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    High contents of proline and anthocyanin increase protective response to salinity in Oryza sativa L. spp. indica
    (2011-09-01) ;
    Cha-um, Suriyan
    ;
    Pigmented rice cultivar is a good candidate for abiotic tolerance especially salt tolerant trait. Physiological and growth parameters of eight indica rice genotypes under control or salt stress conditions were subjected to Hierarchical cluster analysis. From cluster ranking, rice genotypes were classified into two groups, salt-tolerant (Sangyod, Khao Dang, Kulab Dang and TD49) and salt-sensitive genotypes (Klum Sakol, Klum Khonkaen1, Klum Khonkaen2 and Black Sticky Rice). In salt stress (100 mM NaCl), the salt-tolerant genotypes showed the high percentages of proline and anthocyanin more than in the salt-sensitive genotypes. In addition, the photosynthetic pigment stabilizations of stressed-seedlings were positively correlated to the proline (R <sup>2</sup> = 0.98) and anthocyanin (R <sup>2</sup> = 0.70) accumulations. While, the malondialdehyde (MDA) contents of salt-stressed seedlings were negatively correlated to the proline (R <sup>2</sup> = 0.63) and anthocyanin (R <sup>2</sup> = 0.51) accumulations. Moreover, the total antioxidant activity of extraction from stressed-seedlings was assessed using the ABTS (2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) method. The higher percentages of total antioxidant activity were determined in salt-tolerant (125-199%) than salt-sensitive genotypes (106-113%). From the results it can be concluded that accumulation of above mentioned metabolites is associated with cellular protection and salt detoxification in salinity rice seedlings.
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    Increasing plant flavonoid biomaterials in response to UV-A light
    (2013-10-29)
    Kanthang, Supha
    ;
    Flavonoid biomaterials have a protecting function from various stresses. We examined the flavonoid biosynthesis in plant treated under visible light (VL) and additional UV-A light. The transgenic tobacco containing PRODUCTION OF ANTHOCYANIN PIGMENT 1 (PAP1) cDNA, involved in flavonoid biosynthesis from Arabidopsis thaliana, were used for studying the flavonoid biosynthesis under both light conditions comparing to non transgenic tobacco. The flavonoid biomaterials were extracted with acidic methanol and water solvent from treated plant leaves. The absorbance of each biomaterial in the extract was measured under specific wavelength using a spectrophotometer. Additional UV-A radiated to non transgenic and transgenic tobacco affect the increasing of p-coumaric acid, naringenin, apigenin and kaempherol biomaterials from themselves grown under VL (approximately 120-130%). However, PAP1 transgenic tobaccos under additional UV-A radiation enhance the accumulation of these biomaterials up to160-180% higher than non transgenic tobaccos grown under VL condition. Moreover, PAP1 transgenic tobacco radiated with UVA light also significantly increased pelargonidin biomaterial. PAP1 transgenic tobaccos had a similar phenotype with non transgenic tobaccos but the color of fully expanding flower was more pink intensity than non transgenic. © (2013) Trans Tech Publications, Switzerland.
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    Enhancement of plant regeneration efficiency from mature grains of thai indica rice (Oryza sativa L. cv. KDML105)
    In vitro protocol for efficient plant regeneration has been developed from Thai indica rice cultivar (Oryza sativa L. cv. KDML105). Embryogenic callus was initiated from mature seeds on NBMI medium. The highest frequency (96.00%) of callus induction was obtained on NBMI1 medium [2 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) without kinetin]. These calli were sub-cultured on NBM medium supplemented with various concentrations of BA or combination with supporting materials for plant regeneration. Plant regeneration from embryogenic callus was increased in the medium supplemented with 5 g L-1 Phytagel® in the regeneration medium (NBMR9 medium; 30.00%). In addition, the best regenerated frequency in this experiment was obtained by culturing the embryogenic callus to 4:4 ratios of Gro-lux tubes to fluorescent light tubes at 1000 lux light intensity (56.72%). The regenerated plants were successfully transplanted to soil. The results indicated that manipulation of medium supplements and the cultured condition leads to increase plant regeneration efficiency in KDML105 cultivar for genetic transformation in the future.
