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Gibberellins (GAs) are a class of plant hormones that can affect plant growth and development. GA-oxidases are rate-limiting enzymes, which play a direct role in GA accumulation in plants. However, the roles of GA-oxidase on carrot (Daucus carota L.) taproot development are still unclear. In this study, two GA-oxidase genes, DcGA20ox2 and DcGA2ox1, were identified in carrot. Transgenic carrot plants were obtained by using Agrobacterium-mediated genetic transformation method. The results showed that overexpression of DcGA20ox2 significantly promoted the accumulation of active GAs in carrot, increased plant height, generated more branches, and enhanced xylem development. Overexpression of DcGA2ox1 significantly reduced the total contents of active GAs compared with the control group, resulting in a dwarf phenotype and markedly increased lignin content of the transgenic carrot. The expression profiling showed that the genes of GA metabolic pathway responded to the negative feedback regulation mechanism. At the same time, the expression of most genes in lignin biosynthesis and polymerization process was up-regulated, corresponding to the massive accumulation of lignin. These findings indicated that DcGA20ox2 and DcGA2ox1 affected carrot growth and development by regulating the levels of endogenous GAs. The results from current work might shed light on further studies aimed to regulate lignification in carrot and other crops.

IntroductionThe thickening of radish taproots is primarily determined by secondary growth driven by the vascular cambium and is a highly intricate process regulated by plant hormones, transcription factors, and many metabolic pathways. Gibberellin (GA), a plant hormone associated with cell elongation, is essential in secondary growth. However, the mechanism through which exogenous GA3 regulates secondary taproot growth in radishes remains unclear.MethodsIntegrated morphological, anatomical, hormonal, and transcriptomic analyses of taproots in radishes treated with GA3 and its biosynthesis inhibitor paclobutrazol (PBZ) were performed to explore their effects on taproot secondary growth and key regulatory pathways.ResultsGA3 significantly hindered taproot thickening by inhibiting the formation and maintenance of the vascular cambium, and PBZ promoted root development by increasing root length rather than root diameter. Transcriptome analysis revealed 2,014, 948, and 1,831 differentially expressed genes identified from the control vs. GA3, control vs. PBZ, and GA3 vs. PBZ comparisons, respectively. Kyoto Encyclopedia of Genes and Genome pathway enrichment analysis revealed that differentially expressed genes were primarily involved in the biosyntheses of secondary metabolites and metabolic pathways. GA3 significantly increased the levels of endogenous indole-acetic acid and the expression of auxin synthesis and signal transduction genes.DiscussionExogenous GA3 significantly inhibited the expression of genes involved in the maintenance and differentiation of vascular cambium, including WOX14, ER/ERL1, and XCP2. Exogenous GA3 affects root thickening in radishes primarily by regulating hormone signal transduction pathways, vascular cambium activity, and substance and energy metabolisms. Our findings provide insights into the mechanisms underlying taproot thickening in radishes and provide a valuable gene database for future studies.

Betalain has received increased attention because of its high nutritional value and crucial physiological functions. Based on the elucidation of its core biosynthetic pathway, betalain can be produced in additional plants by metabolic engineering. Synthesis of betalain in carrot (Daucus carota L.) can improve its nutritional quality and economic value by extracting betalain from the fleshy root, non-edible part, and processing residue of carrot. In this study, two different constructs, namely, pYB:mCD (AomelOS, BvCYP76AD1S, and BvDODA1S) and pYB:CDD (BvCYP76AD1S, BvDODA1S, and MjcDOPA5GTS), were introduced into carrot for betanin synthesis by Agrobacterium-mediated transformation. Betanin can be synthetized in both transgenic calli, and pYB:mCD-transgenic callus can be used to produce betacyanin by suspension culture. However, pYB:mCD-transgenic seedlings can synthetize betanin only by tyrosine feeding. The pYB:CDD-transgenic lines can synthetize betanin in whole plants. The betanin content in fleshy root of pYB:CDD-transgenic carrot was (63.4 ± 9) μg ·g−1 fresh weight according to quantitative analysis. These betanin-producing carrot plant materials can be used to synthesize betanin for industrial application or consumption as dietary sources.

Headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and electronic nose (E-nose) were performed to investigate change of volatile compounds in celery leaves during storage at 20.0 ℃. The results indicated that a total of 108 volatile compounds were identified by HS-SPME-GC-MS, mainly including monoterpenes (43.2%~52.92%) and phthalides (19.93%~28.97%), in which D-limonene was abundant (6600.64~48566.12 μg/kg). The total volatile substance content of fresh celery was 165682.48 μg/kg, which decreased by 40.92% after 9 days of storage. The percent of monoterpenes and phthalides decreased by 9.72% and increased by 8.98% respectively during storage. In celery leaves, 31 substances were detected according to OAV>1. There were 31 kinds of the same characteristic volatile substances during 0~7 d and the number of characteristic volatile substances decreased to 29 at 9 d. During storage, the total OAV value decreased. The cluster analysis confirmed that the celery leaves were mainly minty and grassy odour during 0~3 d, and sweet, wood and camphorated odour at 7 d. After the 9th day, the aroma of celery leaves was spicy and herbal odour. The PCA of E-nose response data showed that E-nose could distinguish the aroma of celery leaves during different storage period. Based on Pearson correlation analysis, the main substances causing changes in the response value of E-nose were terpenoids, alcohols and aromatic volatile compounds of celery (r≥0.80, P

The plant hormone jasmonic acid plays an important role in plant growth and development, participating in many physiological processes, such as plant disease resistance, stress resistance, organ development, root growth, and flowering. With the improvement in living standards, people have higher requirements regarding the quality of vegetables. However, during the growth process of vegetables, they are often attacked by pests and diseases and undergo abiotic stresses, resulting in their growth restriction and decreases in their yield and quality. Therefore, people have found many ways to regulate the growth and quality of vegetable crops. In recent years, in addition to the role that JA plays in stress response and resistance, it has been found to have a regulatory effect on crop quality. Therefore, this study aims to review the jasmonic acid accumulation patterns during various physiological processes and its potential role in vegetable development and quality formation, as well as the underlying molecular mechanisms. The information provided in this manuscript sheds new light on the improvements in vegetable yield and quality.

Commonly used reporters rely on a single property, such as the fluorescence of GFP and visible color of anthocyanins, therefore these reporters hardly handle the complicated condition in practice. Betaxanthins are a group of plant natural products derived from the amino acid tyrosine. Its visible yellow-orange color and green fluorescence under blue light make it a promising new reporter. Only two enzymatic reactions are required to convert tyrosine into betaxanthins. Here, we synthesized an open reading frame named Bx that contained all the betaxanthins biosynthetic genes and demonstrated its use as a powerful and efficient reporter in tobacco, carrot, and tomato.

This study evaluated the salt tolerance in 40 celery germplasm resources to clarify the different salt tolerances of celery germplasm. A gradient treatment with different concentrations of NaCl solutions (100, 200, and 300 mmol·L−1) was used to simulate salt stress. After 15 days of salt treatment, 14 indicators related to plant growth, physiology, and biochemistry were determined. The results showed that different celery varieties responded differently to salt stress. Notably, there were significant variations in below-ground dry weight, root–crown ratio, antioxidant enzyme activity, and soluble protein content among the accessions under salt stress. Principal component analysis was used to identify important indices for evaluating salt tolerance, including plant height, spread, content of soluble protein, and so on. A comprehensive evaluation was conducted utilizing the salt damage index, principal component analysis, affiliation function analysis, and cluster analysis. The 40 celery germplasms were classified into five highly salt-tolerant, seven salt-tolerant, fifteen moderately salt-tolerant, nine salt-sensitive, and four highly salt-sensitive germplasms. SHHXQ, MXKQ, XBQC, XQ, and TGCXBQ were highly salt-tolerant germplasms, and BFMSGQ, HNXQ, ZQ, and MGXQW were highly salt-sensitive germplasms. The results of this study provide a reference for the variety of celery cultivation in saline areas and lay a foundation for the selection and breeding of salt-tolerant varieties of celery.

