Male Sterile Lines Research Articles

Natural variation of MS2 confers male fertility and drives hybrid breeding in soybean.

A complete and genetically stable male sterile line with high outcrossing rate is a prerequisite for the development of commercial hybrid soybean. It was reported in the last century that the soybean male sterile ms2 mutant has the highest record with seed set. Here we report the cloning and characterization of the MS2 gene in soybean, which encodes a protein that is specifically expressed in the anther. MS2 functions in the tapetum and microspore by directly regulating genes involved in the biosynthesis of secondary metabolites and the lipid metabolism, which is essential for the formation of microspore cell wall. Through comparison of the field performance with the widely used male sterile mutants in the same genetic background, we demonstrated that the ms2 mutant conducts the best in outcrossing rate and makes it an ideal tool in building a cost-effective hybrid system for soybean.

Estimating reliability for evaluating nitrate accumulation in sorghum (Sorghum bicolor L. Moench) inbred A‐lines

AbstractSorghum is an important forage crop in many cropping systems. Nitrate can accumulate in the vegetative plant tissue to levels high enough that result in fatality when fed to ruminant livestock when grown under high nitrate conditions. If variation among inbred lines can be documented for nitrate concentration, it may be possible to reduce nitrate accumulation within forage sorghum hybrids by selecting inbred lines having lower nitrate accumulation. The goal of this study was to evaluate a diverse set of male‐sterile sorghum inbred lines grown under high nitrate conditions, to observe if genotypic variation exists among lines used to produce forage sorghum hybrids. Forty replicates of 20 sorghum A‐lines were grown in a greenhouse and received either 272 kg ha−1 or 408 kg ha−1 of actual nitrogen per plant through applications of ammonium nitrate. Variance components were extracted from linear regression models and reliability was estimated on an entry‐mean basis. Leaf and stem reliability estimates were 0.95 and 0.93, respectively. These reliability estimates indicate the genetic variance comprised most of the phenotypic variance, and that our measurements were consistent. Significant differences were also observed between genotypes for both leaf and stem tissue. Therefore, sufficient genetic variance exists for developing male sterile sorghum inbred lines having leaf nitrate concentrations below the potentially lethal toxicity threshold of 10,000 μg g−1.

Potential candidate genes and pathways related to cytoplasmic male sterility in Dianthus spiculifolius as revealed by transcriptome analysis.

We introduced the candidate gene DsHSP70 into Arabidopsis thaliana, resulting in male gametophyte sterility and abnormal degeneration of sepals and petals. Cytoplasmic male sterility (CMS) is a useful tool for hybrid production. However, the regulatory mechanism of CMS in Dianthus spiculifolius remains unclear. In this study, we investigated whether male-sterile line of D. spiculifolius has a malformed tapetum and fails to produce normal fertile pollen. RNA sequencing technology was used to compare the gene expression patterns of the D. spiculifolius male-sterile line and its male fertility maintainer line during anther development. A total of 12,365 differentially expressed genes (DEGs) were identified, among which 1765 were commonly expressed in the S1, S2 and S3 stages. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses indicated that these DEGs were mainly involved in oxidation-reduction processes, signal transduction and programmed cell death. Additionally, weighted correlation network analysis (WGCNA) showed that three modules may be related to male sterility. A putative regulatory pathway for the male sterility traits was constructed based on the reproductive development network. After introducing the candidate DsHSP70 gene into Arabidopsis thaliana, we found that overexpressing plants showed anther abortion and shorter filaments, and accompanied by abnormal degeneration of sepals and petals. In summary, our results identified potential candidate genes and pathways related to CMS in D. spiculifolius, providing new insights for further research on the mechanism of male sterility.

Combined transcriptome and proteome analyses between the cytoplasmic male sterile line and its maintainer line in Welsh onion (Allium fistulosum)

AbstractCytoplasmic male sterility (CMS) is dispensable for research of heterosis and investigation of nuclear–cytoplasmic interaction in Welsh onion. The molecular mechanism of the cytoplasmic male sterile line and its maintainer line was investigated by combined transcriptome and proteome analyses in Welsh onion. The 410,585 full‐length non‐chimeric (FLNC, full‐length readsnon‐Chimeric) sequences were obtained, and AS events and lncRNAs were identified by full‐length transcriptome. Towards a comprehensive understanding and identification of related genes for male sterility, the transcriptome and proteome sequencing were utilized to explore the differences between the cytoplasmic male sterile line and its maintainer line. The 287 differentially expressed genes (DEGs) and 49 differentially expressed proteins (DEPs) were identified, and we furthermore explored the function of genes and proteins. The heat map preliminarily demonstrated the expression change of the key genes and proteins in two cultivars. CMS‐related genes were screened form differentially expressed genes, and verified by quantitative real‐time quantitative polymerase chain reaction (qRT‐PCR). Our research found the CMS regulatory genes, proteins and related pathways. The transcriptome and proteome datasets contribute to accelerate the development of CMS gene clones and functional genomics research on Welsh onions.

