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Black shank, a devastating disease affecting tobacco production worldwide, is caused by Phytophthora nicotianae. However, few genes related to Phytophthora resistance have been reported in tobacco. Here, we identified NpPP2-B10, a gene strongly induced by P. nicotianae race 0, with a conserved F-box motif and Nictaba (tobacco lectin) domain, in the highly resistant tobacco species Nicotiana plumbaginifolia. NpPP2-B10 is a typical F-box-Nictaba gene. When it was transferred into the black shank-susceptible tobacco cultivar ‘Honghua Dajinyuan’, it was found to promote resistance to black shank disease. NpPP2-B10 was induced by salicylic acid, and some resistance-related genes (NtPR1, NtPR2, NtCHN50, and NtPAL) and resistance-related enzymes (catalase and peroxidase) were significantly upregulated in the overexpression lines after infection with P. nicotianae. Furthermore, we showed that NpPP2-B10 actively regulated the tobacco seed germination rate, growth rate, and plant height. The erythrocyte coagulation test of purified NpPP2-B10 protein showed that NpPP2-B10 had plant lectin activity, and the lectin content in the overexpression lines was significantly higher than that in the WT, which could lead to accelerated growth and improved resistance of tobacco. SKP1 is an adaptor protein of the E3 ubiquitin ligase SKP1, Cullin, F-box (SCF) complex. We demonstrated that NpPP2-B10 could interact with the NpSKP1-1A gene in vivo and in vitro through yeast two-hybrid (Y2H) and bimolecular fluorescence complementation (BiFC), indicating that NpPP2-B10 likely participates in the plant immune response by mediating the ubiquitin protease pathway. In conclusion, our study provides some important insights concerning NpPP2-B10-mediated regulation of tobacco growth and resistance.

Wild loquats (Eriobotrya japonica Lindl.) provide remarkable genetic resources for studying domestication and breeding improved varieties. Herein, we generate the first high-quality chromosome-level genome assembly of wild loquat, with 45 791 predicted protein-coding genes. Analysis of comparative genomics indicated that loquat shares a common ancestor with apple and pear, and a recent whole-genome duplication event occurred in loquat prior to its divergence. Genome resequencing showed that the loquat germplasms can be distinctly classified into wild and cultivated groups, and the commercial cultivars have experienced allelic admixture. Compared with cultivated loquats, the wild loquat genome showed very few selected genomic regions and had higher levels of genetic diversity. However, whole-genome scans of selective sweeps were mainly related to fruit quality, size, and flesh color during the domestication process. Large-scale transcriptome and metabolome analyses were further performed to identify differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) in wild and cultivated loquats at various fruit development stages. Unlike those in wild loquat, the key DEGs and DAMs involved in carbohydrate metabolism, plant hormone signal transduction, flavonoid biosynthesis, and carotenoid biosynthesis were significantly regulated in cultivated loquats during fruit development. These high-quality reference genome, resequencing, and large-scale transcriptome/metabolome data provide valuable resources for elucidating fruit domestication and molecular breeding in loquat.

Huanglongbing (HLB), presumably caused by Candidatus Liberibacter asiaticus (CaLas), is a devastating citrus disease worldwide. While all citrus are affected by HLB, some cultivars display greater tolerance; however, the underlying mechanisms are not fully understood. Here, volatile changes in HLB-tolerant LB8-9 Sugar Belle (SB) and HLB-sensitive Murcott mandarins after CaLas infection were comprehensively compared to determine if specific volatiles are associated with HLB responses and to discern the underlying tolerance mechanisms. These cultivars emitted qualitatively and quantitatively different volatiles in response to HLB induced by artificial graft or natural psyllid inoculation. Increasing amounts of total volatiles and de novo-synthesized new volatiles were two key responses to HLB of both cultivars. Markers potentially associated with HLB and host susceptibility were identified. Terpenoid biosynthetic pathway, green leaf volatile, and thymol metabolic pathways responsive to CaLas infection were dramatically altered. SB mandarin allows simultaneous defense and growth, contributing to its greater HLB tolerance.

