Early Flowering Mutant Research Articles

Functional Characterization of PoEP1 in Regulating the Flowering Stage of Tree Peony.

The tree peony, a traditional flower in China, has a short and concentrated flowering period, restricting the development of the tree peony industry. To explore the molecular mechanism of tree peony flowering-stage regulation, PoEP1, which regulated the flowering period, was identified and cloned based on the transcriptome and degradome data of the early-flowering mutant Paeonia ostii 'Fengdan' (MU) and Paeonia ostii 'Fengdan' (FD). Through bioinformatics analysis, expression pattern analysis, and transgene function verification, the role of PoEP1 in the regulation of tree peony flowering was explored. The open-reading frame of PoEP1 is 1161 bp, encoding 386 amino acids, containing two conserved domains. PoEP1 was homologous to the EP1 of other species. Subcellular localization results showed that the protein was localized in the cell wall and that PoEP1 expression was highest in the initial decay stage of the tree peony. The overexpression of PoEP1 in transgenic plants advanced and shortened the flowering time, indicating that PoEP1 overexpression promotes flowering and senescence and shorten the flowering time of plants. The results of this study provide a theoretical basis for exploring the role of PoEP1 in the regulation of tree peony flowering.

ZmGRAS46 Negatively Regulates Flowering Time in Arabidopsis thaliana

Flowering is an essential process in plant development, and there are six major flowering pathways: the photoperiodic pathway, gibberellin pathway, vernalization pathway, age pathway, autonomous pathway, and temperature pathway. In this study, we screened the transcriptome sequencing of early flowering mutants from the laboratory for the significantly differentially expressed ZmGRAS46, which belongs to the DELLA subfamily of the GRAS family. DELLA is involved in the gibberellin pathway to regulate plant flowering. However, it is not clear whether ZmGRAS46 is involved in the gibberellin pathway which regulates plant flowering; therefore, in this experiment, we investigated the regulatory role of this gene in Arabidopsis flowering by overexpressing ZmGRAS46. It was found that overexpression of ZmGRAS46 in Arabidopsis promotes the formation of rosette leaves and flower buds and delays flowering time in Arabidopsis, and experiments have shown that ZmGRAS46 represses the expression of FLOWERING LOCUS T (FT), SUPPRESSOR OF CONSTANS1 (SOC1), CONSTANS (CO), and LEAFY (LFY). Our results indicated the possibility that ZmGRAS46 represses flowering through the CO-FT-SOC1-mediated photoperiodic flowering pathway. The delayed flowering phenotype of overexpressing ZmGRAS46 Arabidopsis could be rescued by applying GA3. The experimental results indicate that ZmGRAS46 depends on the GA3 pathway to regulate flowering in Arabidopsis.

Identification of early flowering mutant gene in Phalaenopsis amabilis (L.) Blume for sgRNA construction in CRISPR/Cas9 genome editing system.

Phalaenopsis amabilis (L.) Blume commonly called Moth Orchid (Orchidaceae) is a natural orchid species designated as the National Flower of Indonesia for its beautiful flower shape and long-lasting flowering period. Basically, P. amabilis has a long vegetative phase that cause late flowering, about 2 to 3 years for flowering, hence a method to shorten vegetative period is desired. The latest technological approach that can be used to accelerate flowering of P. amabilis is the CRISPR/Cas9 genome editing method to inactivate the GAI (Gibberellic Acid Insensitive) gene as a mutant gene that can accelerate the regulation of FLOWERING TIME (FT) genes flowering biosynthesis pathway. The approach that needs to be taken is to silence the GAI gene with a knockout system which begins with identifying and characterizing the GAI target gene in the P. amabilis which will be used as a single guide RNA. CRISPR/Cas9 mediated knockout efficiency is highly dependent on the properties of the sgRNA used. SgRNA consists of a target sequence, determining its specificity performance. We executed phylogenetic clustering for the PaGAI protein with closely related orchid species such as Dendrobium capra, Dendrobium cultivars and Cymbidium sinensis. SWISS-Model as tool webserver for protein structure homology modeling. Results show that P. amabilis has a specific domain with the occurrence of point mutations in the two conservative domains. Therefore, a single guide RNA reconstruction needs to be implemented.

