Temperature and Twist Sensor Based on the Sagnac Interferometer with Long-Period Grating in Polarization-Maintaining Fiber.

We utilized a CO2 laser to carve long-period fiber gratings (LPFGs) on polarization-maintaining fibers (PMFs) along the fast and slow axes. Based on the spectra of LPFGs written along two different directions, we found that when LPFG was written along the fast axis, the spectrum had lower insertion loss and fewer side lobes. We investigated the temperature and twist characteristics of the embedded structure of the LPFG and Sagnac loop and ultimately obtained a temperature sensitivity of -0.295 nm/°C and a twist sensitivity of 0.87 nm/(rad/m) for the LPFG. Compared to the single LPFG, the embedded structure of the LPFG and Sagnac loop demonstrates a significant improvement in temperature and twist sensitivities. Additionally, it also possesses the capability to discern the direction of the twist. The embedded structure displays numerous advantages, including easy fabrication, low cost, good robustness, a wide range, and high sensitivity. These features make it highly suitable for applications in structural health monitoring and other related fields.

ZjWRKY23 and ZjWRKY40 Promote Fruit Size Enlargement by Targeting and Downregulating Cytokinin Oxidase/Dehydrogenase 5 Expression in Chinese Jujube.

Fruit size is crucial for fruit trees, as it contributes to both quality and yield. However, the underlying mechanism of fruit size regulation remains largely unknown. Taking advantage of using a fruit double-sized bud mutant of Chinese jujube, "Jinkuiwang" and its wild type, "Jinsixiaozao", we carried out a comprehensive study on the mechanism of fruit size development in jujube. Using weighted gene coexpression network analyses, a number of candidate regulators for fruit size including those involved in hormonal signaling pathways, transcription factors, and heat shock proteins were identified. A hub gene named cytokinin oxidase/dehydrogenase 5 (ZjCKX5), responsible for cytokinin degradation, was found to play a negative role in regulating fruit size development, and overexpressing ZjCKX5 in tomato and Arabidopsis resulted in much smaller fruits and dwarf plants. Furthermore, another two hub genes, ZjWRKY23 and ZjWRKY40 transcription factors, were found to participate in fruit size regulation by targeting and downregulating the ZjCKX5 expression. Overexpressing ZjWRKY23 or ZjWRKY40 in tomato led to much larger fruits and promoted plant architecture. Based on these results, a molecular framework for jujube fruit size regulation, namely, ZjWRKY-ZjCKX5 module, was proposed. This study provides a new insight into the molecular networks underlying fruit size regulation.

Dehydrin CaDHN2 Enhances Drought Tolerance by Affecting Ascorbic Acid Synthesis under Drought in Peppers.

Peppers (Capsicum annuum L.), as a horticultural crop with one of the highest ascorbic acid contents, are negatively affected by detrimental environmental conditions both in terms of quality and productivity. In peppers, the high level of ascorbic acid is not only a nutrient substance but also plays a role in environmental stress, i.e., drought stress. When suffering from drought stress, plants accumulate dehydrins, which play important roles in the stress response. Here, we isolated an SK3-type DHN gene CaDHN2 from peppers. CaDHN2 was located in the nucleus, cytoplasm, and cell membrane. In CaDHN2-silenced peppers, which are generated by virus-induced gene silencing (VIGS), the survival rate is much lower, the electrolytic leakage is higher, and the accumulation of reactive oxygen species (ROS) is greater when compared with the control under drought stress. Moreover, when CaDHN2 (CaDHN2-OE) is overexpressed in Arabidopsis, theoverexpressing plants show enhanced drought tolerance, increased antioxidant enzyme activities, and lower ROS content. Based on yeast two-hybrid (Y2H), GST-pull down, and bimolecular fluorescence complementation (BiFC) results, we found that CaDHN2 interacts with CaGGP1, the key enzyme in ascorbic acid (AsA) synthesis, in the cytoplasm. Accordingly, the level of ascorbic acid is highly reduced in CaDHN2-silenced peppers, indicating that CaDHN2 interacts with CaGGP1 to affect the synthesis of ascorbic acid under drought stress, thus improving the drought tolerance of peppers. Our research provides a basis for further study of the function of DHN genes.