Development of alloplasmic male sterile lines and their assessment for heterotic potential in eggplant (Solanum melongena L.)

ABSTRACT The use of Solanum aethiopicum as the female parent with S. melongena led to the development of a cytoplasmic-genic male sterility system (CGMS) in eggplant. The male sterility was complete and expressed as rudimentary anthers with no pollen grains. Ten backcross progenies of male sterile lines in diverse genetic backgrounds of S. melongena were characterised after BC7 generation for 14 horticultural traits. It was observed that male sterile lines and their respective maintainer parents were isogenic for all the traits, except stamen size and days to 50% flowering. The fertility of male sterile lines was restored after hybridising with plants derived from an S. melongena × S. aethiopicum cross. The fertility restoration was governed by a single dominant gene. The maternal cytoplasm of S. aethiopicum in male sterile lines and of S. melongena in maintainers and restorers was confirmed with mitochondria and chloroplast DNA-specific molecular markers. Further, to assess the heterotic potential, 10 cytoplasmic male sterility lines were crossed with three restorers to develop 30 F1 hybrids, out of which D−67A × R−6 and D−104A × R−6 were found consistently better than the standard check for yield plant−1 over two years. In this manuscript a comprehensive study on development and utilisation of an alloplasmic CGMS system in eggplant has been discussed.

A review of rice male sterility types and their sterility mechanisms

Male sterility plays an important role in the utilization of heterosis in rice. The establishment of male sterile lines in rice is one of the key technologies in hybrid rice production systems. The currently widely used male sterile line breeding systems mainly include: three-line hybrid rice based on cytoplasmic male sterility, two-line hybrid rice based on environmental sensitive gene male sterility, and third-generation hybrid rice based on nuclear gene male sterility Seed production system. This study reviewed the types and mechanisms of male sterility in rice, and looked forward to the development direction of hybrid rice.

Irradiated Pollen-Induced Parthenogenesis for Doubled Haploid Production in Sunflowers (Helianthus spp.).

Doubled haploid (DH) technology is a tool used to develop large numbers of inbred lines and increase the rate of genetic gain by shortening the breeding cycles. However, previous attempts to produce DH sunflower plants (Helianthus spp.) have resulted in limited success. In this research, we applied gamma-induced parthenogenesis to assist the production of DH sunflowers. The objectives of the study included (1) identifying optimal gamma ray doses for inducing DH sunflowers using two cytoplasmic male sterility (CMS) lines as female plants and two male pollinators with recognizable morphological markers, (2) selecting new male pollinators from wild sunflower varieties, and (3) testing the efficacy of the selected male pollinators using emasculated non-male sterile sunflower lines as female plants. In these experiments, pollen grains were irradiated with gamma ray doses ranging from 50 to 200 Gy. The optimal gamma ray dose for pollen grain irradiation and DH plant production was identified to be 100 Gy. In addition, a cultivated (G11/1440) and a wild-type (ANN1811) sunflower line can be used as common male pollinators for their distinctive morphological markers and wide capacity for DH induction by gamma-irradiated pollen grains.

Genetic analysis of sunflower fatty acids under optimum and water stressed conditions

Abstract In order to study the genetic control of sunflower fatty acids under optimum and water limited conditions a set of 12 sunflower hybrids were evaluated as randomized complete block design with three replications in two separate experiments during two years (2019 and 2020) in Karaj, Iran. The hybrids were obtained by crossing of four male sterile lines and three restorer lines as testers. Water limitation was imposed by water withholding during reproductive stage. Palmitic acid content was governed by both additive and non-additive effects and linoleic acid content by non-additive gene action under optimum and water limited conditions. Stearic acid content was controlled by both additive and non-additive effects in optimum but by additive effects under water limited condition. Oleic acid content was governed by non-additive factors in optimum, while by both additive and non-additive effects in water limited condition. Oil content and leaf temperature were inherited by additive and seed yield by additive and non-additive effects in both conditions. Relative water content and chlorophyll content were controlled by both additive and non-additive effects under optimum but by non-additive effects in water limited condition. According to these results hybrid breeding is suggested for improvement of main sunflower fatty acids under optimum and water limited condition except that selection-based methods for improvement of stearic acid content.

