A comprehensive understanding of CMS/Rf system enabled by modern omics tools and technologies considerably improves our ability to harness hybrid technology for enhancing the productivity of field crops.
 Harnessing hybrid vigor or heterosis is a promising approach to tackle the current challenge of sustaining enhanced yield gains of field crops. In the context, cytoplasmic male sterility (CMS) owing to its heritable nature to manifest non-functional male gametophyte remains a cost-effective system to promote efficient hybrid seed production. The phenomenon of CMS stems from a complex interplay between maternally-inherited (mitochondrion) and bi-parental (nucleus) genomic elements. In recent years, attempts aimed to comprehend the sterility-inducing factors (orfs) and corresponding fertility determinants (Rf) in plants have greatly increased our access to candidate genomic segments and the cloned genes. To this end, novel insights obtained by applying state-of-the-art omics platforms have substantially enriched our understanding of cytoplasmic-nuclear communication. Concomitantly, molecular tools including DNA markers have been implicated in crop hybrid breeding in order to greatly expedite the progress. Here, we review the status of diverse sterility-inducing cytoplasms and associated Rf factors reported across different field crops along with exploring opportunities for integrating modern omics tools with CMS-based hybrid breeding.

Cytoplasmic male sterility (CMS) in hybrid breeding in field crops

Transcriptome profiling using Illumina- and SMRT-based RNA-seq of hot pepper for in-depth understanding of genes involved in CMV infection

Male sterility is an important mechanism for the production of hybrid seeds in watermelon. Although fruit development has been studied extensively in watermelon, there are no reports on gene expression in floral organs. In this study, RNA-sequencing (RNA-seq) was performed in two near-isogenic watermelon lines (genic male sterile [GMS] line, DAH3615-MS and male fertile line, DAH3615) to identify the differentially expressed genes (DEGs) related to male sterility. DEG analysis showed that 1259 genes were significantly associated with male sterility at a FDR P-value of < 0.01. Most of these genes were only expressed in the male fertile line. In addition, 11 functional clusters were identified using DAVID functional classification analysis. Of detected genes in RNA-seq analysis, 19 were successfully validated by qRT-PCR. In this study, we carried out a comprehensive floral transcriptome sequence comparison of a male fertile line and its near-isogenic male sterile line in watermelon. This analysis revealed essential genes responsible for stamen development, including pollen development and pollen tube elongation, and allowed their functional classification. These results provided new information on global mechanisms related to male sterility in watermelon.

/pdf/transcriptome-profiling-of-differentially-expressed-genes-in-13gc781sxp.pdf

Transcriptome profiling of differentially expressed genes in floral buds and flowers of male sterile and fertile lines in watermelon

Heterosis has historically been exploited in plants; however, its underlying genetic mechanisms and molecular basis remain elusive. In recent years, due to advances in molecular biotechnology at the genome, transcriptome, proteome, and epigenome levels, the study of heterosis in vegetables has made significant progress. Here, we present an extensive literature review on the genetic and epigenetic regulation of heterosis in vegetables. We summarize six hypotheses to explain the mechanism by which genes regulate heterosis, improve upon a possible model of heterosis that is triggered by epigenetics, and analyze previous studies on quantitative trait locus effects and gene actions related to heterosis based on analyses of differential gene expression in vegetables. We also discuss the contributions of yield-related traits, including flower, fruit, and plant architecture traits, during heterosis development in vegetables (e.g., cabbage, cucumber, and tomato). More importantly, we propose a comprehensive breeding strategy based on heterosis studies in vegetables and crop plants. The description of the strategy details how to obtain F1 hybrids that exhibit heterosis based on heterosis prediction, how to obtain elite lines based on molecular biotechnology, and how to maintain heterosis by diploid seed breeding and the selection of hybrid simulation lines that are suitable for heterosis research and utilization in vegetables. Finally, we briefly provide suggestions and perspectives on the role of heterosis in the future of vegetable breeding.

/pdf/molecular-basis-of-heterosis-and-related-breeding-strategies-h0v63tvuvk.pdf

Molecular basis of heterosis and related breeding strategies reveal its importance in vegetable breeding.

