Eriobotrya Japonica Lindl Research Articles

Selection and Validation of Reference Genes for Normalization of RT-qPCR Analysis in Developing or Abiotic-Stressed Tissues of Loquat (Eriobotrya japonica)

Loquat (Eriobotrya japonica Lindl.) is a subtropical evergreen fruit tree that produces fruits with abundant nutrients and medicinal components. Confirming suitable reference genes for a set of loquat samples before qRT-PCR experiments is essential for the accurate quantification of gene expression. In this study, eight candidate reference genes were selected from our previously published RNA-seq data, and primers for each candidate reference gene were designed and evaluated. The Cq values of the candidate reference genes were calculated by RT-qPCR in 31 different loquat samples, including 12 subgroups of developing or abiotic-stressed tissues. Different combinations of stable reference genes were screened according to a comprehensive rank, which was synthesized from the results of four algorithms, including the geNorm, NormFinder, BestKeeper and ΔCt methods. The screened reference genes were verified by normalizing EjLGA1 in each subgroup. The obtained suitable combinations of reference genes for accurate normalization were GAPDH, EF1α and ACT for floral development; GAPDH, UBCE and ACT for fruit setting; EF1α, GAPDH and eIF2B for fruit ripening; ACT, EF1α and UBCE for leaves under heat stress; eIF2B, UBCE and EF1α for leaves under freezing stress; EF1α, TUA and UBCE for leaves under salt stress; ACT, EF1α and eIF2B for immature pulp under freezing stress; ACT, UBCE and eIF2B for immature seeds under freezing stress; EF1α, eIF2B and UBCE for both immature pulp and seeds under freezing stress; UBCE, TUB and TUA for red-fleshed fruits under cold-storage stress; eIF2B, RPS3 and TUB for white-fleshed fruits under cold-storage stress; and eIF2B, UBCE and RPS3 for both red- and white-fleshed fruits under cold-storage stress. This study obtained different combinations of stable reference genes for accurate normalization in twelve subgroups of developing or abiotic-stressed tissues in loquat. To our knowledge, this is the first report to obtain stable reference genes for normalizing gene expression of abiotic-stressed tissues in E. japonica. The use of the three most stable reference genes could increase the reliability of future quantification experiments.

Genome assembly of wild loquat (Eriobotrya japonica) and resequencing provide new insights into the genomic evolution and fruit domestication in loquat.

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.

Genome-Wide Analysis of SAUR Gene Family Identifies a Candidate Associated with Fruit Size in Loquat (Eriobotrya japonica Lindl.).

Fruit size is an important fruit quality trait that influences the production and commodity values of loquats (Eriobotrya japonica Lindl.). The Small Auxin Upregulated RNA (SAUR) gene family has proven to play a vital role in the fruit development of many plant species. However, it has not been comprehensively studied in a genome-wide manner in loquats, and its role in regulating fruit size remains unknown. In this study, we identified 95 EjSAUR genes in the loquat genome. Tandem duplication and segmental duplication contributed to the expansion of this gene family in loquats. Phylogenetic analysis grouped the SAURs from Arabidopsis, rice, and loquat into nine clusters. By analyzing the transcriptome profiles in different tissues and at different fruit developmental stages and comparing two sister lines with contrasting fruit sizes, as well as by functional predictions, a candidate gene (EjSAUR22) highly expressed in expanding fruits was selected for further functional investigation. A combination of Indoleacetic acid (IAA) treatment and virus-induced gene silencing revealed that EjSAUR22 was not only responsive to auxin, but also played a role in regulating cell size and fruit expansion. The findings from our study provide a solid foundation for understanding the molecular mechanisms controlling fruit size in loquats, and also provide potential targets for manipulation of fruit size to accelerate loquat breeding.

Sugar and acid profile of loquat (Eriobotrya japonica Lindl.), enzymes assay and expression profiling of their metabolism-related genes as influenced by exogenously applied boron.