Previous studies indicated that extensive genetic variations could be generated due to polyploidy, which is considered to be closely associated with the manifestation of polyploid heterosis. Our previous studies confirmed that triploid loquats demonstrated significant heterosis, other than the ploidy effect, but the underlying mechanisms are largely unknown. This study aimed to overcome the narrow genetic distance of loquats, increase the genetic variation level of triploid loquats, and systematically illuminate the heterosis mechanisms of triploid loquats derived from two cross combinations. Here, inter-simple sequence repeats (ISSRs) and simple sequence repeats (SSRs) were adopted for evaluating the genetic diversity, and transcriptome sequencing (RNA-Seq) was performed to investigate gene expression as well as pathway changes in the triploids. We found that extensive genetic variations were produced during the formation of triploid loquats. The polymorphism ratios of ISSRs and SSRs were 43.75% and 19.32%, respectively, and almost all their markers had a PIC value higher than 0.5, suggesting that both ISSRs and SSRs could work well in loquat assisted breeding. Furthermore, our results revealed that by broadening the genetic distance between the parents, genetic variations in triploids could be promoted. Additionally, RNA-Seq results suggested that numerous genes differentially expressed between the triploids and parents were screened out. Moreover, KEGG analyses revealed that “photosynthetic efficiency” and “glyco-metabolism” were significantly changed in triploid loquats compared with the parents, which was consistent with the results of physiological indicator analyses, leaf micro-structure observations, and qRT-PCR validation. Collectively, our results suggested that extensive genetic variations occurred in the triploids and that the changes in the “photosynthetic efficiency” as well as “glyco-metabolism” of triploids might have further resulted in heterosis manifestation in the triploid loquats.

Guijie Li,1,* Yujiao Cheng,1,* Tenghui Zhang,2 Yingzhuo Li,3 Leng Han,1 Guolu Liang4 1Citrus Research Institute, Southwest University, Chongqing, People’s Republic of China; 2Chengdu Centre Testing International Group Co., Ltd., Chengdu, People’s Republic of China; 3Chongqing Beibei Agricultural and Rural Committee, Chongqing, People’s Republic of China; 4College of Horticulture and Landscape Architecture, Southwest University, Chongqing, People’s Republic of China*These authors contributed equally to this workCorrespondence: Leng HanCitrus Research Institute, Southwest University, No. 15 Ganjucun, Xiema, Beibei District, Chongqing, 400712, People’s Republic of ChinaEmail hanleng5@cric.cnGuolu LiangCollege of Horticulture and Landscape Architecture, Southwest University, No. 2 Tiansheng Road, Beibei District, Chongqing, 400715, People’s Republic of ChinaEmail lianggl@swu.edu.cnPurpose: Citrus essential oils are widely used for aromatherapy and the alternative treatment of chronic diseases. Beyond the aroma substances, they are known to contain bioactive nonvolatile components; however, little knowledge has been gained about nonvolatiles in the essential oil of pomelo (Citrus grandis Osbeck), the largest citrus fruit. The purpose of this study was to analyze the nonvolatile oxygenated heterocyclic compounds (OHCs) of pomelo essential oils and evaluate their in vitro antioxidant activities for further development.Methods: Cold-pressed essential oil (CPEO) and distilled essential oil (DEO) were obtained from the peel of the Liangping pomelo cultivar. High-performance liquid chromatography (HPLC) coupled with a photodiode array and fluorescence detection method was developed to identify and quantify the OHCs of the two essential oils. Ferric reducing antioxidant power and 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2-phenyl-4,4,5,5-tetramethyl-imidazoline-1-oxyl 3-oxide (PTIO) radical scavenging assays were used to determine the antioxidative capabilities.Results: Thirteen OHCs were identified in CPEO. Coumarins such as meranzin (2.0 mmol L− 1) and furanocoumarins such as isoimperatorin (1.3 mmol L− 1) composed the majority of nonvolatiles in CPEO. These OHCs were characterized by high proportion (58%) of side chain epoxides. Five OHCs, namely, auraptenol, 6ʹ,7ʹ-dihydroxybergamottin (6ʹ,7ʹ-DHB), imperatorin, isoimperatorin and 8-geranyloxypsoralen were first identified in pomelo CPEO. Eight OHCs were detected at trace amounts in pomelo DEO. Antioxidant assays showed that CPEO was multiple times more potent than DEO regarding the total reducing power and radical scavenging capacity. Clearance of PTIO, a stable reactive oxygen species, followed slow kinetics.Conclusion: Coumarins and furanocoumarins, two families of OHCs, constituted most of the nonvolatile components in CPEO. The nonvolatiles contributed significantly to the in vitro antioxidant activity of CPEO. Pomelo CPEO showed good prospects as a potential long-lasting natural antioxidant.Keywords: volatile oil, cold expression, hydrodistillation, psoralen, epoxides, fluorescence, radical scavenging, pummelo, Citrus maxima