Anatomical observation and transcriptome analysis of branch-twisted mutations in Chinese jujube.

BACKGROUND: Plant organs grow in a certain direction and organ twisted growth, a rare and distinctive trait, is associated with internal structure changes and special genes. The twisted branch mutant of Chinese jujube jujube, an important fruit tree native to China and introduced to nearly 50 countries, provides new typical materials for exploration of plant twisted growth. RESULTS: In this study, the cytological characteristics and related genes of twisted branches in Chinese jujube were revealed by microscopy observation and transcriptome analysis. The unique coexistence of primary and secondary structures appeared in the twisted parts of branches, and special structures such as collateral bundle, cortical bundles, and internal phloem were formed. Ninety differentially expressed genes of 'Dongzao' and its twisted mutant were observed, in which ZjTBL43, ZjFLA11, ZjFLA12 and ZjIQD1 were selected as candidate genes. ZjTBL43 was homologous to AtTBL43 in Arabidopsis, which was involved in the synthesis and deposition of cellular secondary wall cellulose. The attbl43 mutant showed significant inflorescence stem bending growth. The transgenic lines of attbl43 with overexpression of ZjTBL43 were phenotypically normal.The branch twisted growth may be caused by mutations in ZjTBL43 in Chinese jujube. AtIQD10, AtFLA11 and AtFLA12 were homologous to ZjIQD1, ZjFLA11 and ZjFLA12, respectively. However, the phenotype of their function defect mutants was normal. CONCLUSION: In summary, these findings will provide new insights into the plant organ twisted growth and a reference for investigation of controlling mechanisms of plant growth direction.

Comparative transcriptome profiling analysis provides insight into the mechanisms for sugar change in Chinese jujube (Ziziphus jujuba Mill.) under rain-proof cultivation.

Chinese jujube (Ziziphus jujuba Mill.) is a globally popular and economically important fruit that is rich in bioactive compounds with strong anti-cancer effects. Rain-proof cultivation is widely used to cultivate Chinese jujube, as it helps avoid rainfall damage during fruit harvest. Although the sugar content of jujube fruits differs between rain-proof and open-field cultivation, the underlying molecular mechanisms are unknown. Here, we analyzed the levels of sugar content, sugar accumulation pattern, and transcriptome profiles of jujube fruits at five developmental stages grown under rain-proof and open-field cultivation modes. The sugar content of jujube fruits was significantly higher under rain-proof cultivation than under open-field cultivation, although the sugar composition and sugar accumulation patterns were comparable. Comparative analysis of transcriptomic profiles showed that rain-proof cultivation enhanced the intrinsic metabolic activity of fruit development. Gene expression and correlation analyses suggested that ZjSPS, ZjSS, ZjHXK, and ZjINV regulate the development-related changes in sugar content in jujube fruits grown under rain-proof cultivation. Temperature, humidity, and moisture conditions were key climatic factors affecting sugar accumulation. Our results provide insights into the molecular mechanisms regulating sugar content and sugar accumulation in Chinese jujube fruits grown under rain-proof cultivation, and we provide genetic resources for studying the development mechanism of Chinese jujube fruit.

Autotetraploidization Alters Morphology, Photosynthesis, Cytological Characteristics and Fruit Quality in Sour Jujube (Ziziphus acidojujuba Cheng et Liu).

