Substitution Lines Research Articles

Construction of Chromosome Segment Substitution Lines and Inheritance of Seed-Pod Characteristics in Wild Soybean.

Genetic populations provide the basis for genetic and genomic research, and chromosome segment substitution lines (CSSLs) are a powerful tool for the fine mapping of quantitative traits, new gene mining, and marker-assisted breeding. In this study, 213 CSSLs were obtained by self-crossing, backcrossing, and marker-assisted selection between cultivated soybean (Glycine max [L.] Merr.) variety Suinong14 (SN14) and wild soybean (Glycine soja Sieb. et Zucc.) ZYD00006. The genomes of these 213 CSSLs were resequenced and 580,524 single-nucleotide polymorphism markers were obtained, which were divided into 3,780 bin markers. The seed-pod-related traits were analyzed by quantitative trait locus (QTL) mapping using CSSLs. A total of 170 QTLs were detected, and 32 QTLs were detected stably for more than 2 years. Through epistasis analysis, 955 pairs of epistasis QTLs related to seed-pod traits were obtained. Furthermore, the hundred-seed weight QTL was finely mapped to the region of 64.4 Kb on chromosome 12, and Glyma.12G088900 was identified as a candidate gene. Taken together, a set of wild soybean CSSLs was constructed and upgraded by a resequencing technique. The seed-pod-related traits were studied by bin markers, and a candidate gene for the hundred-seed weight was finely mapped. Our results have revealed the CSSLs can be an effective tool for QTL mapping, epistatic effect analysis, and gene cloning.

Comparative proteomic analysis of chromosome segment substitution lines of Thai jasmine rice KDML105 under short-term salinity stress.

Heat shock proteins, ROS detoxifying enzymes, and ion homeostasis proteins, together with proteins in carbohydrate metabolism, cell structure, brassinosteroids, and carotenoid biosynthesis pathway were up-regulated in CSSLs under salinity stress. Rice is one of the most consumed staple foods worldwide. Salinity stress is a serious global problem affecting rice productivity. Many attempts have been made to select or produce salinity-tolerant rice varieties. Genetics and biochemical approaches were used to study the salinity-responsive pathway in rice to develop salinity tolerant strains. This study investigated the proteomic profiles of chromosome segment substitution lines (CSSLs) developed from KDML105 (Khao Dawk Mali 105, a Thai jasmine rice cultivar) under salinity stress. The CSSLs showed a clear resistant phenotype in response to 150mM NaCl treatment compared to the salinity-sensitive line, IR29. Liquid chromatography-tandem mass spectrometry using the Ultimate 3000 Nano/Capillary LC System coupled to a Hybrid Quadrupole Q-Tof Impact II™ equipped with a nano-captive spray ion source was applied for proteomic analysis. Based on our criteria, 178 proteins were identified as differentially expressed proteins under salinity stress. Protein functions in DNA replication and transcription, and stress and defense accounted for the highest proportions in response to salinity stress, followed by protein transport and trafficking, carbohydrate metabolic process, signal transduction, and cell structure. The protein interaction network among the 75 up-regulated proteins showed connections between proteins involved in cell wall synthesis, transcription, translation, and in defense responses.

Insights into the Regulation of Rice Seed Storability by Seed Tissue-Specific Transcriptomic and Metabolic Profiling.

Non-dormant seeds are continuously aging and deteriorating during storage, leading to declining seed vigor, which is a challenge for the rice seed industry. Improving the storability of seeds is of great significance to ensure the quality of rice and national food security. Through a set of chromosome segment substitution lines population constructed using japonica rice NIP as donor parent and indica rice ZS97 as recurrent parent, we performed seed storability QTL analysis and selected four non-storable NILs to further investigate the storability regulatory mechanisms underlying it. The seeds were divided into four tissues, which were the embryo, endosperm, aleurone layer, and hull, and tissue-specific transcriptome and metabolome analyses were performed on them. By exploring the common differentially expressed genes and differentially accumulated metabolites, as well as the KEGG pathway of the four non-storable NILs, we revealed that the phenylpropanoid biosynthesis pathway and diterpenoid biosynthesis pathway played a central role in regulating seed storability. Integrated analysis pinpointed 12 candidate genes that may take part in seed storability. The comprehensive analysis disclosed the divergent and synergistic effect of different seed tissues in the regulation of rice storability.

