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  • Correction to: Transcriptome analysis of flowering genes in mango ( Mangifera indica L.) in relation to floral malformation
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2020-01-22
    Ashok Yadav, Pawan K. Jayaswal, K. Venkat Raman, Bhupinder Singh, Nagendra K. Singh, K. Usha

    Correction to: Journal of Plant Biochemistry and Biotechnology

    更新日期:2020-01-23
  • TaZnF, a C3HC4 type RING zinc finger protein from Triticum aestivum is involved in dehydration and salinity stress
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2020-01-02
    Preeti Agarwal, Paramjit Khurana

    C3HC4-type RING finger proteins represent one of the largest transcription factors in the plant kingdom. They are known to play crucial roles in various plant processes like regulation of growth and development, signalling network and abiotic stress etc. The paper attempts to explore and dissect the function of TaZnF in response to abiotic stress like drought and salinity. The gene expression studies revealed the possible role of TaZnF in drought and salt stress response. The expression pattern was found to be similar in root tissue under both drought and salt stress. It was found to increase with the increase in the duration of stress with maximum fold > 3 under drought and > 15 fold in response to salt stress at 8 h. Arabidopsis transgenics overexpressing TaZnF were raised to analyse how the changes in gene expression levels affect plant’s response under stress conditions. TaZnF overexpression conferred increased tolerance to both drought and salt stress as measured by several assays examining their growth and development. The transgenics showed better growth in response to dehydration stress given by both mannitol and Polyethylene glycol (PEG). Further, the transgenics were found to survive when subjected to drought stress for 14 days in contrast to wild type. They also showed high yield under severe drought conditions for one month in comparison to the wild type plants. Similarly the transgenics were found to perform well under salt stress conditions in terms of increased fresh weight, better growth, higher chlorophyll accumulation, higher membrane stability and increased proline content. The expression level of both drought and salt stress related marker genes was found to be higher in transgenics in comparison to wild type plants. Thus these observations clearly indicate that TaZnF functions as a positive regulator of stress response, and can be used as a candidate gene to improve and enhance drought and salt stress tolerance of various crop plants.

    更新日期:2020-01-02
  • Oxygen sequestration by Leghemoglobin is positively regulated via its interaction with another late nodulin, Nlj16 of Lotus japonicus
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-11
    Amit Ghosh, Kaushik Bhar, Anirban Siddhanta

    Nodulin proteins are expressed in legume root cells after infection with rhizobia. Nodulins have been classified as early and late, reflecting the developmental time points of their expression. Leghemoglobin (LegH), which is a classical example of a late nodulin, sequesters oxygen inside the nodule to protect the nitrogenase from oxygen toxicity to sustain symbiotic nitrogen fixation (SNF). Post-translational modification and/or protein–protein interaction are known to regulate activity of proteins. To elucidate the role of post-translational modification of LegH on its oxygen sequestration activity, earlier we have shown that phosphorylation at its S45 imparts most structural disruption of the porphyrin binding pocket responsible for its oxygen binding. In the present report, in an attempt to characterize the protein(s) that may interact with LegH to regulate its activity, it is demonstrated that LegH interact in vitro with Nodulin 16 of Lotus japonicus (Nlj16), another late nodulin. These two interacting proteins resulted in a bigger sized particle which shows higher diffusion coefficient as measured by dynamic light scattering. Interestingly, it was also shown that in vitro oxygen sequestration by LegH is stimulated by this interaction. Furthermore, this interaction is validated by the fact that LegH and Nlj16 could be co-immunoprecipitated from the nodule lysate. Most importantly, fluorescent immunohistochemistry of post-infected nodule sections show perceivable co-localization of these two proteins in the nodule symbiosomes. Thus, this work is a foundation for further investigation on these two interacting late nodulins as one of the plausible regulations for the oxygen sequestration by LegH during SNF.