Background Aneuploidy, a condition caused by an imbalance between the relative dosages of chromosomes, generally produces a novel phenotype specific to the molecular karyotype. Few techniques are currently available for detecting the molecular karyotypes of aneuploids in plants. Results Based on this imbalance in chromosome dosage, a new approach (referred to as ‘SSR-qPCR’) combining simple sequence repeat (SSR) markers and quantitative real-time PCR (qPCR) has been developed and utilized to detect some common aneuploids irrespective of heterozygosity. We screened 17 specific SSR markers covering all loquat linkage groups and redesigned 6 pairs of primers for SSR markers that can detect loquat chromosome aneuploidies. The SSR-qPCR detection results obtained for hybrid progeny and open-pollination progeny of triploid loquat showed diagnostic accuracies of 88.9% and 62.5%, respectively, compared with the chromosome preparation results. Conclusion SSR-qPCR can detect loquat aneuploids and be used to construct the entire molecular karyotypes of aneuploid individuals. Therefore, this method offers a novel alternative for the detection of chromosome aneuploidies.

Loquat (Eriobotrya japonica Lindl.) is an evergreen fruit tree of Chinese origin, and its autumn–winter flowering and fruiting growth habit means that its fruit development is susceptible to low-temperature stress. In a previous study, the triploid loquat (B431 × GZ23) has been identified with high photosynthetic efficiency and strong resistance under low-temperature stress. Analysis of transcriptomic and lipidomic data revealed that the fatty acid desaturase gene EjFAD8 was closely associated with low temperatures. Phenotypic observations and measurements of physiological indicators in Arabidopsis showed that overexpressing-EjFAD8 transgenic plants were significantly more tolerant to low temperatures compared to the wild-type. Heterologous overexpression of EjFAD8 enhanced some lipid metabolism genes in Arabidopsis, and the unsaturation of lipids was increased, especially for SQDG (16:0/18:1; 16:0/18:3), thereby improving the cold tolerance of transgenic lines. The expression of ICE-CBF-COR genes were further analyzed so that the relationship between fatty acid desaturase and the ICE-CBF-COR pathway can be clarified. These results revealed the important role of EjFAD8 under low-temperature stress in triploid loquat, the increase expression of FAD8 in loquat under low temperatures lead to desaturation of fatty acids. On the one hand, overexpression of EjFAD8 in Arabidopsis increased the expression of ICE-CBF-COR genes in response to low temperatures. On the other hand, upregulation of EjFAD8 at low temperatures increased fatty acid desaturation of SQDG to maintain the stability of photosynthesis under low temperatures. This study not only indicates that the EjFAD8 gene plays an important role in loquat under low temperatures, but also provides a theoretical basis for future molecular breeding of loquat for cold resistance.

Volatiles are determinants of fruit aroma and flavor characteristics and also provide valuable information for lemon as ingredient for the food and drinks industry. Volatiles in 'Eureka' lemon and 'Xiangshui' lemon pulps from 130 to 186 days after flowering were enriched by headspace-solid-phase microextraction (HS-SPME), and analyzed by gas chromatography-mass spectrometry (GC-MS).Seventy-seven volatiles of two lemon cultivars at the different ripening stages were identified and divided into six categories. Varieties and ripening stages had significant effects on individual volatiles in each category. The proportion of monoterpenes was found to be higher in 'Eureka' lemon, while 'Xiangshui' lemon had a higher proportion of sesquiterpenes, aldehydes and alcohols. The proportion of monoterpene fluctuation decreased during fruit ripening, while fluctuation of sesquiterpenes, alcohols, aldehydes and esters increased. Among the hydrocarbons, monoterpenes decreased their relative abundance from 91.67% to 81.04% in 'Eureka' lemon, and from 83.01% to 60.04% in 'Xiangshui' lemon; conversely, sesquiterpenes increased from 0.73% to 2.89% in 'Eureka' lemon, and from 3.21% to 8.48% in 'Xiangshui' lemon. Among the oxygenated volatiles, the proportions of alcohols, aldehydes and esters were higher at 186 days after flowering in both two cultivars.The volatile organic compounds during fruit ripening of lemon varieties with different resistance were elucidated. The proportion of oxygenated volatiles increased during fruit ripening, and disease-resistant varieties had a higher proportion. These results provided important theoretical support for the utilization of lemon fruits and the innovation of disease-resistant germplasm resources. © 2021 Society of Chemical Industry.

