Comparative Transcriptomic Analyses of Different Jujube Cultivars Reveal the Co-Regulation of Multiple Pathways during Fruit Cracking.

Fruit cracking is a common physiological disorder in many fruit species. Jujube (Ziziphus jujuba Mill.) is an economically valuable fruit in which fruit cracking seriously affects fruit yield and quality and causes significant economic losses. To elucidate cracking-related molecular mechanisms, the cracking-susceptible cultivars 'Cuizaohong' and 'Jinsixiaozao' and the cracking-resistant cultivar 'Muzao' were selected, and comparative transcriptome analyses of cracking and non-cracking 'Cuizaohong' (CC and NC), cracking and non-cracking 'Jinsixiaozao' (CJ and NJ), and non-cracking 'Muzao' (NM) were conducted. A total of 131 differentially expressed genes (DEGs) were common to the CC vs. NC and CJ vs. NJ comparisons. To avoid passive processes after fruit cracking, we also mainly focused on the 225 gradually downregulated DEGs in the CJ, NJ, and NM samples. The functional annotation of the candidate DEGs revealed that 61 genes related to calcium, the cell wall, the cuticle structure, hormone metabolism, starch/sucrose metabolism, transcription factors, and water transport were highly expressed in cracking fruits. We propose that expression-level changes in these genes might increase the turgor pressure and weaken mechanical properties, ultimately leading to jujube fruit cracking. These results may serve as a rich genetic resource for future investigations on fruit cracking mechanisms in jujube and in other fruit species.

Changes in pectin characteristics of jujube fruits cv "Dongzao" and "Jinsixiaozao" during cold storage.

Softening is one of the main factors affecting market value and consumer preferences for jujubes, and it was closely related to the modification and depolymerization of pectin. Changes in characteristics of three pectins (water-soluble pectin (WSP), sodium carbonate-soluble pectin (SSP) and chelate-soluble pectin (CSP)), including their contents, degree of methylesterification (DM), neutral sugar compositions, the molecular weight (Mw ) distributions and nanostructures, from two jujube fruits cv Dongzao (DZ) and Jinsixiaozao (JS) during cold storage were assessed. The results showed that variation in pectin characteristics during cold storage was similar between DZ and JS. The reduction of firmness corresponded to a conversion of water-insoluble pectin to WSP during cold storage. DM of WSP presented an increase trend in the late storage. Rhamnose (Rha), arabinose (Ara) and glucose (Glc) were the crucial compositions in three pectins, and most neutral sugar compositions in three pectins first increased and then decreased during cold storage. Changes in the ratio of (galactose (Gal)+Ara)/Rha and Ara/Gal represented that the branch chains of rhamnogalacturonan-I in three pectins depolymerized after storage. The high Mw in WSP and SSP of jujubes were solubilized and extensively depolymerized into pectin with lower Mw after storage. AFM images showed an increase in short chains and branch structures of three pectins after storage. Overall, three pectins in DZ and JS depolymerized and solubilized during cold storage. WSP and SSP were more contributed to the softening of jujubes compared to CSP, and they played the critical role for regulating the softening of jujube fruits during cold storage. PRACTICAL APPLICATION: Softening is one of the main factors affecting market value and consumer preferences for jujubes, and it was closely related to the modification and depolymerization of pectin. Changes in characteristics of three pectins (WSP, SSP, CSP), including their contents, degree of methylesterification, neutral sugar compositions, the molecular weight distributions and nanostructures, from two jujube fruits cv Dongzao (DZ) and Jinsixiaozao (JS) during cold storage were assessed. Three pectins in DZ and JS depolymerized and solubilized during cold storage. WSP and SSP were more contributed to the softening of jujubes compared to CSP, and they played the critical role for regulating the softening of jujube fruits during cold storage. This study would elucidate the mechanism of jujube softening and help to regulate the postharvest quality during cold storage.

Genome-wide analysis of the bZIP gene family in Chinese jujube (Ziziphus jujuba Mill.).

