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1.
Biotech Histochem ; 98(1): 29-37, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35775276

RESUMO

Water hyssop (Bacopa monnieri L. Pennel) is a medicinal aquatic herb used to treat diseases in South Asia. Various regeneration protocols have been developed or modified in vitro to ensure the availability of biomass and secondary metabolites of Bacopa. We applied hydrothermally treated titanium dioxide (TiO2) nanoparticles (NPs) (TiO2-NPs) at different concentrations. Three explants, distal portion of half leaf (DPHL), proximal portion of half leaf (PPHL) and full leaf (FL), were used to evaluate response to TiO2. Regeneration from the three explants in vitro was similar except for shoot length. Application of TiO2-NPs exerted significant, but variable, effects on all parameters except percentage of shoot formation, which was 100%. Interactive effects of explant and TiO2-NPs exhibited significant, but variable, effects on fresh weight and percentage of callus formation. All explants produced more shoots using TiO2-NPs compared to control treatments. DPHL explants with application of 8 mg/l TiO2 produced more shoots than controls. Similarly, FL explant treated with 2 mg/l TiO2-NPs produced more shoots/explant than controls. All concentrations of TiO2-NPs produced significantly longer shoots compared to controls. Increased elongation of shoots justifies use of TiO2-NPs for propagation of plants in vitro during acclimatization. Use of TiO2-NPs for rapid elongation of shoots ultimately fosters survival of plants.


Assuntos
Bacopa , Nanopartículas , Plantas Medicinais , Bacopa/metabolismo , Brotos de Planta/fisiologia , Regeneração
2.
Sci Rep ; 12(1): 8507, 2022 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-35596065

RESUMO

Sansevieria trifasciata is used as an indoor plant, in traditional medicine and as a fiber source. Here we characterized fibers of two of varieties of S. trifasciata, Lorentii and Hahnii, and report a protocol for their propagation based on indirect shoot organogenesis. Structural and ribbon fibers were scattered within leaf parenchyma when viewed with confocal laser scanning microscopy. Chemical analysis of the fibers by mass spectrometry and high-performance chromatography revealed higher contents of cellulose and xylose in Lorentii than in Hahnii and significant differences for total lignin between both. A protocol for de novo shoot production was then developed using leaf explants. Time-course histological analyses showed that the first events of transdifferentiation were triggered preferentially in cells surrounding fibers and vascular bundles. Callogenesis and shoot performances were quantified for both varieties, and 2,4-D at 2 and 3 mg·L-1 yielded the best results for primary calli induction and fresh calli mass. The length, number, and mass of shoots produced did not differ significantly between the two cultivars. The fast morphogenic response of S. trifasciata to in vitro culture may be useful for mass propagation or other biotechnological purposes such as metabolite production.


Assuntos
Sansevieria , Cromatografia Gasosa-Espectrometria de Massas , Organogênese , Folhas de Planta , Brotos de Planta/fisiologia , Regeneração
3.
Plant J ; 108(1): 183-196, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34293218

RESUMO

Arbuscular mycorrhizal fungi (AMF) can improve plant nutrient acquisition, either by directly supplying nutrients to plants or by promoting soil organic matter mineralization, thereby affecting interspecific plant relationships in natural communities. We examined the mechanism by which the addition of P affects interspecific interactions between a C4 grass (Bothriochloa ischaemum, a dominant species in natural grasslands) and a C3 legume (Lespedeza davurica, a subordinate species in natural grasslands) via AMF and plant growth, by continuous 13 C and 15 N labelling, combined with soil enzyme analyses. The results of 15 N labelling revealed that P addition affected the shoot uptake of N via AMF by B. ischaemum and L. davurica differently. Specifically, the addition of P significantly increased the shoot uptake of N via AMF by B. ischaemum but significantly decreased that by L. davurica. Interspecific plant interactions via AMF significantly facilitated the plant N uptake via AMF by B. ischaemum but significantly inhibited that by L. davurica under P-limited soil conditions, whereas the opposite effect was observed in the case of excess P. This was consistent with the impact of interspecific plant interaction via AMF on arbuscular mycorrhizal (AM) benefit for plant growth. Our data indicate that the capability of plant N uptake via AMF is an important mechanism that influences interspecific relationships between C4 grasses and C3 legumes. Moreover, the effect of AMF on the activities of the soil enzymes responsible for N and P mineralization substantially contributed to the consequence of interspecific plant interaction via AMF for plant growth.


