inflorescences of haploids of rice.Having been subcultured on solidified subculture media at 26 ℃ for 7 days ,the adventitious buds were loaded into 1.8 mL plastic cryotubes with cryoprotectant and kept on ice for 45 60 min.After cooled at a rate of 1.0℃/min down to -40℃,the samples were kept in liquid nitrogen.The adventitious buds which have been cryopreserved for about 30 days were thawed rapidly in 38 40℃water and then plated on solidified MS medium containing 3 % sucrose,0.5 mg/L NAA and 2.0 mg/L kinetin.After plated, 23% 32 % of adventitious buds resumed growth and 15% 22 % regenerated plantlets.The results of this work indicated that the adventitious buds derived from in vitro cultured young inflorescences is a critical factor for the success and subculturing adventitious buds on MS medium containing 3% sucrose and 4% sorbitol or 20% potato extract is essential to the procedure.The effective cryoprotectant is 10 % DMSO(dimethyl sulfoximide)+0.5 mol/L sorbitol.
基金National Natural Science Foundation of China(32188102,32372177,32025028,and 2023YFD1200800)Central Public-interest Scientific Institution Basal Research Fund(Y2022QC20).
文摘Dear Editor,Doubled haploid(DH)technology is the fastest and most efficient method for producing completely homozygous lines,commonly used in maize breeding(Ren et al.,2017).The discovery of Stock 6,the first maize haploid inducer(HI)reported in 1959,yields 2%–3%maternal haploids during self-pollination or when used as a male in outcrossing.High-efficiency HIs derived from Stock 6,with a haploid induction rate(HIR)of 7%–15%,have significantly contributed to the commercialization of maize DH breeding.Recently,the genetic basis of haploid induction has been identified with two main molecular players in maize:MTL/ZmPLA1/NLD,which triggers haploid production(Gilles et al.,2017;Kelliher et al.,2017;Liu et al.,2017),and DMP,which enhances haploid induction(Zhong et al.,2019).Importantly,MTL is conserved across cereals,and knockout of OsMATL in rice has produced 2%–6%haploids(Yao et al.,2018;Wang et al.,2019;Liu et al.,2024a),presenting promising prospects for rice breeding.However,due to the lack of high-efficiency HIs and limitations in cross-pollination,DH technology based on Osmatl remains an unreliable and inefficient practice for rice breeding.Here,we developed a high-efficiency HI with favorable agronomic performance in rice.We also present a large-scale haploid production approach achieved by combining this HI with photo-thermo-sensitive genic male sterile(PTGMS)lines(Figure 1A).
基金financially supported by the Jiangsu Provincial Key R&D Program(Modern Agriculture)(BE2023313)the National Natural Science Foundation of China(31901535)the China Agricultural Research System(CARS-03).
文摘The identification of haploid induction genes has promoted the advancement of several breeding technologies.Haploid induction genes in wheat,coupled with visual marker,have led to the establishment of a novel in vivo doubled-haploid(DH)technology.When combined with dominant male sterile genes,this innovative DH method presents a promising avenue for high-throughput production of DH lines.Furthermore,the application of haploid induction genes has facilitated the establishment of other innovative breeding technologies,such as HI-Edit and cyto-swapping in creating cytoplasmic male sterility lines,as well as synthetic apomixis.This review summarizes the progress of DH technology in wheat and presents examples of application of haploid induction genes in accelerating breeding practices,aiming to promote the development of these innovative technologies in wheat and enhancing wheat breeding efficiency.
基金supported by the Nanfan Special Project of the Chinese Academy of Agricultural Sciences(Grant Nos.YBXM2320 and YBXM2433)the Project of Sanya Yazhou Bay Science and Technology City,China(Grant No.SCKJ-JYRC-2024-58)the Agricultural Science and Technology Innovation Program,China(Grant No.CAAS-ASTIP-2021-CNRRI).
文摘Doubled haploid(DH)technology has revolutionized crop breeding by enabling the production of homozygous lines in a single generation.In vivo haploid induction(HI)offers a more widely applicable approach that can significantly improve DH breeding efficiency.ToPAR,a parthenogenesis gene,originally identified in dandelion(Taraxacum officinale),has been characterized.Researchers have successfully induced haploid embryo-like structures and haploid offspring in lettuce and foxtail millet,respectively.