CRISPR/Cas9-mediated mutations of FANTASTIC FOUR gene family for creating early flowering mutants in tomato.

Flowering time is of great agricultural importance and the timing and extent of flowering usually determines yield and availability of flowers, fruits and seeds. Identification of genes determining flowering has important practical applications for tomato breeding. Here we demonstrate the roles of the FANTASTIC FOUR (FAF) gene family in regulating tomato flowering time. In this plant-specific gene family, SlFAF1/2a shows a constitutive expression pattern during the transition of the shoot apical meristem (SAM) from vegetative to reproductive growth and significantly influences flowering time. Overexpressing SlFAF1/2a causes earlier flowering compared with the transformations of other genes in the FAF family. SlFAF1/2c also positively regulates tomato flowering, although to a lesser extent. The other members of the SlFAF gene family, SlFAF1/2b, SlFAF3/4a and SlFAF3/4b, are negative regulators of tomato flowering and faf1/2b, faf3/4a and faf3/4b single mutants all display early flowering. We generated a series of early flowering mutants using the CRISPR/Cas9 editing system, and the faf1/2b faf3/4a faf3/4b triple mutant flowering earliest compared with other mutants. More importantly, these mutants show no adverse effect on yield. Our results have uncovered the role of the FAF gene family in regulating tomato flowering time and generated early flowering germplasms for molecular breeding.

EARLY FLOWERING is a dominant gain-of-function allele of FANTASTIC FOUR 1/2c that promotes early flowering in tomato.

Flowering time, an important factor in plant adaptability and genetic improvement, is regulated by various genes in tomato (Solanum lycopersicum). In this study, we characterized a tomato mutant, EARLY FLOWERING (EF), that developed flowers much earlier than its parental control. EF is a dominant gain-of-function allele with a T-DNA inserted 139 bp downstream of the stop codon of FANTASTIC FOUR 1/2c (FAF1/2c). The transcript of SlFAF1/2c was at elevated levels in the EF mutant. Overexpressing SlFAF1/2c in tomato plants phenocopied the early flowering trait of the EF mutant. Knocking out SlFAF1/2c in the EF mutant reverted the early flowering phenotype of the mutant to the normal flowering time of the wild-type tomato plants. SlFAF1/2c promoted the floral transition by shortening the vegetative phase rather than by reducing the number of leaves produced before the emergence of the first inflorescence. The COP9 signalosome subunit 5B (CSN5B) was shown to interact with FAF1/2c, and knocking out CSN5B led to an early flowering phenotype in tomato. Interestingly, FAF1/2c was found to reduce the accumulation of the CSN5B protein by reducing its protein stability. These findings imply that FAF1/2c regulates flowering time in tomato by reducing the accumulation and stability of CSN5B, which influences the expression of SINGLE FLOWER TRUSS (SFT), JOINTLESS (J) and UNIFLORA (UF). Thus, a new allele of SlFAF1/2c was discovered and found to regulate flowering time in tomato.

A 5.5-kb LTR-retrotransposon insertion inside phytochrome B gene (CsPHYB) results in long hypocotyl and early flowering in cucumber (Cucumis sativus L.)

The novel spontaneous long hypocotyl and early flowering (lhef) mutation in cucumber is due to a 5551-bp LTR-retrotransposon insertion in CsPHYB gene encoding PHYTOCHROME B, which plays a major role in regulating photomorphogenic hypocotyl growth and flowering. Hypocotyl length and flowering time are important for establishing high-quality seedlings in modern cucumber production, but little is known for the underlying molecular mechanisms of these two traits. In this study, a spontaneous cucumber long hypocotyl and early flowering mutant was identified and characterized. Based on multiple lines of evidence, we show that cucumber phytochrome B (CsPHYB) is the candidate gene for this mutation, and a 5551-bp LTR-retrotransposon insertion in the first exon of CsPHYB was responsible for the mutant phenotypes. Uniqueness of the mutant allele at CsPHYB was verified in 114 natural cucumber lines. Ectopic expression of the CsPHYB in Arabidopsis phyB mutant rescued the long hypocotyl and early flowering phenotype of phyB-9 mutant. The wild-type CsPHYB protein was localized on the membrane and cytoplasm under white light condition, whereas in the nucleus under red light, it is consistent with its roles as a red-light photoreceptor in Arabidopsis. However, the mutant csphyb protein was localized on the membrane and cytoplasm under both white and red-light conditions. Expression dynamics of CsPHYB and several cell elongation-related genes were positively correlated with hypocotyl elongation; the transcription levels of key positive and negative regulators for flowering time were also consistent with the anthesis dates in the mutant and wild-type plants. Yeast two hybrid and bimolecular fluorescence complementation assays identified physical interactions between CsPHYB and phytochrome interacting factor 3/4 (CsPIF3/4). These findings will provide new insights into the roles of the CsPHYB in cucumber hypocotyl growth and flowering time.