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    Proline accumulation and physiological responses of indica rice genotypes differing in tolerance to salt and drought stresses
    (2010-06-01) ;
    Suriyan, Cha Um
    ;
    The study determined the relationship between proline accumulation and physiological responses of indica rice genotypes to salt and drought stresses in photoautotrophic conditions. Two salt-tolerant genotypes of indica rice [cv. jasmine rice (KDML 105) and Dang], two drought-tolerant genotypes [Rice Department 6 (RD6) and Rice Department 15 (RD15)] and a salt-and drought-sensitive genotype [Pathumthani 1 (PT 1)] were exposed to 100 mM NaCl (salt stress) or 100 mM mannitol (drought stress) for 4 d. The relative water content (RWC) and malondialdehyde (MDA) accumulation in salt-and drought-tolerant genotypes did not significantly change. The proline accumulation in these genotypes greatly increased with stress. On the other hand, the salt-and drought-sensitive genotype which had lower proline accumulation demonstrated a significant decrease in RWC and increase in MDA content. Moreover, our results showed a strong correlation between proline concentrations and free radical scavenging by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The high concentration of proline solution presented high DPPH antioxidant activity. The higher proline accumulation in stressed seedlings of tolerant genotypes may help to maintain the RWC and MDA of rice plant, which could lead to preferable growth performance.
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    An evaluation of water deficit tolerance screening in pigmented indica rice genotypes
    (2012-02-01) ;
    Cha-Um, Suriyan
    ;
    Eight pigmented genotypes of indica subspecies were geminated and then treated by mannitol-induced water deficit stress. A change of growth characteristics, photosynthetic pigments, lipid peroxidation, DNA content, proline content and anthocyanin accumulation in stressed seedling (100 mM mannitol) and control plant (0 mM mannitol) were calculated. Growth performances, photosynthetic pigment concentrations, and DNA contents in all rice genotypes were dropped whereas proline, anthocyanin contents and the lipid peroxidation levels were enriched. The stabilization in total photosynthetic pigment concentrations of stressed-seedlings were positively correlated to the proline or anthocyanin accumulation. In contrast, MDA content, the increases in the percentages of drought-stressed seedlings were negatively correlated to the proline or anthocyanin accumulation. The changes in biochemical, physiological and growth parameters were subjected to Ward's cluster analysis for water deficit tolerance. These cultivars could be classified into two groups, water deficit sensitive, SY, KD, KLD and TD49 and water deficit tolerance, KS, KK1, KK2 and BSR.
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    Effects of modulated concentration of ZnO nanoparticles on enhancing biosynthesis of metabolites and protecting plant membrane
    (2019-01-01)
    Chayaprasert, Wilailack
    ;
    Nowadays, nanoparticles are raising their applications around the world. ZnO nanoparticles are extensively used in the industrial and agricultural processes because of low price and less toxicity. Nevertheless, the pros and cons, effects of ZnO nanoparticles are still under discussing. This research aimed to study the protective effects of ZnO nanoparticles on plant physiology at the cellular level by analyzing biomaterial levels in relation to the stability of cellular membranes. Shoots of wild type and tt8 transgenic tobaccos were grown under tissue culture condition in the medium, adding 0, 10, 20 mg/L ZnO nanoparticles. Under media conditions adding 10, 20 mg/L ZnO nanoparticles, tobacco increased accumulation of plant metabolites (total soluble sugar and flavonoids). In these ZnO conditions, wild type and transgenic tobaccos enhanced total soluble sugar about 30-50% and 20-30%, respectively. The highest content of total soluble sugar was found in 20 mg/L ZnO nanoparticle condition. In similarity to the content of total soluble sugar, tobacco plants showed the highest average contents of flavone, flavonol and anthocyanin after growing in 20 mg/L ZnO nanoparticle conditions, 92.8%, 61.39% and 48.81% in wild type and 73.49%, 52.39% and 81.68% in transgenics, respectively. Tobaccos under 10 mg/L ZnO media condition increased the accumulation of total soluble sugar and flavonoid contents less than those under 20 mg/L ZnO media condition. However, these plants showed the lowest average percentage of cell membrane injury, 19.5% in wild type and 10-18% in transgenic tobaccos, following with plant under 20 mg/L ZnO nanoparticle and non ZnO nanoparticle conditions. Therefore, 10-20 mg/L ZnO nanoparticles in the media showed the induction of plant metabolites, especially in transgenics, and the enhancements of plant protection when compared with the same line tobacco under condition of non ZnO nanoparticles. Ten mg/L ZnO nanoparticle media is the best condition in protecting cellular membrane of tobacco.