Soluble sugars and organic acids are the most abundant components in ripe fruits, and they play critical roles in the development of fruit flavor and taste. Some loquat cultivars have high acid content which seriously affect the quality of fruit and reduce the value of commodity. Consequently, studying the physiological mechanism of sugar-acid metabolism in loquat can clarify the mechanism of their formation, accumulation and degradation in the fruit. Minerals application has been reported as a promising way to improve sugar-acid balance of the fruits. In this study, loquat trees were foliar sprayed with 0.1, 0.2 and 0.3% borax, and changes in soluble sugars and organic acids were recorded. The contents of soluble sugars and organic acids were determined using HPLC-RID and UPLC-MS, respectively. The activities of enzymes responsible forthe metabolism of sugarsand acids were quantified and expressions of related genes were determined using quantitative real-time PCR. The results revealed that 0.2% borax was a promising treatment among other B applications for the increased levels of soluble sugars and decreased acid contents in loquats. Correlation analysis showed that the enzymes i.e., SPS, SS, FK, and HK were may be involved in the regulation of fructose and glucose metabolism in the fruit pulp of loquat. While the activity of NADP-ME showed negative and NAD-MDH showed a positive correlation with malic acid content. Meanwhile, EjSPS1, EjSPS3, EjSS3, EjHK1, EjHK3, EjFK1, EjFK2, EjFK5, and EjFK6 may play an important role in soluble sugars metabolism in fruit pulp of loquat. Similarly, EjPEPC2, EjPEPC3, EjNAD-ME1, EjNAD-MDH1, EjNAD-MDH5-8, EjNAD-MDH10, and EjNAD-MDH13 may have a vital contribution to malic acid biosynthesis in loquat fruits. This study provides new insights for future elucidation of key mechanisms regulating soluble sugars and malic acid biosynthesis in loquats.

Metabolomic analyses reveal potential mechanisms induced by melatonin application for tolerance of water deficit in loquat (Eriobotrya japonica Lindl.)

Melatonin participates in the response to abiotic stressors in plants and enhances tolerance to water stress tolerance in loquat seedlings; however, the underlying mechanism remains unknown. In this study, we conducted physiological and metabolomic analyses to identify the metabolic processes associated with enhanced tolerance of melatonin-treated loquat seedlings under conditions of water deficit. Loquat seedlings were pre-irrigated with either 150 μM of melatonin or water and dehydrated for the next 13 days. Our results showed that water deficit reduced the relative water content and increased the relative electrolyte leakage in the leaves. Compared with untreated plants, melatonin-treated loquat leaves exhibited lower relative conductance and higher relative water content and antioxidant enzyme activity under water deficit conditions. Additionally, after melatonin application, the total phenolic and flavonoid content in the leaves had increased on day 7 but had decreased on day 13. Metabolomic analysis on days 1 and 13 revealed 108 and 196 metabolites, respectively, that were regulated by melatonin in loquat leaves. Importantly, melatonin application enhanced phenylpropanoid biosynthesis and the tricarboxylic acid (TCA) cycle and altered flavone and flavonol biosynthesis and galactose metabolism. Melatonin-treated plants exhibited higher levels of amino acids (threonine, asparagine, glutamate, and valine) and hydroxycinnamic acid derivatives (m-coumaric acid, shikimic acid, ferulic acid, coumaroylquinic acid, and 2-hydroxycinnamic acid). Thus, this study deepens our understanding of melatonin-mediated reprograming of metabolites in loquat to cope with water deficits.

Genome-Wide Identification, In Silico Analysis and Expression Profiling of SWEET Gene Family in Loquat (Eriobotrya japonica Lindl.)