Leaf metabolomics and molecular networking of wild type and mutant genotypes of chia (Salvia hispanica L.)

Salvia hispanica L., commonly named Chia, is a food plant from Central America and Australia, producing seeds whose consumption has been increasing in the last decade. Several articles analysed the seeds metabolite content. However, few is known about Chia leaves. This work is the first report on the whole metabolite profile of chia leaves, determined by spectroscopic methods including NMR, GC-MS and LC-MS coupled with chemometrics analysis. Additionally, molecular networking has been applied to the LC-MS data to determine the flavonoid composition. Different chia sources were compared: one commercial (black) and three early flowering (G3, G8 and G17) mutant genotypes cultivated at two irrigation regimes (50 and 100%). Organic extracts were mainly composed by saturated and mono- and polyunsaturated fatty acids with palmitic being the most abundant followed by oleic and linolenic acids. Aqueous extracts contained glucose, galactose, and fructose as main sugars. Flavonoids were based on vitexin and orientin and their analogues. Chemical composition of early flowering genotypes was quite similar to commercial black chia with the exception of G8 showing significant differences in the polar phase. A generally highest content of omega-9 fatty acids has been found in the early flowering genotypes along with high content of nutraceuticals suggesting them as a potential source of raw materials for the food/feed industry.

Integrated analysis of miRNAome transcriptome and degradome reveals miRNA-target modules governing floral florescence development and senescence across early- and late-flowering genotypes in tree peony.

As a candidate national flower of China, tree peony has extremely high ornamental, medicinal and oil value. However, the short florescence and rarity of early-flowering and late-flowering varieties restrict further improvement of the economic value of tree peony. Specific miRNAs and their target genes engaged in tree peony floral florescence, development and senescence remain unknown. This report presents the integrated analysis of the miRNAome, transcriptome and degradome of tree peony petals collected from blooming, initial flowering, full blooming and decay stages in early-flowering variety Paeonia ostii 'Fengdan', an early-flowering mutant line of Paeonia ostii 'Fengdan' and late-flowering variety Paeonia suffruticosa 'Lianhe'. Transcriptome analysis revealed a transcript ('psu.G.00014095') which was annotated as a xyloglucan endotransglycosylase/hydrolase precursor XTH-25 and found to be differentially expressed across flower developmental stages in Paeonia ostii 'Fengdan' and Paeonia suffruticosa 'Lianhe'. The miRNA-mRNA modules were presented significant enrichment in various pathways such as plant hormone signal transduction, indole alkaloid biosynthesis, arachidonic acid metabolism, folate biosynthesis, fatty acid elongation, and the MAPK signaling pathway. Multiple miRNA-mRNA-TF modules demonstrated the potential functions of MYB-related, bHLH, Trihelix, NAC, GRAS and HD-ZIP TF families in floral florescence, development, and senescence of tree peony. Comparative spatio-temporal expression investigation of eight floral-favored miRNA-target modules suggested that transcript 'psu.T.00024044' and microRNA mtr-miR166g-5p are involved in the floral florescence, development and senescence associated agronomic traits of tree peony. The results might accelerate the understanding of the potential regulation mechanism in regards to floral florescence, development and abscission, and supply guidance for tree peony breeding of varieties with later and longer florescence characteristics.