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    Tobacco randomly inserted TT8 differenly enhance light signals and flavonoid accumulation
    (2015-08-18) ;
    Kanthang, Supha
    The individual lines of tobacco over-expressing TT8, a bHLH gene, were constructed and cultured under tissue culture condition radiating with photosynthetically activation radiation (PAR) or PAR+UVA. They were compared to wild type (WT). Leaf of treated plants was extracted and analyzed for flavonoid accumulations using a spectrophotometer. The extract of TT8 plants significantly contained flavone, flavonol and anthocyanin level, higher than the WT extract did. The petal extracts of mature transgenic under PAR had a similar absorbance profile of each substance, but these extracts had higher flavonoid contents than the leaf extracts did. All flavonoid subgroups and p-coumaric acid biosynthesis were significantly enhanced after the additional UVA radiation to plant. This UVA condition slightly stimulated an accumulation of these substances in normal plant. Some transgenic greatly increased flavonoid accumulation in responding to PAR+UVA, but the others were slightly different compared to WT. The distinct insertion site is directly affected TT8 gene expression. Transgenic seeds had a dark brown color more than WT seed, which indicated high content of polymer flavonoids (proanthocyanins). This over-expressing TT8 in transgenic tobacco may directly or indirectly enhance the signal transductions of PAR and UVA and raise up flavonoid accumulation.
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    A proper concentration of carbon black nanoparticles enhances growth of the regenerated vetiver grass
    (2017-01-01)
    Khunchuay, Chonnikarn
    ;
    Carbon black (CB) nanoparticles were used in enhancing efficiency of the plant regeneration process under tissue culture condition. The calli of vetiver grass (Vetiveria zizanioides L. Nash) were induced by using axillary bud as the explant. These calli were subjected onto plant regeneration medium, Murashige & Skoog (MS) medium supplemented with 1 mgL<sup>-1</sup> α-naphthalene acetic acid (NAA), 2 mgL<sup>-1</sup> 6-benzylaminopurine (BAP) and 2 mgL<sup>-1</sup> kinetin, adding 0-70 mgL<sup>-1</sup> concentrations of carbon black (CB) nanoparticles. After six week incubation, vetiver calli developed on the medium supplemented with 40 mgL<sup>-1</sup> CB nanoparticles showed the rather high frequencies of plant regeneration (93.75%), nevertheless no statistically different with other treatment did. Media containing CB nanoparticles in the range 40-60 mgL<sup>-1</sup> significantly affected on enhancing the average of shoot number per callus more than other CB concentration did. Especially, the average shoot lengths of plantlets derived from 40 and 50 mgL<sup>-1</sup> CB media (1.73 and 2.19 cm) are longer than those from non CB or other CB concentration media (0.88<sup>-1</sup>.05 cm) at the significant level. Adding CB nanoparticles in regeneration medium effects on enhancing electrical conductivity and decreasing the pH value. The concentration of CB nanoparticle effected the ratio of colloidal sizes which was found in the medium. The proper concentration of CB generates the optimum property of the regeneration medium which can promote the average of shoot number per callus, and enhanced shoot elongation of vetiver grass in the regeneration process.