Assuntos
Carbono/metabolismo , Lespedeza/fisiologia , Micorrizas/fisiologia , Nitrogênio/metabolismo , Fósforo/metabolismo , Poaceae/fisiologia , Transporte Biológico , Isótopos de Carbono/análise , Lespedeza/microbiologia , Isótopos de Nitrogênio/análise , Raízes de Plantas/microbiologia , Raízes de Plantas/fisiologia , Brotos de Planta/microbiologia , Brotos de Planta/fisiologia , Poaceae/microbiologia , Solo/química
4.
BMC Plant Biol ; 21(1): 243, 2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34049485

RESUMO

BACKGROUND: Branch angle is a pivotal component of tea plant architecture. Tea plant architecture not only affects tea quality and yield but also influences the efficiency of automatic tea plant pruning. However, the molecular mechanism controlling the branch angle, which is an important aspect of plant architecture, is poorly understood in tea plants. RESULTS: In the present study, three CsLAZY genes were identified from tea plant genome data through sequence homology analysis. Phylogenetic tree displayed that the CsLAZY genes had high sequence similarity with LAZY genes from other plant species, especially those in woody plants. The expression patterns of the three CsLAZYs were surveyed in eight tissues. We further verified the expression levels of the key CsLAZY1 transcript in different tissues among eight tea cultivars and found that CsLAZY1 was highly expressed in stem. Subcellular localization analysis showed that the CsLAZY1 protein was localized in the plasma membrane. CsLAZY1 was transferred into Arabidopsis thaliana to investigate its potential role in regulating shoot development. Remarkably, the CsLAZY1 overexpressed plants responded more effectively than the wild-type plants to a gravity inversion treatment under light and dark conditions. The results indicate that CsLAZY1 plays an important role in regulating shoot gravitropism in tea plants. CONCLUSIONS: The results provide important evidence for understanding the functions of CsLAZY1 in regulating shoot gravitropism and influencing the stem branch angle in tea plants. This report identifies CsLAZY1 as a promising gene resource for the improvement of tea plant architecture.


Assuntos
Camellia sinensis/genética , Genoma de Planta/genética , Gravitropismo/genética , Arabidopsis/genética , Arabidopsis/fisiologia , Camellia sinensis/fisiologia , Filogenia , Brotos de Planta/genética , Brotos de Planta/fisiologia , Caules de Planta/genética , Caules de Planta/fisiologia , Chá
5.
Plant Cell Physiol ; 62(2): 366-377, 2021 May 11.
Artigo em Inglês | MEDLINE | ID: mdl-33399871

RESUMO

Tea (Camellia sinensis [L.] O. Kuntze) tree is a perennial plant in which winter dormancy is an important biological adaptation to environmental changes. We discovered and reported a novel tea tree cultivar that can generate tender shoots in winter several years ago, but the molecular mechanism for this unique phenotype remains unknown . Here, we conducted comparative transcriptomics, proteomics and metabolomics along with phytohormone quantitation between the winter and spring tender shoots to investigate the physiological basis and putative regulatory mechanisms of its evergrowing character during winter. Our multi-omics study has led to the following findings. Gibberellin (GA) levels and key enzymes for GA biosynthesis and the signal transduction pathway were increased in the winter shoots, causing the ABA/GA content ratio to decrease, which might play a key regulatory role in maintaining normal growth during winter. The abundance of proteins, genes and metabolites involved in energy metabolism was all increased in winter shoots, indicating that energy is critical for continuous growth under the relatively weak-light and low-temperature environment. Abiotic resistance-related proteins and free amino acids were also increased in abundance in the winter shoots, which possibly represents an adaptation response to winter conditions. These results allowed us to hypothesize a novel molecular mechanism of adaptation for this unique tender shoot evergrowing in winter.


Assuntos
Camellia sinensis/fisiologia , Brotos de Planta/fisiologia , Adaptação Fisiológica/genética , Adaptação Fisiológica/fisiologia , Camellia sinensis/genética , Camellia sinensis/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Metabolômica , Dormência de Plantas/genética , Dormência de Plantas/fisiologia , Reguladores de Crescimento de Plantas/metabolismo , Reguladores de Crescimento de Plantas/fisiologia , Proteínas de Plantas/classificação , Proteínas de Plantas/metabolismo , Proteínas de Plantas/fisiologia , Brotos de Planta/genética , Brotos de Planta/crescimento & desenvolvimento , Proteômica , Estações do Ano , Transdução de Sinais/fisiologia
6.
Int J Mol Sci ; 21(11)2020 May 30.
Artigo em Inglês | MEDLINE | ID: mdl-32486149