Genome-scale investigation and identification of variations associated with early flowering based on whole genome resequencing and transcriptome integrated analysis in tree peony

Tree peony, a traditional ornamental flower and candidate national flower of China, has thousands of varieties with different flower colors and types after a long history of natural selection and artificial breeding. However, the short and concentrated florescence period and a shortage of early- and late-flowering varieties limit the enhancement of ornamental values. Single nucleotide polymorphisms (SNPs) and insertion/deletion mutations (InDels) are important factors for mutant genotypes generation. Whole genome resequencing and transcriptome integrated analysis was conducted using early-flowering genotype Paeonia ostii ‘Fengdan’ (FD), early-flowering mutant of Paeonia ostii ‘Fengdan’ (MU), and late-flowering genotype Paeonia suffruticosa ‘Lianhe’ (LH) to identify variants related to the early-flowering phenotype. Comparison of FD vs LH, MU vs LH, and MU vs FD revealed 1,488 SNPs, 1,261 InDels, and 1,912 differential expressed genes (DEGs) specific to early-flowering genotype FD and MU, respectively. Additionally, 329 DEGs affected by SNPs and InDels variants were differentially expressed in three tree peony genotypes at the full-blooming stage. Furthermore, 37 DEGs affected by SNP variants and 44 DEGs affected by InDel variants were identified in the early-flowering genotype FD and its corresponding MU at the full-blooming stage. Most of these genes were related to phenylpropanoid biosynthesis, flavonoid biosynthesis, plant hormone signal transduction, RNA degradation, and MAPK signaling pathway. Additionally, four SNPs were validated in FD, MU, and LH by Sanger sequencing. Expression profiles of 25 DEGs containing SNP/Indels associated with the early-flowering trait were verified by RT-qPCR. The abundant genetic variations identified in this study offer valuable genomic resources for ongoing functional and molecular breeding research for the early-flowering breeding of tree peony.

Characteristics of PoVIN3, a Key Gene of Vernalization Pathway, Affects Flowering Time.

The tree peony (Paeonia section Moutan DC.) is the candidate flower in China, with abundant germplasm resources and high ornamental value. However, the short and concentrated flowering period severely restricted the improvement of the economic value of tree peonies. Based on the full-length transcriptome database of tree peonies, the PoVIN3 (GenBank ID: OP341879), involved in the flowering regulation of tree peonies were identified and cloned for the first time. The PoVIN3 was also characterized by bioinformatics methods, quantitative real-time PCR (qRT-PCR), and the establishment of a transgenic system. The expression levels of PoVIN3 in seven different petals developmental stages were the highest at the initial flowering stage of the variant cultivar of Paeonia ostii 'Fengdan,' the initial decay stage of the normal flowering Paeonia ostii 'Fengdan,' and the half opening stage of the late flowering Paeonia suffruticosa 'Lianhe.' Tissue-specific expression analysis showed that the relative expression levels of PoVIN3 were the highest in sepals of both normal flowering Paeonia ostii 'Fengdan' and the late flowering Paeonia suffruticosa 'Lianhe,' and the highest expression was in stamens of early flowering mutant Paeonia ostii 'Fengdan.' In addition, the flowering time of pCAMBIA2300-PoVIN3 transgenic plants was significantly earlier than that of the wild-type, indicating that PoVIN3 could promote plant flowering. The results provide a theoretical basis for exploring the role of PoVIN3 in the regulation of flowering in tree peonies.

Induction of Chlorophyll and Morphological Mutations through Gamma Ray in Traditional Aromatic Cultivar Tulaipanja

The induced mutation of the traditional aromatic cultivar may provide useful alternative or complement to natural variation which may be used directly in mutation breeding or as a source of germ plasm in hybridization programme. Induced mutations irradiated through gamma ray in aromatic cultivar Tulaipanja were studied for chlorophyll and other morphological characters in the M2 generation. The frequency of chlorophyll mutations was high in higher doses. Among the chlorophyll mutants studied, albina was the most frequent, followed by alboxantha, alboviridis, xantha, viridis and striata. The mutation efficiency and the mutagenic effectiveness of the mutagen is more in the lower dose. The semi-dwarf mutants were more prevalent followed by dwarf and semitall-I mutants. The number of height mutants is much more in lower dose than that of higher dose. Among the morphological mutants, a number of mutants with broom stick leaf and few mutants with grassy leaf, rolled leaf, striped leaf were obtained. Besides these, delayed flowering mutants were obtained in low frequency in both the doses while the early flowering mutants were obtained only in the lower dose. The desirable dwarf or semi-dwarf early flowering mutants may be utilized directly or for recombination breeding, whereas the high yielding lines screened may be used directly as aromatic cultivar provided if the performance in the later generation is good.