    更新日期:2019-12-11
  • Functional characterization of Gm ITPK ( myo -inositol: 1, 3, 4 tris phosphate 5/6 kinase) isoforms—‘so different yet so similar’
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-25
    Veda Krishnan, Alkesh Hada, Ashish Marathe, Bhagath Kumar Palaka, Dinakara Rao Ampasala, Shelly Praveen, Markandan Manickavasagam, Archana Sachdev

    The biochemical basis behind the preferential presence and action of GmITPKs (myo-inositol—1, 3, 4 tris phosphate 5/6 kinases) in regulating the phosphorous flux towards phytate synthesis in soybean is an important, still unresolved question. In this study, context specific expression of GmITPKs in developing seeds was analysed, which found to be maximum at early stages (0–4 mm); while isozyme specificity was observed with varied dynamics till maturity. It is commonly believed that similarity at sequence level will infer similarity in structure as well, but the robust correlation between sequence and structure has not yet been proved. Hence full length coding sequences of GmITPKs were cloned and their sequence comparison revealed a low identity score of about 50% among the isoforms. Creative computational tools were employed to predict and comprehend the structural attributes and to explore the inverse correlation between the structural and sequence similarities of GmITPKs. Homology modeling of GmITPKs, their refinement and superposition validated the functional isoformic nature, with 92–97% similarity in spatial configuration. Ligand docking and energy computations showed ADP, IP6 and IP5 as possible ligands, and interactions were studied using docking and simulation. The homology models of GmITPKs were used as the starting point for nanosecond-duration molecular dynamics and were found to be stable during 20 ns simulation period. Existence of situation-specific functional basis for this isoform multiplicity, at structural and expression levels revealed to be a preferred regulatory mechanism to adapt to the metabolic fluctuations in inositol pathway during seed development.

    更新日期:2019-12-11
  • Functional annotation and characterization of hypothetical protein involved in blister blight tolerance in tea ( Camellia sinensis (L) O. Kuntze)
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-19
    Gagandeep Singh, Gopal Singh, Romit Seth, Rajni Parmar, Pradeep Singh, Vikram Singh, Sanjay Kumar, Ram Kumar Sharma

    Tea [Camellia sinensis (L) O. Kuntze], worldwide known source of popular non-alcoholic beverage, is severely affected by various biotic and abiotic stresses. Among these, blister blight (BB) disease caused by the obligate biotrophic fungus Exobasidium vexan is responsible for significant depletion of its yield and quality. Our comparative NGS transcriptomic analysis although elucidated global gene expression pattern of BB defense transitions, yet left with 12,022 transcripts categorized as ‘hypothetical proteins (HPs)’. In this study, efforts were made for assigning functions to HPs derived from RNA-Seq data and successfully identified novel putative candidates involved in BB defense in tea. Domain and family-based characterization identified 9390 HPs representing 2867 protein families and 953 super families. Of these, 213 HPs were assigned with novel putative defense related functional categories (LRR, WRKY, NAC, chitinases and peroxidases). Further, sub-cellular localization (cytosolic,133 HPs; transmembranic, 80 HPs) with abundance of HPs exhibiting of acidic (133) and basic (80) nature suggests their wider functional range. 36 HPs upregulated in tolerant genotype having significant interactions with defense responsive candidates in Protein–Protein Interaction Network analysis, possibly suggests their key regulatory role in BB defense. Interestingly, 12 stereo-dynamically stable structures [LRR (5), NAC (4), WRKY, Chitinase & Peroxidase (1 each)] of HP’s were successfully modelled based on their conserved signature sequences and empirically validated using qRT-PCR analysis, therefore, can be potential novel candidates possibly involved in signal transduction and pathogen recognition during BB defense in tea. Futuristically, novel genes identified in this study can be potentially utilized to expedite genetic improvement efforts in tea. The approach successfully employed in tea can also be adopted for assigning molecular function to the HPs in other plant species.

    更新日期:2019-12-11
  • Transcriptome analysis of Ajowan ( Trachyspermum ammi L.) inflorescence
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-06-05
    Mahboubeh Amiripour, Seyed Ahmad Sadat Noori, Vahid Shariati, Mehdi Soltani Howyzeh

    Trachyspermum ammi (L.) Sprague, commonly known as ‘Ajowan’, belongs to the family ‘Apiaceae’. Ajowan fruits yield 2–6% essential oil, and thymol is the major constituent of the oil (35–60%). T. ammi is of high medicinal value; however, the genomic resources for this medicinal plant are rare. To obtain transcript sequences of ajowan inflorescence, RNA-Seq was applied using Illumina HiSeq 2000 platform. Following de novo assembly, 68051 unigenes were produced, among which 43156 unigenes were annotated against different sequence databases. Blastx results of ajowan unigenes showed that 41950 and 29273 unigenes were hit in the NCBI non-redundant (Nr) protein and UniProt databases, respectively. Gene Ontology (GO) classification showed that 29258 unigenes were categorized into 55 GO terms, and 26002 unigenes were clustered in the Orthologous Group categories. Blastx results against the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway database indicated that 14986 unigenes were represented in 127 pathways. In addition, 8751 sequences containing simple sequence repeats (SSRs) and 1180 putative transcription factor genes were also identified. As far as we know, this repository of genomic information is the first resource currently available for transcriptome characterization, gene discovery, developing SSR molecular markers and future genomic research on ajowan.