RESUMO

The aim of this study is to optimize and evaluate the effectiveness of vitrification, droplet-vitrification, and encapsulation-vitrification techniques in the cryopreservation of Lamprocapnos spectabilis (L.) Fukuhara 'Gold Heart', a popular medicinal and ornamental plant species. In vitro-derived shoot tips were used in the experiments. All three techniques were based on explant dehydration with plant vitrification solution 3 (PVS3; 50% glycerol and 50% sucrose) for 0, 30, 60, 90, 120, 150, or 180 min. The recovered microshoots were subjected to morphometric, biochemical, and molecular analyses (RAPD, ISSR, SCoT). The highest recovery level was reported with the encapsulation-vitrification protocol based on 150 min dehydration (73.1%), while the vitrification technique was the least effective (maximum 25.8% recovery). Explants cryopreserved with the encapsulation-vitrification technique produced the highest mean number of shoots (4.9); moreover, this technique was optimal in terms of rooting efficiency. The highest fresh weight of shoots, on the other hand, was found with the vitrification protocol based on a 30-min PVS3 treatment. The concentrations of chlorophyll a and b were lower in all cryopreservation-derived plants, compared to the untreated control. On the other hand, short dehydration and cryopreservation of non-encapsulated explants stimulated the synthesis of anthocyanins. A small genetic variation in 5% of all samples analyzed was detected by RAPD and ISSR marker systems. Only plants recovered from the encapsulation-vitrification protocol had no DNA sequence alternations.


Assuntos
Criopreservação/métodos , Papaveraceae/fisiologia , Brotos de Planta/fisiologia , Plantas Medicinais/fisiologia , Clorofila/análise , Clorofila A/análise , Crioprotetores , Dessecação , Marcadores Genéticos , Variação Genética , Glicerol , Reação em Cadeia da Polimerase , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sacarose , Vitrificação
7.
Chem Biodivers ; 17(4): e2000043, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32112467

RESUMO

Alternariol and altenuisol were isolated as the major phytotoxins produced by an Alternaria sp. pathogenic fungus of the invasive weed Xanthium italicum. Altenuisol exhibited stronger phytotoxic effect compared with alternariol. At 10 µg/mL, alternariol and altenuisol promoted root growth of the monocot plant Pennisetum alopecuroides by 11.1 % and 75.2 %, respectively, however, inhibitory activity was triggered by the increase of concentration, with root elongation being suppressed by 35.5 % and 52.0 % with alternariol and altenuisol at 1000 µg/mL, respectively. Alternariol slightly inhibited root length of the dicot plant Medicago sativa at 10-1000 µg/mL, whereas altenuisol stimulated root growth by 51.0 % at 10 µg/mL and inhibited root length by 43.4 % at 200 µg/mL. Alternariol and altenuisol did not exert strong regulatory activity on another dicot plant, Amaranthus retroflexus, when tested concentration was low, however, when the concentration reached 1000 µg/mL, they reduced root length by 68.1 % and 51.0 %, respectively. Alternariol and altenuisol exerted similar effect on shoot growth of three tested plants but to a lesser extent. It is noteworthy to mention that this is the first report on the phytotoxicity of altenuisol.


Assuntos
Alternaria/química , Micotoxinas/química , Xanthium/microbiologia , Alternaria/isolamento & purificação , Alternaria/metabolismo , Amaranthus/efeitos dos fármacos , Amaranthus/crescimento & desenvolvimento , Espécies Introduzidas , Lactonas/química , Lactonas/isolamento & purificação , Lactonas/farmacologia , Micotoxinas/isolamento & purificação , Micotoxinas/farmacologia , Pennisetum/efeitos dos fármacos , Pennisetum/crescimento & desenvolvimento , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/fisiologia
8.
New Phytol ; 225(3): 1111-1119, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-31127613

RESUMO

Plants are inherently dynamic. Dynamics minimize stress while enabling plants to flexibly acquire resources. Three examples are presented for plants tolerating saline soil: transport of sodium chloride (NaCl), water and macronutrients is nonuniform along a branched root; water and NaCl redistribute between shoot and soil at night-time; and ATP for salt exclusion is much lower in thinner branch roots than main roots, quantified using a biophysical model and geometry from anatomy. Noninvasive phenotyping and precision agriculture technologies can be used together to harness plant dynamics, but analytical methods are needed. A plant advancing in time through a soil and atmosphere space is proposed as a framework for dynamic data and their relationship to crop improvement.