BACKGROUND: Among several TF families unique to eukaryotes, the basic leucine zipper (bZIP) family is one of the most important. Chinese jujube (Ziziphus jujuba Mill.) is a popular fruit tree species in Asia, and its fruits are rich in sugar, vitamin C and so on. Analysis of the bZIP gene family of jujube has not yet been reported. In this study, ZjbZIPs were identified firstly, their expression patterns were further studied in different tissues and in response to various abiotic and phytoplasma stresses, and their protein-protein interactions were also analyzed. RESULTS: At the whole genome level, 45 ZjbZIPs were identified and classified into 14 classes. The members of each class of bZIP subfamily contain a specific conserved domain in addition to the core bZIP conserved domain, which may be related to its biological function. Relative Synonymous Codon Usage (RSCU) analysis displayed low values of NTA and NCG codons in ZjbZIPs, which would be beneficial to increase the protein production and also indicated that ZjbZIPs were at a relative high methylation level. The paralogous and orthologous events occurred during the evolutionary process of ZjbZIPs. Thirty-four ZjbZIPs were mapped to but not evenly distributed among 10 pseudo- chromosomes. 30 of ZjbZIP genes showed diverse tissue-specific expression in jujube and wild jujube trees, indicating that these genes may have multiple functions. Some ZjbZIP genes were specifically analyzed and found to play important roles in the early stage of fruit development. Moreover, some ZjbZIPs that respond to phytoplasma invasion and abiotic stress environmental conditions, such as salt and low temperature, were found. Based on homology comparisons, prediction analysis and yeast two-hybrid, a protein interaction network including 42 ZjbZIPs was constructed. CONCLUSIONS: The bioinformatics analyses of 45 ZjbZIPs were implemented systematically, and their expression profiles in jujube and wild jujube showed that many genes might play crucial roles during fruit ripening and in the response to phytoplasma and abiotic stresses. The protein interaction networks among ZjbZIPs could provide useful information for further functional studies.

Phytochemical Profiling and Fingerprint Analysis of Chinese Jujube (Ziziphus jujuba Mill.) Leaves of 66 Cultivars from Xinjiang Province.

Foliage of jujube (Ziziphus jujuba Mill.) as a byproduct of agriculture, is a traditional nutraceutical material in China. Previous studies have shown that it is a rich resource of polyphenols. However, information on its complete phenolic profile and the difference between cultivars is still limited. This study investigated and compared phytochemical profiles of leaves of 66 Chinese jujube cultivars. Forty-two compounds, including 22 flavonols, two flavanols, one flavanone, 13 derivatives of phenolic acids, three simple acids, and one unknown hexoside were identified/tentatively identified using high-performance liquid chromatography (HPLC) coupled with high-resolution mass spectrometry. Eight major flavonols were quantified by HPLC coupled with an ultraviolet (UV) detector. The contents of total flavonoids ranged from 2.6-25.1 mg/g dry weight (DW). Differences between cultivars were analyzed by hierarchical cluster analysis (HCA) and principal component analysis (PCA). This study presents a systematic study on the phenolic compounds in Chinese jujube leaves of different cultivars.

New robust sensitive fluorescence spectroscopy coupled with PLSR for estimation of quercetin in Ziziphus mucronata and Ziziphus sativa.

Flavonoids are natural antioxidants derived from plants and commonly found in a variety of foods to sequester free radicals. Quercetin, belonging to flavonol subclass of flavonoids, has received considerable attention because of its wide uses as a nutritional supplement as well as a phytochemical remedy for a number of diseases. In the current study, quantification of quercetin was carried out in two medicinally important flavonoid rich plant Ziziphus mucronata and Ziziphus sativa. Emission spectroscopy was utilized as a new method coupled with Partial Least Squares Regression (PLSR) and the cross validation was done by UV-Visible spectroscopy. The results indicated the higher quercetin content in Z. mucronata (1.50±0.034%) than Z. sativa (1.21±0.052%), and were further verified through Folin-Ciocalteu Colorimetric method (Z. mucronata; 1.41±0.26% and Z. sativa; 1.13±0.136%). In this study the sensitivity was explained in term of slope i.e. Slope=0.9973.

Ziziphus spinosa seeds for insomnia: A review of chemistry and psychopharmacology.