Precise genetic mapping of Rf18(t), a new fertility restorer gene from 'Nipponbare' for wild abortive cytoplasmic male sterility in rice (Oryza sativa L.).

We mapped Rf18(t), a Restorer-of-fertility gene for wild abortive cytoplasmic male sterility from the japonica maintainer 'Nipponbare', to chromosome 1. The best candidate gene, LOC_Os01g71320, is predicted to encode hexokinase. Three-line hybrid rice obtained through cytoplasmic male sterility (CMS) has helped increase the yield of rice globally, and the wild abortive (WA)-type cytoplasm from wild rice (Oryza rufipogon Griff.) is used widely in three-line indica hybrids. The identification and mapping of the Restorer-of-fertility (Rf) genes in maintainer lines aided in uncovering the genetic basis of fertility restoration of WA-type CMS and the development of WA-type hybrids. In this study, we identified a new Rf gene, Rf18(t), for WA-type CMS from the japonica maintainer line 'Nipponbare' using a chromosome segment substitution line population derived from a cross between the indica line 9311 and 'Nipponbare.' Using a substitution mapping strategy, Rf18(t) was delimited to a 48-kb chromosomal region flanked by molecular marker loci ID01M28791 and ID01M28845 on chromosome 1. By comparative sequence analyses, we propose that LOC_Os01g71320 is the most likely candidate gene for Rf18(t), and it is predicted to encode hexokinase. Furthermore, Rf18(t) was found to function in fertility restoration probably by a posttranscriptional mechanism and its function is dependent on the genetic background of 9311. These results broaden our knowledge on the mechanism of fertility restoration of WA-type CMS lines and will facilitate the development of WA-type rice hybrids.

Identification of candidate genes related to soluble sugar contents in soybean seeds using multiple genetic analyses

Soluble sugar content in seeds is an important quality trait of soybean. In this study, 57 quantitative trait loci (QTLs) related to soluble sugar contents in soybean seeds were collected from databases and published papers. After meta-overview-collinearity integrated analysis to refine QTL intervals, eight consensus QTLs were identified. To further verify the consensus QTLs, a population of chromosome segment substitution lines (CSSLs) was analyzed. Two lines containing fragments covering the regions of consensus QTLs and the recurrent parent were selected: one line showed high soluble sugar contents associated with a consensus QTL fragment, and the other line showed low soluble sugar contents. Transcriptome sequencing was conducted for these two lines at the early, middle, and late stages of seed development, which identified 158, 109 and 329 differentially expressed genes, respectively. Based on the analyses of re-sequencing data of the CSSLs and the consensus QTL region, three candidate genes (Glyma.19G146800, Glyma.19G122500, and Glyma.19G128500) were identified in the genetic fragments introduced from wild soybean. Sequence comparisons between the two CSSL parents SN14 and ZYD00006 revealed a single nucleotide polymorphism (SNP) mutation in the coding sequence of Glyma.19G122500, causing a non-synonymous mutation in the amino acid sequence that affected the predicted protein structure. A Kompetitive allele-specific PCR (KASP) marker was developed based on this SNP and used to evaluate the CSSLs. These results lay the foundation for further research to identify genes related to soluble sugar contents in soybean seeds and for future soybean breeding.

Reconstruction of the High Stigma Exsertion Rate Trait in Rice by Pyramiding Multiple QTLs.

Asian cultivated rice is a self-pollinating crop, which has already lost some traits of natural outcrossing in the process of domestication. However, male sterility lines (MSLs) need to have a strong outcrossing ability to produce hybrid seeds by outcrossing with restorer lines of male parents in hybrid rice seed production. Stigma exsertion rate (SER) is a trait related to outcrossing ability. Reconstruction of the high-SER trait is essential in the MSL breeding of rice. In previous studies, we detected eighteen quantitative trait loci (QTLs) for SER from Oryza sativa, Oryza glaberrima, and Oryza glumaepatula using single-segment substitution lines (SSSLs) in the genetic background of Huajingxian 74 (HJX74). In this study, eleven of the QTLs were used to develop pyramiding lines. A total of 29 pyramiding lines with 2–6 QTLs were developed from 10 SSSLs carrying QTLs for SER in the HJX74 genetic background. The results showed that the SER increased with increasing QTLs in the pyramiding lines. The SER in the lines with 5–6 QTLs was as high as wild rice with strong outcrossing ability. The epistasis of additive by additive interaction between QTLs in the pyramiding lines was less-than-additive or negative effect. One QTL, qSER3a-sat, showed minor-effect epistasis and increased higher SER than other QTLs in pyramiding lines. The detection of epistasis of QTLs on SER uncovered the genetic architecture of SER, which provides a basis for using these QTLs to improve SER levels in MSL breeding. The reconstruction of the high-SER trait will help to develop the MSLs with strong outcrossing ability in rice.