Assuntos
Metabolismo Energético , Nitrogênio/metabolismo , Fósforo/metabolismo , Raízes de Plantas/fisiologia , Brotos de Planta/fisiologia , Estresse Fisiológico , Água/metabolismo
9.
Sci Rep ; 9(1): 16580, 2019 11 12.
Artigo em Inglês | MEDLINE | ID: mdl-31719561

RESUMO

Although researchers have determined that attaining high grain yields of winter wheat depends on the spike number and the shoot biomass, a quantitative understanding of how phosphorus (P) nutrition affects spike formation, leaf expansion and photosynthesis is still lacking. A 3-year field experiment with wheat with six P application rates (0, 25, 50, 100, 200, and 400 kg P ha-1) was conducted to investigate this issue. Stem development and mortality, photosynthetic parameters, dry matter accumulation, and P concentration in whole shoots and in single tillers were studied at key growth stages for this purpose. The results indicated that spike number contributed the most to grain yield of all the yield components in a high-yielding (>8 t/ha) winter wheat system. The main stem (MS) contributed 79% to the spike number and tiller 1 (T1) contributed 21%. The 2.7 g kg-1 tiller P concentration associated with 15 mg kg-1 soil Olsen-P at anthesis stage led to the maximal rate of productive T1s (64%). The critical shoot P concentration that resulted in an adequate product of Pn and LAI was identified as 2.1 g kg-1. The thresholds of shoot P concentration that led to the maximum productive ability of T1 and optimal canopy photosynthetic capacity at anthesis were very similar. In conclusion, the thresholds of soil available P and shoot P concentration in whole plants and in single organs (individual tillers) were established for optimal spike formation, canopy photosynthetic capacity, and dry matter accumulation. These thresholds could be useful in achieving high grain yields while avoiding excessive P fertilization.


Assuntos
Fertilizantes , Fósforo/metabolismo , Fotossíntese , Brotos de Planta/fisiologia , Estações do Ano , Solo/química , Triticum/fisiologia , Brotos de Planta/crescimento & desenvolvimento , Triticum/crescimento & desenvolvimento , Água
10.
Sci Rep ; 9(1): 14880, 2019 10 16.
Artigo em Inglês | MEDLINE | ID: mdl-31619728

RESUMO

The positive effects of arbuscular mycorrhizal fungi (AMF) have been demonstrated for plant biomass, and zinc (Zn) and phosphorus (P) uptake, under soil nutrient deficiency. Additionally, a number of Zn and P transporter genes are affected by mycorrhizal colonisation or implicated in the mycorrhizal pathway of uptake. However, a comprehensive study of plant physiology and gene expression simultaneously, remains to be undertaken. Medicago truncatula was grown at different soil P and Zn availabilities, with or without inoculation of Rhizophagus irregularis. Measures of biomass, shoot elemental concentrations, mycorrhizal colonisation, and expression of Zn transporter (ZIP) and phosphate transporter (PT) genes in the roots, were taken. Mycorrhizal plants had a greater tolerance of both P and Zn soil deficiency; there was also evidence of AMF protecting plants against excessive Zn accumulation at high soil Zn. The expression of all PT genes was interactive with both P availability and mycorrhizal colonisation. MtZIP5 expression was induced both by AMF and soil Zn deficiency, while MtZIP2 was down-regulated in mycorrhizal plants, and up-regulated with increasing soil Zn concentration. These findings provide the first comprehensive physiological and molecular picture of plant-mycorrhizal fungal symbiosis with regard to soil P and Zn availability. Mycorrhizal fungi conferred tolerance to soil Zn and P deficiency and this could be linked to the induction of the ZIP transporter gene MtZIP5, and the PT gene MtPT4.


Assuntos
Proteínas de Transporte de Cátions/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Medicago truncatula/efeitos dos fármacos , Fósforo/farmacologia , Proteínas de Plantas/genética , Rhizophoraceae/fisiologia , Zinco/farmacologia , Biomassa , Proteínas de Transporte de Cátions/metabolismo , Humanos , Transporte de Íons/efeitos dos fármacos , Medicago truncatula/crescimento & desenvolvimento , Medicago truncatula/metabolismo , Micorrizas/fisiologia , Fósforo/deficiência , Proteínas de Plantas/metabolismo , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/fisiologia , Solo/química , Simbiose/fisiologia , Zinco/deficiência
11.
BMC Genomics ; 20(1): 708, 2019 Sep 11.
Artigo em Inglês | MEDLINE | ID: mdl-31510936