BACKGROUND: In Chinese medicine, Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou is widely used for the treatment of insomnia. PURPOSE/SECTIONS: This paper summarises the chemistry, psychopharmacology, and compares the pharmaceutical effects of the seeds of Ziziphus jujuba plant, Ziziphus spinosa (ZS) seeds, with benzodiazepines. Whole extracts and constituent compounds have been evaluated in preclinical and clinical studies. CONCLUSIONS: ZS secondary metabolites modulate GABAergic activity and the serotonergic system. The actual therapeutic agents require further confirmation/identification so that new insomnia phytomedicines can be discovered.

Isolation and structure elucidation of cyclopeptide alkaloids from Ziziphus nummularia and Ziziphus spina-christi by HPLC-DAD-MS and HPLC-PDA-(HRMS)-SPE-NMR.

Seven cyclopeptide alkaloids were isolated from the stem bark of Ziziphus nummularia and Ziziphus spina-christi. Three previously undescribed compounds were identified: nummularine-U, spinanine-B and spinanine-C, together with the known compounds mauritine-F, nummularine-D, nummularine-E and amphibine-D. For their purification either semi-preparative HPLC with DAD and ESIMS detection or HPLC-PDA-(HRMS)-SPE-NMR was applied, together with conventional separation methods. Their structures were elucidated by spectroscopic means.

Complete chloroplast genome sequence of a major economic species, Ziziphus jujuba (Rhamnaceae).

Ziziphus jujuba is an important woody plant with high economic and medicinal value. Here, we analyzed and characterized the complete chloroplast (cp) genome of Z. jujuba, the first member of the Rhamnaceae family for which the chloroplast genome sequence has been reported. We also built a web browser for navigating the cp genome of Z. jujuba ( http://bio.njfu.edu.cn/gb2/gbrowse/Ziziphus_jujuba_cp/ ). Sequence analysis showed that this cp genome is 161,466 bp long and has a typical quadripartite structure of large (LSC, 89,120 bp) and small (SSC, 19,348 bp) single-copy regions separated by a pair of inverted repeats (IRs, 26,499 bp). The sequence contained 112 unique genes, including 78 protein-coding genes, 30 transfer RNAs, and four ribosomal RNAs. The genome structure, gene order, GC content, and codon usage are similar to other typical angiosperm cp genomes. A total of 38 tandem repeats, two forward repeats, and three palindromic repeats were detected in the Z. jujuba cp genome. Simple sequence repeat (SSR) analysis revealed that most SSRs were AT-rich. The homopolymer regions in the cp genome of Z. jujuba were verified and manually corrected by Sanger sequencing. One-third of mononucleotide repeats were found to be erroneously sequenced by the 454 pyrosequencing, which resulted in sequences of 1-4 bases shorter than that by the Sanger sequencing. Analyzing the cp genome of Z. jujuba revealed that the IR contraction and expansion events resulted in ycf1 and rps19 pseudogenes. A phylogenetic analysis based on 64 protein-coding genes showed that Z. jujuba was closely related to members of the Elaeagnaceae family, which will be helpful for phylogenetic studies of other Rosales species. The complete cp genome sequence of Z. jujuba will facilitate population, phylogenetic, and cp genetic engineering studies of this economic plant.

Development of Chloroplast Microsatellite Markers and Analysis of Chloroplast Diversity in Chinese Jujube (Ziziphus jujuba Mill.) and Wild Jujube (Ziziphus acidojujuba Mill.).

Ziziphus is an important genus within the family Rhamnaceae. This genus includes several important fruit tree species that are widely planted in China and India, such as the Chinese jujube (Ziziphus jujuba Mill.), the wild jujube (Z. acidojujuba), and the Indian jujube (Z. mauritiana). However, information about their domestication based on the chlorotype diversity of Chinese jujube population is lacking. In this study, chloroplast microsatellite (cpSSR) markers were developed and used to investigate the genetic relationships between and domestication of jujube cultivars and wild jujube populations. Primer sets flanking each of the 46 cpSSR loci in non-coding regions of the chloroplast genome sequence of Z. jujuba Mill. cv. 'Junzao' were designed. In total, 10 markers showed polymorphisms from 15 samples (9 jujube cultivars and 6 wild jujube individuals), of which 8 loci were due to variations in the number of mononucleotide (A/T) repeats and 2 were due to indels. Six cpSSR markers were used in further analyses of 81 additional samples (63 jujube cultivars, 17 wild jujube samples, and 1 Indian jujube). Using these cpSSR markers, the number of alleles per locus ranged from two to four. In general, the Shannon Index (I) for each cpSSR ranged from 0.159 to 0.1747, and the diversity indices (h) and uh were 0.061 to 0.435 and 0.062 to 0.439, respectively. Seven chlorotypes were found; the Indian jujube showed distinct chlorotypes, and both the Chinese and wild jujube had four chlorotypes and shared two chlorotypes. A dominant chlorotype (G) accounted for 53 of 72 jujube cultivars and 13 of 23 wild jujube individuals. All chlorotypes were highly localized along the Yellow River, from the mid- to the lower reaches, suggesting a wide origin of jujube. These cpSSR markers can be applied to population and evolution studies of Chinese jujube and wild jujube.