    更新日期:2019-12-11
  • A modified method for purification of N -acetylgalactosamine specific lectin from Butea monosperma seeds and its effect on human hepatocellular carcinoma cell growth
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-02-23
    Kavita Y. Hiremath, Prajna Hegde, Mamta Sharma, Shashikala R. Inamdar

    Butea monosperma is a medicinal plant extensively used in ayurveda and is known for its medicinal applications. Butea monosperma seed powder is used for treating various health ailments including cancer. Although, most parts of the plant are being used in medicine none of them except for seeds are known to contain lectins. A lectin from seeds has been reported earlier, however it was not tested for its anticancer properties. Since plant lectins are known for their anticancer properties, in order to explore anticancer potential of B. monosperma a legume lectin was purified from seeds in a single step by nine fold, by a modified method using affinity chromatography on lactose-Sepharose 4B column and tested for its anticancer properties against human hepatocellular carcinoma cells. SDS PAGE of BML reveals two bands with Mr of 32 and 34 kDa and a minor band of 67 kDa similar to earlier reported BML. Hapten inhibition assay shows that lectin is specific to GalNAc, galactose, lactose and D-Fucose. BML shows strong binding to Hep G2 cells with MFI of 476. MTT assay showed growth inhibitory effect on Hep G2 cells by BML, PTL1 and PTL2 in a dose and time dependent manner. Growth inhibitory effect of BML on Hep G2 cells was compared with two legume lectins, PTL1 and PTL2 from Psophocarpus tetragonolobus of Fabaceae Family. BML, PTL1 and PTL2 showed growth inhibitory effect on Hep G2 cells. The presence of lectin, in B. monosperma seeds contribute for its anticancer properties.

    更新日期:2019-12-11
  • Cloning and expression analysis of the StCUL1 gene in potato
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-09
    Peng-Xiang Pang, Li Shi, Xiao-Juan Wang, Yan-Nan Chang, Yong-Ping Luo, Jin-Lin Feng, Hemu Eri, Gang Gao

    The Cullin-RING E3 ubiquitin ligase (CRL) complex is the most common E3 ligase, and the SCF complex (CRL1) has the most diverse functions. Cullin1(CUL1) is a scaffolding protein for assembly of the complex. SCF has been shown to participate in the non-biological stress response pathways. In this study, a classic CUL1 protein was identified in Solanum tuberosum, StCUL1. A full-length cDNA of the StCUL1 gene was obtained from ED13 (a potato variety) by Ralstonia solanacearum inoculation using the RACE method. Sequence analysis indicated that the gene comprised 2662 bp, with an open reading frame of 2229 bp encoding 743 amino acids. The expression levels of the StCUL1 gene in potato treated with R. solanacearum and exogenous hormones (such as salicylic acid, jasmonic acid methyl ester and abscisic acid) at different time points were determined by real-time PCR. The results indicated that StCUL1 was induced not only by pathogenic bacteria, but also by exogenous hormones, with sustained high expression. However, there were some differences in the modes of expression. Tissue localization analysis indicated that its expression was tissue specific, and it was mainly in the phloem of the vascular system of stems and leaves.

    更新日期:2019-12-11
  • Agro-morphological description, genetic diversity and population structure of chickpea using genomic-SSR and ESR-SSR molecular markers
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-04-27
    Hiva Seyedimoradi, Reza Talebi, Homayoun Kanouni, Amir Mohammad Naji, Ezzat Karami

    In this study, genetic diversity of agro-morphological traits and population structure of 167 chickpea were assessed using genomic-SSR and EST-SSR molecular markers. Field evaluation of chickpea genotypes showed highly variation for most of measured characteristics such as: number of pods per plant, number of seeds per plant, 100-seed weight and seed yield. Cluster analysis based on morphological traits, grouped chickpea genotypes into four clusters. 37 SSR markers showed highly allelic variation across chickpea genotypes with an average value of 2.95 alleles per locus. Polymorphism information content of SSR locus ranged from 0.27 to 0.80, with an average value of 0.50. SSR-based cluster analysis, grouped 167 chickpea genotypes into two clusters, while cluster I and II divided into five and four sub-clusters. Population structure and discriminant analysis of principal components showed to presence of two clusters within the chickpea, while there was no strong correlation between morphological-based clustering and molecular genetic diversity. Our results showed highly genetic diversity for both agro-morphological and molecular analysis in examined chickpea genotypes. Overall, results of present study can be used for germplasm management and particularly for introducing new genotypes with desirable characteristics for using in breeding programs.