SWEETs (sugars will eventually be exported transporters) have various physiological and biochemical roles in plant growth, including pollen development, seed nourishment, nectar secretion, and longer-distance sugar transportation. The SWEET genes were identified in various plant species, but they have not yet been thoroughly characterized. Here, we discovered 21 putative SWEET genes from the Eriobotrya japonica Lindl. genome. For further elucidation, comprehensive bioinformatics analysis was utilized to determine the physicochemical properties, gene organization, conserved motifs, cis-regulatory elements, gene duplication, and phylogenetic relationships of EjSWEET genes. Most of the SWEET proteins were predicted to be located on the plasma membrane or vacuole. Gene organization and motif analysis showed that the numbers of exons and motifs in each gene ranged strikingly, between 5 and 6 and between 5 and 8, respectively. Synteny analysis showed that the tandem or segmental duplication played a dynamic role in the evolution of SWEET genes in loquat. Likewise, we analyzed the expression patterns of EjSWEET genes in the root, stem, leaf, flower, and fruit of loquat. Some genes exhibited varying expression in loquat tissues, indicating their potential roles in plant development. The relative expression levels of EjSWEET1, EjSWEET3, and EjSWEET16 were noticeably higher in ripened fruits, suggesting their possible role in the transportation and unloading of sugars in fruits. The present study provides initial genome-wide identification and characterization of the SWEET gene family in loquat and lays the foundation for their further functional analysis.

Transcriptomic Analysis Reveals Potential Gene Regulatory Networks Under Cold Stress of Loquat (Eriobotrya japonica Lindl.).

Loquat (Eriobotrya japonica Lindl. ) is one of the most economically important evergreen fruit crops in China, while it often suffered the injury of cold stress in winter and earlier spring, and the annual yield loss of loquat fruits caused by cold or freezing stress was immeasurable. However, knowledge about the physiological response and molecular mechanism under cold stress is still limited. To investigate the potential regulation mechanism pre- and post-cold stress in loquat and the changes in physiological indicators, a comparative transcriptome analysis was performed against a cold-resistant cv. “Huoju” and a cold-sensitive cv. “Ninghaibai”. The results of physiological indicators related to cold resistance indicated that rachis was most sensitive to cold stress and was considered as the representative organ to directly evaluate cold resistance of loquat based on subordinate function analysis. Here, we compared the transcriptome profiles of rachis pre- and under cold stress in “Huoju” and “Ninghaibai”. A total of 4,347 and 3,513 differentially expressed genes (DEGs) were detected in “Ninghaibai” and “Huoju”, among which 223 and 166 were newly identified genes, respectively, most of them were functionally enriched in plant hormone signal transduction (Huoju: 142; Ninghaibai: 200), and there were higher plant hormone content and related DEG expression levels in “Huoju” than that of “Ninghaibai”. Moreover, a total of 3,309 differentially expressed transcription factors (DETFs) were identified, and some DEGs and DETFs were screened to be subjected to co-expression network analysis based on the gene expression profile data. Some candidate DEGs, including UDP-glycosyltransferase (UGT), glycosyltransferase (GT), sugar phosphate/phosphate translocator (SPT), sugar transport protein (STP), proline-rich receptor-like protein kinase (PERK), and peroxidise (POD), were significantly affected by cold stress, and the expression level of these genes obtained from real-time quantitative RT-PCR was consistent with the pattern of transcriptome profile, which suggested that these genes might play the vital roles in cold resistance of loquat. Our results provide an invaluable resource for the identification of specific genes and TFs and help to clarify gene transcription during the cold stress response of loquat.

Effects of Loquat (Eriobotrya japonica Lindl.) Leaf Extract with or without Ascorbic Acid on the Quality Characteristics of Semi-Dried Restructured Jerky during Storage.

Deterioration of jerky during storage is a major concern; this is usually combated with natural or synthetic antioxidants. This study aimed to evaluate the quality characteristics of semi-dried restructured jerky with and without loquat leaf extract (LE) powder and ascorbic acid (AA) during storage for 180 days. The jerkies were formulated with 0%, 0.15%, and 0.3% LE and/or 0.05% AA (Control, no antioxidant; AA, 0.05% AA; LE 0.15, 0.15% loquat LE; LE 0.15-AA, 0.15% loquat LE+0.05% AA; LE 0.3, 0.3% loquat LE; LE0.3-AA, 0.3% loquat LE+0.05% AA). LE is a phenolic compound, whose 1,1-diphenyl-2-picrylhydarzyl radical scavenging activity and metal chelating activity were found to be higher than AA. All antioxidant combinations having higher LE concentration and containing AA were effective in delaying protein and lipid oxidation compared to the control or AA. At the end of storage period, LE 0.15-AA and AA had higher CIE a* and lower shear force than the control. Therefore, the combination of 0.15% LE and 0.05% AA can result in reduced protein and lipid oxidation without any negative effect on the quality characteristics of semi-dried restructured jerky.