基金supported by the Program for Changjiang Scholars and Innovative Research Team in University(Grant No.IRTO453).
文摘Eighteen pairs of diploid-haploid twin-seedlings were identified and screened out fromspecial rice SARII-628 population. Five pairs of themwere selected and randomly designated as A, B, C, Dand E. Simple sequence repeats (SSR) analysisshowed that there was no difference among 310 siteswhich indicated that there was no base mutation onDNA primary structure. DNA methylation plays animportant role in gene expression regulation duringgrowth and development stages in eukaryotes. Amodified AFLP technique (methylation-sensitiveAFLP, MSAP) was employed to detect the DNA me-thylation patterns in the 5′-CCGG sites of the fivepairs of twin-seedlings. Although no methylationmutation was detected among the five diploids,forty-three methylation mutation sites were foundfrom the corresponding haploids. The MSAP ratios,which were the ratios of MSAP sites to the total am-plified sites, in five haploids were 18.79%, 19.35%,18.49%, 18.45% and 18.75%, respectively. And cor-responding full methylation levels (5′-CmCGG indouble strands) of those haploids were 10.58%,11.3%, 10.11%, 10.09% and 10.34%, respectively.Both MSAP and full methylation levels in the fivehaploids were higher than that of their correspondingdiploids, which suggested that hypermethylation oc-curred in some 5′-CCGG sites. Five types of MASPpatterns among the five pairs of twin-seedlings weredetected as follows: (1) no changes, methylation lev-els were the same in both haploids and diploids; (2) demethylation, diploid was methylated but no me-thylation in the same site in haploid; (3) hypermethy-lation, the methylation level in haploid was higher than those in diploid; (4) hypomethylation, methyla-tion in haploid was lower than those in diploid; (5) undecided pattern, change trend of methylation lev-els in haploids was not decided. The bands of 18 sites were reclaimed, then sequenced and searched on website to determine the sites of those sequences on rice chromosomes. The result showed that the methylation mutation involved the whole rice genome and 12 pairs of chromosomes. The mutation was site-related and there were different mutation sites for different haploids. Compared to diploids, the higher methylation level in haploids might be a readjusting reaction to the decrease in ploidy for the sake of sur-vival.
基金the National Natural Science Foundation of China (Grant No. 30771157)the Changjiang Scholars and Innovative Research Team in University (Grant No. IRT0453)
文摘In this study, microarray technique was employed to analyze the gene expression at the RNA level between haploids and corresponding diploids derived from a rice twin-seedling line SARII-628. Differ- ent degrees of expression variations were observed in the plant after haploidization. The main results are as follows: (1) after haploidization, the ratio of the sensitive loci was 2.47% of the total loci designed on chip. Those loci were randomly distributed on the 12 pairs of rice chromosomes and the activated loci were more than the silenced ones. (2) Gene clusters on chromosome were observed for 33 se- quences. (3) GoPipe function classification for 575 sensitive loci revealed an involvement in the bio- logical process, cell component and molecular function. (4) RT-PCR generally validated the result from microarray with a coincidence rate of 83.78%. And for the randomly-selected activated or silenced loci in chip analysis, the coincidence rate was up to 91.86%.
文摘The introduction of alleles into commercial crop breeding pipelines is both time consuming and costly.Two technologies that are disrupting traditional breeding processes are doubled haploid(DH)breeding and genome editing(GE).Recently,these techniques were combined into a GE trait delivery system called HI-Edit(Haploid Inducer-Edit).In HI-Edit,the pollen of a haploid inducer line is reprogrammed to deliver GE traits to any variety,obviating recurrent selection.For HI-Edit to operate at scale,an efficient transformable HI line is needed,but most maize varieties are recalcitrant to transformation,and haploid inducers are especially difficult to transform given their aberrant reproductive behaviors.Leveraging marker assisted selection and a three-tiered testing scheme,we report the development of new Iodent and Stiff Stalk maize germplasm that are transformable,have high haploid induction rates,and exhibit a robust,genetically-dominant anthocyanin native trait that may be used for rapid haploid identification.We show that transformation of these elite‘‘HI-Edit”lines is enhanced using the BABYBOOM and WUSCHEL morphogenetic factors.Finally,we evaluate the HI-Edit performance of one of the lines against both Stiff Stalk and non-Stiff Stalk testers.The strategy and results of this study should facilitate the development of commercially scalable HI-Edit systems in diverse crops.