Differential proteomics analysis to identify proteins and pathways associated with male sterility of soybean using iTRAQ-based strategy

Background 
The use of cytoplasmic male sterility (CMS) in F1 hybrid seed production of chili pepper is increasingly popular. However, the molecular mechanisms of cytoplasmic male sterility and fertility restoration remain poorly understood due to limited transcriptomic and genomic data. Therefore, we analyzed the difference between a CMS line 121A and its near-isogenic restorer line 121C in transcriptome level using next generation sequencing technology (NGS), aiming to find out critical genes and pathways associated with the male sterility.
Results 
We generated approximately 53 million sequencing reads and assembled de novo, yielding 85,144 high quality unigenes with an average length of 643 bp. Among these unigenes, 27,191 were identified as putative homologs of annotated sequences in the public protein databases, 4,326 and 7,061 unigenes were found to be highly abundant in lines 121A and 121C, respectively. Many of the differentially expressed unigenes represent a set of potential candidate genes associated with the formation or abortion of pollen.
Conclusions 
Our study profiled anther transcriptomes of a chili pepper CMS line and its restorer line. The results shed the lights on the occurrence and recovery of the disturbances in nuclear-mitochondrial interaction and provide clues for further investigations.

/pdf/transcriptome-sequencing-and-de-novo-analysis-of-a-wqcvt5jtmn.pdf

Transcriptome Sequencing and De Novo Analysis of a Cytoplasmic Male Sterile Line and Its Near-Isogenic Restorer Line in Chili Pepper (Capsicum annuum L.)

C-repeat binding factors/dehydration responsive element binding factors 1 (CBFs/DREB1s) are a small family of transcription factors that play important roles in plant resistance to various external stresses. However, functional characterization of tea plant (Camellia sinensis L.) CBF gene (CsCBF) was still seldom reported. Here, functional study of the cold-responsive CBF gene (CsCBF2) was done. Results showed that CsCBF2 had conserved AP2 DNA-binding domain and the typical PKK/RPAGRxKFxETRHP and DSAWR signature sequences of CBF/DREB1. Yeast one-hybrid and transcription activation assays revealed that the activation domain of CsCBF2 could activate the reporter gene expression, and the N terminal of CsCBF2 displayed an inhibitory effect. Although CsCBF2 was conserved to bind the C-repeat/dehydration-responsive element (CRT/DRE), intact CsCBF2 protein preferred the CRT cis element. Under normal growth conditions, CsCBF2-overexpressing tobacco plants (CsCBF2-OX) exhibited lighter green leaf color, growth retardation, and dwarfism. Smaller leaf of CsCBF2-OX was only seen in eight weeks after been sown in soil. Under cold, salinity, or drought stress, CsCBF2-OX displayed better growing with longer roots, heavier fresh weight, higher germination rate, and accumulated more proline and sugar contents, but lower electrolyte leakage. The results demonstrated that CsCBF2 enhanced plant tolerance to multiple abiotic stresses.

/pdf/functional-characterization-of-tea-plant-camellia-sinensis-l-3sky985i.pdf

Functional Characterization of Tea Plant (Camellia sinensis L.) CsCBF2 Gene Involved in Multiple Abiotic Stress Response in Tobacco (Nicotiana tabacum L.)

Class B gene APETALA3 (AP3) plays a key role in the development of petals and stamens. Here, we investigated the expression pattern of PAP3 gene (genbank accession number: HM104635) in the buds of cytoplasmic male sterility line 121A and its near-isogenic restorer line 121C at four developmental stages and analyzed the possible association between Class B genes and cytoplasmic male sterility of pepper. Semi-quantitative PCR and quantitative real-time RT-PCR (qRT-PCR) as well as RNA in situ hybridization showed increased expression of PAP3 at late phase of anther development and its higher expression in restorer line compared with sterility line indicating PAP3's role at late developmental stage of anther and suppressed expression in sterility line. RNA in situ hybridization showed Class B gene features: high abundance in stamen and petal; lower expression in pistil; no expression in sepal. Results of transient expression in onion epidermal cells also showed PAP3 localized in the nucleus, which is consistent with the expression pattern of transcription factors of MADS-box gene family.

Expression pattern of Class B gene PAP3 in flower development of pepper.

DNA (deoxyribose nucleic acid) fragment and application thereof in preparation of male sterile hot pepper

PAP3 Regulates Stamen but Not Petal Development in Capsicum annuum L.

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Papers

Transcriptome Sequencing and De Novo Analysis of a Cytoplasmic Male Sterile Line and Its Near-Isogenic Restorer Line in Chili Pepper (Capsicum annuum L.)

Functional Characterization of Tea Plant (Camellia sinensis L.) CsCBF2 Gene Involved in Multiple Abiotic Stress Response in Tobacco (Nicotiana tabacum L.)

Expression pattern of Class B gene PAP3 in flower development of pepper.

DNA (deoxyribose nucleic acid) fragment and application thereof in preparation of male sterile hot pepper

PAP3 Regulates Stamen but Not Petal Development in Capsicum annuum L.