First De novo whole genome sequencing and assembly of mutant Dendrobium hybrid cultivar 'Emma White'.

The Dendrobium hybrid cultivar 'Emma White' is an ornamental, successfully commercialised orchid. We used a gamma ray-induced early flowering mutant and the Illumina HiSeqX10 sequencing platform to generate the first draft de novo whole genome sequence and assembly. The draft sequence was 678,650,699bp in length, comprising 447,500 contigs with an N50 of 1423 and 33.48% GC content. Comparing 95,529 predicted genes against the Uniprot database revealed 60,741 potential genes governing molecular functions, biological processes and cellular components. We identified 216,232 simple sequence repeats and 138,856 microsatellite markers. Chromosome-level genome assembly of Dendrobium huoshanense was used to RagTag-scaffold available contigs of the mutant, revealing a total length of 687,254,899bp with an N50 of 2096. The longest final contiguous length was 18,000,059bp from 30,571bp. BUSCO genome completeness was 93.6%. This study is valuable for investigating the mechanisms of mutation, and developing Dendrobium hybrid cultivars using mutation breeding.

Development of Ethyl Methanesulfonate Mutant Edamame Soybean (Glycine max (L.) Merr.) Populations and Forward and Reverse Genetic Screening for Early-Flowering Mutants.

Induced mutation is a viable breeding strategy that is widely utilized in the development of elite plant varieties. We aimed to improve a variety of edamame by constructing novel mutant populations using the ethyl methanesulfonate in soybeans (Glycine max (L.) Merr.). In the M2 population, the flowering stage showed a considerable standard deviation compared to the wild type, confirming that the mutant populations had the expected DNA mutations. To identify the DNA mutations in the mutant populations, we used the targeting induced local lesions in genomes (TILLING) method, which is a reverse genetic method, to search for soybean flowering-related gene mutants. A total of 30 mutants from E1, E3, E4, and PhyA1 genes, which are known to be highly effective genes, or their homologous gene for flowering and maturation found in soybean quantitative trait locus analyses were isolated from our TILLING screening. Among these mutants, there were eleven nonsynonymous substitution mutants, one nonsense mutant, and two single nucleotide deletion mutants that could be expected to reduce or eliminate gene function. The e1, e3, and e4 mutants obtained in this study flowered considerably earlier than the wild type. In particular, the e1 mutant with a nonsynonymous substitution flowered approximately 1 month after sowing regardless of the sowing date, and its harvest date was approximately 1 month earlier than that of the wild type. Mutations identified using the TILLING method could not only be used as gel-based DNA markers with the same manipulation method, but the mutations could also be detected as DNA markers by the high-resolution melting method. These results indicate that mutations achieved without chromosome modification by crossbreeding are effective for early and practical improvement of superior varieties and that efficient selection of mutants by reverse genetics is an effective method for the identification of genetic modifications. The edamame mutant populations developed in this study are believed to possess various useful alleles which may be applicable in the search for mutations that lead to improved edamame yield and eating quality beyond the flowering stage.

Identification and Characterization of the MIKC-Type MADS-Box Gene Family in Brassica napus and Its Role in Floral Transition.