Loquat (Eriobotrya japonica (Thunb) Lindl.): Evaluation of nutritional value, polyphenol composition, antidiabetic effect, and toxicity of leaf aqueous extract

Ethnopharmacological relevanceLoquat (Eriobotrya japonica (Thunb.) Lindl.) is an evergreen tree native to China, which is introduced in many Mediterranean countries. As in many ancient medical systems, loquat leaves have been used in Moroccan traditional medicine to treat diabetes and its complications. Aim of the studyThis study aims to determine the nutritional and polyphenol composition and to evaluate the in vivo antidiabetic, and antihyperlipidemic properties and oral toxicity of a leaf aqueous extract (LLE) derived from loquat grown in Morocco. Materials and methodsEnergy value and fiber, fatty acids, minerals, vitamins, total carbohydrate, sugar, lipid, and protein contents were determined according to international methods committee guidelines. Polyphenol profiling was carried out using the HPLC-DAD method. Mice fed a high-fat and high-glucose (HFG) diet for 10 weeks were used as a model to assess the antidiabetic and antihyperlipidemic effects of a daily administration of LLE at three different doses (150, 200, 250 mg/kg body weight (BW)/day), in comparison with metformin (50 mg/kg BW/day) and pravastatin (20 mg/kg BW/day). The oral toxicity was determined following OECD 425 Guideline. ResultsLoquat leaves were found to be rich in fiber, minerals (potassium, calcium, magnesium, iron, and sodium), and vitamins (B2, B6, and B12) and lower in energy, sugar, and fat. Ten different phenolic compounds were characterized. Naringenin, procyanidin C1, epicatechin, and rutin were the more abundant compounds in LLE. The administration of the LLE dose-dependently ameliorated hyperglycemia, insulin resistance, oxidative stress, and hyperlipidemia in HFG diet-fed mice. The median lethal dose of LLE was higher than 5000 mg/kg BW. ConclusionsLoquat leaves are a potential source of micronutrients and polyphenols with beneficial effects on diabetes and its complications.

Self-compatibility of ‘Zaohuang’ loquat is attributed to the lack of a 52 bp fragment in the S10-RNase promoter

Loquat (Eriobotrya japonica Lindl.) is a typical gametophytic self-incompatible (GSI) subtropical evergreen fruit tree, but there are also some self-compatible varieties. In this study, the fruit set rate of ‘Zaohuang (S2S10)’ and ‘Baiyu (S2S10)’ loquats were investigated after self-pollination and cross-pollination, respectively. The analysis demonstrated that ‘Zaohuang’ (‘ZH’) was self-compatible (SC) and ‘Baiyu’ (‘BY’) was self-incompatible (SI), while they were cross-compatible. The expression levels of S-RNase in the pistils of ‘ZH’ and ‘BY’ were analyzed by qPCR, indicating that the expression of S2-RNase was similar in the two cultivars, but the expression of S10-RNase in the pistils of ‘ZH’ was significantly lower than that in ‘BY’. In addition, cloning of the full-length open reading frame (ORF) sequence and promoter of S10-RNase in ‘ZH’ and ‘BY’ by TAIL-PCR showed that the full-length ORF sequence of S10-RNase was completely consistent in two loquat cultivars. However, the promoter of S10-RNase in ‘ZH’ (ZH-S10-RNase-P) lacks 52 bp fragment comparing with that in ‘BY’ (BY-S10-RNase-P). The deletion fragment was located at 1755 bp upstream to the start code ATG of S10-RNase, containing a TGACG-motif and a CGTCA-motif. To detect the activities of ZH-S10-RNase-P and BY-S10-RNase-P, we constructed ZH-S10-RNase-P::GFP and BY-S10-RNase-P::GFP plasmids and introduced them into tobacco pistils by transient expression method. The fluorescence of GFP was much lower in ZH-S10-RNase-P::GFP transformed tobacco pistils than that in BY-S10-RNase-P::GFP transformed pistils. These results indicated that the 52 bp deletion in the promoter of S10-RNase results in reducing the expression of S10-RNase and eventually leads to self-compatibility of ‘ZH’ loquat. Our research provided a new direction for screening loquat SC varieties with S10-RNase promoter.