RESUMO

BACKGROUND: Bermudagrass (Cynodon dactylon L.) is an important turfgrass species with two types of stems, shoots and stolons. Despite their importance in determining the morphological variance and plasticity of bermudagrass, the intrinsic differences between stolons and shoots are poorly understood. RESULTS: In this study, we compared the proteomes of internode sections of shoots and stolons in the bermudagrass cultivar Yangjiang. The results indicated that 376 protein species were differentially accumulated in the two types of stems. Pathway enrichment analysis revealed that five and nine biochemical pathways were significantly enriched in stolons and shoots, respectively. Specifically, enzymes participating in starch synthesis all preferentially accumulated in stolons, whereas proteins involved in glycolysis and diverse transport processes showed relatively higher abundance in shoots. ADP-glucose pyrophosphorylase (AGPase) and pyruvate kinase (PK), which catalyze rate-limiting steps of starch synthesis and glycolysis, showed high expression levels and enzyme activity in stolons and shoots, respectively, in accordance with the different starch and soluble sugar contents of the two types of stems. CONCLUSIONS: Our study revealed the differences between the shoots and stolons of bermudagrass at the proteome level. The results not only expand our understanding of the specialization of stolons and shoots but also provide clues for the breeding of bermudagrass and other turfgrasses with different plant architectures.


Assuntos
Cynodon/metabolismo , Brotos de Planta/metabolismo , Proteômica , Cynodon/enzimologia , Cynodon/fisiologia , Brotos de Planta/fisiologia , Caules de Planta/metabolismo , Caules de Planta/fisiologia , Proteínas Quinases/metabolismo , Solubilidade , Amido/metabolismo , Açúcares/química , Açúcares/metabolismo
12.
New Phytol ; 221(2): 764-777, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30267566

RESUMO

Many Proteaceae are highly phosphorus (P)-sensitive and occur exclusively on old nutrient-impoverished acidic soils (calcifuge), whilst a few also occur on young calcareous soils (soil-indifferent) that are higher in available calcium (Ca) and P. Calcium increases the severity of P-toxicity symptoms, but its underlying mechanisms are unknown. We propose that Ca-enhanced P toxicity explains the calcifuge habit of most Proteaceae. Four calcifuge and four soil-indifferent Proteaceae from South-Western Australia were grown in hydroponics, at a range of P and Ca concentrations. Calcium increased the severity of P-toxicity symptoms in all species. Calcifuge Proteaceae were more sensitive to Ca-enhanced P toxicity than soil-indifferent ones. Calcifuges shared these traits: low leaf zinc concentration ([Zn]), low Zn allocation to leaves, low leaf [Zn]:[P], low root : shoot ratio, and high seed P content, compared with soil-indifferent species. This is the first demonstration of Ca-enhanced P toxicity across multiple species. Calcium-enhanced P toxicity provides an explanation for the calcifuge habit of most Proteaceae and is critical for the management of this iconic Australian family. This study represents a major advance towards an understanding of the physiological mechanisms of P toxicity and its role in the distribution of Proteaceae.


Assuntos
Baías , Cálcio/farmacologia , Fósforo/toxicidade , Proteaceae/fisiologia , Solo/química , Biomassa , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/fisiologia , Brotos de Planta/efeitos dos fármacos , Brotos de Planta/fisiologia , Proteaceae/efeitos dos fármacos
13.
Physiol Plant ; 166(4): 996-1007, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30515843

RESUMO

The fragmentary information on phosphorus (P) × zinc (Zn) interactions in plants warrants further study, particularly in plants known for their high P and Zn requirements, such as cotton (Gossypium hirsutum L.). The objective of this study was to investigate the effect of P × Zn interactions in a modern cultivar of cotton grown hydroponically. Biomass, mineral nutrition and photosynthetic parameters were monitored in plants receiving contrasting combinations of P and Zn supply. Root biomass, length and surface area were similar in plants with low P and/or low Zn supply to those in plants grown with high P and high Zn supply, reflecting an increased root/shoot biomass quotient when plants lack sufficient P or Zn for growth. Increasing P supply and reducing Zn supply increased shoot P concentrations, whilst shoot Zn concentrations were influenced largely by Zn supply. A balanced P × Zn supply (4 mM P × 4 µM Zn) enabled greatest biomass accumulation, while an imbalanced supply of these nutrients led to Zn deficiency, P toxicity or Zn toxicity. Net photosynthetic rate, stomatal conductance, transpiration rate and instantaneous carboxylation efficiency increased as P or Zn supply increased. Although increasing P supply reduced the P-use efficiency in photosynthesis (PUEP) and increasing Zn supply reduced the Zn-use efficiency in photosynthesis (ZnUEP), increasing Zn supply at a given P supply increased PUEP and increasing P supply at a given Zn supply increased ZnUEP. These results suggest that agricultural management strategies should seek for balanced mineral nutrition to optimize yields and resource-use efficiencies.