Increasing rapeseed yield has always been a primary goal of rapeseed research and breeding. However, flowering time is a prerequisite for stable rapeseed yield and determines its adaptability to ecological regions. MIKC-type MADS-box (MICK) genes are a class of transcription factors that are involved in various physiological and developmental processes in plants. To understand their role in floral transition-related pathways, a genome-wide screening was conducted with Brassica napus (B. napus), which revealed 172 members. Using previous data from a genome-wide association analysis of flowering traits, BnaSVP and BnaSEP1 were identified as candidate flowering genes. Therefore, we used the CRISPR/Cas9 system to verify the function of BnaSVP and BnaSEP1 in B. napus. T0 plants were edited efficiently at the BnaSVP and BnaSEP1 target sites to generate homozygous and heterozygous mutants with most mutations stably inherited by the next generation. Notably, the mutant only showed the early flowering phenotype when all homologous copies of BnaSVP were edited, indicating functional redundancy between homologous copies. However, no changes in flowering were observed in the BnaSEP1 mutant. Quantitative analysis of the pathway-related genes in the BnaSVP mutant revealed the upregulation of SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) and FLOWERING LOCUS T (FT) genes, which promoted early flowering in the mutant. In summary, our study created early flowering mutants, which provided valuable resources for early maturing breeding, and provided a new method for improving polyploid crops.

The APETALA2 homolog CaFFN regulates flowering time in pepper

Flowering time is an important agronomic trait that contributes to fitness in plants. However, the genetic basis of flowering time has not been extensively studied in pepper. To understand the genetics underlying flowering time, we constructed an F2 population by crossing a spontaneous early flowering mutant and a late-flowering pepper line. Using bulked segregant RNA-seq, a major locus controlling flowering time in this population was mapped to the end of chromosome 2. An APETALA2 (AP2) homolog (CaFFN) cosegregated with flowering time in 297 individuals of the F2 population. A comparison between the parents revealed a naturally occurring rare SNP (SNP2T > C) that resulted in the loss of a start codon in CaFFN in the early flowering mutant. Transgenic Nicotiana benthamiana plants with high CaFFN expression exhibited a delay in flowering time and floral patterning defects. On the other hand, pepper plants with CaFFN silencing flowered early. Therefore, the CaFFN gene acts as a flowering repressor in pepper. CaFFN may function as a transcriptional activator to activate the expression of CaAGL15 and miR156e and as a transcriptional repressor to repress the expression of CaAG, CaAP1, CaSEP3, CaSOC1, and miR172b based on a qRT-PCR assay. Direct activation of CaAGL15 by CaFFN was detected using yeast one-hybrid and dual-luciferase reporter assays, consistent with the hypothesis that CaFFN regulates flowering time. Moreover, the CaFFN gene association analysis revealed a significant association with flowering time in a natural pepper population, indicating that the CaFFN gene has a broad effect on flowering time in pepper. Finally, the phylogeny, evolutionary expansion and expression patterns of CaFFN/AP2 homologs were analyzed to provide valuable insight into CaFFN. This study increases our understanding of the involvement of CaFFN in controlling flowering time in pepper, thus making CaFFN a target gene for breeding early maturing pepper.

Early flowering, good grain quality mutants through gamma rays and EMS for enhancing per day productivity in rice (Oryza sativa L.)

Purpose Rice is the prominent crop of world and it frames the important component of food chain. But, long duration of highly preferred rice varieties requires more water and are prone to biotic and abiotic stresses. Short duration rice varieties serve as a promising alternative in such cases. Hence, the present study was taken up to induce early flowering mutants in popular rice variety Bapatla 2231 (BPT 2231), a long duration medium slender rice variety using gamma rays and Ethyl Methane Sulfonate (EMS) treatment combinations. Materials and methods Five hundred well filled and viable grains of optimum moisture content (12%) were subjected to irradiation with 8 doses comprising 100 Gy, 200 Gy, 300 Gy, 350 Gy, 400 Gy, 450 Gy, 500 Gy and 600 Gy treatments. For combination treatments, the irradiated seeds of gamma rays were further treated with 30 mM concentrations of Ethyl Methane sulfonate (EMS). The irradiated population were raised up to M3 generation for the identification of early maturing mutants. The isolated early maturing mutants in M3 generation were characterized for phenotypic, biochemical and grain qualities. Results The frequency of desirable early maturing mutants was observed more in combination treatments than the individual treatments of gamma rays. A total of 34 early maturing mutants (M) with variation for grain quality and biochemical traits were obtained. The mutants M-1, M-3, M-5, M-32, and M-34 were high yielding with fine grain type. The mutants M-4, M-17, M-18, M-19, M-20, and M-26 were high yielding with bold grain type. The mutants M-22, M-23, M-24, M-27, and M-28 were high in nutrient content. The early mutants matured 40−50 days earlier than control. Conclusion In this variety, the combination treatments showed higher mutation frequency, mutagenic effectiveness and efficiency than the gamma rays in induction of both chlorophyll and viable mutants. This study revealed that the combination treatments were more effective in inducing early mutants than the gamma ray treatments alone. The early maturing mutants can be released as variety after evaluation under different yield trials and the mutants with increased nutrient content and varied grain quality can be utilized in hybridization/heterosis breeding programs for rice crop improvement.