Assay of phenolic compounds from four species of ber (Ziziphus mauritiana L.) fruits: comparison of three base hydrolysis procedure for quantification of total phenolic acids.

The present study was undertaken to investigate the flavonoid profile in four species of ber (Ziziphus mauritiana Lamk.) fruit. The 12 flavonoids identified were quercetin 3-O-robinobioside, quercetin 3-O-rutinoside, quercetin 3'-O-galactoside, quercetin 3'-O-glucoside, quercetin 3'-O-rhamnoside, quercetin 3'-O-pentosylhexoside, quercetin 3-O-6'malonylglucoside, quercetin 3'-O-malonylglucoside, luteolin 7-O-6'malonylglucoside, luteolin 7-O-malonylglucoside, myricetin 3-O-galactoside, and naringenin tri glycoside. This is the first report on extraction of nine additional flavonoids from the ber fruits. In addition, we also compared the impact of three different base hydrolysis techniques namely ultrasonic assisted base hydrolysis (UABH), microwave assisted base hydrolysis (MWABH), and pressurised liquid assisted base hydrolysis (PLABH) for the quantification of total phenolic acids. Nine phenolic acids, protocatechuic acid, p-hydroxybenzoic acid, ferulic acid, chlorogenic acid, vanillic acid, caffeic acid, vanillin, ortho- and para-coumaric acids, were identified and quantified. The three major phenolic acids identified in all four ber species were p-coumaric acid, vanillin and ferulic acids. Higher amounts (p<0.05) of total phenolic acids in all cultivars were obtained with the PLABH technique as compared to other two procedures (UABH and MWABH).

[Discrimination of the fresh jujube varieties and dehiscent fruit by NIR spectroscopy].

There are many fresh jujube varieties. The different variety has different quality. In addition, dehiscent fruit easily rot and the rotten fruit contaminated the full fruit very rapidly. It is necessary to discriminate the jujube varieties and dehiscent fruit to reduce the storage loss. The objective is to discriminate varieties and dehiscent Fruit of fresh jujube using near infrared (NIR) spectroscopy. Two jujube varieties were investigated. Smoothing, multiplicative scatter correction, the first derivative and second derivative methods were adopted to pretreat the spectra. The regression coefficient and principal component analysis were used to select wavenumber. Multilayer perceptron artificial neural network was used to build varieties and dehiscent fruit qualitative discrimination model. The results showed that the varieties and dehiscent fruit could be correctly discriminated and both the discrimination accuracy rates were 100%. Hence, near infrared spectroscopy could achieve to identify the variety of Dongzao and Lizao, and dehiscent fruit and intact fruit.

Textural characteristic, antioxidant activity, sugar, organic acid, and phenolic profiles of 10 promising jujube (Ziziphus jujuba Mill.) selections.