To clarify the impact of continuous dark stress on lignin and cellulose synthesis in celery, shade-tolerant celery varieties were screened. Yellow celery variety ‘Qianhuang No.1’ and green celery variety ‘Qianlv No.1’ were separately grown in vegetable greenhouses. Dark treatments were applied using PVC shading sleeves for 4, 8, 12, and 16 d after celery had grown 10–13 true leaf blades. This study aimed to investigate the impact of varying periods of dark treatment on the morphological characteristics, lignin accumulation, and cellulose accumulation in celery. The results showed that dark treatment led to celery yellowing, a reduced stem thickness, and an increased plant height. Analysis of lignin and cellulose contents, as well as the expression of related genes, showed that dark treatment caused down-regulation of AgLAC, AgC3′H, AgCCR, AgPOD and AgCAD genes, leading to changes in lignin accumulation. Dark treatment inhibited the expression of the AgCesA6 gene, thus affecting cellulose synthesis. Under dark conditions, the expression of AgF5H and AgHCT genes had little effect on lignin content in celery, and the expression of the AgCslD3 gene had little effect on cellulose content. Analysis of morphological characteristics, lignin accumulation and cellulose accumulation after different lengths of dark treatment demonstrated that ‘Qianlv No.1’ is a shade-tolerant variety in contrast to ‘Qianhuang No.1’.

The circadian clock refers to the formation of a certain rule in the long-term evolution of an organism, which is an invisible ‘clock’ in the body of an organism. As one of the largest TF families in higher plants, the MYB transcription factor is involved in plant growth and development. MYB is also inextricably correlated with the circadian rhythm. In this study, the transcriptome data of the tea plant ‘Baiyeyihao’ were measured at a photoperiod interval of 4 h (24 h). A total of 25,306 unigenes were obtained, including 14,615 unigenes that were annotated across 20 functional categories within the GO classification. Additionally, 10,443 single-gene clusters were annotated to 11 sublevels of metabolic pathways using KEGG. Based on the results of gene annotation and differential gene transcript analysis, 22 genes encoding MYB transcription factors were identified. The G10 group in the phylogenetic tree had 13 members, of which 5 were related to the circadian rhythm, accounting for 39%. The G1, G2, G8, G9, G15, G16, G18, G19, G20, G21 and G23 groups had no members associated with the circadian rhythm. Among the 22 differentially expressed MYB transcription factors, 3 members of LHY, RVE1 and RVE8 were core circadian rhythm genes belonging to the G10, G12 and G10 groups, respectively. Real-time fluorescence quantitative PCR was used to detect and validate the expression of the gene transcripts encoding MYB transcription factors associated with the circadian rhythm.

High temperature stress is regarded as one of the significant abiotic stresses affecting the composition and distribution of natural habitats and the productivity of agriculturally significant plants worldwide. The HSF family is one of the most important transcription factors (TFs) families in plants and capable of responding rapidly to heat and other abiotic stresses. In this study, 29 AgHSFs were identified in celery and classified into three classes (A, B, and C) and 14 subgroups. The gene structures of AgHSFs in same subgroups were conserved, whereas in different classes were varied. AgHSF proteins were predicted to be involved in multiple biological processes by interacting with other proteins. Expression analysis revealed that AgHSF genes play a significant role in response to heat stress. Subsequently, AgHSFa6-1, which was significantly induced by high temperature, was selected for functional validation. AgHSFa6-1 was identified as a nuclear protein, and can upregulate the expression of certain downstream genes (HSP98.7, HSP70-1, BOB1, CPN60B, ADH2, APX1, GOLS1) in response to high-temperature treatment. Overexpression of AgHSFa6-1 in yeast and Arabidopsis displayed higher thermotolerance, both morphologically and physiologically. In response to heat stress, the transgenic plants produced considerably more proline, solute protein, antioxidant enzymes, and less MDA than wild-type (WT) plants. Overall, this study revealed that AgHSF family members perform a key role in response to high temperature, and AgHSFa6-1 acts as a positive regulator by augmenting the ROS-scavenging system to maintain membrane integrity, reducing stomatal apertures to control water loss, and upregulating the expression level of heat-stress sensitive genes to improve celery thermotolerance.