Artificially induced polyploidization is one of the most effective techniques for improving the biological properties and creating new cultivars of fruit trees. Up to now, systematic research on the autotetraploid of sour jujube (Ziziphus acidojujuba Cheng et Liu) has not been reported. 'Zhuguang' is the first released autotetraploid sour jujube induced with colchicine. The objective of this study was to compare the differences in the morphological, cytological characteristics, and fruit quality between diploid and autotetraploid. Compared with the original diploid, 'Zhuguang' showed dwarf phenotypes and decreased tree vigor. The sizes of the flowers, pollen, stomata, and leaves of 'Zhuguang' were larger. Perceptible darker green leaves were observed in 'Zhuguang' trees owing to increased chlorophyll contents, which led to higher photosynthesis efficiency and bigger fruit. The pollen activities and the contents of ascorbic acid, titratable acid, and soluble sugar in the autotetraploid were lower than those in diploids. However, the cyclic adenosine monophosphate content in autotetraploid fruit was significantly higher. The sugar/acid ratio of autotetraploid fruit was higher than that of diploid fruit, which made the autotetraploid fruit taste different and better. The results indicated that the autotetraploid we generated in sour jujube could greatly meet the goals of our multi-objective optimized breeding strategies for improving sour jujube, which includes tree dwarfing, increased photosynthesis efficiency, and better nutrient values and flavors as well as more bioactive compounds. Needless to say, the autotetraploid can also serve as material for generating valuable triploids or other types of polyploids and are also instrumental in studying the evolution of both sour jujube and Chinese jujube (Ziziphus jujuba Mill.).

Large-scale analysis of putative Euphorbiaceae R2R3-MYB transcription factors identifies a MYB involved in seed oil biosynthesis.

BACKGROUND: MYB transcription factors are widely distributed in the plant kingdom and play key roles in regulatory networks governing plant metabolism and biochemical and physiological processes. RESULTS: Here, we first determined the R2R3-MYB genes in five Euphorbiaceae genomes. The three Trp (W) residues from the first MYB domain (R2) were absolutely conserved, whereas the first W residue from the second MYB domain (R3) was preferentially mutated. The R2R3-MYBs were clustered into 48 functional subfamilies, of which 34 had both R2R3-MYBs of Euphorbiaceae species and AtMYBs, and four contained only Euphorbiaceae R2R3-MYBs. The whole-genome duplication (WGD) and/or segmental duplication (SD) played key roles in the expansion of the R2R3-MYB family. Unlike paralogous R2R3-MYB family members, orthologous R2R3-MYB members contained a higher selective pressure and were subject to a constrained evolutionary rate. VfMYB36 was specifically expressed in fruit, and its trend was consistent with the change in oil content, indicating that it might be involved in oil biosynthesis. Overexpression experiments showed that VfMYB36 could significantly provide linolenic acid (C18:3) content, which eventually led to a significant increase in oil content. CONCLUSION: Our study first provides insight into understanding the evolution and expression of R2R3-MYBs in Euphorbiaceae species, and also provides a target for the production of biomass diesel and a convenient way for breeding germplasm resources with high linolenic acid content in the future.

Genome-wide screening of the RNase T2 gene family and functional analyses in jujube (Ziziphus jujuba Mill.).

BACKGROUND: Ribonuclease (RNase T2) plays crucial roles in plant evolution and breeding. However, there have been few studies on the RNase T2 gene family in Ziziphus jujuba Mill., one of important dried fruit tree species. Recently, the released sequences of the reference genome of jujube provide a good chance to perform genome-wide identification and characterization of ZjRNase gene family in the jujube. RESULTS: In this study, we identified four members of RNase T2 in jujube distributed on three chromosomes and unassembled chromosomes. They all contained two conserved sites (CASI and CASII). Analysis of the phylogenetic relationships revealed that the RNase T2 genes in jujube could be divided into two groups: ZjRNase1 and ZjRNase2 belonged to class I, while ZjRNase3 and ZjRNase4 belonged to class II. Only ZjRNase1 and ZjRNase2 expression were shown by the jujube fruit transcriptome analysis. So ZjRNase1 and ZjRNase2 were selected functional verification by overexpression transformation of Arabidopsis. The overexpression of these two genes led to an approximately 50% reduction in seed number, which deserve further attention. Moreover, the leaves of the ZjRNase1 overexpression transgenic lines were curled and twisted. Overexpression of ZjRNase2 resulted in shortened and crisp siliques and the production of trichomes, and no seeds were produced. CONCLUSION: In summary, these findings will provide new insights into the molecular mechanisms of low number of hybrid seeds in jujube and a reference for the future molecular breeding of jujube.

Genome-wide identification and expression profile analysis of SWEET genes in Chinese jujube.