UNLABELLED: Ten promising jujubes were analyzed for textural characteristic (hardness), antioxidant activity, sugar, organic acid, and phenolic profiles. Total phenolic content (TP) measured by Folin-Ciocalteu method ranged from 275.6 to 541.8 mg of gallic acid equivalents per 100 g fresh weight (mg GAE/100 g FW). Four sugars (rhamnose, fructose, sucrose, and glucose), 3 organic acids (malic, citric, and succinic acids), and 11 phenolic compounds (gallic acid, protocatechuic acid, cinnamic acid, chlorogenic acid, caffeic acid, ferulic acid, ellagic acid, catechin, epicatechin, rutin, and quercetin) were identified and quantified by high-performance liquid chromatography in jujube fruits. The results showed that ascorbic acid and proanthocyanidins contents were positively correlated with hardness, and antioxidant activity was well correlated with TP content. Among 10 jujube cultivars, Zizyphus jujuba cv. Qingjianmuzao is good for direct consumption containing high levels of total soluble solids, total sugars, fructose, and glucose, while Zizyphus jujuba cv. Jiaxianmuzao could be an important dietary source of natural antioxidants. PRACTICAL APPLICATION: Genotype is the main factor influencing the composition of bioactive compounds in jujubes. Zizyphus jujuba cv. Qingjianmuzao is good for direct consumption, while Zizyphus jujuba cv. Jiaxianmuzao could be an important dietary source of natural antioxidants for prevention of diseases caused by oxidative stress.

[Species differentiation and quality assessment of Ziziphus jujuba var. spinosa based on the HPLC fingerprint and quantitative analysis].

OBJECTIVE: To establish an HPLC method of a characteristical chemical fingerprint analysis in combination with simultaneous determination of four bioactive components for species differentiation and quality assessment of Ziziphus jujuba var. spinosa. METHODS: The chromatographic separation was performed on an Agilent TC-C18 BDS (250 mm x 4.6 mm, 5 microm) column. The mobile phase consisted of acetonitrile and water in a linear gradient elution procedure. The evaporator tube temperature of ELSD was set at 110.5 degrees C with the nebulizing gas flow rate of 3.1 mL/min and the flow rate of mobile phase was 1.0 mL/min. The column was maintained at 30 degrees C. The injection volume was 20 microL. RESULTS: HPLC methodology for both chemical fingerprint analysis and quantitative determination of four active ingredients were validated, respectively. According to the contents of the four ingredients and the chemical fingerprints of Ziziphus jujuba var. spinosa using principal component analysis, Ziziphus jujuba var. spinosa was different from the fake derived from the seeds of Ziziphus mauritiana. CONCLUSION: The developed HPLC method is exclusive and repetitive for the species identification and quality evaluation of Ziziphus jujuba var. spinosa.

Chemotaxonomic variability in Zizyphus mouritiana varieties and its pharmacological properties in relation to human health.

Twenty varieties of ber (Zizyphus mouritiana), namely umaran, katha, bilayati, kaithli, ZG-3, gola, safeda rohtak, takadi, tikari, banarasi karaka, seo, sonaur-2, sonaur-3, ilaichi, mundia murahra, pathan, kakrola gola, seb, golden yellow and chhuhara, were investigated for the presence of phenolic acids in stem bark, leaves and fruits using high performance liquid chromatograph. Results indicated the presence of tannic (retention time (Rt.) 2.76 min), gallic (Rt. 2.86 min), caffeic (Rt. 3.12 min), vanillic (Rt. 3.26 min), ferulic (Rt. 3.42 min), chlorogenic (Rt. 4.16 min) and cinnamic acids (Rt. 4.45 min) in varying amounts in different parts in of these varieties. In fruits of seven varieties, namely, kaithly, sonaur-2, sonaur-3, mundia murahra, pathan, golden yellow and chhuhara, oxalic acid (Rt. 3.00 min) was also detected. Pharmacological properties of phenolic acids of fruits in relation to human health and the possible implications of different phenolic acids in chemotaxonomy of different varieties of ber are discussed.

[Studies on HPLC-FPS of the saponins from Semen Ziziphi Spinosae].

OBJECTIVE: To establish a method of HPLC-fingerprint spectrum (HPLC-FPS) for the active part in Semen Ziziphi Spinosae (SZS) in order to control the quality of SZS from different places. METHODS: The gradient elution mode was applied in chromatographic separation, and data were analysed by" Similarity Evaluation software" to compare the HPLC-FPS of SZS from different places. RESULTS: The conditions for HPLC analysis of SZS were established and the FPS of samples from different habitats showed some differences. CONCLUSION: All components in the spectrum were separated well and the HPLC fingerprint method is repeatable. The method can be used in quality assessment of SZS.