The genus Apium, belonging to the family Apiaceae, comprises roughly 20 species. Only two species, Apium graveolens and Apium leptophyllum, are available in China and are both rich in nutrients and have favorable medicinal properties. However, the lack of genomic data has severely constrained the study of genetics and evolution in Apium plants. In this study, Illumina NovaSeq 6000 and Nanopore sequencing platforms were employed to identify the mitochondrial genomes of A. graveolens and A. leptophyllum. The complete lengths of the mitochondrial genomes of A. graveolens and A. leptophyllum were 263,017 bp and 260,164 bp, respectively, and contained 39 and 36 protein-coding genes, five and six rRNA genes, and 19 and 20 tRNA genes. Consistent with most angiosperms, both A. graveolens and A. leptophyllum showed a preference for codons encoding leucine (Leu). In the mitochondrial genome of A. graveolens, 335 SSRs were detected, which is higher than the 196 SSRs found in the mitochondrial genome of A. leptophyllum. Studies have shown that the most common RNA editing type is C-to-U, but, in our study, both A. graveolens and A. leptophyllum exhibited the U-C editing type. Furthermore, the transfer of the mitochondrial genomes of A. graveolens and A. leptophyllum into the chloroplast genomes revealed homologous sequences, accounting for 8.14% and 4.89% of the mitochondrial genome, respectively. Lastly, in comparing the mitochondrial genomes of 29 species, it was found that A. graveolens, A. leptophyllum, and Daucus carota form a sister group with a support rate of 100%. Overall, this investigation furnishes extensive insights into the mitochondrial genomes of A. graveolens and A. leptophyllum, thereby enhancing comprehension of the traits and evolutionary patterns within the Apium genus. Additionally, it offers supplementary data for evolutionary and comparative genomic analyses of other species within the Apiaceae family.

Celery (Apium graveolens L.) is an important vegetable crop cultivated worldwide for its medicinal properties and distinctive flavor. Volatile organic compound (VOC) analysis is a valuable tool for the identification and classification of species. Currently, less research has been conducted on aroma compounds in different celery varieties and colors. In this study, five different colored celery were quantitatively analyzed for VOCs using HS-SPME, GC-MS determination, and stoichiometry methods. The result revealed that γ-terpinene, d-limonene, 2-hexenal,-(E)-, and β-myrcene contributed primarily to the celery aroma. The composition of compounds in celery exhibited a correlation not only with the color of the variety, with green celery displaying a higher concentration compared with other varieties, but also with the specific organ, whereby the content and distribution of volatile compounds were primarily influenced by the leaf rather than the petiole. Seven key genes influencing terpenoid synthesis were screened to detect expression levels. Most of the genes exhibited higher expression in leaves than petioles. In addition, some genes, particularly AgDXS and AgIDI, have higher expression levels in celery than other genes, thereby influencing the regulation of terpenoid synthesis through the MEP and MVA pathways, such as hydrocarbon monoterpenes. This study identified the characteristics of flavor compounds and key aroma components in different colored celery varieties and explored key genes involved in the regulation of terpenoid synthesis, laying a theoretical foundation for understanding flavor chemistry and improving its quality.

Protoplast culture and transformation technology offer a novel method for developing new plant varieties. Nonetheless, the effective preparation of protoplasts and transformation technology specific to celery has yet to be achieved. This study utilized celery seedling leaves as the primary materials to examine the key factors influencing protoplast isolation. The aim was to prepare leaf protoplasts with a high yield and of high quality and subsequently conduct transient gene transformation and expression. The findings indicated that the most effective procedure for isolating and purifying protoplasts was enzymatic digestion using an enzyme solution consisting of 2.0% cellulase, 0.1% pectolase, and 0.6 M mannitol for a duration of 8 h. Subsequently, the protoplasts were filtered through a 400-mesh sieve and purified through centrifugation at 200× g. Within this system, the overall protoplast yield was exceptionally high, reaching a viability rate of up to 95%. The transient transformation system yielded a maximum transformation efficiency of approximately 53%, as evaluated using the green fluorescent protein (GFP) as a reporter gene. The parameters of the transient transformation system were as follows: a protoplast concentration of 5 × 105 cells·mL−1, exogenous DNA concentration of 500 μg·mL−1, final concentration of PEG4000 at 40%, and transformation duration of 15 min. The transient transformation system was also utilized to further analyze the protein localization characteristics of the celery transcription factor AgMYB80. The findings indicated that AgMYB80 predominantly localizes in the nucleus, thereby confirming the reliability and effectiveness of the transient transformation system. This study successfully established an efficient system for isolating, purifying, and transforming celery protoplasts, and will serve as a basis for future studies on molecular biology and gene function.