    更新日期:2019-12-11
  • Sneak peek of Hypericum perforatum L.: phytochemistry, phytochemical efficacy and biotechnological interventions
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-02-12
    Mohammad Yaseen Mir, Saima Hamid, Azra N. Kamili, Qazi P. Hassan

    The phytochemistry of Hypericum perforatum (St. John’s wort) has been elaborated extensively owing to its immense application in medicinal chemistry. Illustrated pharmacological activities like antidepressant, antiviral, and antibacterial effects demonstrated its substantiation for numerous of the conventional purposes reported for St John’s wort. St. John’s wort is herbal remedy extensively used in mild to moderate depression. Most of pharmacological activities were assigned to presence of photosensitive naphthodianthrone; hypericin and other allied flavonoid constituents. Escalating demands of hyperforin as antidepressant added more lure to H. perforatum L. The crude extracts of H. perforatum containing phloroglucinols were used for free radical scavenging and against DNA damage. To meet increasing demands of this drug, researchers need to tailor out the biosynthetic pathways to improve secondary metabolite. This necessitated advancement in biotechnological intervention to improve phytochemical potential of this growing herb. This review will brief out ecology, chemistry and phytochemical efficacy with respective phytoconstituents. Further, review will emphasize biotechnological interventions including both conventional and modern contrivances that have been implemented to augment the glory of this species.

    更新日期:2019-12-11
  • Isolation and characterization of biogenic calcium oxalate crystals from Amorphophallus paeoniifolius
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-02-25
    S. Subashini, K. Sathish Kumar

    Amorphophallus paeoniifolius known as elephant foot yam is an underutilized tuberous food crop grown primarily in Africa, South Asia, and tropical Pacific islands. Aim of the present work was to isolate, identify and study various physicochemical characteristics of calcium oxalate crystals present in elephant foot yam. The calcium oxalate crystals from elephant foot yam were isolated using freeze thawing followed by aqueous extraction method. The optimum isolation conditions for maximum calcium oxalate yield was found as grinding time of 12.5 min, solid to liquid ratio of 1:2, temperature of 50 °C, grinding speed of 2000 rpm and 3 extraction cycles. Morphological characteristics of isolated calcium oxide crystals were studied using Scanning Electron Microscope and the presence of calcium oxalate crystals in the form of whewellite was confirmed. The purity of isolated calcium oxalate crystals was confirmed using Energy Dispersive x-ray Spectroscopy. The spherical and oval shapes of calcium oxalate crystals were confirmed using Transmission Electron Microscope. X-Ray Diffraction analysis has confirmed the tetragonal phase structure of isolated calcium oxalate crystals with good crystal stability. Calcium oxalate crystals from elephant foot yam were found stable at 175 °C using Thermal Gravimetric and Differential Scanning Calorimetric analysis.

    更新日期:2019-12-11
  • Promoter and methylation status analysis revealed the importance of PkGES gene in picroside biosynthesis in Picrorhiza kurrooa
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-18
    Bharati Lalhal Barsain, Sudesh Kumar Yadav, Vipin Hallan

    Picrorhiza kurrooa Royle ex Benth., an important high altitude plant produces picrosides, the monoterpenoids of high therapeutic value. Production of geraniol in the iridoid branch of the picroside biosynthetic pathway is a key step in the biosynthesis of picrosides. Geraniol synthase, catalyzes the conversion of geranyl pyrophosphate into geraniol. In view of this, here we report the identification and sequence analysis of PkGES promoter. The PkGES promoter sequence was isolated following a directional genome walking approach. Presence of similar cis elements such as GT1 consensus elements, LTREs, DREs, WRKY binding elements, PREATPRODH motif, BIHD1OS motif, SEBFCONSSTPR10A motif and MeJA responsive elements in the promoter sequence of PkGES documented its multiple stress responsive characteristic signifying its role in adaptation. Further, PkGES showed differential expression pattern in leaves, rhizome and roots. To elucidate whether the differential expression pattern of PkGES correlates with its epigenetic regulation, DNA methylation analysis of promoter as well as coding region of PkGES was conducted. The DNA methylation levels in the promoter sequence of PkGES gene were found to be dominated at CG followed by CHH and CHG sequence contexts. Epigenetic regulation (DNA methylation) of promoter as well as coding region of PkGES suggested its role in the regulation of PkGES transcripts in diverse tissues. The present study has revealed the presence of several stress inducible cis elements in PkGES promoter sequence and its tissue specific epigenetic regulation in P. kurrooa.

