Jasmonic Acid and Ethylene Participate in the Gibberellin-Induced Ovule Programmed Cell Death Process in Seedless Pear ‘1913’ (Pyrus hybrid)

Seedless fruit is a feature appreciated by consumers. The ovule abortion process is highly orchestrated and controlled by numerous environmental and endogenous signals. However, the mechanisms underlying ovule abortion in pear remain obscure. Here, we found that gibberellins (GAs) have diverse functions during ovules development between seedless pear ‘1913’ and seeded pear, and that GA4+7 activates a potential programmed cell death process in ‘1913’ ovules. After hormone analyses, strong correlations were determined among jasmonic acid (JA), ethylene and salicylic acid (SA) in seedless and seeded cultivars, and GA4+7 treatments altered the hormone accumulation levels in ovules, resulting in significant correlations between GA and both JA and ethylene. Additionally, SA contributed to ovule abortion in ‘1913’. Exogenously supplying JA, SA or the ethylene precursor 1-aminocyclopropane-1-carboxylic acid promoted ‘Bartlett’ seed death. The regulatory mechanism in which ethylene controls ovule death has been demonstrated; therefore, JA’s role in regulating ‘1913’ ovule abortion was investigated. A further study identified that the JA signaling receptor MYC2 bound the SENESCENCE-ASSOCIATED 39 promoter and triggered its expression to regulate ovule abortion. Thus, we established ovule abortion-related relationships between GA and the hormones JA, ethylene and SA, and we determined their synergistic functions in regulating ovule death.

Pollen Grain Number and Viability Contribute to Variation in Effective Ovule Number Among Oilseed Rape Genotypes

While no significant differences in initial ovule number were found among oilseed rape genotypes, there was a large variation in effective ovule number (EON), which determines the final seeds per silique (SPS), a critical component of yield. In this study, we selected 18 oilseed rape genotypes with contrasting nitrogen utilization efficiency (NUtE) to unravel the main factors responsible for different EON and determine the critical period of EON formation under both a field and a pot experiments from 2016-2018. The high NUtE genotypes displayed significantly higher NUtE by 14.3%, along with greater yield per plant (29.4%) and SPS (21.1%) than the low NUtE genotypes. The greater productivity of the high NUtE genotypes was associated with 44.1% higher pollen grain number, 23.5% greater pollen viability, and 39.3% lower ovule abortion rate, compared to the low NUtE genotypes. In addition, the heart stage was the critical ovule development period for delineating the variability of EON among contrasting NUtE oilseed rape genotypes, when the high NUtE genotypes displayed higher silique net photosynthetic rate, surface area, biomass, and RNA expression levels. Taken together, this study indicated the pollen grain number, pollen viability and ovule abortion rate contributed to the final variation in EON and the heart stage was the critical period of determining the EON differences among contrasting NUtE genotypes. Increasing pollen grain number and pollen viability, and decreasing ovule abortion rate before heart stage should be the prerequisite for breeders to improve yield and NUtE of oilseed rape genotypes.

Morphological observation and protein expression of fertile and abortive ovules in Castanea mollissima

Chinese chestnuts (Castanea mollissima Blume.) contain 12–18 ovules in one ovary, but only one ovule develops into a seed, indicating a high ovule abortion rate. In this study, the Chinese chestnut ‘Huaihuang’ was used to explore the possible mechanisms of ovule abortion with respect to morphology and proteomics. The morphology and microstructure of abortive ovules were found to be considerably different from those of fertile ovules at 20 days after anthesis (20 DAA). The fertile ovules had completely formed tissues, such as the embryo sac, embryo and endosperm. By contrast, in the abortive ovules, there were no embryo sacs, and wide spaces between the integuments were observed, with few nucelli. Fluorescence labelling of the nuclei and transmission electron microscopy (TEM) observations showed that cells of abortive ovules were abnormally shaped and had thickened cell walls, folded cell membranes, condensed cytoplasm, ruptured nuclear membranes, degraded nucleoli and reduced mitochondria. The iTRAQ (isobaric tag for relative and absolute quantitation) results showed that in the abortive ovules, low levels of soluble protein with small molecular weights were found, and most of differently expressed proteins (DEPs) were related to protein synthesis, accumulation of active oxygen free radical, energy synthesis and so on. These DEPs might be associated with abnormal ovules formation.

Combined Transcriptome Analysis Reveals the Ovule Abortion Regulatory Mechanisms in the Female Sterile Line of Pinus tabuliformis Carr.

Ovule abortion is a common phenomenon in plants that has an impact on seed production. Previous studies of ovule and female gametophyte (FG) development have mainly focused on angiosperms, especially in Arabidopsis thaliana. However, because it is difficult to acquire information about ovule development in gymnosperms, this remains unclear. Here, we investigated the transcriptomic data of natural ovule abortion mutants (female sterile line, STE) and the wild type (female fertile line, FER) of Pinus tabuliformis Carr. to evaluate the mechanism of ovule abortion during the process of free nuclear mitosis (FNM). Using single-molecule real-time (SMRT) sequencing and next-generation sequencing (NGS), 18 cDNA libraries via Illumina and two normalized libraries via PacBio, with a total of almost 400,000 reads, were obtained. Our analysis showed that the numbers of isoforms and alternative splicing (AS) patterns were significantly variable between FER and STE. The functional annotation results demonstrate that genes involved in the auxin response, energy metabolism, signal transduction, cell division, and stress response were differentially expressed in different lines. In particular, AUX/IAA, ARF2, SUS, and CYCB had significantly lower expression in STE, showing that auxin might be insufficient in STE, thus hindering nuclear division and influencing metabolism. Apoptosis in STE might also have affected the expression levels of these genes. To confirm the transcriptomic analysis results, nine pairs were confirmed by quantitative real-time PCR. Taken together, these results provide new insights into ovule abortion in gymnosperms and further reveal the regulatory mechanisms of ovule development.