Advances in optical sensing technology have led to new approaches to monitoring and determining crop seed vigor. In order to improve crop performance to secure reliable yield and food supply, calibrating seed vigor, purity, germination rate, and clarity is very critical to the future of the agriculture/horticulture industry. Traditional methods of seed vigor determination are lengthy in process, labor intensive, and sometimes inaccurate, which can lead to false yield prediction and faulty decision-making. Optical sensing technology offers rapid, accurate, and non-destructive calibration methods to help the industry develop accurate decisions for seed usage and agronomic evaluation. In this review, we hope to provide a summary of current research in the optical sensing technology used in seed vigor assessments.

Carica papaya maintains an abnormally small but complete NLR family while showing weak disease resistance. To better understand their origin, evolution, and biological function, we identified 59 NLR genes via a customized RGAugury and investigated their characteristics, evolutionary history, and expression profiles based on the improved papaya genome and large-scale RNA-seq data. The results indicated that duplication is a major evolutionary force driving the formation of the papaya NLR family. Synteny analyses of papaya and other angiosperms showed that both insertion and inheritance-derived NLRs are present in papaya. Transcriptome-based expression and network analyses revealed that NLRs are actively involved in biotic stress responses. For example, a papaya-specific inserted TNL was up-regulated strongly by the fungal infection. Both transcriptome and qRT-PCR analyses confirmed the expression divergence of an RNL and an RCNL, a pair of tandem duplication genes involved in different co-expression modules. Furthermore, we observed an inserted gene cluster composed of five duplicated CNLs, showing dosage effects and functional differentiation of disease-resistance genes during evolution. This research will enhance our knowledge of the special NLR family in papaya, which may serve as a model plant for disease-resistance genetic studies.

Persimmon: Interstock alters water flow and hormone metabolism, prompting dwarfism The use of an interstock from a dwarf cultivar lowers the water conductance and affects hormone signal metabolism and transport of the sweet persimmon (Diospyros kaki Thunb.), resulting in smaller and more manageable fruit-bearing trees. Shenchun Qu from Nanjing Agricultural University, China, and colleagues compared ‘Kanshu’ persimmon scions grafted on a ‘Diospyros lotus’ rootstock with and without an interstock from a rare dwarf cultivar called ‘Nantong-xiaofangshi’. The interstock-grafted plants moved water less efficiently through their stems. The same plants also exhibited gene activity patterns that correlated with lower levels of the key growth hormones auxin and gibberellic acid. Expressing one of those putative hormone-repressor genes in transgenic tobacco caused the plants to grow shorter. By helping elucidate the mechanisms of interstock-induced dwarfing, the findings could help improve future persimmon cultivation.

The TALE gene family is a subfamily of the homeobox gene family and has been implicated in regulating plant secondary growth. However, reports about the evolutionary history and function of the TALE gene family in bamboo are limited. Here, the homeobox gene families of moso bamboo Olyra latifolia and Bonia amplexicaulis were identified and compared. Many duplication events and obvious expansions were found in the TALE family of woody bamboo. PhTALEs were found to have high syntenies with TALE genes in rice. Through gene co-expression analysis and quantitative real-time PCR analysis, the candidate PhTALEs were thought to be involved in regulating secondary cell wall development of moso bamboo during the fast-growing stage. Among these candidate PhTALEs, orthologs of OsKNAT7, OSH15, and SH5 in moso bamboo may regulate xylan synthesis by regulating the expression of IRX-like genes. These results suggested that PhTALEs may participate in the secondary cell wall deposition in internodes during the fast-growing stage of moso bamboo. The expansion of the TALE gene family may be implicated in the increased lignification of woody bamboo when divergent from herbaceous bamboos.