Novel QTL for lodging resistance, PRL4, improves physical properties with high non-structural carbohydrate accumulation of basal culms in rice (Oryza sativa L.)

Resistance to lodging, an important problem in rice production, has three types: low plant height, strong culm, and high strength of the lower part of the plant. The determinants of strength of the lower part remains unclear, compared with plant height and culm strength. This study identified a new genetic factor involved in the strength of the lower part, as assessed by pushing resistance, using chromosomal segment substitution lines (CSSLs) to clarify the determinants of strength of the lower part by functional analysis of the CSSL and the introgression line (IL) harboring the identified quantitative trait locus (QTL). QTL analysis identified the QTL for increasing pushing resistance on chromosome 4, PRL4, which was not related to days to heading. The CSSL with PRL4 showed increased pushing resistance and physical strength of the basal culm, but decreased filled grain ratio and grain weight. The IL with PRL4, developed by backcrossing this CSSL, improved pushing resistance and the strain of culm until breaking under compression, and did not decrease yield traits. These lines with PRL4 increased the accumulation of non-structural carbohydrate (NSC) in the basal culm at the fully ripe stage. Thus, the genetic control of NSC accumulation in culms by PRL4 may improve the strength of the lower part by enhancing culm toughness with strength and ductility.

Localization of salt-tolerant QTL in rice germination stage under different salinity concentrations

Salt stress is an important abiotic stress, which has seriously affected the reproductive development of rice in many parts of the world. Therefore, it is particularly important to understand the genetic mechanism of salt tolerance in rice. In this study, we preliminarily located some quantitative trait loci (QTL) for root length, bud length, and survival percent under different salinity conditions (0, 100, 200 and 400 mM NaCl), using a population of chromosome segment substitution lines (CSSLs) constructed by Nipponbare and 9311. A total of 18 QTLs were identified, which explained the phenotypic variation of 4.76–37.59%, among which 13 QTLs were detected under salt stress condition. These salt tolerance related QTLs were divided into two categories, QTL expressed both under control and salt stress conditions (qSP3, qBL8), and QTL expressed just under salt stress condition (qBL3, qRL3, qSP4, qRL5, qSP7, qSP9, qRL10, qBL11-1, qRL11-1, qRL11, qSP12). qSP3, qSP4, qRL5, qSP7, qRL10, qRL10-1, qRL11, qRL11-1, qBL11, qBL11-1 and qSP12 were reported to be related with salt stress for the first time. These QTLs identified under salt stress may be valuable genetic factors for improving salt tolerance of rice by molecular markers technology, which will help to further understand the genetic mechanism of salt tolerance of rice.

Molecular Cytogenetic Identification of the Wheat–Dasypyrum villosum T3DL·3V#3S Translocation Line with Resistance against Stripe Rust

The annual species Dasypyrum villosum possesses several potentially valuable genes for the improvement of common wheat. Previously, we identified a new stripe rust-resistant line, the Chinese Spring (CS)–D. villosum 3V#3 (3D) substitution line (named CD-3), and mapped its potential rust resistance gene (designated as YrCD-3) on the 3V#3 chromosome originating from D. villosum. The objective of the present study was to further narrow down the YrCD-3 locus to a physical region and develop wheat-3V#3 introgression lines with strong stripe rust resistance. By treating CD-3 seeds with 60Co γ-irradiation, two CS-3V#3 translocation lines, T3V#3S.3DL and T3DS.3V#3L (termed 22-12 and 24-20, respectively), were identified from the M4 generation through a combination of non-denaturing fluorescence in situ hybridization (ND-FISH) and functional molecular markers. Stripe rust resistance tests showed that the line 22-12 exhibited strong stripe rust resistance similarly to CD-3, whereas 24-20 was susceptible to stripe rust similarly to CS, indicating that YrCD-3 is located on the short arm of 3V#3. The line 22-12 can potentially be used for further wheat improvement. Additionally, to trace 3V#3 in the wheat genetic background, we produced 30 3V#3-specific sequence tag (EST) markers, among which, 11 markers could identify 3V#3S. These markers could be valuable in fine-mapping YrCD-3.