A straight-forward seed production technology system for foxtail millet (Setaria italica).

For autogamous crops, a precondition for using heterosis is to produce sufficient pure male-sterile female parents that can be used to produce hybrid seeds. To date, cytoplasmic male sterility (CMS) and environment-sensitive genic male sterility (EGMS) have been used commercially to exploit heterosis for autogamous species. However, neither CMS nor EGMS has been established for foxtail millet (Setaria italica). Here, we report on the establishment and application of a seed production technology (SPT) system for this crop. First, we established a DsRed-based SPT system, but found that it was unsuitable because it required the use of a fluorescent device for seed sorting. Instead, we constructed an SPT system with de novo betalain biosynthesis as the selection marker. This allowed us to distinguish transgenic seeds with the naked eye, thereby facilitating the identification of SPT maintainer line seeds. In this system, a seed sorter was not required to obtain sufficient seeds. The key point of the strategy is that the seed pool of the SPT maintainer line is propagated by artificial identification and harvesting of male-fertile individuals in the field, and the male-sterile line seed pool for hybrid production is produced and propagated by free pollination of male-sterile plants with the SPT maintainer line. In a field experiment, we obtained 423.96 kg male-sterile line seeds per acre, which is sufficient to plant 700.18 acres of farmland for hybrid seed production or male-sterile line reproduction. Our study therefore describes a powerful tool for hybrid seed production in foxtail millet, and demonstrates how the SPT system can be used for a small-grained crop with high reproduction efficiency.

Gibberellin confers to the expression of TaGA-6D and negatively regulates the fertility of wheat with Aegilops juvenalis cytoplasm

As the trace signal molecules widely existing in plants, plant hormones can regulate physiological responses of plants at low concentrations. At present, the effect of plant endogenous hormones on wheat male fertility has attracted attention, but the molecular mechanism underlying fertility regulation is unclear. Given this, the anthers of five isonuclear alloplasmic male sterile lines and their maintainer line were RNA-sequenced. A gene TaGA-6D encoding gibberellin (GA) regulated protein was isolated, which was located to the nucleus, cell wall and/or cell membrane, and predominantly highly expressed in the anther of Ju706A, a male sterile line with Aegilops juvenalis cytoplasm. By spraying assay of GA with different concentrations on fertility line Ju706R, it was found that with the increase of exogenous GA concentration, the content of endogenous GA and expression level of TaGA-6D in anther gradually increased, and the fertility decreased. However, silencing of TaGA-6D partially restore the fertility of Ju706R sprayed with 1000 ng/μl GA, and indicating that gibberellin can promote the expression of TaGA-6D and negatively regulates the fertility of wheat with Aegilops juvenalis cytoplasm, which providing new insights for understanding hormone regulation of male fertility in wheat.

Industrial chicory genome gives insights into the molecular timetable of anther development and male sterility.

Industrial chicory (Cichorium intybus var. sativum) is a biannual crop mostly cultivated for extraction of inulin, a fructose polymer used as a dietary fiber. F1 hybrid breeding is a promising breeding strategy in chicory but relies on stable male sterile lines to prevent self-pollination. Here, we report the assembly and annotation of a new industrial chicory reference genome. Additionally, we performed RNA-Seq on subsequent stages of flower bud development of a fertile line and two cytoplasmic male sterile (CMS) clones. Comparison of fertile and CMS flower bud transcriptomes combined with morphological microscopic analysis of anthers, provided a molecular understanding of anther development and identified key genes in a range of underlying processes, including tapetum development, sink establishment, pollen wall development and anther dehiscence. We also described the role of phytohormones in the regulation of these processes under normal fertile flower bud development. In parallel, we evaluated which processes are disturbed in CMS clones and could contribute to the male sterile phenotype. Taken together, this study provides a state-of-the-art industrial chicory reference genome, an annotated and curated candidate gene set related to anther development and male sterility as well as a detailed molecular timetable of flower bud development in fertile and CMS lines.

Evaluation of Cytoplasmic Male Sterile Lines and Restorer Lines in Moricandia System for Heterosis Breeding in Indian Mustard [Brassica juncea (L.) Czern & Coss.