Effects of Tray-Drying on the Physicochemical, Microbiological, Proximate, and Sensory Properties of White- and Red-Fleshed Loquat (Eriobotrya Japonica Lindl.) Fruit

Loquat fruits, highly valued by consumers for their characteristic aroma and pleasant taste, have a short post-harvest life and are susceptible to mechanical damage, loss of firmness, and initial organoleptic characteristics. The aim of this work was to develop a drying method suitable for storing loquat fruits in polyamide/polyethylene (PA/PE) bags containing two gaseous mixtures (treatments): MAPN2 (100% N2) and MAPP (21% O2 and 0.04% CO2), at room temperature (20 ± 1 °C) for at least 2 months. The effects of these conditions on the physico-chemical, microbiological, proximate, and sensory properties of fruit stored over a 50-day time interval were studied. The results showed that convective tray dehydration treatment at 70° for 12 h had good drying efficiency for loquat slices. In addition, the MAPN2 packaging limited the browning of the slices, keeping the microbial groups below the detection limits, with a clear positive effect on some minerals and vitamins, which were higher in concentration compared to the MAPP-packed samples. From an applicative point of view, the tray drying method for loquat fruits is useful on a small scale but could also be easily industrialized.

Morphological and Developmental Features of Stone Cells in Eriobotrya Fruits.

Some members of the Rosaceae family, particularly pear, contain stone cells in their fruits. Although stone cells in pear fruits are well studied, relatively little attention has been given to loquat stone cells. Only a few reports have suggested a relationship between stone cell traits and storage and transport tolerance of loquat fruits. Previously, we generated the variety JT8 from the interspecific hybrid of the loquat cultivar Jiefangzhong (JFZ; Eriobotrya japonica Lindl. cv. Jiefangzhong, female parent) and wild Taiwanese loquat (TL; E. deflexa Nakai, male parent). The JT8 fruits had a granular feel, similar to that of pear fruits, due to the presence of stone cells. In this study, the shape, size, development, and distribution dynamics of stone cells of Eriobotrya plants were thoroughly investigated. The results showed that loquat stone cells are brachysclereids and often contain typical branching pits. Loquat stone cells were distributed as both single stone cells and in stone cell clusters (SCCs), and the density of the stone cells near the core was higher than that near the peel. Stone cell density first increased and then decreased during fruit development. These traits noted in Eriobotrya were very similar to those observed in pear, indicating a close relationship between loquat and pear. Moreover, the contents, density dynamics, and aggregation traits of stone cells of the interspecific hybrid JT8 were derived from the male parent (TL). Transgressive segregation was likely exhibited in the content of stone cells and the size of the SCCs. More specifically, the content of stone cells reached 1.61% (w/w). In extreme cases, SCCs of JT8 exceeded 1,000 μm in length and 500 μm in width. This demonstrated that stone cell traits could be transmitted from parent to progeny through interspecific hybridization. The density dynamics of stone cells in two loquat cultivars with different storage and transport tolerances were also investigated, which indicated that the cultivar with more stone cells was more tolerant to storage and transport. We suggest that wild loquat genetic resources containing stone cells in Eriobotrya plants can be used to gradually improve the storage and transport tolerance of loquat fruits.