Plants with alien genomic components (alien chromosomes / chromosomal fragments / genes) are important materials for genomic research and crop improvement. To date, four strategies based on trait observation, chromosome analysis, specific proteins, and DNA sequences have been developed for the identification of alien genomic components. Among them, DNA sequence-based molecular markers are mainly used to identify alien genomic components. This review summarized several molecular markers for identification of alien genomic components in wheat, cabbage and other important crops. We also compared the characteristics of nine common molecular markers, such as simple sequence repeat (SSR), insertion-deletion (InDel) and single nucleotide polymorphism (SNP). In general, the accuracy of using a combination of different identification methods is higher than using a single identification method. We analyzed the application of different combination of identification methods, and provided the best combination for wheat, brassica and other crops. High-throughput detection can be easily achieved by using the new generation molecular markers such as InDel and SNP, which can be used to determine the precise localization of alien introgression genes. To increase the identification efficiency, other new identification methods, such as microarray comparative genomic hybridization (array-CGH) and suppression subtractive hybridization (SSH), may also be included.

Background Ploidy manipulation is effective in seedless loquat breeding, in which flesh color is a key agronomic and economic trait. Few techniques are currently available for detecting the genotypes of polyploids in plants, but this ability is essential for most genetic research and molecular breeding. Results We developed a system for genotyping by quantitative PCR (qPCR) that allowed flesh color genotyping in multiple tetraploid and triploid loquat varieties (lines). The analysis of 13 different ratios of DNA mixtures between two homozygous diploids (AA and aa) showed that the proportion of allele A has a high correlation (R2 = 0.9992) with parameter b [b = a1/(a1 + a2)], which is derived from the two normalized allele signals (a1 and a2) provided by qPCR. Cluster analysis and variance analysis from simulating triploid and tetraploid hybrids provided completely correct allelic configurations. Four genotypes (AAA, AAa, Aaa, aaa) were found in triploid loquats, and four (AAAA, AAAa, AAaa, Aaaa; absence of aaaa homozygotes) were found in tetraploid loquats. DNA markers analysis showed that the segregation of flesh color in all F1 hybrids conformed to Mendel's law. When tetraploid B431 was the female parent, more white-fleshed triploids occurred among the progeny. Conclusions qPCR can detect the flesh color genotypes of loquat polyploids and provides an alternative method for analyzing polyploid genotype and breeding, dose effects and allele-specific expression.

Triploid loquat (2n = 3x = 51) has stronger growth vigor and larger leaves, flowers, and fruit compared with its diploid parental plant (2n = 2x = 34), but the effects of triploidization on the contents of flavonoids and phenolics in leaves and flowers, which are the most important antioxidant compounds for pharmacological applications, have not been reported. In this report, 58 triploid loquat genotypes and seven corresponding diploid parental cultivars were used to evaluate the effects of triploidization on the contents of total flavonoids and phenolics and the antioxidant activities of leaves and flower buds. The results showed that the contents of total flavonoids and phenolics and their corresponding antioxidant activities were higher in most of the triploid loquat genotypes than their diploid parents. The antioxidant activities of leaves and flower buds were significantly correlated with the total flavonoids and phenolics contents in both diploid loquat and triploid loquat. It could be inferred that triploidization could increase the contents of flavonoids and phenolics in leaves and flower buds of loquat. Notably, the contents of total flavonoids and phenolics of leaves in triploid genotype ‘H3/24’ were the highest, reaching 212.00 mg rutin equivalent (RE)/g DW and 93.06 mg gallic acid equivalents (GAE)/g DW, respectively, which were significantly higher than those previously reported. Such a valuable trait may be stacked with other triploid traits that are already established, such as larger vegetative organs and better tolerance to various stresses, as a feasible strategy for breeding loquat cultivars with high pharmaceutical potency.