Characterization of a naturally occurring early-flowering rice mutant resulting from a novel variation in the Ghd7 locus

Rice is one of the main staple food crops and is consumed by more than half of the global population. Optimizing the flowering time may help to maximize grain production. In this study, we characterized a rice mutant, Zixiangnuo early-flowering mutant (ZXN-E). Compared with the wild-type variety, Zixiangnuo (ZXN), the mutant plants flowered earlier, were shorter and produced smaller panicles. Bulked segregant analysis revealed a 7-bp deletion in the first exon of the Grain number, plant height, and heading date 7 (Ghd7) locus. The allele with the 7-bp deletion was not detected in the rice varieties included in the 3 000 Rice Genomes Project. Thus, the 7-bp deletion identified in the naturally occurring mutant ZXN-E is a novel variation. Our findings suggest that ZXN-E may be a useful germplasm for breeding new rice varieties.

Molecular Analysis Associated with Early Flowering Mutant in Brassica napus

Flowering time in crops is an important agronomic character and is closely related to yield. However, the molecular mechanisms of flowering regulation in rapeseed remain elusive. In the current study, we report a rapeseed mutant with an early flowering (eaf) phenotype. The results of field observation showed that the flowering time of the mutant was about 18 days earlier than that of the wild type (WT, Zhongshuang 11), but there were no significant differences in other agronomic characters. Proteomic and transcriptomic analyses revealed that flowering and hormone-related proteins and genes were significantly differentially expressed between WT and eaf, indicating that the differential expression of flowering and hormone-related genes may be the main reason for early flowering in the mutant. Besides, the up-regulation of stress-related genes in the eaf mutant may be another possible reason of the mutant’s early flowering. Taken together, our findings provide a clue for dissecting the molecular mechanisms of early flowering in the mutant, and also potential materials for rapeseed molecular biology research.

Character Identification of an Early Flowering Mutant

The concept of gene-function-genetic trait was introduced to explore the effects of early flowering on the growth and development of maize at the jointing stage and to obtain early flowering mutants using ethyl methanesulfonate mutagenesis. First, we studied gene expression, phytohormones, and lignin content to explore the physiological peculiarities of the early flowering mutant. Then we analyzed the genetic features of the mutants during the jointing stage by measuring physiological and biochemical indices of drought tolerance. The results showed that the photosynthetic rate of the mutant was significantly higher than that of the control and the rate of accumulation of dry matter was rapid. In addition, the lignin content increased while drought resistance diminished. Therefore, we concluded that early flowering leads to faster overall growth and development.

Exogenous ascorbic acid delayed leaf senescence of early flowering rice mutant FTL10

FTL10 is an early flowering mutant of OsFTL10-suppressed transgenic rice (Oryza sativa L.) with premature senescence phenotype. Early leaf senescence can cause negative effects on rice yield, therefore delaying leaf senescence and prolonging the leaf functional stage is one of the important approaches to increase the rice yield. It is well known that ascorbic acid (AsA) is involved in regulating plant growth. To explore the effect of AsA on leaf senescence of FTL10, we treated rice leaves with 0.28 mM AsA. Results showed that total antioxidant capacity was higher and reactive oxygen species were lower in the AsA-treated group. The expression of senescence-associated genes was higher in the control group. Exogenous AsA can stabilize chlorophyll and Rubisco protein contents, delay leaf senescence, and maintain net photosynthetic rate (PN) of rice leaves. Our results suggest that exogenous application of AsA can delay leaf senescence, increase PN, and then increase the rice yield.