Cytomixis in wheat male meiosis: influence analysis of the substitution of chromosome 1A, 2D, 5A, or 5D

ABSTRACT Cytomixis is a process of intercellular nuclear migration (INM) often observable in plant male meiosis. The genes responsible for this phenomenon have not been discovered yet. In an attempt to identify a “cytomictic chromosome,” we studied the male meiosis of wheat–rye single-chromosome substitution lines. Markers of the chromatin state were analyzed to figure out the nature of INM in cereal plants: is it a rather normal process or a consequence of random injury/deviation? There were no obvious correlations between the loss of wheat chromosome 1A, 2D, 5A, or 5D by hybrid plants and the frequency and cytological picture of INM in their meiosis. Acquisition of rye chromosome 1 Rv, 2 R, or 5 R did not correlate with any changes in the manifestation of INM either. Some unusual cytological patterns of the substituted-chromosome behavior were observed during INM. Chromosomes 1Rv, 2 R, and 5 R, which are alien to the wheat genome, were found to migrate preferentially between meiocytes. Analysis of post-translational histone modifications and synaptonemal-complex formation revealed that the migrating chromatin does not differ from the intact one. It is not inactivated, is properly condensed, and forms bivalents. Our findings suggest that none of the studied chromosomes carry “cytomictic genes.” At the same time, INM in male meiosis of cereals appears to be a normal process that may be involved in hybrid-genome rearrangements.

QTL Analysis of Z414, a Chromosome Segment Substitution Line with Short, Wide Grains, and Substitution Mapping of qGL11 in Rice

Most agronomic traits of rice (Oryza sativa), such as grain length, are complex traits controlled by multiple genes. Chromosome segment substitution lines (CSSLs) are ideal materials for dissecting these complex traits. We developed the novel rice CSSL ‘Z414’, which has short, wide grains, from progeny of the recipient parent ‘Xihui 18’ (an indica restorer line) and the donor parent ‘Huhan 3’ (a japonica cultivar). Z414 contains four substitution segments with an average length of 3.04 Mb. Z414 displays seven traits that significantly differ from those of Xihui 18, including differences in grain length, width, and weight; degree of chalkiness; and brown rice rate. We identified seven quantitative trait loci (QTL) that are responsible for these differences in an F2 population from a cross between Xihui 18 and Z414. Among these, six QTL (qPL3, qGW5, qGL11, qRLW5, qRLW11, and qGWT5) were detected in newly developed single-segment substitution lines (SSSLs) S1–S6. In addition, four QTL (qGL3, qGL5, qCD3, and qCD5) were detected in S1 and S5. Analysis of these SSSLs attributed the short, wide grain trait of Z414 to qGL11, qGL3, qGL5, and qGW5. Substitution mapping delimited qGL11 within an 810-kb interval on chromosome 11. Sequencing, real time quantitative PCR, and cell morphology analysis revealed that qGL11 might be a novel QTL encoding the cyclin CycT1;3. Finally, pyramiding qGL3 (a = 0.43) and qGL11 (a = − 0.37) led to shorter grains in the dual-segment substitution line D2 and revealed that qGL11 is epistatic to qGL3. In addition, S1 and D2 exhibited different grain sizes and less chalkiness than Z414. In conclusion, the short grain phenotype of the CSSL Z414 is controlled by qGL11, qGL3, and qGL5. qGL11 might be a novel QTL encoding CycT1;3, whose specific role in regulating grain length was previously unknown, and qGL11 is epistatic to qGL3. S1 and D2 could potentially be used in hybrid rice breeding.