The novel sugar transporter known as SWEET (sugars will eventually be exported transporter) is involved in the transport and distribution of photosynthesis products in plants. The SWEET protein is also involved in pollen development, nectar secretion, stress responses, and other important physiological processes. Although SWEET genes have been characterized and identified in model plants, such as Arabidopsis and rice, little is known about them in jujube. In this study, the molecular characteristics of the SWEET gene family in the Chinese jujube (Ziziphus jujuba Mill.) and their expression patterns in different organs, at different fruit developmental stages, and under abiotic stress were analyzed. A total of 19 ZjSWEET genes were identified in jujube through a genome-wide study; these were classified into four sub-groups based on their phylogenic relationships. The gene structure analysis of ZjSWEET genes showed that all the members had introns. The expression patterns of different ZjSWEET genes varied significantly in different organs (root, shoot, leave, flower, fruit), which indicated that ZjSWEETs play different roles in multiple organs. According to the expression profiles by quantitative real-time PCR analysis during fruit development, the expression levels of the two genes (ZjSWEET11, ZjSWEET18) gradually increased with the development of the fruit and reached a high level at the full-red fruit stage. A prediction of the cis-acting regulatory elements indicated that the promoter sequences of ZjSWEETs contained nine types of phytohormone-responsive cis-regulatory elements and six environmental factors. In addition, the expression profiles by quantitative real-time PCR analysis showed that some of the ZjSWEETs responded to environmental changes; ZjSWEET2 was highly induced in response to cold stress, and ZjSWEET8 was significantly up-regulated in response to alkali and salt stresses. This study showed that the functions of the ZjSWEET family members of jujube are different, and some may play an important role in sugar accumulation and abiotic stress in jujube.

Three Novel Adenylate Cyclase Genes Show Significant Biological Functions in Plant.

Adenylate cyclase is the key enzyme solely synthesizing cAMP which participates in cell metabolism regulations and functions as an intracellular second messenger. However, the biological functions of plant ACs have not been elucidated clearly for their poor conservative sequences and low detectable cAMP. We performed a systematic study of plant ACs by using Chinese jujube, whose fruit exhibits the highest cAMP content among plants. Three novel ACs were identified from Chinese jujube, and two types of methods including in vitro and in vivo were used to certificate ZjAC1-3 which can catalyze the conversion of ATP into cAMP. The biological functions of significant accelerations of seed germination, root growth, and flowering were found via overexpression of these AC genes in Arabidopsis, and these functions of ACs were further demonstrated by treating the AC-overexpressing transgenic lines and wild type Arabidopsis with bithionol and dibutyryl-cAMP. At last, transcriptome data revealed that the underlying mechanism of the biological functions of ACs might be regulation of the key genes involved in the circadian rhythm pathway and the hormone signal transduction pathway. This research established a foundation for further investigating plant AC genes and provided strong evidence for cAMP serving as a signaling molecule in plants.

Genome-Wide Identification and Phylogenetic Analysis of TRP Gene Family Members in Saurian.

The transient receptor potential plays a critical role in the sensory nervous systems of vertebrates in response to various mechanisms and stimuli, such as environmental temperature. We studied the physiological adaptive evolution of the TRP gene in the saurian family and performed a comprehensive analysis to identify the evolution of the thermo-TRPs channels. All 251 putative TRPs were divided into 6 subfamilies, except TRPN, from the 8 saurian genomes. Multiple characteristics of these genes were analyzed. The results showed that the most conserved proteins of TRP box 1 were located in motif 1, and those of TRP box 2 were located in motif 10. The TRPA and TRPV in saurian tend to be one cluster, as a sister cluster with TRPC, and the TRPM is the root of group I. The TRPM, TRPV, and TRPP were clustered into two clades, and TRPP were organized into TRP PKD1-like and PKD2-like. Segmental duplications mainly occurred in the TRPM subfamily, and tandem duplications only occurred in the TRPV subfamily. There were 15 sites to be under positive selection for TRPA1 and TRPV2 genes. In summary, gene structure, chromosomal location, gene duplication, synteny analysis, and selective pressure at the molecular level provided some new evidence for genetic adaptation to the environment. This result provides a basis for identifying and classifying TRP genes and contributes to further elucidating their potential function in thermal sensors.