Previous observation in our laboratory found that growth vigor of triploid loquats was greater than in diploids and tetraploids despite the existence of ploidy effects, but it is still lack of systematic studies (i.e., histological, physiological). In addition, the underlying mechanisms of the heterosis are largely unknown due to the fact that the origin and genetic pedigree are unclear. In this study, we sought to further confirm the heterosis of triploid loquat systematically and elucidate the relationship between heterosis and circadian rhythms in triploid loquat by using trees with defined genetic pedigree. Two sets of triploid loquat were generated by crossing a tetraploid loquat with two wild diploid loquats (GC-1 and GC-23 from Guizhou Province, China (GC). Heterosis was systematically evaluated by comparing various histological, physiological and disease resistance metrics to the parental types. The results showed that the two sets of triploid loquat generated here exhibited pronounced degree of heterosis in all three categories and this was consistent with our previous observation. To determine if heterosis was also affected by the circadian clock, three circadian clock genes (EjLHY, EjTOC1 and EjGIGANTEA) were screened out, and the expression level of three circadian clock genes and 7 clock output genes were also analyzed. Phylogenetic analysis of three circadian genes showed that they were highly homologous with any other species in Rosaceae family. QRT-PCR analysis of the three circadian clock genes showed that all of them expressed above-high-parent (AHP) in almost all of the hybrids. Surprisingly, the 7 clock output genes also demonstrated AHP expression pattern in almost all the hybrids, and this was consistent with the results from our histological, physiological and disease resistance analysis. Our results indicated that triploid loquat exhibits pronounced heterosis compared with the diploid and tetraploid loquats, and the triploid loquat heterosis may be related to the altered amplitude of circadian rhythms.

The WRKY gene family, which is one of the largest transcription factor (TF) families, plays an important role in numerous aspects of plant growth and development, especially in various stress responses. However, the functional roles of the WRKY gene family in loquat are relatively unknown. In this study, a novel WRKY gene, EjWRKY17, was characterized from Eriobotrya japonica, which was significantly upregulated in leaves by melatonin treatment during drought stress. The EjWRKY17 protein, belonging to group II of the WRKY family, was localized in the nucleus. The results indicated that overexpression of EjWRKY17 increased cotyledon greening and root elongation in transgenic Arabidopsis lines under abscisic acid (ABA) treatment. Meanwhile, overexpression of EjWRKY17 led to enhanced drought tolerance in transgenic lines, which was supported by the lower water loss, limited electrolyte leakage, and lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Further investigations showed that overexpression of EjWRKY17 promoted ABA-mediated stomatal closure and remarkably up-regulated ABA biosynthesis and stress-related gene expression in transgenic lines under drought stress. Overall, our findings reveal that EjWRKY17 possibly acts as a positive regulator in ABA-regulated drought tolerance.

The WRKY gene family, which is one of the largest transcription factor (TF) families, plays an important role in numerous aspects of plant growth and development, especially in various stress responses. However, the functional roles of the WRKY gene family in loquat are relatively unknown. In this study, a novel WRKY gene, EjWRKY17, was characterized from Eriobotrya japonica, which was significantly upregulated in leaves by melatonin treatment during drought stress. The EjWRKY17 protein, belonging to group II of the WRKY family, was localized in the nucleus. The results indicated that overexpression of EjWRKY17 increased cotyledon greening and root elongation in transgenic Arabidopsis lines under abscisic acid (ABA) treatment. Meanwhile, overexpression of EjWRKY17 led to enhanced drought tolerance in transgenic lines, which was supported by the lower water loss, limited electrolyte leakage, and lower levels of reactive oxygen species (ROS) and malondialdehyde (MDA). Further investigations showed that overexpression of EjWRKY17 promoted ABA-mediated stomatal closure and remarkably up-regulated ABA biosynthesis and stress-related gene expression in transgenic lines under drought stress. Overall, our findings reveal that EjWRKY17 possibly acts as a positive regulator in ABA-regulated drought tolerance.

Loquat (

Additional file 6: Table S1. Primers used for reverse transcription quantitative PCR (RT-qPCR). Table S2. Color parameters in GB_Pe, WF_Pe, and YF_Pe. Table S3. Differentially-accumulated anthocyanins in WF_Pe vs GB_Pe, YF_Pe vs GB_Pe and/or YF_Pe vs WF_Pe. Table S4. Throughput and quality of L. japonica transcriptome data. Table S5. Summary of L. japonica de novo transcriptome assembly. Table S6. Mapping results of L. japonica unigenes to various databases.