Fine mapping and analysis of candidate genes for qFT7.1, a major quantitative trait locus controlling flowering time in Brassica rapa L.

qFT7.1, a major QTL for flowering time in Brassica rapa was fine-mapped to chromosome A07 in a 56.4-kb interval, in which the most likely candidate gene is BraA07g018240.3C. In Brassica rapa, flowering time (FT) is an important agronomic trait that affects the yield, quality, and adaption. FT is a complicated trait that is regulated by many genes and is affected greatly by the environment. In this study, a chromosome segment substitution line (CSSL), CSSL16, was selected that showed later flowering than the recurrent parent, a rapid-cycling inbred line of B. rapa (RcBr). Using Bulked Segregant RNA sequencing, we identified a late flowering quantitative trait locus (QTL), designated as qFT7.1, on chromosome A07, based on a secondary-F2 population derived from the cross between CSSL16 and RcBr. qFT7.1 was further validated by conventional QTL mapping. This QTL explained 39.9% (logarithm of odds = 32.2) of the phenotypic variations and was fine mapped to a 56.4-kb interval using recombinant analysis. Expression analysis suggested that BraA07g018240.3C, which is homologous to ATC (encoding Arabidopsis thaliana CENTRORADIALIS homologue), a gene for delayed flowering in Arabidopsis, as the most promising candidate gene. Sequence analysis demonstrated that two synonymous mutations existed in the coding region and numerous bases replacements existed in promoter region between BraA07g018240.3C from CSSL16 and RcBr. The results will increase our knowledge related to the molecular mechanism of late flowering in B. rapa and lays a solid foundation for the breeding of late bolting B. rapa.

QTL mapping for plant architecture in rice based on chromosome segment substitution lines

<p id="C3">Plant architecture is a compound trait integrated with multiple morphological and physiological traits, and it is closely related to rice yield. Deciphering excellent plant architecture alleles or QTLs is of great significance for high-yield rice breeding. In this study, we constructed a set of Changhui 121/Koshihikari chromosome segment substitution lines (CSSLs) with the size of 208 in our laboratory. QTLs controlling plant height, flag leaf morphology, and tiller numbers were detected under three environments. A total of 35 QTLs for rice architecture were identified on 11 chromosomes except chromosome 9, and the range of the phenotypic variation explaining was 2.00%-22.86%. It was worth noting that <italic>qPH-1-1</italic>, <italic>qFLW-6</italic>, and <italic>qFLA-3</italic> could be detected in three environments, among which <italic>qFLW-6</italic> was a newly identified QTL of the flag leaf width. Phenotypic identification verified that the additive effects and environmental stability of the two locus alleles by the replacement lines carrying <italic>qPH-1-1</italic> and sites. The results of this study laid the foundation for further fine mapping and cloning of QTLs for rice plant architecture and the molecular marker-assisted selection (MAS) in rice breeding.

Root system architecture analysis and genome-wide association study of root system architecture related traits in cotton

<p id="C3">Root is the main organ of plants that absorbs water and nutrients. The growth of the root system will directly affect the absorption and utilization of nutrients, the resistance to abiotic stress and finally the yield of cotton. In this study, a natural population of 220 upland cotton accessions and a chromosomal segment substitution line (CSSL) population with 325 lines, derived from the crossing and backcrossing of <italic>Gossypium barbadense</italic> acc. 3-79 with <italic>G. hirsutum</italic> cv. ‘Emian 22’ were selected to collect the major root phenotypic traits. Four major traits, namely main root length (MRL), root fresh weight (RFW), root dry weight (RDW) and lateral root angle (LRA) were investigated, and genome-wide association analysis (GWAS) was performed for four root traits in natural populations in combination with genome resequencing. The results showed that the four root traits of the natural population are all in line with normal distribution, and the CSSL population were all in skewed distribution. The mean value of root index of CSSL population was higher than that of natural population. A total of 2,714,140 SNP was obtained from the resequencing data of the natural population. The principal component analysis (PCA) showed that the RFW and MRL could be used as two indexes for cotton root classification, through which the cotton root could be divided into nine types in each population. The analysis of population structure demonstrated that the natural population can be divided into five subgroups. Genome-wide association analysis rrevealed that two association sites were simultaneously associated by RFW and RDW in natural populations. The results of this study provide a theoretical basis for further research on root system architecture (RSA) and its genetic mechanism, and it is also of great significance to cotton breeding of abiotic stress resistance.