    更新日期:2019-12-11
  • Genetically engineered rice with appA gene enhanced phosphorus and minerals
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-04-26
    Sananda Bhattacharya, Shinjini Sengupta, Aritra Karmakar, Sailendra Nath Sarkar, Gaurab Gangopadhyay, Karabi Datta, Swapan Kumar Datta

    Phytic acid is the major source of phosphorus and other mineral bound compounds in many plant tissues especially seeds and bran of cereals. During germination, phytase enzyme degrades phytic acid and bound phosphate and minerals are released. The monogastric animal cannot digest phytate due to lack of the phytase enzyme. Considering that, we have generated low phytate rice by over expressing appA gene cloned from E. coli under the aleurone-specific promoter of maize zein gene. Molecular analysis confirmed the stable integration of transgene and plants were grown up to T3 generation. The T3 seeds showed around 45% decrease in seed phytate content with a fourfold increase of inorganic phosphorus (Pi) level. The enhanced iron and zinc was twofold and threefold respectively in polished seeds of transgenic plants. There was no change in germination behaviour and other morphological traits in transgenic seeds. Thus, this result provides evidence that tissue-specific expression of bacterial phytase can lead to the reduction of phytic acid in rice seeds without hampering its other physiological processes and phenotypic cost.

    更新日期:2019-12-11
  • Expression of the Arabidopsis SWEET genes during rice false smut infection in the transgenic Arabidopsis thaliana containing increased levels of ATP and sucrose
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-05-27
    Mebeaselassie Andargie, Jianxiong Li

    Ustilaginoidea virens is a causal agent of rice false smut disease which hampers the global rice production. This pathogen is able to continually reach into the central vascular tissues and is able to hijack nutrients from the host plants. It is known that during the later stage of U. virens colonization, the floral region particularly the basal area which is having conducive micro-environment in terms of nutrients and moisture content witnessed the presence of a dense mass of mycelia. Nevertheless, the mechanism which underlies U. virens infection in relation to sugar transport is not well established using a molecular approach. Here, we analyzed contents of sugar that are present in leaves pre and post U. virens infection using a transgenic Arabidopsis thaliana leaves containing increased levels of ATP and sucrose. Glucose and sucrose contents in the leaves of the transgenic Arabidopsis lines were significantly increased compared to WT after U. virens infection, suggesting that U. virens growth and translocation of sugar are directly linked. From the four A. thaliana SWEET genes tested, two AtSWEETs which belong to Clade III were up-regulated significantly upon U. virens infection particularly in the OE7 transgenic line. Our studies suggest that active sugar translocation between the sugar producing tissues probably was triggered by U. virens infection, and the identified SWEET family genes are directly involved in facilitating this process.

    更新日期:2019-12-11
  • Brassica rapa expansin-like B1 gene ( BrEXLB1 ) regulate growth and development in transgenic Arabidopsis and elicits response to abiotic stresses
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-18
    Panneerselvam Krishnamurthy, Muthusamy Muthusamy, Jin A. Kim, Mi-Jeong Jeong, Soo In Lee

    The functional role of expansin-like A and B family members remains unclear in plants. In this study, we investigate the functional role of Brassica rapa expansin-like B1 (BrEXLB1) by overexpressing BrEXLB1 sense, antisense and BrEXLB1 specific promoter: GUS in Arabidopsis. The overexpressors of sense BrEXLB1, antisense BrEXLB1 showed variable and unstable pleiotropic effects on leaf growth. Interestingly, overexpressors of sense BrEXLB1 enhances plant height than antisense overexpressors and wild types. GUS reporter-aided analysis of BrEXLB1 promoter showed their activity prominently in seeds, roots and carpels. This is further confirmed by relative quantification of BrEXLB1 among various tissue samples of B. rapa by semi-quantitative Reverse transcription-polymerase chain reaction assay (RT-PCR). We found that BrEXLB1 promoter has several cis-acting elements including light-responsive, phytohormone-responsive, environmental/stress-responsive and tissue-specific elements in promoter sequences. Hence, we analysed the promoter activity of BrEXLB1 by GUS reporter-aided approach, in which stress, phytohormones and other factors regulated changes in GUS transcripts were measured using quantitative RT-PCR. This study suggests that promoter activity is inducible to exogenous application of phytohormones such as indole-3-acetic acid, jasmonic acid, and other factors such as white light and drought stress. These results suggest that BrEXLB1 under its specific promoters may participate in regulation of leaf, plant growth and responds to hormone availability, light quality, dark periods, developmental stages, and drought conditions.