The interaction of ABA and ROS in plant growth and stress resistances.

The plant hormone ABA (abscisic acid) plays an extremely important role in plant growth and adaptive stress, including but are not limited to seed germination, stomatal closure, pathogen infection, drought and cold stresses. Reactive oxygen species (ROS) are response molecules widely produced by plant cells under biotic and abiotic stress conditions. The production of apoplast ROS is induced and regulated by ABA, and participates in the ABA signaling pathway and its regulated plant immune system. In this review, we summarize ABA and ROS in apoplast ROS production, plant response to biotic and abiotic stresses, plant growth regulation, ABA signal transduction, and the regulatory relationship between ABA and other plant hormones. In addition, we also discuss the effects of protein post-translational modifications on ABA and ROS related factors.

Divergence of flowering-related genes to control flowering in five Euphorbiaceae genomes.

Reproductive growth and vegetative growth are a pair of main contradictions in the process of plant growth. Flowering, as part of reproductive growth, is a key switch in the life cycle of higher plants, which affects the yield and economic benefits of plants to a certain extent. The Euphorbiaceae species, including castor bean (Ricinus communis), physic nut (Jatropha curcas), tung tree (Vernicia fordii), cassava (Manihot esculenta), and rubber tree (Hevea brasiliensis), have important economic values because they are raw materials for the production of biodiesel, rubber, etc. The flowering mechanisms are still excluded in the Euphorbiaceae species. The flowering-related genes of Arabidopsis thaliana (Arabidopsis) were used as a reference to determine the orthologs of these genes in Euphorbiaceae genomes. The result showed that 146, 144, 114, 114, and 149 of 207 A. thaliana genes were respectively matched to R. communis, V. fordii, J. curcas, H. brasiliensis, and M. esculenta. These identified genes were clustered into seven pathways including gibberellins, floral meristem identity (FMI), vernalization, photoperiod, floral pathway integrators (FPIs), and autonomous pathways. Then, some key numbers of flowering-related genes are widely conserved in the Euphorbiaceae genomes including but not limited to FPI genes LFY, SOC1, FT, and FMI genes AG, CAL, and FUL. However, some genes, including FRI, FLC, and GO, were missing in several or all five Euphorbiaceae species. In this study, we proposed the putative mechanisms of flowering-related genes to control flowering and provided new candidate flowering genes for using marker-assisted breeding to improve variety quality.

Evolution and function of ubiquitin-specific proteases (UBPs): Insight into seed development roles in plants.

The tung oil produced by the tung tree (Vernicia fordii) provides resources for the manufacture of biodiesel. Ubiquitin-specific proteases (UBPs) are the largest group of deubiquitinases and play key roles in regulating development and stress responses. Here, 21 UBPs were identified in V. fordii, roughly one-half the number found in Manihot esculenta and Hevea brasiliensis. Most UBP duplications are produced from whole-genome duplication (WGD), and significant differences in gene retention existed among Euphorbiaceae. The great majority of UBP-containing blocks in V. fordii, V. montana, Ricinus communis, and Jatropha curcas exhibited extensive conservation with the duplicated regions of M. esculenta and H. brasiliensis. These blocks formed 14 orthologous groups, indicating they shared WGD with UBPs in M. esculenta and H. brasiliensis, but most of these UBPs copies were lost. The UBP orthologs contained significant functional divergence which explained the susceptibility of V. fordii to Fusarium wilt and the resistance of V. montana to Fusarium wilt. The expression patterns and experiments suggested that Vf03G1417 could affect the seed-related traits and positively regulate the seed oil accumulation. This study provided important insights into the evolution of UBPs in Euphorbiaceae and identified important candidate VfUBPs for marker-assisted breeding in V. fordii.

Rapid identification of flaxseed oil based on portable fiber optic Raman spectroscopy combined with an oil microscopy method.