Although several results have been obtained in triploid loquat heterosis (i.e., leaf size of triploid loquat) studies in the past years, the underlying mechanisms of the heterosis are still largely unknown, especially the regulation effects of one specific gene on the corresponding morphology heterosis. In this study, we sought to further illustrate the regulatory mechanisms of one specific gene on the leaf size heterosis of triploid loquats. A leaf size development-related gene (EjGIF1) and its promoter were successfully cloned. Ectopic expression ofEjGIF1inArabidopsisshowed that the leaf size of transgenic plantlets was larger than that of WTs, and the transgenic plantlets had more leaves than WTs. Quantitative Reverse Transcription PCR (qRT-PCR) showed that the expression level ofEjGIF1showed an AHP expression pattern in most of the hybrids, and this was consistent with our previous phenotype observations. Structure analysis ofEjGIF1promoter showed that there were significantly more light-responsive elements than other elements. To further ascertain the regulatory mechanisms ofEjGIF1on triploid loquat heterosis, the methylation levels ofEjGIF1promoter in different ploidy loquats were analyzed by using bisulfite sequencing. Surprisingly, the total methylation levels ofEjGIF1promoter in triploid showed a decreasing trend compared with the mid-parent value (MPV), and this was also consistent with the qRT-PCR results ofEjGIF1. Taken together, our results suggested thatEjGIF1played an important role in promoting leaf size development of loquat, and demethylation ofEjGIF1promoter in triploid loquats causedEjGIF1to exhibit over-dominance expression pattern and then further to promote leaf heterosis formation. In conclusion,EjGIF1played an important role in the formation of triploid loquat leaf size heterosis.

BackgroundPlants have remarkable diversity in petal colour through the biosynthesis and accumulation of various pigments. To better understand the mechanisms regulating petal pigmentation inLonicera japonica, we used multiple approaches to investigate the changes in carotenoids, anthocyanins, endogenous hormones and gene expression dynamics during petal colour transitions, i.e., green bud petals (GB_Pe), white flower petals (WF_Pe) and yellow flower petals (YF_Pe).ResultsMetabolome analysis showed that YF_Pe contained a much higher content of carotenoids than GB_Pe and WF_Pe, with α-carotene, zeaxanthin, violaxanthin and γ-carotene identified as the major carotenoid compounds in YF_Pe. Comparative transcriptome analysis revealed that the key differentially expressed genes (DEGs) involved in carotenoid biosynthesis, such asphytoene synthase,phytoene desaturaseandζ-carotene desaturase, were significantly upregulated in YF_Pe. The results indicated that upregulated carotenoid concentrations and carotenoid biosynthesis-related genes predominantly promote colour transition. Meanwhile, two anthocyanins (pelargonidin and cyanidin) were significantly increased in YF_Pe, and the expression level of ananthocyanidin synthasegene was significantly upregulated, suggesting that anthocyanins may contribute to vivid yellow colour in YF_Pe. Furthermore, analyses of changes in indoleacetic acid, zeatin riboside, gibberellic acid, brassinosteroid (BR), methyl jasmonate and abscisic acid (ABA) levels indicated that colour transitions are regulated by endogenous hormones. The DEGs involved in the auxin, cytokinin, gibberellin, BR, jasmonic acid and ABA signalling pathways were enriched and associated with petal colour transitions.ConclusionOur results provide global insight into the pigment accumulation and the regulatory mechanisms underlying petal colour transitions during the flower development process inL. japonica.