This study was undertaken with the objectives of Cytoplasmic Male Sterility (CMS) assessment and study of pollen fertility restoration using five Mori based Cytoplasmic Male Sterile lines viz., Mori ‘A’ SKM 109, Mori ‘A’ SKM 125, Mori ‘A’ SKM 201, Mori ‘A’ SKM 219, Mori ‘A’ SKM 303 and eight diverse Mori based fertility restorer lines viz., Mori ‘R’ GM 2, Mori ‘R’ GM 3, Mori ‘R’ SKM 9033, Mori ‘R’ SKM 301, Mori ‘R’ Pusa Bahar, Mori ‘R’ Vardan, Mori ‘R’ Bio 902, Mori ‘R’ 1-14 to identify good fertility restorer line and stain ability of pollen grains of sterile and fertile lines to Moricandia arvensis, cytoplasmic background of converted cytoplasmic male sterile lines of Indian Mustard [Brassica juncea (L.) Czern & Coss.] by repeated backcrossing at the Main Castor-Mustard Research station, Sardarkrushinagar Dantiwada Agricultural University, Sardarkrushinagar-385506 Banaskantha. The pollen stain ability, plant fertility status and percentage siliquae set per plant was recorded for the morphological characterization and to confirm stability of true Cytoplasmic Male Sterile (CMS) line. The experimental material comprised of fifty-four genotypes consisting of five diverse CMS lines and eight Fertility Restorer lines were crossed in line X tester mating design and resultant forty hybrids along with their thirteen parents and standard check variety (Kranti) were evaluated in randomized block experimental design. The morphological characters were studied viz., pollen fertility, Number of siliquae set per plant, per cent siliquae set per plant in field as well as laboratory tests were conducted with 2 % Aceto-carmine to confirm pollen stain ability and purity of CMS (A-lines) for male sterility and pollen fertility restorability of R-lines. There were visual differences observed for the parents (male sterile lines and fertility restorer lines), all the F1 crosses and standard check parent for pollen fertility. The male sterile lines exhibited (100%) pollen sterility and the pollen fertility restorer lines varied from 80.05 % (Mori ‘R’ Pusa Bahar) to 97.97 %(Mori ‘R’ SKM 9033).

Floral nectary structure, nectar presentation and morpho-anatomical analysis of male‐fertile and male‐sterile anthers in onion (Allium cepa, Amaryllidaceae)

Background and aims: The onion (Allium cepa var. cepa) is an allogamous species in which male-sterile lines have been developed. However, the structure of the flower and floral nectaries have only been briefly described. The objective of this study is to update the floral anatomy data, comparing the gynoecium and androecium of male-fertile and male-sterile lines, and to analyze the three-dimensional structure of the floral nectary. M&M: Conventional optical and scanning electron microscopy techniques were used. Results: Fertile and sterile male flowers have the same structural organization. The septal or gynopleural nectary is organized in three zones: 1) production area formed by the glandular tissue arranged radially in the septa of the ovary, 2) discharge zone: a small channel at the apex connects the nectariferous tissue with the pocket formed between the ovary wall and the overlying ridge, and 3) presentation and harvest zone, where the excreted nectar accumulates, formed between the widened base of the inner staminal filaments opposite the ovary. Conclusions: The only anatomical differences in the flowers sterile male lines are the lack of pollen production and the absence of dehiscence, despite having a normal stomium and anther wall. The floral nectaries of onion are a clear example of secondary presentation of nectar.

Development of Specific Molecular and Phenotypic Marker-Based Haploid Inducers in Rice

Doubled haploid (DH) technology is an efficient strategy for producing completely homozygous lines for breeding programs. Mutations in the MATRILINEAL (MTL) phospholipase trigger intraspecific haploid induction in cereals. Although an in vivo haploid induction system based on OsMTL-edited plants has been established in rice (Oryza sativa), DH technology is still limited by other factors, such as haploid identification, which is one of the essential steps required for DH technology. In the study, we addressed this technical challenge by integrating specific molecular and phenotypic markers into rice haploid inducers. We first generated large fragment insertion or deletion mutations within the OsMTL gene and designed a pair of primers flanking the mutational sites to be used as the specific and universal molecular markers between wild-type and Osmtl plants. Next, we screened for hairy leaf as a single dominant trait and integrated it into specific molecular marker-based haploid inducers using the cross and self-cross method. When crossing cytoplasmic male sterile lines with these haploid inducers, we utilized the specific InDel marker and hairy leaf phenotypic marker to identify putative haploids (or double haploids). These putative haploids were further confirmed through ploidy and phenotypic analysis, demonstrating the high efficiency of haploid identification using these markers. The haploid induction rate (HIR) of the developed specific molecular and phenotypic marker-based haploid inducers ranged from 3.7% to 12.5%. We have achieved successful integration of distinct molecular and phenotypic markers into rice haploid inducers. Our advanced marker-based system has significantly enhanced the accuracy of haploid identification, thereby expediting the adoption of DH technology in rice breeding.