    更新日期:2019-12-11
  • Molecular detection and in silico characterization of cold shock protein coding gene ( csp A) from cold adaptive Pseudomonas koreensis
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-03-27
    Srikant Awasthi, Anjney Sharma, Pragya Saxena, Jagriti Yadav, K. Pandiyan, M. Kumar, Arjun Singh, Hillol Chakdar, Arpan Bhowmik, Prem L. Kashyap, Alok K. Srivastava, Anil K. Saxena

    Cold shock proteins (CSPs) are greatly conserved family of structurally related DNA binding proteins which are produced during temperature drift. A 213 bp long cspA gene was cloned and sequenced from Pseudomonas koreensis P2 in the present study. The expression analysis of the cspA showed > 2.5 folds increase in the mRNA level at 15 °C while the expression was almost on par at 30 °C and 5 °C indicating its role in moderately low temperature. In silico analyses of the gene showed that the gene codes for 7.69 kDa protein which was phylogenetically very similar to CspA present in Pseudomonads. Amino acid composition of the CspA from P. koreensis was different from that of mesophilic Pseudomonas and tiny/small amino varied significantly between CspA of cold adaptive and mesophilic species. The CspA from P. koreensis P2 contained RNP motifs involved in binding of DNA and RNA. Phylogenetic analyses revealed that the CspA protein of P. koreensis P2 was more close to CspA of distant subgroups of Pseudomonas like P. fluorescens and P. putida subgroup indicating a possible intra-specific gene transfer.

    更新日期:2019-12-11
  • Isolation and functional analysis of FLOWERING LOCUS T orthologous gene from Vanda hybrid
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-01-29
    Kanokwan Panjama, Eriko Suzuki, Masahiro Otani, Masaru Nakano, Norikuni Ohtake, Takuji Ohyama, Weenun Bundithya, Kuni Sueyoshi, Soraya Ruamrungsri

    Vanda is an important ornamental tropical orchid in Thailand and shows great potential in international markets. Since there has been limited research on flowering processes and regulations, flower production programs cannot be properly managed. To understand the genetic mechanisms in the Vanda flowering process, the ortholog of the FLOWERING LOCUS T (FT) gene (VaFT) was isolated and characterized by using Vanda ‘Ratchaburi-Fusch Katsura’. An open reading frame of 531 bp, translating a protein of 176 amino acids (AAs), was obtained from VaFT. The AA sequence alignment of VaFT indicated that it contains a conserved domain, distinctive to the phosphatidylethanolamine-binding proteins (PEBPs) superfamily, and shares high homology with other orchid FT proteins: 93% of PaFT from Phalaenopsis aphrodite, 91% of CgFT from Cymbidium goeringii and 89% of OnFT from Oncidium Gower Ramsey. Ectopic expression of VaFT in transgenic Arabidopsis resulted in activation of floral meristem identity gene APETALA1 (AP1) and early flowering with fewer rosette leaves than non-transgenic Arabidopsis. Our data suggests that VaFT is apparently a PEBPs gene in orchids that conducts the transition of flowering.

    更新日期:2019-12-11
  • Expression profiling of miRNAs indicates crosstalk between phytohormonal response and rhizobial infection in chickpea
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-11-30
    Manish Tiwari, Sabhyata Bhatia

    Legumes develop root nodules in which bacteria fix nitrogen for plants. The phytohormones auxin and cytokinin regulate nodule organogenesis by recruiting various genes to effect symbiosis. Moreover, these genes are regulated by the action of microRNAs also. To understand the complex regulatory network involving miRNAs in response to phytohormones and rhizobial interactions in chickpea roots, a miRNA expression profiling was performed. Indole acetic acid and 6-benzylaminopurine at concentrations of 0.1, 1 and 10 µM were used for auxin and cytokinin exogenous treatment and Mesorhizobium ciceri to study rhizobial interaction with chickpea root. Expression profiling of a set of 11 miRNAs was performed. Further, the targets of the candidate miRNAs were identified, followed by functional annotation. This analysis revealed that cat-miR160, cat-miR164, cat-miR396 and cat-miR398 were responsive to auxin and cytokinin. cat-miR319 was found to be only auxin responsive and is known to regulate auxin signalling by targeting TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) which interacts with auxin inducible genes. Further, cytokinin elicited a response at very low concentration of 0.1 µM, and most of the miRNAs investigated were responsive to cytokinin. Interactome analysis revealed that cat-miR164 and cat-miR168 work in conjunction to regulate auxin signalling. Interestingly, cat-miR169 and cat-miR482 were low expressing during auxin treatment and M. ciceri infection but their expression spiked during cytokinin treatment, indicating a cytokinin mediated mode of action. The miRNA expression profiling in response to phytohormones and rhizobia and the reported function of their target genes suggested a crosstalk among the phytohormonal responses during chickpea nodulation.