Fine Mapping and Candidate Gene Analysis of a Major Locus Controlling Ovule Abortion and Seed Number Per Silique in Brassica Napus L.

Key message A major QTL controlling ovule abortion and SN was fine-mapped to a 80.1-kb region on A8 in rapeseed, and BnaA08g07940D and BnaA08g07950D are the most likely candidate genes.Abstact The seed number per silique (SN), an important yield determining trait of rapeseed, is the final consequence of a complex developmental process including ovule initiation and the subsequent ovule/seed development. To elucidate the genetic mechanism regulating the natural variation of SN and its related components, quantitative trait locus (QTL) mapping was conducted using a doubled haploid (DH) population derived from the cross between C4-146 and C4-58B, which showed significant differences in SN and aborted ovule number (AON), but no obvious differences in ovule number (ON). QTL analysis identified 19 consensus QTLs for six SN-related traits across three environments. A novel QTL on chromosome A8, un.A8, which pleiotropically controls all these traits except for ON, was stably detected across the three environments.This QTL explained more than 50% of the SN, AON and percentage of aborted ovule (PAO) variations as well as a moderate contribution on silique length (SL) and thousand seed weight (TSW). The C4-146 allele at the locus increases SN and SL but decreases AON, PAO and TSW. Further fine mapping narrowed down this locus into a 80.1-kb interval flanked by markers BM1668 and BM1672, and six predicted genes were annotated in the delimited region. Expression analyses and DNA sequencing showed that two homologs of Arabidopsis photosystem I subunit F (BnaA08g07940D) and zinc transporter 10 precursor (BnaA08g07950D) were the most promising candidate genes underlying this locus. These results provide a solid basis for cloning un.A8 to reduce the ovule abortion and increase SN in the yield improvement of rapeseed.

Single molecular real-time sequencing revealing the ovule abortion regulatory mechanisms in the female-sterile line of Pinus tabuliformis Carr.

Background Ovule abortion is a common phenomenon in plants, which has an essential significance in seed production. The development of the female gametophyte (FG) is one of the crucial processes of the life cycle, and FG dysplasia is a significant cause contribute to ovule abortion in many species. However, because it is difficult to acquire the mutant about ovule development in gymnosperms, previous studies of the ovule and FG development are mainly focused on angiosperms, especially in Arabidopsis thaliana. Thus the investigation on this field of gymnosperms remains unclear. Results In this study, we investigated the transcriptome data of wild-type (female fertile line, FER) and natural ovule abortion mutant (female sterile line, STE) of Pinus tabuliformis Carr. to evaluate the mechanism of ovule abortion during the process of free nuclear mitosis (FNM). Using single molecule real-time (SMRT) sequencing and next-generation sequencing (NGS), we obtained 202,869 (FER), 197,977 (STE) mapped full-length non-chimeric (FLNC) reads and analyzed 18 cDNA libraries. Based on the SMRT sequencing, we found that both GO annotation and KEGG pathway terms results were similar in FER and STE. NGS analysis further showed altogether 99 differentially expressed genes (DEGs) with opposite expression patterns during the FNM process in FER and STE. According to SMRT sequencing, we found that the number of isoforms and alternative splicing (AS) patterns are variable between FER and STE. Moreover, 5,530 and 5,096 potential simple sequence repeats (SSRs) were identified in FER and STE, respectively. Functional annotation results demonstrated that genes involved in energy metabolism, signal transduction, cell division, and stress response were differentially expressed in different lines, 9 pairs of which were confirmed by quantitative real-time PCR. Conclusion Taken together, these results provide some new insights about the ovule abortion in gymnosperms and further reveal the regulatory mechanisms of ovule development.

Pollen number and vigor contribute to effective ovule numbers among contrasting nitrogen utilization efficiency oilseed rapes genotypes before heart stage

Background: While no significant differences in initial ovule number were found among oilseed rape genotypes, there was a large variation in effective ovule number (EON), which determines the final seeds per silique (SPS), a critical component of yield. Up to date, on study has been focused on unraveling the pre-flowering main factors to restrict EON and identifying the critical period of EON formation among contrasting nitrogen utilization efficiency (NUtE) oilseed rape genotypes.Results: In this study, we selected 18 oilseed rape genotypes with different NUtE to identify the main factors that contribute to EON, and determine if genotypes differed in the critical period of EON formation under both field and pot experiments from 2016-2018. Our results showed the high NUtE genotypes also showed 14.3% higher NUtE, accompanied with 29.4% higher yield per plant and 21.1% higher SPS. The greater productivity of the high NUtE oilseed rape genotypes was associated with 44.1% greater pollen number, 23.5% higher pollen vigor, and 39.3% lower ovule abortion rate, compared to the low NUtE genotypes. In addition, at the heart stage, the high NUtE genotypes displayed higher silique net photosynthetic rate, surface area, biomass, and RNA expression levels, compared to the low NUtE ones. Taken together, this study indicated the pollen number, pollen vigor and ovule abortion rate contributed to the final EON of diverse oilseed rape genotypes; the critical period of determining EON among contrasting NUtE genotypes was at the heart stage.Conclusion: Increasing pollen number and vigor, and decreasing ovule abortion rate before the heart stage should be the prerequisite for breeders to improve yield and NUtE of oilseed rape genotypes.