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    Synergistic effects of BAP and kinetin media additives on regeneration of vetiver grass (Vetiveria zizanioides L. Nash)
    (2016-05-01) ;
    Khunchuay, Chonnikarn
    Regeneration efficiency was enhanced by adding auxin and cytokinin at different concentrations to the media. It was found that the two highest frequencies-90 and 85%-of callus induction occured when the buds were cultured in an MS medium supplemented with 2 mgL<sup>-1</sup> a-naphthalene acetic acid (NAA) plus 1 mgL<sup>-1</sup> N<sup>6</sup>-benzylaminopurine (BAP) (CIM2) and in the same medium supplemented with 2 mgL<sup>-1</sup> NAA plus 1 mgL<sup>-1</sup> kinetin (CIM5), respectively. When cultured further in a plant regeneration medium supplemented with 1 mgL<sup>-1</sup> NAA and 2 mgL<sup>-1</sup> BAP (PRM1), these calli regenerated with high efficiencies, 80% for calli grown in CIM2 and 85% for calli grown in CIM5. The low BAP concentration in CIM1 (2 mgL<sup>-1</sup> NAA and 0.5 mgL<sup>-1</sup> BAP) resulted in a low regeneration efficiency and fewer shoots per callus even though the PRM3 regeneration medium had a high concentration of cytokinins (2 mgL<sup>-1</sup> BAP and 2 mgL<sup>-1</sup> kinetin) . However, the calli cultured in CIM2 without kinetin but with a suitable level of BAP still regenerated with high efficiencies in PRM2 and PRM3 that contained 1 and 2 mgL<sup>-1</sup> kinetin, respectively. Similarly, when calli cultured in CIM5 without BAP but with kinetin were regenerated in PRM1 with BAP alone, they also regenerated with a high regeneration efficiency. These results showed that culturing a specific tissue in a medium with suitable concentrations of NAA and BAP or kinetin resulted in a large number of multiple shoots, and BAP and kinetin worked synergistically to produce a high plant regeneration efficiency.
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    Tobacco expressing PAP1 increases the responses to PAR and UV-A by enhancing soluble sugars and flavonoids and elevating plant protections
    (2015-04-01) ;
    Kanthang, Supha
    Five lines of transgenic tobacco over-expressing Production of Anthocyanin Pigment 1 (PAP1) cDNA were analysis of metabolic response against the radiation and their protection of the plant under tissue culture condition. PAP1 transgenic and wild type (WT) plants were treated with the radiations of photosynthetically activate radiation (PAR) or PAR combined with UV-A. All lines of transgenic significantly increased in amounts of p-coumaric acid, naringenin apigenin more than WT under both treatments. Additional UV-A radiating to plant rose up kaempferol content in WT plant (1.5 times) and in PAP1 transgenics (1.8 times). These transgenic plants treated under both conditions had also increased anthocyanin substances (pelargonidin) with significant value after compared to WT. Content of total soluble sugar (TSS) was related to the content of total flavonoids in transgenic. PAR combined with UV-A had a lower induction of the electrolyte leakage percentage and malondialdehyde (MDA) level in the transgenic leaf tissue compared to WT tissue. The metabolic substance levels were considered on its protection of plant cells. In transgenic tissue, the enhancement of apigenin level strongly diminished the increase level of electrolyte leakage while the levels of TSS, p-coumaric acid and naringinin less affected. Moreover, the increase levels of kaempferol and pelargonidin associated with the decrease level of MDA, while the TSS level reversely responded. The PAP1 transgenic increased response of light by adaptation of their metabolites (TSS, p-coumaric acid and flavonoids) consequently enhance parameter indicating protections of the cell.