    更新日期:2019-12-11
  • Phosphoinositide-specific phospholipase C signalling mediates expression of two phenylalanine ammonia lyase genes induced by salicylic acid in Capsicum chinense cells
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-11-19
    Beatriz A. Rodas-Junco, Víctor González-Mendoza, Armando Muñoz-Sánchez, Wilbert Poot-Poot, Felipe Vázquez-Flota, S. M. Teresa Hernández-Sotomayor

    The role of salicylic acid (SA) in the plant stress response is mediated by phenylalanine ammonia lyase (PAL), a key enzyme in the phenylpropanoid pathway in Capsicum chinense cells. However, the relationship between PAL expression in response to SA and the precise involvement of the phosphoinositide-phospholipase C (PI-PLC) transduction pathway linking these effects remains unclear. A pharmacological approach was used to investigate the role of the PI-PLC pathway in the transcriptional regulation of two different PAL gene members in C. chinense suspension cells exposed to SA. Two putative C. chinense PAL genes (CchPAL1 and CchPAL5) were identified after clustering as an independent group in a phylogenetic tree built with 67 Solanaceae genes, corresponding to six species. The expression of CchPAL1 and CchPAL5 showed differential patterns in C. chinense cells exposed to salicylic acid. Moreover, PLC inhibitors, such as neomycin and U73122, differentially modified the expression profiles. These findings suggest that the PI-PLC pathway participates in the activation of preferred PAL genes in response to SA in Capsicum cells.

    更新日期:2019-12-11
  • A protocol for functional study of genes in Brassica juncea by Agrobacterium -mediated transient expression: applicability in other Brassicaceae
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-11-19
    Madhumanti Das, Haraprasad Naiya, Ananya Marik, Gairik Mukherjee, Anindita Seal

    Agrobacterium-mediated transient expression in plant organs is a quick and reliable method for studying gene functions. Due to the significance of transient transformation, substantial efforts have been dedicated to developing such protocols in various plants including the model Arabidopsis thaliana. Despite the importance, a reliable protocol is still lacking in Brassicaceae due to their recalcitrance towards Agrobacterium-mediated transient transformation. We have developed protocols for transient expression in Brassica juncea (PI 211000) and tested three other Brassica sp. for the suitability of the protocol. Co-infiltration of a bacteria-derived avirulence protein AvrPto1 significantly improved expression in B. juncea cotyledonary leaves. The protocol was used successfully in studying protein localization, protein–protein interaction by co-immunoprecipitation assay and transient silencing in B. juncea indicating it to be an excellent model system for transient expression. The efficiency of the protocol varied between Brassica sp. and depended highly on the Agrobacterium strain used. The protocol would be useful in designing functional analyses of genes using transient expression in Brassicaceae, including Arabidopsis and enable inclusion of mutant lines for such studies.

    更新日期:2019-12-11
  • GFP tagging based method to analyze the genome editing efficiency of CRISPR/Cas9-gRNAs through transient expression in N. benthamiana
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-11-15
    Swapnil S. Thakare, Navita Bansal, S. Vanchinathan, G. Rama Prashat, Veda Krishnan, Archana Sachdev, Shelly Praveen, T. Vinutha

    The CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats/CRISPR—associated proteins 9) is simple and highly efficient technology applied to functional studies of genes and genetic crop improvement. In this study, we have demonstrated the utility of green fluorescent protein (GFP) marker to detect the targeting efficiency of gRNAs. As a proof of concept, Glycine max De-Etiolated 1 (GmDET1) gene was chosen and tagged with GFP to rapidly analyze genome editing efficiency of gRNAs. Results showed weaker GFP fluorescence signal in the N. benthamiana leaves co-infiltrated with GmDET1-GFP overexpression (OE) + DET1 gRNA1 constructs as compared to the stronger GFP florescence signal in the leaves co-infiltrated with DET1 gRNA2 and gRNA3 constructs, thus indicating the highest of DET1 gRNA1. These results were further confirmed by the detection of the mutation frequencies through T7 endonuclease (T7E1) assay and sequencing; the highest mutation rate of 38.46% in GmDET1 targeted by DET1 gRNA1 to that of DET1 gRNA2 (7.69%) and gRNA3 (15.38%) was observed. Thus our studies showed “GFP tagging” as the most reliable and rapid method-one can apply to minimize the generation of non-edited transgenic plants resulting from inefficient gRNAs.