Male sterility and hybrid breeding in soybean

Hybrid breeding can help us to meet the challenge of feeding a growing world population with limited agricultural land. The demand for soybean is expected to grow; however, the hybrid soybean is still in the process of commercialization even though considerable progress has been made in soybean genome and genetic studies in recent years. Here, we summarize recent advances in male sterility-based breeding programs and the current status of hybrid soybean breeding. A number of male-sterile lines with cytoplasmic male sterility (CMS), genic-controlled photoperiod/thermo-sensitive male sterility, and stable nuclear male sterility (GMS) have been identified in soybean. More than 40 hybrid soybean varieties have been bred using the CMS three-line hybrid system and the cultivation of hybrid soybean is still under way. The key to accelerating hybrid soybean breeding is to increase the out-crossing rate in an economical way. This review outlines current problems with the hybrid soybean breeding systems and explores the current efforts to make the hybrid soybean a commercial success.

Heterosis and combining ability vis‐à‐vis association for green fruit yield and component traits involving male sterile lines in chilli (Capsicum annuum) under wet temperate zone of North Western Himalayas

AbstractFourteen genetically diverse inbred lines comprising of four genic male sterile lines and 10 often pollinated lines as testers were crossed following ‘line × tester’ mating design to generate 40 F1s. These F1s, parents and check ‘CH‐27’ were evaluated in α‐lattice square design, replicated thrice during summer seasons of 2020 and 2021. The male sterile line DPChMS 9‐2 was good general combiner for marketable fruit yield and eight other component traits while testers namely, DPCh 10, PBC 535, VVG, Him Palam Mirch‐2 and DPCh 40 showed significant GCA for majority of yield related traits. The hybrids DPChMS 9‐2 × HPM‐2, DPChMS 9‐2 × DPCh 40, DPChMS 9‐2 × DPCh 101, DPChMS 9‐2× VVG and DPChMS 26‐1× HPM‐1 were the most desirable based on specific combining ability (SCA) and economic heterosis for fruit yield and other important traits. Heterosis was positively correlated with SCA indicating prediction of potential hybrids can be made based on both these parameters simultaneously. Present study provides an opportunity to exploit top hybrids for stability and commercial cultivation.

Low temperature compensates for defective tapetum initiation to restore the fertility of the novel TGMS line ostms15.

In rice breeding, thermosensitive genic male sterility (TGMS) lines based on the tms5 locus have been extensively employed. Here, we reported a novel rice TGMS line ostms15 (Oryza sativa ssp. japonica ZH11) which show male sterility under high temperature and fertility under low temperature. Field evaluation from 2018 to 2021 revealed that its sterility under high temperature is more stable than that of tms5 (ZH11), even with occasional low temperature periods, indicating its considerable value for rice breeding. OsTMS15 encodes an LRR-RLK protein MULTIPLE SPOROCYTE1 (MSP1) which was reported to interact with its ligand to initiate tapetum development for pollen formation. In ostms15, a point mutation from GTA (Val) to GAA (Glu) in its TIR motif of the LRR region led to the TGMS phenotype. Cellular observation and gene expression analysis showed that the tapetum is still present in ostms15, while its function was substantially impaired under high temperature. However, its tapetum function was restored under low temperature. The interaction between mOsTMS15 and its ligand was reduced while this interaction was partially restored under low temperature. Slow development was reported to be a general mechanism of P/TGMS fertility restoration. We propose that the recovered protein interaction together with slow development under low temperature compensates for the defective tapetum initiation, which further restores ostms15 fertility. We used base editing to create a number of TGMS lines with different base substitutions based on the OsTMS15 locus. This work may also facilitate the mechanistic investigation and breeding of other crops.