Assuntos
Gossypium/metabolismo , Fósforo/metabolismo , Zinco/metabolismo , Biomassa , Gossypium/fisiologia , Fotossíntese/fisiologia , Raízes de Plantas/metabolismo , Raízes de Plantas/fisiologia , Brotos de Planta/metabolismo , Brotos de Planta/fisiologia , Estômatos de Plantas/metabolismo , Estômatos de Plantas/fisiologia
14.
Methods Mol Biol ; 1815: 457-474, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29981142

RESUMO

Hairy root (HR) culture is considered as "green factory" for mass production of bioactive molecules with pharmaceutical relevance. As such, HR culture has an immense potential as a valuable platform to elucidate biosynthetic pathways and physiological processes, generate recombinant therapeutic proteins, assist molecular breeding, and enhance phytoremediation efforts. However, some plant species appear recalcitrant to the classical Agrobacterium rhizogenes transformation techniques. Sonication-assisted Agrobacterium-mediated transformation (SAArT) is a highly effective method to deliver bacteria to target plant tissues that includes exposure of the explants to short periods of ultrasound in the presence of the bacteria.Nuclear magnetic resonance (NMR)-based metabolomics is one of the most powerful and suitable platforms for identifying and obtaining structural information on a wide range of compounds with a high analytical precision. In terms of plant science, NMR metabolomics is used to determine the phytochemical variations of medicinal plants or commercial cultivars in certain environments and conditions, including biotic stress and plant biotic interaction, structural determination of natural products, quality control of herbal drugs or dietary supplements, and comparison of metabolite differences between plants and their respective in vitro cultures.In this chapter, we attempt to summarize our knowledge and expertise in induction of hairy roots from rare and recalcitrant plant species by SAArT technique and further methodology for extraction of secondary metabolites of moderate to high polarity and their identification by using NMR-based metabolomics.


Assuntos
Espectroscopia de Ressonância Magnética/métodos , Metabolômica/métodos , Raízes de Plantas/genética , Técnicas de Cultura de Tecidos/métodos , Transformação Genética , Agrobacterium/metabolismo , Análise de Dados , Metaboloma , Análise Multivariada , Brotos de Planta/fisiologia , Sementes/fisiologia , Sonicação
15.
Methods Mol Biol ; 1815: 475-492, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29981143

RESUMO

Pentalinon andrieuxii is a species used in Mayan traditional medicine due to its biological properties. Recent studies indicate that it produces a pentacyclic triterpene-denominated betulinic acid, which presents various biological activities: antibacterial, antifungal, antiplasmodial, anti-inflammatory, antimalarial, anticancer, leishmanicidal, and antiviral, as well as steroids and sterols with leishmanicidal properties. A recent study also reported the presence of urechitol A and B in the roots; these are secondary metabolites whose biochemical function is as yet unknown. This plant therefore represents a natural source of metabolites with potential application in the pharmaceutical industry. In this chapter, a protocol is described for obtaining transgenic plants, at the reporter gene of the ß-glucuronidase (GUS) via Agrobacterium tumefaciens from hypocotyl and root explants. The protocol established herein could be employed for the manipulation of the genes involved in the biosynthesis of isoprenoids or secondary metabolites of interest. To our knowledge, this is the first report of stable transformation of Pentalinon andrieuxii via Agrobacterium tumefaciens.