文摘The technique of producing doublehaploid of wheat by distant hybridization between wheat and maize has characterized with better inducing effect, shorter in- ducing period, easy operation, and so on. At present, it is the most efficient and has great potential of application in breeding of wheat. This article reviewed princi- ple and production process of the technique, research situation of the three key in- dicators of the technology（embryo rate, seedling rate and success rate of doubling）in recent years, and application of the technology in breeding, genetics, germplasm improvement of wheat. At last, both the achievements and the direction of further improvement and development of the technology in our program were discussed.
基金Supported by Major Research Project for Innovation of Main Crop Varieties in the " Twelfth Five-Years" Plan of Chongqing City "Breeding of Maize Germplasm and Creation of Breeding Materials" (cstc2012ggC80004) "Study on New Methods and New Technologies for Maize Breeding" (cstc2012ggC80003)+3 种基金 "Breeding of New Varieties of High-quality Drought-resistant and Barren-tolerant Maize" (cstc2012ggC80006)Rural Science and Technology Project in the "Twelfth Five-Years" Plan of China "Study on DH Engineering Breeding Techniques and New Variety Breeding of Maize" (2012AA101203) "Creation and Application of Strong Heterosis Maize Hybrid in Southwest Mountain" (2011AA10A103)Major Research Project in Chongqing City "Creation of DH Lines of High-quality Stress-resistant Maize" (CSTC2011ggB80057)~~
文摘This study aimed to investigate the induction effect on different genotypes of maize of haploid inducers RWS, Stock6, CAU Inducer and hybridization-derived breeding lines. The results showed that 2005 had the highest induction rate （2.85%） when using inducer Stock6 as male parent material; haploid induction rates were significantly different among different genotypes of maize when using RWS, Stock6, CAU Inducer and their crossbreeding materials as the male parents; inducers 2061, 2062, 2058 and 2059 led to relatively high induction rates, which could be emphatically utilized. Furthermore, during the screening of induction materials, different female parent materials should be selected based on the breeding objective.
基金Supported by Innovation Center of Agricultural Sciences,HubeiProvince~~
文摘[Purpose] The paper is to discuss the high-efficient breeding technology platform of dwarf male-sterile wheat.[Method] Conclusion was made upon the development of the creation and application of dwarf male-sterile wheat,and the conception of introducing haploid breeding technology into the technological system of high-efficient breeding of dwarf male-sterile wheat was proposed.[Result] Dwarf male-sterile wheat not only retains the properties of total male abortion of Taigu Genetic Sterile Wheat,steady sterility,and high hybrid rate after open pollination,but also has the characteristic of high dwarfing ability of Aibian No.1,becoming a comparatively ideal population modification means.At present,the application of dwarf male-sterile wheat mainly includes germplasm resources platform of population modification by using dwarf male-sterile wheat,technology platform of constructing new high-efficient breeding system by using dwarf male-sterile wheat,and production platform of using dwarf male-sterile wheat to breed new varieties.Through the introduction of haploid breeding into the already established wheat high-efficient breeding system,a new system of high-efficient biological breeding technology of dwarf male-sterile wheat was constructed theoretically.[Conclusion] The study provides references to the further study and application of dwarf male-sterile wheat.