Integration of genomics, transcriptomics and metabolomics identifies candidate loci underlying fruit weight in loquat.

Fruit weight is an integral part of fruit quality and directly influences the commodity value and economic return of fruit crops. Despite the importance of fruit weight, its underlying molecular mechanisms remain understudied, especially for perennial fruit tree crops such as cultivated loquat (Eriobotrya japonica Lindl.). Auxin is known to regulate fruit development, but its role and metabolism during the development of loquat fruit remain obscure. In this study, we used a multi-omics approach, integrating whole-genome resequencing-based quantitative trait locus (QTL) mapping with an F1 population, population genomics analysis using germplasm accessions, transcriptome analysis, and metabolic profiling to identify genomic regions potentially associated with fruit weight in loquat. We identified three major loci associated with fruit weight, supported by both QTL mapping and comparative genomic analysis between small- and large-fruited loquat cultivars. Comparison between two genotypes with contrasting fruit weight performance by transcriptomic and metabolic profiling revealed an important role for auxin in the regulation of fruit development, especially at the fruit enlargement stage. The multi-omics approach identified homologs of ETHYLENE INSENSITIVE 4 (EjEIN4) and TORNADO 1 (EjTRN1) as promising candidates for the control of fruit weight. Three single nucleotide polymorphism (SNP) markers were also closely associated with fruit weight. Results from this study provide insights into the genetic and metabolic controls of fruit weight in loquat from multiple perspectives. The candidate genomic regions, genes, and sequence variants will facilitate our understanding of the molecular basis of fruit weight and lay a foundation for future breeding and manipulation of fruit weight in loquat.

Effect of Paper and Aluminum Bagging on Fruit Quality of Loquat (Eriobotrya japonica Lindl.).

Bagging regulates the fruit microenvironment and improves the quality and market value of fruits. It is a safe and ecofriendly technique to protect fruits from insect/pest infestation and multiple biotic and abiotic stresses. In the current study, the influence of fruit bagging was evaluated on the development and quality of loquat fruits. Fruits from a healthy loquat orchard (Cv. Zaozhong No.6), located in Fujian, China, were enveloped in paper (T1), aluminum (T2), and aluminum–polyethylene bags (T3), while unbagged fruits were maintained as control (T0). In general, fruit bagging improved fruit quality in terms of fruit physiological and biochemical attributes and protected fruits from physical damage. In particular, aluminum–polyethylene bagging enhanced fruit weight, length, and width by 1.37-, 1.18-, and 1.13-fold, respectively. Loquat fruits bagged with paper bags exhibited the maximum soluble sugar and lowest titratable acid content. Fruits treated with paper and aluminum–ethylene bags showed twofold higher sugar–acid ratio as compared to control. Aluminum–polyethylene bagging caused 66.67%, 55.56%, and 33.33% reductions in skin burn, fruit rotting, and black spot of loquat. The fruits bagged in aluminum and aluminum–polyethylene did not show insect or bird damage, while unbagged fruits had 14.70% and 17.65% insect and bird damage, respectively. Overall, the results suggest that paper, aluminum, and aluminum–polyethylene bagging improved fruit health by 75%, 131%, and 144%, respectively, as compared to control. To delineate bagging type-dependent effects, principal component analysis was performed. Paper bagging was positively correlated with fruit firmness, rotting, soluble sugars, sugar–acid ratio, and proline content. Aluminum bagging was highly associated with improvements in titratable acids, cystine, and methionine. Aluminum–polyethylene bags were correlated with fruit weight, size, peel thickness, edible rate, and certain amino acids.