Celery is an important leafy vegetable that can grow during the cool season and does not tolerate high temperatures. Heat stress is widely acknowledged as one of the main abiotic stresses affecting the growth and yield of celery. The morphological and physiological indices of celery were investigated in the present study to explore the physiological mechanisms in response to high temperatures. Results showed that the antioxidant enzyme activity, proline, relative conductivity, and malondialdehyde were increased, while chlorophyll and the water content of leaves decreased under high-temperature conditions. Short-term heat treatment increased the stomatal conductance to cool off the leaves by transpiration; however, long-term heat treatment led to stomatal closure to prevent leaf dehydration. In addition, high temperature caused a disordered arrangement of palisade tissue and a loose arrangement of spongy tissue in celery leaves. Combined metabolomic and transcriptomic analyses were further used to reveal the regulatory mechanisms in response to heat stress at the molecular level in celery. A total of 1003 differential metabolites were identified and significantly enriched in amino acid metabolism and the tricarboxilic acid (TCA) cycle. Transcriptome sequencing detected 24,264 different genes, including multiple transcription factor families such as HSF, WRKY, MYB, AP2, bZIP, and bHLH family members that were significantly upregulated in response to heat stress, suggesting that these genes were involved in the response to heat stress. In addition, transcriptional and metabolic pathway analyses showed that heat stress inhibited the glycolysis pathway and delayed the TCA cycle but increased the expression of most amino acid synthesis pathways such as proline, arginine, and serine, consistent with the results of physiological indicators. qRT-PCR further showed that the expression pattern was similar to the expression abundance in the transcriptome. The important metabolites and genes in celery that significantly contributed to the response to high temperatures were identified in the present study, which provided the theoretical basis for breeding heat-resistant celery.

Quantitative real-time reverse-transcriptase PCR (qRT-PCR) has been frequently used for detecting gene expression. To obtain reliable results, selection of suitable reference genes is a fundamental and necessary step. Garlic (Allium sativum), a member from Alliaceae family, has been used both as a food flavoring and as a traditional medicine. In the present study, garlic plants were exposed to salt stress (200 mM NaCl) for 0, 1, 4 and 12 h, and garlic roots, bulbs, and leaves were harvested for subsequent analysis. The expression stability of eight candidate reference genes, eukaryotic translation initiation factor 4α (eIF-4α), actin (ACTIN), tubulin β-7 (TUB7), TAP42-interacting protein of 41 kDa (TIP41), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), SAND family protein (SAND), elongation factor 1 alpha (EF-1α), and protein phosphatase 2A (PP2A) were evaluated by geNorm, NormFinder, and BestKeeper. All genes tested displayed variable expression profiles under salt stress. In the leaf and root group, ACTIN was the best reference gene for normalizing gene expression. In garlic clove, ACTIN and SAND were the least variable, and were suitable for gene expression studies under salt stress; these two genes also performed well in all samples tested. Based on our results, we recommend that it is essential to use specific reference genes in different situations to obtain accurate results. Using a combination of multiple stable reference genes, such as ACTIN and SAND, to normalize gene expression is encouraged. The results from the study will be beneficial for accurate determination of gene expression in garlic and other plants.

The success of quantitative real-time reverse transcription polymerase chain reaction (RT-qPCR) to quantify gene expression depends on the stability of the reference genes used for data normalization. To date, systematic screening for reference genes in persimmon (Diospyros kaki Thunb) has never been reported. In this study, 13 candidate reference genes were cloned from 'Nantongxiaofangshi' using information available in the transcriptome database. Their expression stability was assessed by geNorm and NormFinder algorithms under abiotic stress and hormone stimulation. Our results showed that the most suitable reference genes across all samples were UBC and GAPDH, and not the commonly used persimmon reference gene ACT. In addition, UBC combined with RPII or TUA were found to be appropriate for the "abiotic stress" group and α-TUB combined with PP2A were found to be appropriate for the "hormone stimuli" group. For further validation, the transcript level of the DkDREB2C homologue under heat stress was studied with the selected genes (CYP, GAPDH, TUA, UBC, α-TUB, and EF1-α). The results suggested that it is necessary to choose appropriate reference genes according to the test materials or experimental conditions. Our study will be useful for future studies on gene expression in persimmon.