Identification and Cloning of a Putative Male Fertility Gene Encoding an Oxidosqualene Cyclase in Qingke

Anther development is crucial for controlling crop fertility. To elucidate the underlying mechanisms of reproductive development of highland barley (called Qingke in Chinese), two main Qingke cultivars, Zangqing 2000 and Ximala 22, were utilized. Transcriptome analysis showed that lipid, sugar and phenylpropane metabolisms might be the major pathways associated with Qingke male fertility by analyzing the possible common DEGs before anther maturation in both varieties. Additionally, 26 genes related to crop genic male sterility were screened to identify homologous genes for Qingke male sterile lines. Among them, HORVU.MOREX.r3.7HG0634780 (HvtOSC12), an oxidosqualene cyclase, was highlighted as a candidate gene for affecting Qingke male fertility, as it is highly and especially expressed before Qingke anther maturation. Furthermore, HvtOSC12 (including promoter sequence) was cloned by homology-based cloning. The further bioinformatic analysis deduced that MYBs might be one of the transcription factors affecting expression of HvtOSC12 by responding to environmental changes. These results might lay a foundation for the potential applications for the creation of environment-sensitive genic male sterility in Qingke.

Assessment of Cytomorphological Differences in Sorghum Fertility Restoration

Sorghum (Sorghum bicolor L. Moench) is ranked fifth as a cereal crop after maize, rice, wheat, and barley based on global cultivation area. However, heat and drought stresses cause improper fertility restoration and inefficient pollination, severely affecting sorghum productivity. The discovery of cytoplasmic male sterility (CMS) is a milestone for commercializing hybrids. This study assessed the pollen fertility and in vitro pollen germination percentage of male and female lines and F1 hybrids of sorghum using two years of pooled data with multivariate analysis. The principal component analysis (PCA) of female and male lines showed that PC1 represented 82.8% of the variation, whereas PCA of hybrids revealed a significant genetic divergence of 97.1%. Agglomerative hierarchical clustering marked that cluster II genotypes have a high pollen fertility contribution, which can generate superior and high-yielding hybrids. Three male-sterile lines exhibited 100% pollen sterility, with morphological attributes, viz., pinpointed, flattened, low anther extrusion, and starch-digested pollens. Pollen fertility restoration behavior revealed that nine hybrids were fully fertile, eighteen were partially fertile, and three were completely sterile amongst thirty hybrids. The findings of this study will facilitate the identification of potential restorers for the exploitation of high-yielding hybrids in sorghum breeding programs.

Registration of grain sorghum seed (A/B) and pollinator (R) parent lines for chilling and drought tolerance

AbstractFour sorghum [Sorghum bicolor (L.) Moench] seed parents, male sterile A and four male fertile B maintainer lines (KS148A [Reg. no. PL‐321, PI 701488], KS148B [Reg. no. PL‐322, PI 701489], KS149A [Reg. no. PL‐323, PI 701490], KS149B [Reg. no. PL‐324, PI 701491], KS150A [Reg. no. PL‐325, PI 701492], KS150B [Reg. no. PL‐326, PI 701493], KS151A [Reg. no. PL‐327, PI 701494], KS151B [Reg. no. PL‐328, PI 701495]), and six pollinator parents [KS152R (Reg. no. PL‐329, PI 701496), KS153R (Reg. no. PL‐330, PI 701497), KS154R (Reg. no. PL‐331, PI 701498), KS155R (Reg. no. PL‐332, PI 701499), KS156R (Reg. no. PL‐333, PI 701500), and KS157R (Reg. no. PL‐334, PI 701501) from the Kansas State University, Agricultural Research Center, Hays, KS, were approved for release by the Kansas State University in July 2021. These 10 lines were from different populations and were developed through recurrent selection (random mating) followed by pedigree and backcross breeding methods between the selected germplasms (PI 574562, PI 550610, PI 574599, PI 574560, PI 550607, and IS 1269C), adapted breeding lines and different genetic male sterile ms3 lines (KP 8B, N310B, B06‐41701‐167, B9614, BHF14, BH8606‐6) in different combinations. Parent lines KS148A&B, KS149A&B, KS152R, and KS153R are tolerant to chilling stress and KS150A&B, KS151A&B, KS154R, KS155R, KS156R, and KS157R are tolerant to drought stress and are recommended to develop hybrids respectively for early (end of April to first week of May) and regular (end of May to first week of June) planting for the central Great Plains. Preliminary screening of all 10 lines showed tolerance to nine postemergence herbicides. These 10 parent lines are early to medium maturity, three‐dwarf (dw1, Dw2, dw3, dw4) in height, photoperiod‐insensitive, and possess unique combinations of plant and seed color. All 10 parent lines are tannin free with acceptable grain quality, short peduncle length, compact or semi‐compact panicles, and combining ability for yield and standability.