基金supported by the National Key Research and Development Program of China(2016YFD0101200)the China Agriculture Research System(CARS-02)+1 种基金2020 Research Program of Sanya Yazhou Bay Science and Technology City(SKJC-2020-02-03)the National Natural Science Foundation of China(91935303,32001554)。
文摘Doubled haploid(DH) technology is important in modern maize breeding. Haploid inducers determine the efficiency of both haploid induction and identification. It has taken decades to improve the efficiency,haploid induction rate(HIR), from the ~2% of the ancestor haploid inducer, stock6, to the ~10% of modern haploid inducers. Improvement of kernel oil content(KOC) would further enhance haploid identification efficiency. Using molecular marker-assisted selection, in combine with the number of haploids per ear as phenotypic criterion, we developed a new high-oil haploid inducer line, CHOI4, with a mean HIR of 15.8%and mean KOC of 11%. High KOC of CHOI4 can achieve a mean accuracy greater than 90% in identification of haploids of different backgrounds, with reduced false discovery rates and false negative rates in comparison with the previous high-oil haploid inducer line, CHOI3. Comparison of phenotypic selection strategies suggested that the number of haploids per ear can be used as a phenotyping criterion during haploid inducer line development. CHOI4 could further increase the efficiency of large-scale DH breeding programs with lower cost.
基金The study was funded by the National 863 Program, China （2002AA207012-1-3, 2001AA241121-9）the National Natural Science Foundation of China （3980453）.
文摘Pepper hybrid seeds production using male sterility could lower cost by reducing time and labour, and increase the genetic purity of the F1 seeds. To investigate the genetics of fertility restoration of the Peterson cytoplasmic sterility in pepper, a doubled haploid population of 115 pepper lines obtained from anther culture of the F1 hybrid between Yolo Wonder （sterility maintainer line） and Perennial （fertility restorer line） and the parental lines were test-crossed by 77013A （a strict cytoplasmic-genic male sterile line）. The fertility of the test-crossed lines was assessed in greenhouse and open field with the following three criteria： pollen index （PI, visual estimation of pollen amount per flower）, pollen number （PN, pollen counting under microscope）, and seed number （SN, the number of seeds per fruit in open pollination）. Correlations between the each couple of criteria within, as well as between the cultivation methods ranged from 0.55 to 0.84. Analysis of variance showed that the genotype （DH line） and environment were the significant sources of variation of the fertility. Narrow sense of heritance of fertility restoration ranged from 0.38 to 0.92, depending on the criteria and environment. The distribution of the progeny was continuous between the parental genotypes indicating the quantitative inheritance of fertility restoration. Inferred from segregation according to Snape et al.（1984）, the number of segregating genes was estimated to be that three to four genetic factors were involved in pollen traits （PI and PN） and five to eight genetic factors in seed production （SN）. The heredity analysis of the CMS will be helpful for understanding of the genetic mechanism of the fertility restoration and the exploitation of the CMS in hybrid seed production.
文摘Field trials with a set of 108 doubled haploid lines(DHs) derived from a cross between the Chinese winter wheat cvs.CA9613 and H1488 were run at Beijing(China).Phenotypic data were recorded for major agronomic yield traits,i.e.grain weight per ear,grain number per ear and thousand grain weight(Tgw) in two field trials at Beijing.Based on the phenotypic data and a genetic map comprising 168 SSR markers,an analysis of quantitative trait loci(QTL) was carried out for yield and yield parameters using the composite interval mapping(CIM) approach.A total of 14 QTL were detected for these traits across two environments.Four of these QTL located on chromosomes 1A and 2B,respectively,exhibited pleiotropic effects.Loci showing pleiotropic effects will be very useful for understanding the homeologous relationships of QTL and designing an appropriate marker-assisted selection programme by multi-trait selection in order to accumulate("pyramide") favorable alleles at different loci.
基金Supported by National High Technology Research and Development Program of China(863 Program)(2011AA10A106)Yunnan Provincial Fund for Applied Basic Researches(2010CC001)Key New Product Development Plan of Yunnan Province(2012BB015)~~
文摘A DH population derived from C49S-87/01Y1-1069 was used to study the inheritance of wheat haploid embryo production frequency（EPF） in wheat × maize cross with the mixed major gene and polygene inheritance model of quantitative traits. The results showed that the EPF of wheat × maize cross was controlled by two dominant epistatic genes and polygene with gene effects of 1.95 for the first major gene, 6.69 for the second one and 2.80 for the polygene. The inheritability of major genes was as high as 72.09%, suggesting that the differences in EPF among wheat materials were mainly influenced by genotype. However, non-genetic factors were still important, especially for wheat materials with low EPF.