Assuntos
Apocynaceae/genética , Técnicas de Cultura de Tecidos/métodos , Transformação Genética , Adaptação Fisiológica , Agrobacterium tumefaciens/metabolismo , Meios de Cultura/química , Genes Reporter , Germinação/efeitos dos fármacos , Glucuronidase/metabolismo , Hipocótilo/crescimento & desenvolvimento , Canamicina/farmacologia , Brotos de Planta/fisiologia , Plasmídeos/metabolismo , Reação em Cadeia da Polimerase , Sementes/fisiologia
16.
Plant Mol Biol ; 97(4-5): 347-355, 2018 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-29869190

RESUMO

KEY MESSAGE: AFP improved cryopreservation efficiency of potato (Solanum tuberosum cv. Superior) by regulating transcript levels of CBF1 and DHN1. However, the optimal AFP concentration required for strong induction of the genes was dependent on the type of vitrification solution to which the AFP was added: This finding suggests that AFP increased cryopreservation efficiency by transcriptional regulation of these genes, which might protect plant cell membranes from cold stress during cryopreservation. Despite the availability of many studies reporting the benefits of anti-freeze protein III (AFP III) as a cryoprotectant, the role of AFP III in this process has not been well demonstrated using molecular analysis. In addition, AFP III has not been exploited in the cryopreservation of potato thus far. Therefore, we studied the effects of AFP III on the cryopreservation of potato (Solanum tuberosum cv. Superior). We found that CBF1 and DHN1 genes are low temperature-inducible in potato leaves (S. tuberosum cv. Superior). Transcript levels of these genes expressed in shoot tips cryopreserved with AFP III were higher than those of the controls. However, the optimal AFP III concentration required for strong induction of the genes was dependent on the type of cryoprotection solution to which the AFP III was added: 500 ng/mL worked best for PVS2, while 1500 ng/mL was optimal for LS. Interestingly, the involvement of AFP III in the cryoprotection solutions improved cryopreservation efficiency as compared to the control, and expression levels of the detected genes were associated with cryopreservation efficiency. This finding suggests that AFP III increased cryopreservation efficiency by transcriptional regulation of these genes, which might protect plant cell membranes from cold stress during cryopreservation. Therefore, we expect that our findings will lead to the successful application of AFP III as a potent cryoprotectant in the cryopreservation of rare and commercially important plant germplasms.


Assuntos
Crioprotetores/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Solanum tuberosum/fisiologia , Temperatura Baixa , Criopreservação , Proteínas de Plantas/genética , Brotos de Planta/genética , Brotos de Planta/fisiologia , Solanum tuberosum/genética
17.
Plant Physiol ; 177(4): 1382-1395, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29871979

RESUMO

Efforts to understand the genetic and environmental conditioning of plant morphology are hindered by the lack of flexible and effective tools for quantifying morphology. Here, we demonstrate that persistent-homology-based topological methods can improve measurement of variation in leaf shape, serrations, and root architecture. We apply these methods to 2D images of leaves and root systems in field-grown plants of a domesticated introgression line population of tomato (Solanum pennellii). We find that compared with some commonly used conventional traits, (1) persistent-homology-based methods can more comprehensively capture morphological variation; (2) these techniques discriminate between genotypes with a larger normalized effect size and detect a greater number of unique quantitative trait loci (QTLs); (3) multivariate traits, whether statistically derived from univariate or persistent-homology-based traits, improve our ability to understand the genetic basis of phenotype; and (4) persistent-homology-based techniques detect unique QTLs compared to conventional traits or their multivariate derivatives, indicating that previously unmeasured aspects of morphology are now detectable. The QTL results further imply that genetic contributions to morphology can affect both the shoot and root, revealing a pleiotropic basis to natural variation in tomato. Persistent homology is a versatile framework to quantify plant morphology and developmental processes that complements and extends existing methods.


Assuntos
Estudos de Associação Genética , Modelos Teóricos , Folhas de Planta/fisiologia , Raízes de Plantas/fisiologia , Solanum/fisiologia , Processamento de Imagem Assistida por Computador , Folhas de Planta/anatomia & histologia , Raízes de Plantas/anatomia & histologia , Brotos de Planta/fisiologia , Locos de Características Quantitativas , Solanum/genética
18.
Tree Physiol ; 38(11): 1655-1671, 2018 11 01.
Artigo em Inglês | MEDLINE | ID: mdl-29688561