    更新日期:2019-12-11
  • Expression and characterization of a thermostable l -aminoacylase in transgenic rice
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2019-11-14
    Parawee Kanjanaphachoat, I-Wen Wang, Kun-Ting Hsieh, Ching-Shan Tseng, Liang-Jwu Chen

    The gene encoding a thermostable l-aminoacylase (LAA) from Deinococcus radiodurans BCRC12827 was isolated and expressed in transgenic rice under the control of a rice actin gene promoter or a seed-specific promoter, Ose705. The recombinant LAA in the transgenic line Ose705:LAA was specifically detected in rice grains, but not in leaves, and its identity was confirmed by a LC/MS/MS assay. Furthermore, was efficiently purified via affinity chromatography using a nickel column. Enzymatic activity of this rice-produced LAA was determined by HPLC and a maximum activity at pH 8.0 and 45 °C in a phosphate buffer supplemented with the divalent metal ion Co2+ using NAc-l-HPA as a substrate was obtained, similar to its host counterpart. This rice-produced LAA maintained approximately 50% enzyme activity after 48 h of incubation under 45 °C and maintained approximately 90% activity compared to a freshly prepared sample after being stored in rice seeds for 4 years. The present study indicated that seed-specific protein production in transgenic rice is a good and safe source for mass production of LAA, and this system can be useful for the production of other biomedical proteins as well.

    更新日期:2019-12-11
  • The first draft of the pigeonpea genome sequence.
    J. Plant Biochem. Biot. (IF 1.038) Pub Date : 2012-01-01
    Nagendra K Singh,Deepak K Gupta,Pawan K Jayaswal,Ajay K Mahato,Sutapa Dutta,Sangeeta Singh,Shefali Bhutani,Vivek Dogra,Bikram P Singh,Giriraj Kumawat,Jitendra K Pal,Awadhesh Pandit,Archana Singh,Hukum Rawal,Akhilesh Kumar,G Rama Prashat,Ambika Khare,Rekha Yadav,Ranjit S Raje,Mahendra N Singh,Subhojit Datta,Bashasab Fakrudin,Keshav B Wanjari,Rekha Kansal,Prasanta K Dash,Pradeep K Jain,Ramcharan Bhattacharya,Kishor Gaikwad,Trilochan Mohapatra,R Srinivasan,Tilak R Sharma

    Pigeonpea (Cajanus cajan) is an important grain legume of the Indian subcontinent, South-East Asia and East Africa. More than eighty five percent of the world pigeonpea is produced and consumed in India where it is a key crop for food and nutritional security of the people. Here we present the first draft of the genome sequence of a popular pigeonpea variety 'Asha'. The genome was assembled using long sequence reads of 454 GS-FLX sequencing chemistry with mean read lengths of >550 bp and >10-fold genome coverage, resulting in 510,809,477 bp of high quality sequence. Total 47,004 protein coding genes and 12,511 transposable elements related genes were predicted. We identified 1,213 disease resistance/defense response genes and 152 abiotic stress tolerance genes in the pigeonpea genome that make it a hardy crop. In comparison to soybean, pigeonpea has relatively fewer number of genes for lipid biosynthesis and larger number of genes for cellulose synthesis. The sequence contigs were arranged in to 59,681 scaffolds, which were anchored to eleven chromosomes of pigeonpea with 347 genic-SNP markers of an intra-species reference genetic map. Eleven pigeonpea chromosomes showed low but significant synteny with the twenty chromosomes of soybean. The genome sequence was used to identify large number of hypervariable 'Arhar' simple sequence repeat (HASSR) markers, 437 of which were experimentally validated for PCR amplification and high rate of polymorphism among pigeonpea varieties. These markers will be useful for fingerprinting and diversity analysis of pigeonpea germplasm and molecular breeding applications. This is the first plant genome sequence completed entirely through a network of Indian institutions led by the Indian Council of Agricultural Research and provides a valuable resource for the pigeonpea variety improvement.

    更新日期:2019-11-01
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