Correlation of the free radical and antioxidant activities of Eriobotrya Japonica Lindl. with phenolic and flavonoid contents

This study was conducted to compare the correlations between antioxidant activity and free radicals. Total Polyphenol and Flavonoid contents were the highest 31.97 ± 1.29 mg TAE/g and 96.1 ± 0.31 mg CE/g in hot water extract, respectively. FRAP activity was highest at 559.17 ± 2.69 mg TE/g in 60% EtOH extract. The RC50 values of DPPH and ABTS radical scavenging activity were lowest at 61.96 ± 5.01 μg/mL in 80% EtOH, 73.81 ± 0.16 μg/mL in 60% EtOH, respectively. The RC50 values of hydrogen peroxide scavenging activity and NO radical scavenging activity were lowest at 265.33 ± 3.88 μg/mL in 60% EtOH and 52.51 ± 8.54 μg/mL in hot water extract, respectively. The total polyphenol and flavonoid contents had significant correlation with the free radical scavenging ability and electron donating ability. On the other hand, there was confirmed significant correlation of free radical and antioxidant activities.

Study morphology of loquat plants (Eriobotrya Japonica Lindl.) from Karo, Dairi, and Simalungun Districts, North Sumatra

Abstrak. Loquat (Eriobotrya japonica Lindl.) is a flowering plant that belongs to the Rosacea family. The loquat has many health benefits. Cultivation and information about loquat plants in Indonesia are still limited, so they are rarely found and known by the public. Limited information and data regarding loquat plants is also an obstacle to the development of loquat plants. Research on loquat plants aims to analyze the morphological characters in three districts, namely, Karo, Dairi, and Simalungun districts. This research was conducted using a descriptive method. The analysis of the morphological characteristics of loquat plants using morphological data scoring into binary data. The similarity between individuals was analyzed using clusters with the NTSYS program version 2.0 with the UPGMA method of the SimQual function. Morphological Observation Results Loquat plants (Eriobotrya japonica Lindl.) in Karo, Dairi, and Simalungun Districts have uniform characters in the morphology of stems, leaves, and flowers. However, the observed fruit and seed morphology showed different characters. Different characters exist in the shape of the fruit and seeds. The morphological similarity level of loquat plants was grouped at a similarity coefficient value of 95%. Clusters I and II have the highest similarity with a coefficient value of 100%. Cluster III has the lowest similarity with a coefficient value of 97%.

Genome-Wide Identification, Characterization and Expression Profiling of Aluminum-Activated Malate Transporters in Eriobotrya japonica Lindl.

Aluminum-activated malate transporters (ALMTs) have multiple potential roles in plant metabolism such as regulation of organic acids in fruits, movement of guard cells and inducing tolerance against aluminum stress. However, the systematic characterization of ALMT genes in loquat is yet to be performed. In the current study, 24 putative ALMT genes were identified in the genome of Eriobotrya japonica Lindl. To further investigate the role of those ALMT genes, comprehensive bioinformatics and expression analysis were performed. In bioinformatics analysis, the physiochemical properties, conserved domains, gene structure, conserved motif, phylogenetic and syntenic analysis of EjALMT genes were conducted. The result revealed that the ALMT superfamily domain was conserved in all EjALMT proteins. EjALMT proteins were predicted to be localized in the plasma membrane. Genomic structural and motif analysis showed that the exon and motif number of each EjALMT gene ranged dramatically, from 5 to 7, and 6 to 10, respectively. Syntenic analysis indicated that the segmental or whole-genome duplication played a vital role in extension of the EjALMT gene family. The Ka and Ks values of duplicated genes depicted that EjALMT genes have undergone a strong purifying selection. Furthermore, the expression analysis of EjALMT genes was performed in the root, mature leaf, stem, full-bloom flower and ripened fruit of loquat. Some genes were expressed differentially in examined loquat tissues, signifying their differential role in plant growth and development. This study provides the first genome-wide identification, characterization, and relative expression of the ALMT gene family in loquat and provides the foundation for further functional analysis.

The Putative Role of the NAC Transcription Factor EjNACL47 in Cell Enlargement of Loquat (Eriobotrya japonica Lindl.)