Development of Wide-Compatible Indica Lines by Pyramiding Multiple Neutral Alleles of Indica-Japonica Hybrid Sterility Loci.

Since the development of indica hybrid rice in the 1970s, great success has been achieved in hybrid rice production in China and around the world. The utilization of inter-subspecific indica–japonica hybrid rice has always been considered due to its stronger heterosis characteristics. However, indica–japonica hybrids face a serious problem of sterility, which hinders the exploitation of their heterosis. In the past decades, the genetic basis of indica–japonica hybrid sterility has been well studied. It was found that in sterile indica–japonica hybrids, female sterility was mainly controlled by the S5 locus and male sterility by the Sa, Sb, Sc, Sd, and Se loci. In this study, we developed wide-compatible indica lines (WCILs) by pyramiding multiple neutral (n) alleles of the hybrid sterility loci. First, we identified Sn alleles of the loci in single-segment substitution lines (SSSLs) in the genetic background of indica Huajingxian 74 (HJX74). Then, the Sn alleles of S5, Sb, Sc, Sd, and Se loci in SSSLs were pyramided in the HJX74 genetic background. The WCILs carrying Sn alleles at the S5, Sb, Sc, Sd, and Se loci showed wide compatibility with indica and japonica rice varieties. Therefore, the WCILs will be used to develop inter-subspecific indica–japonica hybrid rice with normal fertility.

Genetic Mapping and Candidate Gene Prediction of a QTL Related to Early Heading on Wild Emmer Chromosome 7BS in the Genetic Background of Common Wheat

Heading date (HD) is an essential agronomic objective in wheat conventional breeding. Field experiments from several years and locations indicated that the chromosome arm substitution line (CASL) of wild emmer chromosome 7BS in the genetic background of common wheat var. Chinese Spring (CS) always showed a substantially earlier HD than CS planted in different seasons; usually about 8 d earlier than CS grown under a normal autumn sowing season. CASL7BS consistently showed a much earlier HD than CS when treated for vernalization under a long or short photoperiod and then grown under a short or long photoperiod in the growth room. CASL7BS showed faster spike development than CS at the stages before the glume stage when grown under long days, and depicted relatively rapid growth at all stages when grown under short days. To map the early gene in CASL7BS, F2 plants from the cross of CASL7BS and CS were planted in the field and growth room, forming two mapping populations (P1 and P2, respectively). According to the HD distribution of P1, the HD was most likely regulated by a dominant gene. A QTL was detected consistently in the distal region of about 8.94 cM flanked by C268 and C309 with LOD scores of 5–8, explaining 9.14 and 12.35% of the phenotypic variation in the two mapping populations. The QTL was further narrowed down to an interval between ZAFU058724 and ZAFU061354 of 58–61 Mb based on the HD and genotype of F3 and F4 families. A total of 41 genes were located in this region, and eleven of them were thought to be the candidate genes based on the gene functions. According to the HD and mapping location, the QTL identified in this study was a new gene associated with flowering, which will be helpful in understanding the mechanism of wheat flowering and for breeding an early wheat variety.

Identification of Candidate Genes for Lint Percentage and Fiber Quality Through QTL Mapping and Transcriptome Analysis in an Allotetraploid Interspecific Cotton CSSLs Population.