Non-heading Chinese cabbage {Brassica campestris ssp. chinensis Makino) is one of the most commonly consumed vegetables in China, and has recently been introduced into other countries in Asia, North America, and Europe. In this study, we obtained tetraploid, non-heading Chinese cabbage by treating the apical portion of diploid seedlings with various concentrations of colchicine for different treatment times. Treatments included a total of two, four, or six applications of 1.5 g l-1 or 2.0 g l-1 colchicine, performed twice a day at 09.00 h and 15.00 h. Tetraploid plants were selected and identified based on their morphological, anatomical, agronomic, and cytological characteristics, and on their DNA content, as measured by flow cytometry. The highest rate of tetraploidy (8.6%) occurred in seedlings treated six-times with 1.5 g l-1 colchicine. Compared to diploid plants (untreated controls), the tetraploid plants had wider leaves, larger petals, longer siliques, larger seeds, larger stomata, and fewer chloroplasts per guard cell. HPLC showed that tetraploidisation could increase the level of ascorbic acid (AsA) in this crop. Quantitative real-time PCR (qRT-PCR) showed that the levels of expression of genes encoding enzymes involved in the AsA-L-galactose biosynthetic pathway increased in tetraploid plants. This is the first study to compare levels of expression of genes involved in the biosynthetic pathway of AsA between diploid and tetraploid non-heading Chinese cabbage plants.. [ABSTRACT FROM AUTHOR]

Background The WRKY transcription factor is an important member of the stress-related transcription factors, which mediate diverse abiotic stresses in many plants. However, up until now, the number of WRKY members, and the regulatory mechanisms involved in abiotic stress responses in Pak-choi (Brassica campestris ssp. chinensis), remained unknown. Results We isolated and identified 56 full-length WRKY cDNAs from a Pak-choi stress-induced cDNA library. The 56 putative BcWRKY proteins were divided into three groups based on structural and phylogenetic analyses. A subcellular localization prediction indicated that the putative BcWRKY proteins were enriched in the nuclear region. Experiments involving BcWRKY25 and BcWRKY40 confirmed the prediction. A total of 22 BcWRKYs were differentially expressed in response to at least one stress condition (abscisic acid, cold, salinity, heat, or osmosis) tested on Pak-choi leaves, and a co-expression analysis indicated stress-inducible BcWRKYs co-regulated multiple abiotic stresses. BcWRKY33, BcWRKY40, BcWRKY53, and BcWRKY70 acted as key regulators and played dominant roles within coregulatory networks of stress-inducible BcWRKYs. Conclusions We first isolated and characterized the 56 stress-inducible WRKY transcription factor family members. A total of 22 stress-inducible BcWRKYs found in leaves can co-regulate multiple environmental stresses by integrating the potential mutual interactions of WRKYs in Pakchoi. This information will be valuable when exploring the molecular mechanisms of WRKYs in response to abiotic stresses in plants. [ABSTRACT FROM AUTHOR]

Abstract  A novel NAC (NAM, ATAF1/2, CUC2) homologous gene, OsNAC52, was isolated from Oryza sativa L using RACE (rapid amplification of cDNA ends). Analysis of the amino acid sequence of OsNAC52 revealed a highly conserved NAC domain and a potential nuclear localization sequence in its N-terminus, and the transcriptional activation motif in the C-terminal region. Transgenic plants over-expressing OsNAC52 were highly sensitive to ABA (Abscisic acid), and the growth of the 35S-OsNAC52 transgenic seedlings was significantly more restrained by ABA treatment than those of the wild-type seedlings. Furthermore, over-expression of OsNAC52 activated the expression of downstream genes in transgenic Arabidopsis, resulting in enhanced tolerance to drought stresses but not growth retardation. The results from this study indicate that this novel rice OsNAC52 gene functions as an important transcriptional activator in ABA-inducible gene expression and may be useful in improving plant tolerance to abiotic stress. [ABSTRACT FROM AUTHOR]

A putative transcription factor, named RAMY, that binds to the 20-bp O2S sequences of the regulatory region of the Amy2 gene promoter has been identified using the yeast one-hybrid system from a rice library. The full length RAMY cDNA clone encodes a 218-amino acid protein and is homologous to the late embryogenesis-abundant protein (LEA5). In vitro mutagenesis and electrophoretic mobility shift assays confirmed that RAMY can bind with O2S specifically through an unusual zinc finger with a CXCX4CX2H consensus sequence. Low levels of RAMY mRNAs were detected in rice leaves and roots by Northern blot hybridization. The plant hormone gibberellin (GA) induces expression of both RAMY and Amy2 genes, as performed by Northern blot hybridization, but the increase in RAMY mRNA level occurs prior to that of the Amy2 mRNA level in the GA-treated aleurone tissues. These data suggest that RAMY may act as a trans-acting protein and is probably involved in the GA-induced expression of the rice α-amylase gene. [ABSTRACT FROM AUTHOR]