RESUMO

Green tea has attracted an increasing number of consumers worldwide due to its multiple health benefits. With the increase in global warming, more frequent cold spells in the spring often cause more serious damage to green tea production because of the young leaves used. We recorded the changes in climatic conditions during a typical cold spell and the damage symptoms caused by the cold spell in different tea cultivars and breeding lines. By simulating the low temperature of a cold spell under controlled conditions, comparative transcriptome and metabolic analyses were performed with sprouting shoots. Many pathways and genes were regulated differentially by the cold spell conditions. Taking into account the metabolic analysis, the results suggested that the mitogen-activated protein kinase (MAPK)-dependent ethylene and calcium signalling pathways were two major early cold-responsive mechanisms involved in sprouting shoots and were followed by the induction of the Inducer of CBF Expressions (ICE)-C-repeat binding factors (CBF)-cold-responsive (COR) signalling pathway to augment cold tolerance. During the cold shock, growth, photosynthesis and secondary metabolism-mainly involving flavonoid biosynthesis-were remarkably affected. Notably, the increased starch metabolism, which might be dependent on the high expression of ß-amylase3 (BAM3) induced by CBF, played crucial roles in protecting young shoots against freezing cold. A schematic diagram of cold spell response mechanisms specifically involved in the sprouting shoots of the tea plant is ultimately proposed. Some essential transcriptional and metabolic changes were further confirmed in the plant materials under natural cold spell conditions. Our results provide a global view of the reprograming of transcription and metabolism in sprouting tea shoots during a cold spell and meaningful information for future practices.


Assuntos
Camellia sinensis/fisiologia , Temperatura Baixa , Brotos de Planta/fisiologia , Camellia sinensis/genética , Metaboloma/fisiologia , Modelos Biológicos , Estresse Fisiológico , Transcriptoma/fisiologia
19.
J Exp Bot ; 69(11): 2837-2846, 2018 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-29514292

RESUMO

Crop biomass and yield are tightly linked to how the light signaling network translates information about the environment into allocation of resources, including photosynthates. Once activated, the phytochrome (phy) class of photoreceptors signal and re-deploy carbon resources to alter growth, plant architecture, and reproductive timing. Most of the previous characterization of the light-modulated growth program has been performed in the reference plant Arabidopsis thaliana. Here, we use Brassica rapa as a crop model to test for conservation of the phytochrome-carbon network. In response to elevated levels of CO2, B. rapa seedlings showed increases in hypocotyl length, shoot and root fresh weight, and the number of lateral roots. All of these responses were dependent on nitrogen and polar auxin transport. In addition, we identified putative B. rapa orthologs of PhyB and isolated two nonsense alleles. BrphyB mutants had significantly decreased or absent CO2-stimulated growth responses. Mutant seedlings also showed misregulation of auxin-dependent genes and genes involved in chloroplast development. Adult mutant plants had reduced chlorophyll levels, photosynthetic rate, stomatal index, and seed yield. These findings support a recently proposed holistic role for phytochromes in regulating resource allocation, biomass production, and metabolic state in the developing plant.


Assuntos
Brassica rapa/fisiologia , Dióxido de Carbono/metabolismo , Fitocromo B/metabolismo , Brassica rapa/crescimento & desenvolvimento , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/fisiologia , Brotos de Planta/crescimento & desenvolvimento , Brotos de Planta/fisiologia , Plântula/crescimento & desenvolvimento , Plântula/fisiologia
20.
Plant Cell Environ ; 41(7): 1579-1592, 2018 07.
Artigo em Inglês | MEDLINE | ID: mdl-29574982

RESUMO

Root phenes and phene states that reduce the metabolic cost of soil exploration may improve plant growth under low phosphorus availability. We tested the hypothesis that under low phosphorus, reduced living cortical area (LCA) would increase soil exploration, phosphorus capture, biomass, and grain yield. Maize genotypes contrasting in LCA were grown in the field and in greenhouse mesocosms under optimal and suboptimal phosphorus regimes. Percent LCA in nodal roots ranged from 25% to 67%. Plants with 0.2 mm2 less LCA under low phosphorus had 75% less root segment respiration, 54% less root phosphorus content, rooted 20 cm deeper, allocated up to four times more roots between 60 and 120 cm depth, had between 20% and 150% more biomass, 35-40% greater leaf phosphorus content, and 60% greater grain yield compared with plants with high LCA. Low-LCA plants had up to 55% less arbuscular mycorrhizal colonization in axial roots, but this decrease was not correlated with biomass or phosphorus content. The LCA components cortical cell file number and cortical cell size were important for biomass and phosphorus content under low phosphorus. These results are consistent with the hypothesis that root phenes that decrease the metabolic cost of soil exploration are adaptive under phosphorus stress.


Assuntos
Fósforo/deficiência , Raízes de Plantas/metabolismo , Zea mays/crescimento & desenvolvimento , Grão Comestível/crescimento & desenvolvimento , Micorrizas/metabolismo , Fósforo/metabolismo , Raízes de Plantas/anatomia & histologia , Raízes de Plantas/fisiologia , Brotos de Planta/metabolismo , Brotos de Planta/fisiologia , Zea mays/metabolismo
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