A triploid was obtained from loquat (Eriobotrya japonica cv. ‘Ninghaibai’) (2n = 2x = 34) × bronze loquat (Eriobotrya deflexa, 2n = 2x = 34). The leaves of the triploid had no petioles, similar to the female parent, and its stipules were obvious and dark green, similar to those of the male parent. There were signals of male parent DNA on ten chromosomes of the triploid according to genomic in situ hybridization (GISH) when DNA of bronze loquat was used as probe; the diploid hybrid also had ten chromosomes with signals of male parent DNA. All five selected simple sequence repeats (SSR) primers amplified specific loci of both parents. In particular, primer CH04c06 amplified both specific loci of the female parent but only amplified one specific locus of the male parent. The results of GISH and SSR marker analysis indicated that the 2n gamete which formed the triploid was from first-division restitution or pre-meiotic doubling of the female parent cv. ‘Ninghaibai’. The allotriploid can be used to breed seedless loquat germplasm with different flowering times from loquat directly or to breed new germplasm containing a few genomic components from bronze loquat, such as monosomic alien addition lines, monosomic alien substitution lines, and introgression lines. This is the first report of allotriploid in the genus Eriobotrya and it will provide a basis for breeding new loquat germplasms by distant hybridization and provide an ideal material for breeding new loquat varieties with different flowering time or time to maturity.

Background: Plants have remarkable diversity in petal color through the biosynthesis and accumulation of various pigments. To better understand the mechanisms regulating the petal pigmentation, we used multiple approaches to investigate the changes of carotenoids, gene expression dynamics and endogenous hormones in Lonicera japonica during petal color transitions, i.e. green bud petal (GB_Pe), white flower petal (WF_Pe) and yellow flower petal (YF_Pe). Results: Metabolome analysis showed that YF_Pe had a much higher content of carotenoids than white petals, with violaxanthin identified as the major carotenoid compound in YF_Pe. Comparative transcriptome analysis revealed that the key differentially expressed genes (DEGs) involved in carotenoid biosynthesis, such as phytoene synthase, phytoene desaturase and ζ-carotene desaturase, were significantly up-regulated in YF_Pe. However, expression levels of the carotenoid degradation-related genes, including abscisic-aldehyde oxidase 3 and carotenoid cleavage dioxygenase 4, were significantly down-regulated in YF_Pe. The results indicated that upregulated carotenoids concentrations and carotenoids biosynthesis-related genes promote the color transition. Furthermore, enrichment analysis of DEGs was mainly associated with the metabolic pathways of hormone signal transduction during petal color transitions. The DEGs were mainly involved in the auxin, cytokinin, gibberellin (GA), brassinosteroid (BR), jasmonic acid and abscisic acid (ABA) signal pathways. Accordingly, analyses of changes in indoleacetic acid, zeatin riboside, GA, BR, methyl jasmonate and ABA levels indicated that the color transitions are regulated by endogenous hormones. Conclusion: Our results provide new insight into the regulatory mechanisms underlying the petal color transitions during flower development process in L. japonica.

Flower development is a vital developmental process in the life cycle of woody perennials, especially fruit trees. Herein, we used transcriptomic, proteomic, and hormone analyses to investigate the key candidate genes/proteins in loquat (Eriobotrya japonica) at the stages of flower bud differentiation (FBD), floral bud elongation (FBE), and floral anthesis (FA). Comparative transcriptome analysis showed that differentially expressed genes (DEGs) were mainly enriched in metabolic pathways of hormone signal transduction and starch and sucrose metabolism. Importantly, the DEGs of hormone signal transduction were significantly involved in the signaling pathways of auxin, gibberellins (GAs), cytokinin, ethylene, abscisic acid (ABA), jasmonic acid, and salicylic acid. Meanwhile, key floral integrator genes FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS1 (SOC1) and floral meristem identity genes SQUAMOSA PROMOTER BINDING LIKE (SPL), LEAFY (LFY), APETALA1 (AP1), and AP2 were significantly upregulated at the FBD stage. However, key floral organ identity genes AGAMOUS (AG), AP3, and PISTILLATA (PI) were significantly upregulated at the stages of FBE and FA. Furthermore, transcription factors (TFs) such as bHLH (basic helix-loop-helix), NAC (no apical meristem (NAM), Arabidopsis transcription activation factor (ATAF1/2) and cup-shaped cotyledon (CUC2)), MYB_related (myeloblastosis_related), ERF (ethylene response factor), and C2H2 (cysteine-2/histidine-2) were also significantly differentially expressed. Accordingly, comparative proteomic analysis of differentially accumulated proteins (DAPs) and combined enrichment of DEGs and DAPs showed that starch and sucrose metabolism was also significantly enriched. Concentrations of GA3 and zeatin were high before the FA stage, but ABA concentration remained high at the FA stage. Our results provide abundant sequence resources for clarifying the underlying mechanisms of the flower development in loquat.