To explore a fast, simple, and accurate method to identify adulteration in flaxseed oil, the Raman spectral data of 130 samples containing flaxseed, canola, cottonseed, and adulterated oils were obtained using a portable fiber optic Raman spectrometer. The Raman spectral results showed that the Raman spectra of the flaxseed and canola oils had noticeable peak shifts, whereas the peak positions of the flaxseed and cottonseed oils were relatively similar. Clear peak intensity differences were observed in the flaxseed, cottonseed, and canola oils, mainly at 868 cm-1 , 1022 cm-1 , 1265 cm-1 , and 1655 cm-1 , with Raman shift intensities in the following order: Iflaxseed oil  > Icottonseed oil  > Icanola oil . Similarly, the peak intensity of the flaxseed and adulterated oils also exhibited certain differences (at 868 cm-1 , 1022 cm-1 , 1265 cm-1 , and 1655 cm-1 ), and the Raman shift intensity tended to decrease gradually with the increasing content of canola and cottonseed oils in the flaxseed oil. Additionally, the results of Raman spectroscopy combined with the "oil microscopy" method exhibited large variations in the radar patterns of the flaxseed, canola, and cottonseed oils, whereas the radar patterns of the flaxseed and adulterated oils closely resembled each other. The results indicated that Raman spectroscopy in combination with oil microscopy more effectively revealed the subtle differences in the Raman shift intensity, serving as a more visual and comprehensive approach for differentiating the quality variations between pure flaxseed oil and other oil species and adulterated oil. PRACTICAL APPLICATION: This study analyzed the Raman spectra of flaxseed, canola, cottonseed, and adulterated oils using fiber optic Raman spectroscopy. Combined with the oil microscopy method for comprehensive evaluation and analysis, it is feasible to effectively identify the quality differences among flaxseed, canola, cottonseed, and adulterated oils.

The Crosstalk of the Salicylic Acid and Jasmonic Acid Signaling Pathways Contributed to Different Resistance to Phytoplasma Infection Between the Two Genotypes in Chinese Jujube.

Jujube witches' broom disease (JWB), one of the most serious phytoplasma diseases, usually results in the destruction of Chinese jujube (Ziziphus jujuba Mill.). Although most jujube cultivars are sensitive to JWB, we found a few genotypes that are highly resistant to JWB. However, the molecular mechanism of phytoplasma resistance has seldom been studied. Here, we used Chinese jujube "T13," which has strong resistance to JWB, and a typical susceptible cultivar, "Pozao" ("PZ"), as materials to perform comparative transcriptome, hormone, and regulation analyses. After phytoplasma infection, the differential expression genes (DEGs) were detected at all three growth phases (S1, S2, and S3) in "PZ," but DEGs were detected only at the first growth phase in "T13." Meanwhile, no phytoplasma was detected, and the symptoms especially witches' broom caused by JWB were not observed at the last two growth phases (S2 and S3) in "T13." Protein-protein interaction analysis also showed that the key genes were mainly involved in hormone and reactive oxygen species (ROS) signaling. In addition, during the recovered growth phase in "T13" from S1 to S2, the level of hydrogen peroxide (H2O2) was significantly increased and then decreased from S2 to S3. Moreover, jasmonic acid (JA) was significantly accumulated in "PZ" diseased plants, especially at the S2 phase and at the S2 phase in "T13," while the content of salicylic acid (SA) decreased significantly at the S2 phase of "T13" compared to that in "PZ." The changes in H2O2 and JA or SA were consistent with the changes in their key synthesis genes in the transcriptome data. Finally, exogenous application of an SA inhibitor [1-aminobenzotriazole (ABT)] rescued witches' broom symptoms, while the contents of both JA and MeJA increased after ABT treatment compared to the control, demonstrating that exogenous application of an SA inhibitor rescued the symptoms of jujube after phytoplasma infection by decreasing the contents of SA and increasing the contents of JA and MeJA. Collectively, our study provides a new perspective on the transcriptional changes of Chinese jujube in response to JWB and novel insights that the crosstalk of JA and SA signaling communicated together to contribute to "T13" JWB resistance.

AHLs' life in plants: Especially their potential roles in responding to Fusarium wilt and repressing the seed oil accumulation.