NAC transcription factors (TFs) are plant-specific TFs that play essential roles in plant development; however, the function of NAC TFs in loquat development remains unknown. The natural triploid loquat (Eriobotrya japonica Lindl.), Longquan No.1. B355, has larger organs than its corresponding diploid loquat (B2). Here, we cloned an NAC-like TF (EjNACL47 (NAC-like 47)) from the cDNA of triploid loquat B355 flowers. EjNACL47 has a conserved domain of NAC TFs and is homologous to AtNAC47. Transient expression in tobacco leaves revealed that EjNACL47 localized to the nucleus, and yeast-two-hybrid screening confirmed that the C-terminus displayed transcriptional activity. Interestingly, real-time qRT-PCR indicated that the expression levels of EjNACL47 in leaves and flower organs in triploid loquat (B355) were higher than those in diploid loquat (B2), implying that EjNACL47 might be associated with the larger organ size in B355. Moreover, Arabidopsis lines ectopically expressing EjNACL47 presented obviously larger leaves, flowers, and siliques than the wild-type variant, suggesting that EjNACL47 plays a positive role in Arabidopsis organ enlargement. These results offer insight into the molecular mechanism of NAC TFs involved in regulating organ size in loquat.

The Antifungal Activity of Loquat (Eriobotrya japonica Lindl.) Leaves Extract Against Penicillium digitatum.

This study was performed to determine the antifungal activity of loquat (Eriobotrya japonica Lindl) leaf extract (LLE) against the citrus postharvest pathogen Penicillium digitatum (P. digitatum). The LLE exhibited an antifungal activity against P. digitatum, with a minimum inhibitory concentration (MIC) of 0.625 mg/ml and a minimum fungicidal concentration (MFC) of 1.25 mg/ml. Significant inhibitory effects of LLE on mycelial growth and spore germination of P. digitatum were seen in a dose-dependent manner. Simultaneously, to investigate possible antifungal mechanisms by LLE, we analyzed their influence on morphological changes, cell membrane permeability, cell wall and cell membrane integrity, and adenosine phosphates (ATP, ADP, and AMP) levels. Alterations, such as sunken surface and malformation, occurred in the LLE-treated P. digitatum spores. Furthermore, intracellular inclusion content decreased after LLE treatment, indicating an increase in cell membrane permeability. Besides, the LLE treatment induced a significant decline in the level of adenosine monophosphate (AMP), adenosine diphosphate (ADP), and adenosine triphosphate (ATP) with a noticeable addition of extracellular ATP, ADP, and AMP during the entire treatment period. Overall, the results manifested that the antifungal activity of LLE against P. digitatum can be attributed to the derangement of cell membrane permeability and disordered energy metabolism. This is the first report on the mechanism of antifungal activity of LLE and could be useful in the development of targeted fungicides from natural origin.

The efficacy of acetylsalicylic acid, spermidine and calcium preharvest foliar spray applications on yield efficiency, incidence of physiological disorders and shelf-life performance of loquat fruit

The effect of preharvest foliar spray applications at fruit colour breaker stage with acetylsalicylic acid (synthetic salicylate), spermidine (polyamine), their combination and a commercial calcium supplement on loquat (Eriobotrya japonica Lindl cv. ‘Karantoki’) fruit at successive harvests and subsequently during maintenance at room temperature (shelf-life) for 3 and 6 days respectively, were investigated. Yield efficiency, qualitative attributes, textural properties and the incidence of physiological disorders, namely purple spot and fruit cracking, were determined. Loquat fruit quality parameters were affected by spraying treatments, harvesting day and shelf-life periods applied. Acetylsalicylic acid is recommended as an efficient and cost-effective treatment that needs to be further explored towards production of loquat fruit with enhanced quality properties. However, spermidine application alone, or in combination with acetylsalicylic acid, did not show any beneficial and/or synergistic effect on loquat fruit quality parameters to justify its application. All foliar applications did not affect fruit growth and yield efficiency. The current study shed some light on the potential use of foliar spraying with acetylsalicylic acid, alone or in combination with a calcium supplement, towards enhancement of fruit quality properties of an added value product as loquat. To this aim, further studies need to be implemented to test their efficacy under different environmental conditions that may accelerate the incidence of physiological disorders and/or multiple applications over the growing season.