Upland cotton (Gossypium hirsutum) has long been an important fiber crop, but the narrow genetic diversity of modern G. hirsutum limits the potential for simultaneous improvement of yield and fiber quality. It is an effective approach to broaden the genetic base of G. hirsutum through introgression of novel alleles from G. barbadense with excellent fiber quality. In the present study, an interspecific chromosome segment substitution lines (CSSLs) population was established using G. barbadense cultivar Pima S-7 as the donor parent and G. hirsutum cultivar CCRI35 as the recipient parent. A total of 105 quantitative trait loci (QTL), including 85 QTL for fiber quality and 20 QTL for lint percentage (LP), were identified based on phenotypic data collected from four environments. Among these QTL, 25 stable QTL were detected in two or more environments, including four for LP, eleven for fiber length (FL), three for fiber strength (FS), six for fiber micronaire (FM), and one for fiber elongation (FE). Eleven QTL clusters were observed on nine chromosomes, of which seven QTL clusters harbored stable QTL. Moreover, eleven major QTL for fiber quality were verified through analysis of introgressed segments of the eight superior lines with the best comprehensive phenotypes. A total of 586 putative candidate genes were identified for 25 stable QTL associated with lint percentage and fiber quality through transcriptome analysis. Furthermore, three candidate genes for FL, GH_A08G1681 (GhSCPL40), GH_A12G2328 (GhPBL19), and GH_D02G0370 (GhHSP22.7), and one candidate gene for FM, GH_D05G1346 (GhAPG), were identified through RNA-Seq and qRT-PCR analysis. These results lay the foundation for understanding the molecular regulatory mechanism of fiber development and provide valuable information for marker-assisted selection (MAS) in cotton breeding.

Emergence of micronuclei as polymorphism induced genomic instability in Fennel (Foeniculum vulgare Mill.).

Micronuclei (MN) are extra-nuclear bodies containing damaged chromosomal fragments as well as complete chromosomes that are not otherwise incorporated into the nucleus after cell division. The present experiment was conducted to assess mutagenic effects of ethyl methane sulphonate (EMS) on the meiotic behavior of pollen mother cells of fennel (Foeniculum vulgare Mill.). Cytological analysis revealed that micronuclei were incited at all the three concentrations of EMS. A peculiar correlation of chromosomal abnormalities, mainly bridge and laggard at anaphase I and II with this nuclear polymorphism were experienced for genomic instability. Pollen viability was additionally inspected through pollen fertility. Thus, the MN formations bring about incomplete elimination of the genome, which can be proficiently use as the light of hope inbreeding programmers to produce addition and substitution lines.

Development and Characterization of Chromosome Segment Substitution Lines Derived from Oryza rufipogon in the Background of the Oryza sativa indica Restorer Line R974

Dongxiang wild rice (DXWR) (O. rufipogon Griff.), which has the northernmost worldwide distribution of a wild rice species, is a valuable genetic resource with respect to improving stress tolerance in cultivated rice (Oryza sativa L.). In the three-line hybrid rice breeding system, restorer lines play important roles in enhancing the tolerance of hybrid rice. However, restorer lines have yet to be used as a genomic background for development of substitution lines carrying DXWR chromosome segments. We developed a set of 84 chromosome segment substitution lines (CSSLs) from a donor parent DXWR × recurrent parent restorer line R974 (Oryza sativa indica) cross. On average, each CSSL carried 6.27 introgressed homozygous segments, with 93.37% total genome coverage. Using these CSSLs, we identified a single QTL, qDYST-1, associated with salt stress tolerance on chromosome 3. Furthermore, five CSSLs showing strong salt stress tolerance were subjected to whole-genome single-nucleotide polymorphism chip analyses, during which we detected a common substitution segment containing qDYST-1 in all five CSSLs, thereby implying the validity and efficacy of qDYST-1. These novel CSSLs could make a significant contribution to detecting valuable DXWR QTLs, and provide important germplasm resources for breeding novel restorer lines for use in hybrid rice breeding systems.

Wheat-Thinopyrum Substitution Lines Imprint Compensation Both From Recipients and Donors.

Even frequently used in wheat breeding, we still have an insufficient understanding of the biology of the products via distant hybridization. In this study, a transcriptomic analysis was performed for six Triticum aestivum-Thinopyrum elongatum substitution lines in comparison with the host plants. All the six disomic substitution lines showed much stronger “transcriptomic-shock” occurred on alien genomes with 57.43–69.22% genes changed expression level but less on the recipient genome (2.19–8.97%). Genome-wide suppression of alien genes along chromosomes was observed with a high proportion of downregulated genes (39.69–48.21%). Oppositely, the wheat recipient showed genome-wide compensation with more upregulated genes, occurring on all chromosomes but not limited to the homeologous groups. Moreover, strong co-upregulation of the orthologs between wheat and Thinopyrum sub-genomes was enriched in photosynthesis with predicted chloroplastic localization, which indicates that the compensation happened not only on wheat host genomes but also on alien genomes.