Members of the AT-hook motif nuclear localized (AHL) family contain diverse but poorly understood biological functions. We identified 371 AHLs in 20 land plants, varying from the early diverging lycophyte Selagineila moellendorfi to a variety of higher plants. The AHLs were divided into two clades (Clade-A and Clade-B) with three different types (Type-I, Type-II, and Type-III AHLs). The divergence between Clade-A and Clade-B likely occurred before the separation of S. moellendorfi from the vascular plant lineages. Members of the AHLs family expanded with the specific whole-genome duplication (WGD)/segmental duplication in some genomes, such as Hevea brasiliensis. The ortholog (Vf00G1914/Amo018442) exhibited opposite expression patterns between two Vernicia species (V. fordii and V. montana), indicating that it was implicated in resistance to Fusarium wilt disease. The expression of Vf09G2138 exhibited a negative correlation with lipid biosynthesis in V. fordii seeds during different stages of development, suggesting that this gene might repress the seed oil accumulation. The core AT-hook motif and PPC domain were responsible for guiding the localization of AHL in the nucleus. This study helps us to understand the evolution of AHLs in multiple plants, further highlight their functions during V. fordii seed development and response to Fusarium wilt disease.

The regulation of cell wall lignification and lignin biosynthesis during pigmentation of winter jujube.

Fruit lignification is due to lignin deposition in the cell wall during cell development. However, there are few studies on the regulation of cell wall lignification and lignin biosynthesis during fruit pigmentation. In this study, we investigated the regulation of cell wall lignification and lignin biosynthesis during pigmentation of winter jujube. The cellulose content decreased, while the lignin content increased in the winter jujube pericarp during pigmentation. Safranin O-fast green staining showed that the cellulose content was higher in the cell wall of winter jujube prior to pigmentation, whereas the lignin in the cell wall increased after pigmentation. The thickness of the epidermal cells decreased with pericarp pigmentation. A combined metabolomics and transcriptomics analysis showed that guaiacyl-syringyl (G-S) lignin was the main lignin type in the pericarp of winter jujube, and F5H (LOC107424406) and CCR (LOC107420974) were preliminarily identified as the key genes modulating lignin biosynthesis in winter jujube. Seventeen MYB and six NAC transcription factors (TFs) with potential regulation of lignin biosynthesis were screened out based on phylogenetic analysis. Three MYB and two NAC TFs were selected as candidate genes and further studied in detail. Arabidopsis ectopic expression and winter jujube pericarp injection of the candidate genes indicated that the MYB activator (LOC107425254) and the MYB repressor (LOC107415078) control lignin biosynthesis by regulating CCR and F5H, while the NAC (LOC107435239) TF promotes F5H expression and positively regulates lignin biosynthesis. These findings revealed the lignin biosynthetic pathway and associated genes during pigmentation of winter jujube pericarp and provide a basis for further research on lignin regulation.

A Chinese White Pear (Pyrus bretschneideri) BZR Gene PbBZR1 Act as a Transcriptional Repressor of Lignin Biosynthetic Genes in Fruits.

BZR transcription factors play essential roles in plant growth and environmental stimuli, and they are also the positive regulators of Brassinosteroid (BR) signal transduction in diverse plants. In addition, BZR TFs, as crucial regulators of BR synthesis, may have multiple stress-resistance functions and their related regulatory mechanisms have been well illustrated in model plants. Here, we carried out a genome-wide identification of BZR members in Chinese pear (Pyrus bretschneideri) and identified 13 members. By comparative analysis in five Rosaceae genomes, BZR members in the pear genome may have undergone large-scale duplication events during evolution. Purifying selection played an important role in almost all of the orthologous and paralogous gene pairs. According to the expression analysis of the PbBZRs during fruit development, three PbBZRs were selected for detailed analysis. Transcriptional activation assays presented that PbBZR1 repressed the promoters of P. bretschneideri lignin biosynthetic genes, such as PbCES9, PbCOMT3, and PbHCT6. Our study traces the evolution of BZR gene family members in Rosaceae genomes and illustrates that the rates of gene loss and gain are far from equilibrium in different species. At the same time, our results suggest that PbBZR1 may be involved in the negative regulation of lignin biosynthesis.